Synthesis, molecular docking and evaluation of 1,5-diarylpyrazole/oxime hybrids targeting EGFR and JNK-2 as antiproliferative agents

Synthesis, molecular docking and evaluation of 1,5-diarylpyrazole/oxime hybrids targeting EGFR and JNK-2 as antiproliferative agents | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Synthesis, molecular docking and evaluation of 1,5-diarylpyrazole/oxime hybrids targeting EGFR and JNK-2 as antiproliferative agents Kamal S. Abdelrahman, Heba A. Hassan, Salah A. Abdel-Aziz, Adel A. Marzouk, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2772431/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract A series of new 1,5-diarylpyrazole oxime hybrid derivatives ( scaffold A and B ) were designed, synthesized, and their structures were examined for purity with different spectroscopic techniques. All the synthesized compounds ( 7a-j ), ( 8a-j ), ( 9a-c ) and ( 10a-c ) were biologically evaluated for their in vitro cytotoxicity against a panel of five cancer cell lines known to express EGFR and JNK-2, namely human colorectal adenocarcinoma cell line DLD-1, human cervical cancer cell line Hela, human leukemia cell line K562, human pancreatic cell line SUIT-2 and human hepatocellular carcinoma cell line HepG2. The oxime containing compounds ( 8a-j ) and ( 10a-c ) were more active as antiproliferative agents than their non-oxime congeners ( 7a-j ) and ( 9a-c ). Compounds 8d, 8g, 8i, and 10c inhibited EGFR with IC 50 values ranging from 8 to 21 µM when compared to sorafenib. Compound 8i inhibited JNK-2 as effectively as sorafenib, with an IC 50 of 1.00 µM. Furthermore, compound 8g showed cell cycle arrest at the G2/M phase in the Hela cell line cell cycle analysis, whereas compound 8i showed combined S phase and G2 phase arrest. Docking studies revealed that oxime derivatives fit well at the EGFR binding site, with binding free energies ranging from -12.98 to 32.30 kcl/mol for compounds 8d, 8g, 8i, and 10c , while compounds 8d and 8i had binding free energies ranging from -9.16 to -12.00 kcl/mol at the JNK-2 binding site. Pyrazole EGFR JNK-2 Docking Anti-proliferative Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 1. Introduction Cancer is the second-leading cause of death throughout the world. Hence, the incidence rate of cancer mortality is becoming increasingly important on a global scale [ 1 ]. Chemotherapy, which uses drugs that target cell division and angiogenesis or that induces cancer cell death via various signaling pathways, is one of the cancer treatment strategies. However, because of chemotherapy side effects and the development of drug resistance in cancer cells, there is an urgent need for the design, synthesis, and development of effective and safe chemotherapy[ 2 ]. EGFR is a tyrosine kinase receptor that plays an important role in cellular signaling activities including cell growth, division, differentiation, metabolism, adhesion, and death [ 3 ]. Four tyrosine kinase-related receptors (EGFR, HER2, HER3, and HER4) have been classified into the HER family. Deregulation of HER family signals enhances proliferation, invasion, metastasis, angiogenesis, and cancer cell survival [ 4 ]. EGFR receptors are over-expressed in different human tumors such as breast, ovarian, prostate, colon, renal, pancreatic, hepatocellular carcinoma, cervical cancer, non-small cell lung cancer (NSCLS) and leukemia [ 4 – 11 ]. Thus, EGFR inhibition has been developed as one of the most efficient strategies for cancer therapy. Several small molecules targeting EGFR are now clinically available such as gefitinib, erlotinib, lapatinib and dacomitinib [ 12 ]. JNK-2 is a member of the MAP kinase family involved in signaling pathways that has been implicated in several diseases like cancer and inflammatory diseases [ 13 ]. Due to the important key roles of JNK-2 in cancer progression through control of proliferation, differentiation, survival and migration, JNK-2 becomes an appealing oncogenic target for cancer therapy due to its high expression in a variety of cancers, including colorectal adenocarcinoma, cervical cancer, pancreatic cancer, hepatocellular carcinoma, and leukemia.[ 14 – 20 ]. JNK signaling is apparently involved in cancer development and progression as in lymphoma cancer cells through protection it from apoptosis by decreasing ROS accumulation. Also, JNK regulates micro RNA-92a and glucose regulating protein 78 (GRP78) in human pancreatic cancer, which promotes cell proliferation and survival. In hepatocellular carcinoma (HCC) JNK pathway is responsible for its development and progression and becomes the target for therapeutic treatment of HCC. Otherwise, blocking of the JNK pathway leading to inhibition of proliferation human B lymphoma cell due to downregulation of early growth response gen-1(Egr-1) protein. Moreover, the relation between the JNK pathway and other pathways like kappa B(NF- KB) and p38, which are acting together for the regulation of cell proliferation and survival, Also, there is a close relation between JNK and immune evasion regulatory factors such as transforming growth factor-β (TGF-β) and interferon-γ (IFN-γ) mediate cell survival. In addition, JNK can promote cancer cells survival by autophagy to counteract apoptosis. To date, the majority of JNK inhibitors target the highly conserved ATP- binding site, while number of this inhibitors were proven in vivo in animal model, but not applied therapeutically until now due to lack of its specificity and its side effect, in addition, to increase in the concentration of ATP decrease its efficacy[ 21 ]. Pyrazoles are an important class of heterocyclic compounds and exhibit broad range of biological activities including anticancer [ 22 ], anti-inflammatory [ 23 ], antimicrobial [ 24 ], antiviral [ 25 ] and antitubercular activities [ 26 ]. Biological evaluation revealed that some pyrazole-containing compounds, such as compounds I and II (Fig. 1 ) exhibited antiproliferative activity against the human cervical cancer cell line Hela by inhibiting cell migration and potent EGFR tyrosine kinase inhibitory activity with IC 50 values of 0.07 and 0.06 µM, respectively, in comparison to the positive control erlotinib (IC 50 = 0.03 µM) [ 27 ]. Moreover, the selective COX-2 inhibitor compound SC-236 (Fig. 1 ) showed antitumor activity through blocking of tumor promotor-induced activator protein-1 (AP1) activation as result of suppression of JNK expression and used to treat hepatocellular cancer in conjunction with doxorubicin [ 28 ]. Furthermore, compound SC74102 (Fig. 1 ) displayed JNK-2 inhibitory activity with IC 50 of 1.35 µ mol/l as well as its p38α inhibitory activity. Additionally, diarypyrazoles have been reported to have STAT3 inhibitory activity as in compound MNS1-Leu V [ 29 ] and heat shock protein inhibitory activity as in compound CCT072453 VI (Fig. 1 ) [ 30 ]. Oximes became with high interest in pharmaceutical chemistry as well. The oxime moiety can hydrogen bind with amino acid residues in the active site of many enzymes and is easy to coordinate with metal ions. Thus, the oxime moiety can improve the binding of the whole molecule to its binding site. Oximes can also release nitric oxide free radicals, which have a cytostatic and cytotoxic effect on cancer cells. It can prevent cancer cells from spreading and help macrophages to kill cancer cells. Several targets have been reported to combine NO with cancer therapy, including either the synergistic effect between anticancer drugs and nitric oxide, increasing the flow of anticancer therapy by NO to intracellular compartments, or increasing the efficiency of cytostatic therapy and overcome of resistance to anticancer agents.[ 31 ]. On the other hand, the introduction of an oxime group into an appropriate chemical backbone is a reasonable approach for the preparation of cytotoxic agents, and many oxime derivatives have been reported to have therapeutic activity for cancer[ 32 – 34 ]. Also, the introduction of oxime moiety in some natural compounds such as psammaplin A (Fig. 2 ) analog was responsible for high anticancer activity [ 35 ]. Triterpene-derived acylated oximes have demonstrated cytotoxic or antiproliferative action against numerous cancer cell lines [ 36 ] and several indirubin oxime showed anticancer activity better than natural alkaloid indirubin (Fig. 2 ). Furthermore, the oxime derivative of natural alkaloid tryptanthrin (Fig. 2 ) showed JNK1/2/3 inhibition [ 37 ]. Finally. Oximes have been employed in the development of several kinase inhibitors, including those for JNK [ 37 , 38 ], phosphorylase kinase (PhK), and phosphatidyl inositol 3-kinase (PI3K)[ 39 ]. Indirubin oximes, for example, have a high affinity for binding to the ATP-binding site of protein kinases involved in the development of tumors, such as cyclin-dependent kinases (CDK), glycogen synthase kinase 3 (GSK) 1, vascular endothelial growth factor receptor 2 (VEGFR-2), c-Src, and casein kinase 2(CK2). Many of these kinases could serve as molecular targets for drugs that combat cancer. [ 40 ]. Based on the information mentioned above and in continuing of our efforts to find small compounds with potential anticancer activity, the aim of this work was to construct a hybrid series of 1,5-diarylpyrazole derivatives (Scaffold A and B ) (Fig. 3 ) that target EGFR and JNK-2 and contain oxime as a NO release moiety to enhance anti-cancer activity. To have the crucial pharmacophoric characteristics of EGFR/JNK-2 inhibitors, Scaffolds A and B were designed using the ester (Scaffold A) or amide moiety (Scaffold B), as well as oxime moiety to produce hydrogen bonding connections. Moreover, vicinal 1,5-diarylpyrazole seems to be more flexible in occupying both enzymes allosteric hydrophobic sites. Different substitutions (electron donating and withdrawing groups) were used in order to study their SAR. Additionally, the distinction between scaffolds A and B guarantees that the optimal pharmacophore with the best replacement for enzyme binding is preserved. This hybridization was carried out to produce a synergistic impact, increase anticancer effectiveness, and/or lessen adverse effects, if any. 2. Results And Discussion 2.1. Chemistry Claisen condensation of different substituted acetophenone 1a-e with diethyl oxalate in the presence of sodium ethoxide to gives 1,3-dicarbonyl compounds (β-diketoester) gave compounds 2a-e in a good yield. 4-Hydrazinylbenzenesulfonamide hydrochloride 4b was synthesized by diazotization of sulfanilamide with sodium nitrite and hydrochloric acid followed by reduction with sodium sulfite in the presence of sodium hydroxide and hydrochloric acid [ 41 ]. 1,5-Diarylpyarzole carboxylate derivatives 5a-j were synthesized by condensation of 1,3-dicarbonyl compounds (β-diketoester) 3a-e with phenyl hydrazine 4a directly or in the presence of sodium acetate as in compound 4b. Hydrolysis of 1,5-diarylpyrazole ester 5a-j derivatives with alcoholic potassium hydroxide yielded 1,5-diarylpyrazole carboxylic acid derivatives 6a-j [ 42 ]. Compounds 7a-j were synthesized according to Steglich esterification by coupling of 1,5-diarylpyarzole carboxylic acid derivatives 6a-j with 4-hydroxy-3-methoxy acetophenone using EDC as coupling agent and HOBt as additives in presence of DIPEA as base (Scheme 1 ). The structures of the synthesized compounds 7a-j were confirmed by IR, 1 H-NMR, 13 C-NMR and HRMS (ESI) spectroscopy. The IR spectra showed significant stretching bands at 1658–1746 cm -1 related to carbonyl of ester group ( CO O-Ph) and at 1162–1164 cm -1 for compounds 7f-j related to (SO 2 NH 2 ) group. The 1 H-NMR spectra of compounds 7a-j showed two common singlet peaks at δ 3.71–3.85 ppm related to methoxy group of acetophenone and at δ 2.51–2.75 ppm attributed to (CO- CH 3 ). The 13 C-NMR spectra of compounds 7a-j showed significant signals related to carbonyl carbon of ketone ( CO -CH 3 ) appeared at δ 197.11–197.80 ppm. Peak at δ 159.99-167.77 ppm was related to carbonyl carbon of ester ( CO O-Ph), at δ 54.96–57.14 ppm attributed to carbon of methoxy group that attached to acetophenone moiety and at δ 26.65–28.85 ppm attributed to carbon of methyl group (CO- CH 3 ). HRMS (ESI) data for compounds 7a-j further confirmed their assigned structure. The m/z value of molecular ion peak [M + 1] + or [M + Na] + was close to calculated one in all target compounds. The target oxime derivatives 8a-j were prepared by refluxing a mixture of ketone intermediates 7a-j and hydroxylamine hydrochloride in absolute ethanol. The chemical structure of the prepared compounds was elucidated by IR, 1 H-NMR, 13 C-NMR and HRMS spectroscopy. The IR spectra of compounds 8a-j were characterized by the appearance of intense broad bands at 3139–3681 cm -1 related to OH group, in addition to (COO-Ph) that exhibited stretching vibration at 1717–1756 cm -1 . A characteristic feature of the 1 H-NMR spectra for oximes 8a-j is the appearance of downfield singlets in the range δ 10.36–11.34 ppm related to the hydroxyl group. The resonances of CH 3 protons were observed in the expected regions at δ 1.90–2.28 ppm and appeared to be more upfield shifted than the CH 3 protons of the corresponding ketones by 0.40–0.60 ppm due to the low electronegativity of N atom of the oxime relative to O atom of the ketone. Also, all the aromatic protons appeared in their expected chemical shift. One of the characteristic features of 13 C-NMR spectra of compounds 8a-j is the disappearance of ketonic carbonyl due to its conversation to ketoxime group (C = N-OH), which appeared at δ 150.82-159.99 ppm. Also, the methyl group attached to ketoxime appeared at δ 11.65–14.71 ppm. HRMS (ESI) data for compounds 8a-j further confirmed their assigned structure. The m/z value of molecular ion peak [M + 1] + or [M + Na] + was close to calculated one in all cases. Compounds 9a-c were synthesized by activating 1,5-diarypyrazole carboxylic acid derivatives 6f , 6h and 6j with thionyl chloride in benzene to obtain acyl chloride derivatives, which were coupled with 4-aminoacetophenone by heating in dry DMF in presence of triethylamine as base ( Scheme 2 ). The structure of the synthesized compounds 9a-c was confirmed by IR, 1 H-NMR, 13 C-NMR and HRMS (ESI) spectroscopy. The IR spectra showed significant stretching bands at 1669–1681 cm -1 assigned to ( CO NH) and at 1160–1161 cm -1 related to (SO 2 NH 2 ). In the 1 H-NMR spectra for compounds 9a-c two singlet peaks are common and appeared at δ 10.53–11.18 ppm related to the amidic (NH) proton and at δ 2.47–2.53 ppm related to (CO- CH 3 ). The 13 C-NMR spectra of compounds 9a-c showed significant signals related to carbonyl carbon of ketone ( CO -CH 3 ) appeared at δ 195.96-197.09 ppm, peak at δ 164.12-167.91 ppm related to carbonyl carbon of amide ( CO NH) and at δ 26.61–26.95 ppm related to carbon of methyl (CO- CH 3 ). HRMS (ESI) data for compounds 9a-c confirmed their assigned structure. The m/z value of molecular ion peak [M-1] - or [M + Na] + was close to the calculated one in all cases . Oxime derivatives 10a-c were synthesized by refluxing a mixture of ketone intermediates 9a-c and hydroxylamine hydrochloride in absolute ethanol. The chemical structure of the prepared compounds was elucidated by IR, 1 H-NMR, 13 C-NMR and HRMS spectroscopy. The IR spectra of compounds 10a-c was characterized by the appearance of intense broad bands at 3220–3681 cm -1 related to OH and NH groups, in addition to ( CO -NH) and (SO 2 NH 2 ) groups that exhibited stretching vibration at 1677–1680 cm -1 and 1161–1162 cm -1 , respectively. A characteristic feature of the 1 H-NMR spectra for oximes 10a-c was the appearance of downfield singlets in the range δ 8.75–10.9 ppm related to the hydroxyl group. The resonances of NH and CH 3 protons were observed in the expected regions at δ 10.30-10.36 ppm and δ 2.06–2.08 ppm, respectively. The CH 3 protons appeared to be more upfield shifted than the CH 3 protons of the corresponding ketones by 0.40–0.45 ppm due to the low electronegativity of the N atom of the oxime relative to O atom of the ketone. One of the characteristic features of 13 C-NMR spectra of compounds 10a-c is the disappearance of ketonic carbonyl due to its conversation to ketoxime group (C = N-OH), which appeared at δ 153.24-159.18 ppm. Also, the methyl group attached to ketoxime appeared at δ 12.14–17.34 ppm. The HRMS (ESI) data for compounds 10a-c further confirmed their assigned structure. The m/z value of the molecular ion peak [M-1] - or [M + 1] + was close to calculated one in all target compounds. 2.2 Biology 2.2.1. In vitro antiproliferative screening activities. Compounds 7a-j, 8a-j, 9a-c , and 10a-c were evaluated for in vitro anticancer activities against different five cancer cell lines namely, human colorectal adenocarcinoma cell lines DLD-1, human cervical cancer cell line Hela, human pancreatic cancer cell line SUIT-2, human myelogenous leukemia cell line K562 and human hepatocellular carcinoma cell line HepG2 using WST-8 assay at concentration of 100 µM to investigate the growth inhibition percent (GI%) of each compound using daunorubicin as reference drug [ 43 ] (Table 1 ). Table 1 Antiproliferative activity of compounds 7a-j , 8a-j , 9a-c, 10a-c and daunorubicin against DLD-1, Hela, K562, SUIT-2 and HepG2 cell lines at 100 µM using WST-8 assay. Compound no. Growth inhibition (GI%) DLD-1 Hela K562 SUIT-2 HepG2 7a 0 55 0 101 5 7b 81 83 70 103 85 7c 0 52 48 68 0 7d 72 104 50 104 31 7e 59.40 73 0 101 14 7f 18.65 16 24 75 10 7g 96.70 30 77 92 66 7h 0 0 0 73 0 7i 46.00 101 101 101 63 7j 0.10 10 11 81 0 8a 25.70 33 94 77 0 8b 92.00 60 99 102 107 8c 0 78 67 61 51 8d 72.00 48 95 77 98 8e 29.20 77 93 83 99 8f 72.90 107 76 95 80 8g 85.80 78 87 96 93 8h 52.30 7 78 73 63 8i 34.00 100 97 98 90 8j 9.60 53 34 79 60 9a 60.70 9 66 60 36 9b 42.15 0 31 45 19 9c 16.80 9 57 42 22 10a 99.00 0 84 88 72 10b 72.50 2 84 50 2 10c 92.40 75 87 103 52 Daunorubicin 82.45 100 100 92 100 From the screening results in Table 1 , compounds 7a-j and 9a-c that are 1,5-diarylpyrazole acetophenone derivatives displayed considerable cytotoxicity towards the pancreatic cell line SUIT-2 with GI% ranging from 68–104% for compounds 7a-j and 42–60% for compounds 9a-c . Compounds 7b ( R 1 = p -CH 3 , R 2 = H) demonstrated moderate to high cytotoxicity against five cancer cell lines with GI% ranging from 46–103%, while compound 7d ( R 1 = p -Cl, R 2 = H) established an excellent cytotoxicity towards DLD-1, Hela, and SUIT-2 with GI% of 72%, 104%, and 104%, respectively, and moderate cytotoxicity against K562 cell line with GI% 50. Furthermore, compound 7i ( R 1 = p -Cl, R 2 = SO 2 NH 2 ) displayed superior antiproliferative activity against Hela, K562 and SUIT-2 cell lines with GI% of 101%,101% and 101% and moderate activity against HepG2 cell line with GI% of 63%. Importantly, all oxime derivatives 8a-j and 10a-c exhibited marked antiproliferative activity in comparison to ketone derivatives 9a-c and 11a-c as a result of the role of oxime moiety in cytotoxicity. Compounds 8b-j demonstrated a significant cytotoxicity against SUIT-2 and HepG2 cell lines with GI% ranging from 51–107%. Compounds 8b ( R 1 = p -CH 3 , R 2 = H), 8f ( R 1 = H, R 2 = SO 2 NH 2 ) and 8g ( R 1 = p -CH 3 , R 2 = SO 2 NH 2 ) exhibited broadness cytotoxicity in five cancer cell lines with GI% ranging from 60–107%. Moreover, compounds 8a-i exhibited high antiproliferative activity against leukemia cell line K562 with GI% ranging from 67–99%. Compound 8e ( R 1 = 3,4-OCH 3 , R 2 = H) displayed remarked cytotoxicity in Hela, K562, SUIT-2 and HepG2 with GI% of 77%, 93%, 83% and 99% respectively, while compound 8i ( R 1 = p -Cl, R 2 = SO 2 NH 2 ) exhibited high antiproliferative activity against Hela, K562, SUIT-2 and HepG2 with GI% of 100%, 97%, 98% and 90% respectively. Furthermore, compounds 10a-c showed moderate to high cytotoxicity in pancreatic cancer cell line SUIT-2 with GI% of 88%, 50% and 103% respectively, only compound 10c ( R = 3,4-OCH 3 ) demonstrated antiproliferative activity in the five cancer cell lines with GI% ranging from 52–103%. So as a conclusion on the SAR study of those compounds as antiproliferative agents, oxime moiety potentiated the anticancer activity and electron donating groups on R 1 played an important role on the activity. Moreover, the sulfamoyl moiety on R 2 seems to be of potential role in the activity of the prepared compounds. 2.2.2 In vitro cytotoxicity measurements (IC 50 ) against five cancer cell lines. For further investigation, compounds with committed antiproliferative activity against five cancer cell lines (DLD-1, Hela, K562, SUIT-2 and HepG2) at 100 µM were selected to measure growth inhibition percentage using WST-8 assay at different six concentrations of 1, 10, 20, 50, 80, 100 µM for calculating their IC 50 using the daunorubicin as reference. All selected compounds and daunorubicin were recorded as the minimum concentration required to inhibit half cell growth (IC 50 ) and results were listed in Table 2 . Table 2 In vitro antiproliferative activity of the most active compounds expressed as IC 50 values using WST-8 assay against DLD-1, Hela, K562, SUIT-2 and HepG2 cancer cell lines. The results recorded as IC 50 (µM) using daunorubicin as reference. Compound no. IC 50 (µM) DLD-1 Hela K562 SUIT-2 HepG2 7b 13 55 NT 45 45 7d 81 15 NT 43 NT 7g 13 NT NT 31 NT 7i ND 25 93 92 NT 8a NT NT 22 NT NT 8b 10 32 13 27 35.7 8d 14.4 57 9 NT 23.3 8e NT 22 20 NT 4.7 8f NT 22 15.6 NT 22.3 8g 32.3 8 7.6 19 12.3 8h NT 74 21 26 NT 8i ND 13 71 62 ND 10a 26 NT 16 NT ND 10b 36 NT ND NT NT 10c NT 5 29 13 NT Daunorubicin 30 0.097 13.30 9 22 a: ND means not determined, b: NT means not tested The target oxime derivatives exhibited promising antiproliferative activity against five cancer cell line as listed in Table 2 more than the corresponding ketone, such as compounds 8b ( R 1 = p -CH 3 , R 2 = H), 8d ( R 1 = p -Cl, R 2 = H), 8g ( R 1 = p -CH 3 , R 2 = SO 2 NH 2 ), 10a ( R = H), and 10b ( R = p - OCH 3 ) that showed remarkable cytotoxicity against DLD-1 cell line with IC 50 of 10, 14.4, 32.30, 26 and 36 µM, respectively in comparison to daunorubicin (IC 50 = 30 µM). Also, compounds 8g , 8i ( R 1 = p -CH 3 , R 2 = SO 2 NH 2 ) and 10c ( R 1 = 3,4-di-OCH 3 ) demonstrated high antiproliferative activity against Hela cell line with IC 50 of 8, 13, and 5 µM respectively, while compounds 8b, 8d, 8e ( R 1 = 3,4-OCH 3 , R 2 = H), 8f ( R 1 = H, R 2 = SO 2 NH 2 ) and 8h ( R 1 = p -OCH 3 , R 2 = SO 2 NH 2 ) showed a moderate antiproliferative activity with IC 50 of 32, 57, 22, 22 and 74 µM in comparison with daunorubicin. Moreover, compounds 8b , 8d , 8f , 8g and 10a established excellent anticancer activity in comparison to daunorubicin (IC 50 = 13 µM) against the human myelogenous leukemia cell line K562 with IC 50 of 13, 9, 15.6, 7.6 and 16 µM. Compounds 8a ( R 1 = H, R 2 = H), 8e , 8h , and 10c exhibited good anticancer activity at the same cell line with IC 50 of 22, 20, 21 and 29 µM. Furthermore, compounds 8b , 8g , 8h and 10c showed a significant anticancer activity against the human pancreatic cancer cell line SUIT-2 with IC 50 of 27, 19, 26 and 13 µM, respectively. Compounds 8e and 8g demonstrated excellent antiproliferative activity better than daunorubicin (IC 50 = 22 µM) against the hepatocellular carcinoma cell line HepG2 with IC 50 of 4.7 and 12.3 µM, respectively. In addition, compounds 8d and 8f showed equal anticancer activity to daunorubicin at the same cell line with IC 50 of 23.3 and 22.3 µM, respectively. Unlike to anticancer activity of ketone derivatives 7a-j , compounds 7b ( R 1 = p -CH 3 , R 2 = H) and 7g ( R 1 = p -CH 3 , R 2 = SO 2 NH 2 ) displayed excellent antiproliferative activity in comparison with daunorubicin against human colorectal adenocarcinoma DLD-1 with IC 50 of 13 µM. Also, compound 7d established remarkable anticancer activity towards the human cervical cancer cell line Hela with IC 50 of 15 µM. Substituents on the terminal phenyl ring of the 1,5-diarylpyrazole part, showed a significant effect on the biological profile of anticancer activity. Compounds 8b , 8f, 8g, 8h and 8i , which are considered the most potent anticancer oxime derivatives showed substituents on R 1 = H, p -CH 3 and p -Cl, but when R 1 = OCH 3 , moderate anticancer activity was observed as in compound 8h , which indicated the presence of a lipophilic group (Cl, CH 3 ) improving anticancer activity Also, the sulfamoyl group at the para position of the phenyl ring of the diarylpyrazole part is essential for the broadness of anticancer activities such as compounds 8g , 8i , 8h and compound 10c as results of hydrogen bonding formation on the active site. Meanwhile, compound 10c , R 1 = 3,4-di OCH 3 established good anticancer activity with (IC 50 = 5–29 µM) and the remaining compounds in scaffold B showed weak anticancer activity. The difference between the hydrophilic and hydrophobic substitutions in scaffold A, as well as the presence of the methoxy group in the 4-hydroxy-3-methoxyl acetophenone carrying oxime moiety, led to scaffold A superior anticancer efficacy compared to scaffold B. The anticancer properties of 3,4-di-OCH3-containing compounds, however, were goo in both scaffolds. 2.2.3. Evaluation of EGFR inhibitory activity Being over-expressed in a variety of human cancers and connected to cancer proliferation, angiogenesis, and metastasis, EGFR has received substantial study and clinical validation as a target for cancer treatment. Most of these medications are designed to bind to the ATP active site of EGFR-TK. The structural study of previously reported instances of tyrosine kinase anticancer medications served as the basis for the introduction of such drugs [ 4 ]. Herein, the human EGFR-TK Elisa kit assay was performed to evaluate the in vitro EGFR-inhibitory potency of the more active anticancer compounds 8b , 8d , 8g , 8i and 10c using the multi-target kinase inhibitor drug sorafenib as a reference using quantitative sandwich enzyme immunoassay technology [ 44 ]. The test investigated the potential of the test compounds to bind to EGFR, resulting in suppression of epidermal growth factor from binding to EGFR that led to inhibition of receptor dimerization and tyrosine autophosphorylation and suppression of cancer cell proliferation. Screening results of EGFR inhibitory activity of tested compounds and sorafenib expressed as IC 50 in µM and recorded in Table 3 . Results showed that compound 8g ( R 1 = p- CH 3 , R 2 = SO 2 NH 2 ), 8i ( R 1 = p- Cl, R 2 = SO 2 NH 2 ) and 10c (R = 3,4-di OCH 3 ) exhibited moderate EGFR inhibitory activity with IC 50 of 18, 21 and 12 µM, respectively. On the other hand, compound 8d ( R 1 = p- Cl, R 2 = H) displayed successful EGFR inhibitory activity with IC 50 of 8 µM in comparison to positive control drug sorafenib (IC 50 = 3.5 µM). These findings revealed that lipophilic substitutions on vicinal 1,5- diarylpyarzole, together with the oxime moiety as in compound 8d , have good fitting and binding on EGFR, making it an effective EGFR inhibitor. Table 3 EGFR-TK inhibitory activity of target compounds represented as IC 50 using sorafenib as reference. Compound no IC 50 (µM) 8b > 1000 8d 8 8g 18 8i 21 10c 12 sorafenib 3.5 2.2.4. Evaluation of JNK-2 inhibitory activity JNK-2 is a member of the MAP kinase family involved in signaling pathways, which has been implicated in several diseases like cancer and inflammatory diseases [ 13 ]. So, this family is widely considered for targeting by small molecule therapeutics [ 13 ]. JNK-2 inhibitors are classified to two types: Type I inhibitors target the DFG-in conformation (open conformation), while type II inhibitors target the DFG-out conformation (closed conformation) [ 13 , 14 ]. Based on the vital role of JNK-2 in human malignancies as a result of its contribution to multiple cancer-related pathways and the reported role of celecoxib as JNK inhibitor, we examined the JNK-2 inhibitory activity of compounds 8d , 8g and 8i using the multitarget kinase drug sorafenib as reference [ 45 , 46 ]. The JNK-2 inhibitory activity of the selected compounds was investigated in vitro using a simple step ELISA kit for quantitative measurement of JNK-2 (pT138/Y185) protein in human cells, which investigated the possible binding of tested compounds in the ATP binding site of JNK-2, leading to inhibition of substrate binding on JNK-2 enzyme and inhibition of the JNK-2 pathway that could explain the antiproliferative activity of these compounds [ 47 ]. Screening results of JNK-2 inhibitory activity of tested compounds and sorafenib expressed as IC 50 in µM and recorded in Table 4 . Results showed that the oxime derivative 8i ( R 1 = p -Cl, R 2 = SO 2 NH 2 ) is a potent inhibitor of JNK-2 with an IC 50 of 1 µM same as the activity of the multi-target kinase sorafenib. These results indicated that the anticancer activity of compound 8i was due to the dual inhibition of EGFR and JNK-2. Also, compound 8d ( R 1 = p -Cl, R 2 = H) showed moderate JNK-2-inhibiting activity with an IC 50 of 49 µM, which indicated its anticancer activity because of dual inhibition of EGFR and JNK-2. It was clear from these data that the oxime moiety and p -Cl substitution improved the anticancer activity of two compounds in addition to the sulfamoyl moiety that makes compound 8i more potent as JNK-2 inhibitor. Table 4 JNK-2 inhibitory activity of target compounds represented as IC 50 using sorafenib as reference. Compound no IC 50 (µM) 8d 49 8g > 200 8i 1 Sorafenib 1 2.2.5. Cell cycle analysis and apoptosis detection 2.2.5.1. Cell cycle analysis This analysis was applied to investigate the effects of tested compounds on cell cycle distribution and on cell-death associated DNA fragmentation. The Hela cell line was analyzed flow cytometrically after propidium iodide (PI) staining following treatment of compounds 8g and 8i . As shown in Fig. 4 , compound 8g established G2/M arrest of Hela cancer cells indicating cell death and DNA fragmentation and the percentage of Hela cells at G2 phase increased from 7.3% (DMSO treated Hela cell) to 16.5% after 24h and 20.5% after 48h. While compound 8i showed a combined S phase and G2 phase arrest and the percentage of Hela cells at S phase increased from 8.47% (control) to 59.4%, while the percentage of Hela cells at G2 phase raised from 7.3% (control) to 18.2%. 2.2.5.2. Apoptosis assay For further investigation of the anticancer activity of compounds 8g and 8i , studies of apoptotic changes after treatment of Hela cells with these inhibitors were examined using fluorescent microscope and flowcytometry. Moreover, to discriminate between apoptosis and necrosis after treatment of Hela cell with two inhibitors 8g and 8i at different concentrations, a fluorescent microscope was used to distinguish between apoptosis and necrosis based on their characteristic difference in morphology using annexin V and PI [ 48 ]. As shown in Fig. 5 , compounds 8g and 8i established remarkable apoptosis at low concentrations that was marked in green because of the binding of annexin-v with phosphatidylserine. This was exposed in the cell membrane of Hela cells after treatment with two the inhibitors due to plasma membrane sprouting and chromatin concentration. Prolonged incubation of a Hela cells with two the inhibitors directed cells to necrosis, which was marked in red because of PI staining of necrotic cells as a result of losing their dye-excluding ability owing to loss of plasma membrane integrity and dissolution of nuclear chromatin [ 49 ]. Subsequently, flowcytometric analysis of the effect of compounds 8g and 8i on Hela cell apoptosis at concentration double the IC 50 of each compound for 24h using Annexin v/PI were investigated and apoptotic marker changes for each compound on Hela cells were analyzed in comparison to control untreatable Hela cells. As appeared in Figs. 6 and 7 the parentage of apoptotic Hela cells was increased significantly after treatment with compounds 8g and 8i from 1.82% for control untreatable Hela cells to 34.7% for compound 8g and 17.3% for compound 8i . These results attributed that those apoptotic cells were increased after treatment of these compounds as result of antiproliferative activity not due to cytotoxicity. 2.2.6. Evaluation of cytotoxicity towards the normal cell line PC12 Evaluation of normal cell viability is fundamental to analyze the efficacy of target compounds and is often used in conjunction with cytotoxicity tests to help understand how target compounds toxicity affects normal cell health. To investigate the selectivity of the target compounds towards cancer cells, the cytotoxicity of compounds 8g and 8i was measured against the normal cell line PC12 (rat adrenal-derived pheochromocytoma cells) using a WST-8 assay at six concentrations 1, 10, 20, 50, 80, 100 µM to calculate CC 50 in comparison to daunorubicin as reference drug as recorded in Table 5 . The results listed in Table 5 showed that no cytotoxicity was observed with compound 8i against PC12 cell line with CC 50 > 100 µM. While compound 8g showed a moderate cytotoxicity with CC 50 of 16.1 µM in comparison to daunorubicin, which established high cytotoxicity towards PC12 cell line with percentage growth inhibition of 113% at 100µM, 107% at 50µM and 81% at 1µM. These results indicated that these target compounds had no or little cytotoxicity against the normal cell line and demonstrated selectivity towards cancer cells. Table 5 Cytotoxicity of compounds 8g, 8i , and daunorubicin against the PC12 cell line represented as CC 50 . Compound no CC 50 (µM) 8g 16.1 8i > 100 Daunorubicin NA 2.3. Measurement of nitric oxide release Griess colorimetric method was used for the indirect determination of NO, which includes spectrophotometry measurements of the stable decomposition products NO 2 − and NO 3 − . This method requires that NO 3 − first reduced to NO 2 − and then NO 2 − is determined by the Griess reaction that includes two steps, the first step is a diazotization reaction where the NO-derived nitrogen agent, dinitrogen trioxide (N 2 O 3 ) resulting from the spontaneous oxidation of NO with sulfanilamide to produce the diazonium ion. The second step is the coupling of diazonium with N -(1-napthyl)ethylenediamine dihydrochloride (NEDD) to form a strongly absorbed azo colorimetric product at λ max 546 nm. In order to evaluate thiol-induced NO generation from the appropriate compounds including NO-donating oximes 8a-b, 8d-i and 10a-c , they were incubated in aqueous phosphate buffer of pH 7.4 in the presence of excess N -acetylcysteine, which serves as a source of thiols that are essential for release of NO from oximes [ 50 ]. The signal intensity of the dye is proportional to the amount of NO released. To quantify the amount of NO released, a standard curve was made by measuring the change in absorbance of various concentration of standard sodium nitrite solutions treated by the same way. The results expressed as amount of NO released (mol/mol) and listed in Table 6 . The obtained results indicated that the NO-donating oximes 8a-b, 8d-i and 10a-c achieved maximum amount of NO released after 2 hours. Table 6 The amount of NO released from compounds 8a-b, 8d-i and 10a-c in phosphate buffer of pH = 7.4. Compound No. Amount of NO released (mol/mol) 1h 2h 3h 4h 8a 0.095 ± 0.036 0.121 ± 0.047 0.108 ± 0.042 0.056 ± 0.022 8b 0.102 ± 0.039 0.159 ± 0.019 0.105 ± 0.025 0.095 ± 0.030 8d 0.096 ± 0.036 0.106 ± 0.040 0.101 ± 0.038 0.079 ± 0.030 8e 0.064 ± 0.022 0.120 ± 0.046 0.081 ± 0.030 0.048 ± 0.018 8f 0.096 ± 0.036 0.116 ± 0.045 0.079 ± 0.030 0.048 ± 0.018 8g 0.083 ± 0.035 0.154 ± 0.044 0.141 ± 0.039 0.089 ± 0.019 8h 0.099 ± 0.029 0.158 ± 0.014 0.104 ± 0.033 0.094 ± 0.0022 8i 0.105 ± 0.029 0.115 ± 0.018 0.109 ± 0.046 0.087 ± 0.035 10a 0.054 ± 0.023 0.108 ± 0.043 0.069 ± 0.024 0.036 ± 0.012 10b 0.095 ± 0.035 0.114 ± 0.046 0.079 ± 0.031 0.031 ± 0.018 10c 0.075 ± 0.036 0.151 ± 0.049 0.114 ± 0.030 0.081 ± 0.020 The data recorded in Table 6 indicated that the NO-donating oximes 8a, 8b, 8d, 8e, 8f, 8g, 8h, 8i and 10a-c achieved maximum amount of NO release after 2 hours. At the first hour the amount of nitric oxide released is increased in compounds 8i , 8b and 8h , while maximum release of nitric oxide occurred in compounds 8b and 8h was at 2 hours. Then, the amount of nitic oxide released was declined, which may explain the biological important of oxime moiety as source of nitic oxide release in comparison to ketone intermediates 7a-j and 9a-c as shown in anticancer activity of oxime derivatives. 2.4. Docking 2.4.1. In silico molecular docking study into EGFR For mechanistic investigation of antiproliferative activity of compounds 8d, 8g, 8i and 10c , the in silico simulation studies targeting EGFR tyrosine kinase domain (Fig. 8 ) using sorafenib, a multitarget kinase inhibitor drug, as reference was done. For the Epidermal Growth Factor Receptor (EGFR), the structural models of the selected ligands were built against the human EGFR complexed with AZD9291 inhibitor (2.80 Å; PDB ID: 4ZAU)[ 49 ]. Since the docking simulation scored the ligands based on the structural compatibility and the electrostatic potential, the ligands differentially bound the active site of EGFR at different affinities, as illustrated in Table 7. Table 7 Calculated binding properties of test compounds and sorafenib against EGFR. Test compounds Binding score Hydrogen bonding energy (Kcal/mol) Hydrophobic interaction energy (Kcal/mol) Sorafenib -38.13 -2.52 -7.12 8d -32.33 -1.19 -7.97 8g -16.46 -2.25 -7.62 8i -12.98 -2.47 -7.50 10c -23.74 -3.36 -7.35 As recorded in Table 7 , the predicted binding affinities, in turn, come with the evaluated inhibitory values of the EGFR enzymatic activity (Table 3 ). Sorafenib and oxime ligands ( 8d, 8g, 8i and 10c ) bound the active site by hydrogen bonding and hydrophobic interactions with respect to sorafenib, which had the highest affinity to the active site. Importantly, the amine group of M793 residue acts as a hydrogen donor that forms hydrogen bonding with the carbonyl groups of the ligands and sorafenib. Like sorafenib, compound 8d was shown to occupy the active site with a binding affinity higher than the other compounds due to its flexibility to fit the active site ( Figs. 9 and 10 ) . The sandwiching effect was achieved by the hydrophobic contacts formed by the non-polar residues L718, V726, A743 and L792. Compound 10c showed a relatively similar binding score but without Van der Waals forces ( Fig. 13 ) . Compounds 8g and 8i showed the same affinity levels to bind the active site. ( Figs. 11 and 12 ) . 2.3.2 . In silico molecular docking study on JNK-2 For further mechanistic investigation of the antiproliferative activity, compounds 8d and 8i were investigated for their in silico simulation studies targeting the JNK-2 binding pocket (Fig. 14 ) using sorafenib as reference drug. JNK-2 inhibitors are classified to two types: Type I inhibitors target the DFG-in conformation (open conformation), while type II inhibitors target the DFG-out conformation (closed conformation) [ 13 , 14 ]. Based on open confirmation, we docked the selected compounds against a pre-defined open conformation of JNK-2 (Fig. 14 ) [ 13 ]. Sorafenib is a common MAP kinase inhibitor, and it was shown to have a higher binding affinity to JNK-2 active site compared to the other ligands (Table 8 ). Sorafenib extends at the active site from the hinge region (L110 and M111) to the DFG conformation (D169), which makes it structurally compatible to fit the active site (Fig. 14 ). Sorafenib binds to the ATP site of JNK-2 and forms two hydrogen bonds: one with M111 of the hinge and the other with K55 of the N -terminal b3-strand. In addition, the binding of sorafenib is supported by Van der Waals forces with the hinge residues E109 and Q117 as well as hydrophobic interactions. Like sorafenib, compound 8i is bound to the active site by the same machinery, forming hydrogen bonds with M111 and K55 ( Figs. 15 and 16 ). However, the weaker affinity of compound 8i than that of sorafenib comes from the lesser extent of compound 8i to the DFG conformation, resulting in weaker hydrophobic interactions. Furthermore, the sulfonamide group of compounds 8i was shown to be extended outside the binding pocket near the hinge region, describing the group's lower interactivity. Compound 8i binding is supported by Van der Waals forces with the hinge region (E109, N114 and Q117). However, the binding energy is lower than that of sorafenib. Compound 8i binds to the hinge region by only one hydrogen bond with M111 supported by Van der Waals force with Q117 ( Fig. 16) . On the other hand, compound 8d hydrophobically binds with the hinge region with extension to the N -terminal b3-strand, where it forms a hydrogen bond with K55 and a Van der Waals contact with E109 ( Fig. 17 ). The structural incompatibility of compound 8d results in a weaker binding score, which in turn, results in weaker binding energy and consequently, a weaker inhibitory effect. Table 8 Calculated binding properties of test compounds andsorafenib against JNK-2. Test compounds Binding score Hydrogen bonding energy (Kcal/mol) Hydrophobic interaction energy (Kcal/mol) Van der Waals interaction energy (Kcal/mol) Sorafenib -21.48 -2.72 -8.46 -35.60 8d -9.16 -0.99 -7.97 -33.70 8i -12.00 -3.19 -7.66 -32.59 3. Conclusion A series of 1,5-diarylpyrzole derivatives targeting EGFR and JNK-2 were developed and synthesized and biologically evaluated for their anticancer activity against a panel of five cancer cell lines, namely human colorectal adenocarcinoma cell line DLD-1, human cervical cancer cell line Hela, human leukemia cell line K562, human pancreatic cell line SUIT-2 and human hepatocellular carcinoma cell line HepG2. Oxime derivative compounds 8a-j and 10a-c showed better anticancer activity than their corresponding ketones. Regarding substituents on the terminal phenyl ring of the 1,5-diarylpyrazole part, they showed a significant effect on the biological profile of anticancer activity especially when R 1 = H, p -CH 3 and p -Cl, such as compounds 8b , 8f, 8g, 8h and 8i , which are considered the most potent anticancer oxime derivatives. Also, oxime derivatives 8g , 8i and 10c exhibited moderate EGFR inhibitory activity with IC 50 of 18, 21 and 12 µM respectively, while compound 8d displayed good EGFR inhibitory activity with IC 50 of 8 µM. Moreover, compound 8i showed potent JNK-2 inhibitory activity with IC 50 = 1.00 µM, similar to the positive reference drug sorafenib. Selectivity of compounds 8i and 8g towards cancer cells rather than normal cells was evaluated and compound 8i observed no cytotoxicity against the PC12 cell line with CC 50 > 100 µM, while compound 8g showed moderate cytotoxicity with CC 50 of 16.1 µM in comparison to reference drug daunorubicin. Furthermore, compound 8g exhibited cell cycle arrest at the G2/M phase in the cell cycle analysis of the Hela cell line, while compound 8i showed combined S phase and G2 phase arrest. Additionally, Hela cell apoptotic changes after treatments of compounds 8g and 8i were investigated by fluorescent microscope and flowcytometry as results of their antiproliferative activity. Lastly, in silico molecular docking studies showed a good fitting of compounds 8d , 8g , 8i and 10c at the EGFR binding site with binding free energies in the range of -12.98 to 32.30 kcl/mol, while compounds 8d and 8i showed binding free energies in range of -9.16 to -12.00 kcl/mol at the JNK-2 binding site. 4. Experimental 4.1. Chemistry Material and equipment All chemicals used for the preparation of the target compounds are of analytical grade and can be used without further purification. Solvents were purified and freshly distilled before use according to the standard procedures. Reaction progress was monitored by thin layer chromatography (Merck Silica gel 60 F254) on glass plates and visualized with a UV lamp (254 nm). Column chromatography was performed using spherical, neutral silica gel of diameter 40–100 µm (Kanto chemical co. Inc., Tokyo, Japan). Melting points were recorded at a ATM-02 (AS ONE, Tokyo, Japan). IR spectra were recorded at FT/IR-Spectrum Two (PerkinElmer, MA, USA) at the Faculty of Engineering, Yamagata University, Yonezawa, Japan. 1 H-NMR (400 or 500 MHz) and 13 C-NMR (100 or 125 MHz) spectra were recorded on either a JNM-ECX400 or JNM-ECX500 (JEOL, Tokyo, Japan) in Faculty of Engineering, Yamagata University, Yonezawa, Japan. Chemical shifts are reported in ppm relative to tetramethylsilane (0 ppm), chloroform (7.26 ppm: 1H, 77.1 ppm: 13C) and dimethyl sulfoxide (2.50 ppm: 1H, 39.6 ppm: 13C). Coupling constant ( J ) is measured in hertz (Hz). Multiplicity was designated as: s, singlet; d, doublet; t, triplet; q, quartet; p, pentet; dd, doublet of doublet; and m for multiplet. Mass spectra (ESI-MS) were carried out using the AccuTOF JMS-T100LC (JEOL, Tokyo, Japan) at the Faculty of Engineering, Yamagata University, Yonezawa, Japan. 4.1.1. General procedure for the synthesis of ethyl 4-(substituted phenyl)-2-hydroxy-4-oxobut-2-enoates (2a-e). A mixture of diethyl oxalate (2.92 g, 0.02 mol) and substituted acetophenone derivatives (0.01 mol) in ethanol (50 mL) was added to previously prepared sodium ethoxide (sodium, 0.46 g, 0.02 mol, ethanol 100 mL) at 50°C. The reaction mixture was heated under reflux for 2–3 h. After cooling, the solvent was removed, and the residue was taken up in water (200 mL) and acidified with concentrated HCl (1 mL). The aqueous mixture was extracted with ethyl acetate (3x150 mL). The combined extracts were washed with brine (100 mL), dried (MgSO 4 ), and concentrated. The obtained solid was recrystallized from methanol to give compounds 2a-e and the produced compounds used in the next step without further purification [ 51 – 53 ]. 4.1.2. General procedure for synthesis of 4-hydrazinylbenzenesulfonamide hydrochloride 4b. A cold, stirred mixture of sulfanilamide (3.42 g, 0.02 mol), hydrochloric acid (10 mL) and crushed ice (200 g) was diazotized by the dropwise addition of sodium nitrite (1.4 g, 0.02 mol) in water (25 mL) over 30 min. The cold diazonium salt solution thus formed was rapidly added to a well-cooled solution of sodium sulfite (2.52 g) and sodium hydroxide (0.800g) in water (50 mL) with vigorous stirring, and the resulting mixture was left in the ice bath for 15 min, followed by acidification with 10 mL HCl and concentrated. The precipitated 4-hydrazineylbenzenesulfonamide hydrochloride 4b was collected and dried: white crystals; mp :225°C (lit. mp: 225°C); yield 3.9 g (88%) [ 41 ]. 4.1.3. General procedure for the synthesis of ethyl 1,5 diarypyarzole-3-carboxylate (5a-j). A mixture of diketoesters 2a-c (0.01mole) and phenylhydrazine 4a (0.01mole) was dissolved in suitable amount of absolute ethanol (40 mL) and refluxed for 5 h to afford compounds 5a-c . A mixture of diketoesters 2d-f and 4-hydrazinylbenzenesulfonamide hydrochloride 4b was refluxed in absolute ethanol for 5 h in the presence of sodium acetate (0.02 mole) to afford compounds 5d-f. The reaction process was monitored by TLC using chloroform: methanol (98: 2) solvent system for compounds 5a-c and chloroform: methanol (95: 5) solvent system for compounds 5d-f . The content of reaction mixture was evaporated under vacuum and the crude product was purified by column chromatography [ 42 ]. Ethyl 1,5-diphenyl-1H-pyrazole-3-carboxylate (5a) : Reddish brown solid; yield (75%); mp: 85–87°C (lit. 86°C)[ 54 ]. Ethyl 1-phenyl-5-(p-tolyl)-1H-pyrazole-3-carboxylate (5b) : Reddish solid; yield (80%), mp: 87–88°C (lit. 84–86°C)[ 55 ]. Ethyl 5-(4-methoxyphenyl)-1-phenyl-1H-pyrazole-3-carboxylate(5c) : Reddish brown solid; yield (81%); mp: 97–99°C (lit. 97°C)[ 51 ]. Ethyl 5-(4-chlorophenyl)-1-phenyl-1H-pyrazole-3-carboxylate (5d) : Reddish brown solid; yield (89%); mp: 92–94°C (lit. 95–97°C)[ 51 ] . Ethyl 5-(3,4-dimethoxyphenyl)-1-phenyl-1H-pyrazole-3-carboxylate (5e) : Brownish solid, yield (63%); mp: 174–176°C (lit. 177°C)[ 56 ]. Ethyl 5-phenyl-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylate (5f) : Reddish powder; yield (66%), mp: 192–194°C (lit. 192)[ 57 ]. Ethyl 1-(4-sulfamoylphenyl)-5-(p-tolyl)-1 H -pyrazole-3-carboxylate (5g) : Reddish brown; yield (75%), mp: 227–228°C (lit. 227°C)[ 58 ]. Ethyl 5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylate (5h) : Reddish brown powder; yield (71%), mp: 207–209°C (lit. 205–207°C)[ 59 ]. Ethyl 5-(4-chlorophenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylate (5i) : Reddish brown powder; yield (80%), mp: 107–109°C (lit. 108°C)[ 59 ]. Ethyl 5-(3,4-dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1H-pyrazole-3-carboxylate (5j) : Reddish brown powder; yield (64%); mp: 214–215°C[ 60 ]. 4.1.4. General procedure for the synthesis of 1,5-diarypyrazole carboxylic acids 6a-j. A mixture of methanolic solution of compounds 5a-j (4 mmol), potassium hydroxide (KOH, 20%, 10 mL) was stirred at 60°C for 4 h. After cooling, the mixture solution was poured into water and acidified with hydrochloric acid solution (1 M) to pH = 3. The aqueous mixture was extracted with ethyl acetate (3x50 mL) and the aqueous layer was discarded. The combined organic extracts were dried with anhydrous MgSO 4 . The organic solvent was evaporated under vacuum to obtain solid products 6a-j [ 61 ]. 1,5-Diphenyl-1 H -pyrazole-3-carboxylic acid (6a) : Brown powder; yield (84%); mp: 180–182°C (lit. 182–183)[ 52 ]. 1-Phenyl-5-(p-tolyl)-1 H -pyrazole-3-carboxylic acid (6b) : Reddish powder; yield (87%); mp: 171–172°C[ 62 ]. 5-(4-Methoxyphenyl)-1-phenyl-1 H -pyrazole-3-carboxylic acid (6c) : Reddish brown powder; yield (79%); mp: 192–195°C (lit. 196–197°C)[ 60 ]. 5-(4-Chlorophenyl)-1-phenyl-1 H -pyrazole-3-carboxylic acid (6d) : Yellowish brown powder; yield (78%); mp: > 300°C[ 58 ]. 5-(3,4-Dimethoxyphenyl)-1-phenyl- 1H -pyrazole-3-carboxylic acid (6e) : Brown powder; yield (84%); mp: 213–214°C; 1 H-NMR (400 MHz, DMSO- d 6 ) δ (ppm): 7.94 7.52 (m, 5H, Ar-H), 7.39 (s, 1H, pyrazole-H), 6.94–6.68 (m, 3H, Ar-H), 3.79 (s, 3H, OCH 3 ), 3.77 (s, 3H, OCH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 163.36, 160.36, 145.81, 144.09, 142.55, 130.71, 128.76, 127.45, 126.47, 125.40, 122.05, 120.40, 115.26, 110.45, 56.23, 56.12; ESI-MS m/z [M + H] + for C 18 H 17 N 2 O 4 calculated: 325.1183, found: 325.256. 5-Phenyl-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylica acid(6f) : Yellowish brown powder; yield (78%), mp: 184–186°C (lit. 188°C)[ 61 ]. 1-(4-Sulfamoylphenyl)-5-( p -tolyl)-1 H -pyrazole-3-carboxylic acid (6g) : Reddish brown powder; yield (88%); mp: 194–195°C[ 52 ]. 5-(4-Methoxyphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylic acid (6h) : Brownish powder; yield (73%), mp: 197–198°C[ 63 ]. 5-(4-Chlorophenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylic acid (6i) : Yellowish brown powder; yield (84%); mp: 212–214°C[ 64 ]. 5-(3,4-Dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylic acid (6j) : Reddish brown powder; yield (77%); mp: 206–208°C; 1 H-NMR (400 MHz, DMSO- d 6 ) δ (ppm): 10.76 (s, 1H, OH), 7.99 (s, 1H, Ar-H), 7.91(d, J = 8.00 Hz, 2H, Ar-H), 7.79 (d, J = 8.00 Hz, 2H, Ar-H), 7.65–7.60 (m, 4H, 2Ar-H, SO 2 NH 2 ), 7.43 (s, 1H, pyrazole-H), 3.89 (s, 6H, 2 OCH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 163.36, 159.98, 147.36, 145.23, 141.21, 131.47, 130.17, 128.82, 126.87, 125.54, 122.87, 119.99, 115.14, 107.53, 56.38, 56.21; ESI-MS m/z [M + H] + for C 18 H 18 N 3 O 6 S calculated: 404.0911, found: 404.0271. 4.1.5. General procedure for synthesis of 4-acetyl-2-methoxyphenyl 5-(4-subistituted-phenyl) 1-(4-substituted-phenyl)-1 H -pyrazole-3-carboxylate 7a-j. A mixture of pyrazole carboxylic acid derivatives 6a-j (0.001 mol), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (0.384 g, 0.002 mol), 1-hydroxybenzotriazole (HOBt) (0.306 g, 0.002 mol), were stirred in dry DMF (5 mL) for 30 min, then N,N- diisopropylethylamine (DIPEA) (0.258 g, 0.002 mol) and 4-hydroxy-3-methoxyacetophenone (0.002 mol) were added to the mixture and stirred for 12 h. 20 mL Distilled water was added followed by acidification with dil. HCl. Extraction twice with ethyl acetate and purification were performed by using column chromatography with chloroform as eluent for compounds 7a-e and chloroform: methanol 98:2 for compounds 7f-j. 4-Acetyl-2-methoxyphenyl 1,5-diphenyl-1 H -pyrazole-3-carboxylate (7a) : Yellowish brown solid; yield (75%); mp: 98–100°C; IR (ATR) cm − 1 ; 1748 ( CO O-Ph), 1725 ( Co- CH 3 ), 1574 (C = C); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 7.65 (d, J = 8.50 Hz, 1H, Ar-H), 7.46 (d, J = 8.00 Hz, 2H, Ar-H), 7.43–7.39 (m, 3H, Ar-H), 7.38 (s, 1H, Ar-H), 7.34–7.32 (m, 3H, Ar-H), 7.30 (s, 1H, pyrazole-H), 7.28–7.27 (m, 2H, Ar-H), 6.84 (d, J = 8.50 Hz, 1H, Ar-H), 3.75 (s, 3H, OCH 3 ), 2.57 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.35, 160.07, 151.86, 147.61, 146.33, 145.04, 143.48, 142.86, 141.18, 139.68, 136.44, 129.91, 129.45, 129.11, 126.55, 123.97, 122.69, 114.98, 111.62, 56.74, 27.17; ESI-MS m/z [M + Na] + for C 25 H 20 N 2 NaO 4 calculated: 435.13208, found: 435.13095. 4-Acetyl-2-methoxyphenyl-1-phenyl 5- p -tolyl-1 H -pyrazole-3-carboxylate (7b) : Yellowish solid; yield (81%); mp: 110–112°C; IR (ATR) cm − 1 ; 1743 ( C OO-Ph), 1710 ( CO -CH 3 ), 1575 (C = C); 1 H-NMR (500 MHz, CDCl 3 ) δ (ppm): 7.65 (s, 1H, Ar-H), 7.53–7.50 (m, 6H, Ar-H), 7.36–7.35 (m, 2H, Ar-H), 7.15 (s, 1H, pyrazole-H), 6.92–6.94 (m, 3H, Ar-H), 3.88 (s, 3H, OCH 3 ), 2.62 (s, 3H, CH 3 ), 2.33 (s, 3H, CH 3 ); 13 C-NMR (100MHz, CDCl 3 ) δ (ppm): 197.40, 159.67, 150.10, 146.64, 143.95, 142.51, 139.46, 138.90, 136.03, 130.16, 129.85, 128.80, 125.35, 124.00, 122.97, 121.94, 111.97, 109.93, 56.48, 26.65, 21.49; ESI-MS m/z [M + Na] + for C 26 H 22 N 2 NaO 4 calculated: 449.1477, found: 449.14830. 4-Acetyl-2-methoxyphenyl − 5-(4-methoxyphenyl) 1-phenyl-1 H -pyrazole- 3-carboxylate (7c) : Yellowish brown solid; yield (85%); mp: 69–71°C; IR (ATR) cm − 1 ; 1743 ( CO O-Ph), 1725 ( CO -CH 3 ), 1577 (C = C); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 7.64 (d, J = 7.00 Hz, 1H, Ar-H), 7.46 (d, J = 8.50 Hz, 2H, Ar-H), 7.42 (s, 1H, Ar-H), 7.38 (d, J = 7.00 Hz, 1H, Ar-H), 7.34 (d, J = 7.50 Hz, 2H, Ar-H), 7.22 (s, 1H, pyrazole-H), 7.17 (d, J = 8.50 Hz, 2H, Ar-H), 6.88–6.85 (m, 3H, Ar-H), 3.84 (s, 3H, OCH 3 ), 3.79 (s, 3H, OCH 3 ), 2.58 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.73, 159.99, 151.77, 148.69, 145.56, 143.22, 142.20, 139.78, 136.74, 130.95, 130.16, 129.55, 125.76, 124.02, 122.28, 116.09, 115.07, 111.96, 110.36, 57.17, 55.74, 26.65; ESI-MS m/z [M + Na] + for C 26 H 22 N 2 NaO 5 calculated: 465.1426, found: 465.14282. 4-Acetyl-2-methoxyphenyl-5-(4-chlorophenyl) 1-phenyl-1 H -pyrazole- ---3-carboxylate (7d) : Yellowish brown solid; yield (73%); mp: 85–87°C; IR (ATR) cm − 1 ; 1739 ( CO O-Ph), 1728 ( CO- CH 3 ), 1575 (C = C); 1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 7.60 (s, 1H, Ar-H), 7.56 (d, J = 6.00 Hz, 1H, Ar-H), 7.50–7.46 (m, 4H, Ar-H), 7.32–7.35 (m, 2H, Ar-H), 7.17–7.15 (m, 2H, Ar-H), 7.14 (s, 1H, pyrazole-H), 6.90 (d, J = 6.50 Hz, 2H, Ar-H), 3.88 (s, 3H, OCH 3 ), 2.51 (s, 3H, CH 3 ); 13 C-NMR (100MHz, CDCl 3 ) δ (ppm): 197.11, 160.00, 150.43, 147.01, 146.31, 143.53, 143.23, 138.32, 135.31, 133.65, 129.23, 128.88, 127.17, 126.08, 125.74, 123.01, 113.97, 111.69, 109.80, 56.14, 27.26; ESI-MS m/z [M + H] + for C 25 H 20 ClN 2 O 4 calculated: 447.1106, found: 447.10859. 4-Acetyl-2-methoxyphenyl-5-(3,4-dimethoxyphenyl) 1-phenyl-1 H -pyrazole-3-carboxylate (7e) : Reddish yellow solid; yield (71%); mp: 74–76°C; IR (ATR) cm − 1 ; 1740 ( CO O-Ph), 1715 ( CO -CH 3 ), 1589 (C = C); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 7.65 (s, 1H, Ar-H), 7.63 (s, 1H, Ar-H), 7.56 (d, J = 8.50 Hz, 1H, Ar-H), 7.46 (d, J = 8.50 Hz, 2H, Ar-H), 7.38–7.41 (m, 3H, Ar-H), 7.30 (s, 1H, pyrazole-H), 6.97 (d, J = 8.50 Hz, 1H, Ar-H), 6.85–6.83 (m, 2H, Ar-H), 3.84 (s, 3H, OCH 3 ), 3.76 (s, 3H, OCH 3 ), 3.71 (s, 3H, OCH 3 ), 2.59 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO-d6) δ (ppm): 197.38, 160.00, 153.54, 151.71, 149.03, 147.99, 147.03, 142.92, 140.14, 136.74, 130.54, 130.34, 129.55, 127.07, 123.73, 122.65, 121.95, 115.07, 112.67, 112.01, 110.57, 57.15, 56.48, 55.44, 26.95; ESI-MS m/z [M + H] + for C 27 H 25 N 2 O 6 calculated: 473.1707, found: 473.17144. 4-Acetyl-2-methoxyphenyl-5-phenyl 1-(4-sulfamoylphenyl)-1 H -pyrazole-3 -carboxylate (7f) : Yellowish solid; yield (65%); mp: 69–72°C; IR (ATR) cm − 1 ; 1735 ( CO O-Ph), 1724 ( CO -CH 3 ), 1589 (C = C aromatic), 1162 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 7.91 (s, 1H, Ar-H), 7.87 (d, J = 8.50 Hz, 2H, Ar-H), 7.80 (d, J = 8.00 Hz, 1H, Ar-H), 7.68–7.64 (m, 3H, Ar-H), 7.55 (d, J = 8.50 Hz, 2H), 7.53 (s, 2H, SO 2 NH 2 ), 7.41–7.39 (m, 2H, Ar-H), 7.35 (s, 1H, pyrazole-H), 7.31 (d, J = 8.00 Hz, 1H, Ar-H), 3.85 (s, 3H, OCH 3 ), 2.57 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.75, 167.77, 159.18, 151.90, 144.65, 144.14, 142.86, 141.18, 135.65, 132.33,129.48, 129.40, 127.04, 125.27, 124.50, 123.58, 122.00, 112.02, 111.12, 56.74, 28.85; ESI-MS m/z [M + Na] + for C 25 H 21 N 3 Na O 6 S calculated: 514.1049, found: 514.10449. 4-Acetyl-2-methoxyphenyl-1-(4-sulfamoylphenyl) 5- p -tolyl-1 H -pyrazole-3-carboxylate (7g) : Yellowish solid; yield (78%), mp: 83–85°C; IR (ATR) cm − 1 ; 1746 ( COO-Ph ), 1718 ( CO -CH 3 ), 1595 (C = C), 1162 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 7.95 (d, J = 8.50 Hz, 1H, Ar-H), 7.86 (d, J = 8.50 Hz, 2H, Ar-H), 7.69–7.63 (m, 2H, Ar-H), 7.55 (d, J = 8.50 Hz, 2H, Ar-H), 7.52 (s, 2H, SO 2 NH 2 ), 7.39 (d, J = 8.50 Hz, 2H, Ar-H), 7.29 (s, 1H, pyrazole-H), 7.19 (d, J = 8.50 Hz, 2H, Ar-H), 3.84 (s, 3H, OCH 3 ), 2.60 (s, 3H, CH 3 ), 2.28 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.74, 163.64, 151.71, 146.50, 144.25, 143.93, 142.21, 139.63, 130.19, 129.12, 128.82, 128.45, 127.77, 126.14, 125.04, 123.75, 122.31, 112.64, 110.31, 56.44, 27.25, 21.47; ESI-MS m/z [M + Na] + for C 26 H 23 N 3 NaO 6 S calculated: 528.1205, found: 528.12058. 4-Acetyl-2-methoxyphenyl 5-(4-methoxyphenyl) 1-(4-sulfamoylphenyl)-1 H - pyrazole-3-carboxylate (7h) :Yellowish solid; yield (55%), mp: 80–83°C; IR (ATR) cm − 1 ; 1750 ( CO O-Ph), 1720 ( CO -CH 3 ), 1585 (C = C), 1164 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 7.86 (d, J = 9.00 Hz, 2H, Ar-H), 7.66 (s, 1H, Ar-H), 7.64 (d, J = 7.50 Hz, 2H, Ar-H), 7.55 (d, J = 9.00 Hz, 2H, Ar-H), 7.51 (s, 2H, SO 2 NH 2 ), 7.39 (d, J = 8.00 Hz, 1H, Ar-H), 7.13 (s, 1H, pyrazole-H), 7.26–7.23 (m, 3H, Ar-H), 3.84 (s, 3H, OCH 3 ), 3.74 (s, 3H, OCH 3 ), 2.60 (s, 3H, CH 3 ); 13 C NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.35, 160.08, 151.86, 148.90, 144.65, 143.35, 141.58, 136.44, 130.40, 129.61, 127.43, 126.55, 123.98, 123.58, 121.01, 115.47, 113.69, 111.12, 57.53, 55.85, 26.68; ESI-MS m/z [M + Na] + for C 26 H 23 N 3 Na O 7 S calculated: 544.1154, found: 544.11530. 4-Acetyl-2-methoxyphenyl-5-(4-chlorophenyl) 1-(4-sulfamoylphenyl)-1 H - pyrazole-3-carboxylate (7i) : Yellowish brown solid; yield (76%); mp: 71–74°C; IR (ATR) cm − 1 ; 1744 ( CO O-Ph), 1726 ( CO -CH 3 ), 1591 (C = C), 1162 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm):, 7.89 (d, J = 8.00 Hz, 2H, Ar-H), 7.86 (s, 1H, Ar-H), 7.80 (d, J = 9.00 Hz, 1H, Ar-H), 7.65 (d, J = 9.00 Hz, 1H, Ar-H), 7.57 (d, J = 10.00 Hz, 2H, Ar-H), 7.52 (s, 2H, SO 2 NH 2 ), 7.47 (d, J = 8.00 Hz, 2H, Ar-H), 7.38 (s, 1H, pyrazole-H), 7.33 (d, J = 10.00 Hz, 2H, Ar-H), 3.84 (s, 3H, OCH 3 ), 2.69 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.80, 162.85, 159.53, 144.43, 143.25, 143.15, 141.69, 136.56, 132.33, 129.46, 127.90, 127.43, 126.56, 126.09, 123.83, 122.31, 120.09, 112.31, 111.90, 57.14, 27.14; ESI-MS m/z [M-H] − for C 25 H 20 ClN 3 NaO 6 S calculated: 524.0689, found: 524.06882. 4-Acetyl-2-methoxyphenyl-5-(3,4-dimethoxyphenyl) 1-(4-sulfamoylphenyl) -1 H -pyrazole-3-carboxylate (7j) : Yellowish brown solid; yield (52%); mp: 75–78°C; IR (ATR) cm − 1 ; 1749( CO O-Ph), 1727 ( CO -CH 3 ), 1579 (C = C), 1162 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 8.05 (d, J = 8.50 Hz, 2H, Ar-H), 7.96 (d, J = 7.50 Hz, 1H, Ar-H), 7.80 (s, 1H, Ar-H), 7.74 (d, J = 8.50 Hz, 2H, Ar-H), 7.69 (s, 2H, SO 2 NH 2 ), 7.56 (d, J = 7.50 Hz, 1H, Ar-H), 7.50 (s, 1H, Ar-H), 7.10 (d, J = 7.50 Hz, 1H, Ar-H), 7.05 (s, 1H, pyrazole-H), 6.95 (d, J = 7.50 Hz, 1H, Ar-H), 3.99 (s, 3H, OCH 3 ), 3.89 (s, 3H, OCH 3 ), 3.75 (s, 3H, OCH 3 ), 2.75 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.75, 163.93, 151.47, 149.29, 148.89, 145.80, 144.68, 143.33, 142.86, 142.08, 136.04, 129.13, 127.41, 126.55, 125.27, 123.58, 122.29, 121.00, 112.90, 111.12, 109.84, 57.13, 56.74, 54.96, 27.17; ESI-MS m/z [M + Na] + for C 27 H 25 N 3 NaO 8 S calculated: 574.1260, found: 574.12616. 4.1.6. General procedure for synthesis of ( E )-4-(1-(hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-substituted phenyl)-1-(4-substituted phenyl)-1 H -pyrazole-3-carboxylate 8a-j. A mixture of the appropriate ketone derivatives 7a-j (0.001 mol) and hydroxylamine hydrochloride (0.138 g, 0.002 mol) in 30 mL of absolute ethanol was heated under reflux for 8–12 h and then left to cool to room temperature. The separated solid was filtered off, washed with 10% ammonia solution, then with distilled water, dried, and crystallized from absolute ethanol, affording the target products 8a-j . ( E )-4-(1-(Hydroxyimino)ethyl)2-methoxyphenyl 1,5-diphenyl-1 H -pyrazole-3-carboxylate (8a) : Yellowish brown solid; yield (67%); mp: 167–170°C; IR (ATR) cm − 1 ; 3191 (OH), 1756 ( C = O ), 1594 (C = C aromatic); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 11.30 (s, 1H, OH), 7.47 (s, 1H, Ar-H), 7.41–7.37 (m, 4H, Ar-H), 7.33–7.30 (m, 3H, Ar-H), 7.28–7.23 (m, 3H, Ar-H), 7.20 (s, 1H, pyrazole-H), 7.00 (d, J = 8.00 Hz, 1H, Ar-H), 6.78 (d, J = 8.00 Hz, 1H, Ar-H), 3.73 (s, 3H, OCH 3 ), 2.06 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 160.07, 152.75, 148.50, 148.01, 144.05, 142.67, 139.72, 136.71, 132.58, 130.41, 129.62, 128.72, 127.84, 126.55,, 123.37, 119.35, 118.92, 116.26, 109.46, 56.24, 12.14; ESI-MS m/z [M + Na] + for C 25 H 21 N 3 NaO 4 calculated: 450.1430, found: 450.14238. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 1-phenyl-5- p -tolyl- 1 H -pyrazole-3-carboxylate (8b) : Yellowish green powder; yield (75%); mp: 123–125°C; IR (ATR) cm − 1 ; 3138 (OH), 1736 ( C = O ), 1593 (C = C aromatic); 1 H-NMR (500 MHz, DMSO-d6) δ (ppm): 10.36 (s, 1H, OH), 7.40–7.35 (m, 3H, Ar-H), 7.29 (s, 1H, Ar-H), 7.15–7.05 (m, 5H, 4 Ar-H, pyrazole-H), 6.96 (d, J = 7.50 Hz, 2H, Ar-H), 6.75 (d, J = 7.50 Hz, 2H, Ar-H), 3.74 (s, 3H, OCH 3 ), 2.21 (s, 3H, Ph- CH 3 ), 2.02 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 160.73, 153.20, 152.18, 151.13, 147.97, 147.97, 142.01, 139.43, 136.85, 129.81, 129.06, 128.64, 126.07, 124.01, 119.19, 116.09, 115.42, 111.97, 108.89, 56.78, 21.85, 11.83; ESI-MS m/z [M + H] + for C 26 H 24 N 3 O 4 calculated: 442.1761, found: 442.17438. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-methoxyphenyl) 1 -phenyl-1 H -pyrazole-3-carboxylate (8c) : Yellowish brown powder; yield (75%); mp: 132–135°C; IR (ATR) cm − 1 ; 3400 (OH), 1739( C = O ), 1590 (C = C aromatic); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.43 (s, 1H, OH), 7.47 (d, J = 8.50 Hz, 1H, Ar-H), 7.34 (d, J = 10.00 Hz, 2H, Ar-H), 7.25 (s, 1H, Ar-H), 7.18 (d, J = 10.00 Hz, 2H, Ar-H), 7.11–7.08 (m, 2H, Ar-H), 7.04 (s, 1H, pyrazole-H), 7.95 (d, J = 8.50 Hz, 1H, Ar-H), 6.80–6.77 (m, 3H, Ar-H), 3.68 (s, 3H, OCH 3 ), 3.62 (s, 3H, OCH 3 ), 2.02 (s, 3H, CH 3 ); 13 C NMR (100MHz, DMSO- d 6 ) δ (ppm): 162.15, 159.58, 150.58, 147.22, 145.43, 143.76, 142.47, 140.80, 136.88, 130.90, 129.61, 127.83, 126.55, 123.18, 121.16, 120.61, 115.87, 114.57, 109.43, 56.74, 54.96, 14.71; ESI-MS m/z [M + H] + for C 26 H 24 N 3 O 5 calculated: 458.17160, found: 458.17154. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-chlorophenyl)- 1-phenyl-1 H -pyrazole-3-carboxylate (8d) : Yellowish brown powder; yield (65%); mp: 106–109°C; IR (ATR) cm − 1 ; 3228 (OH), 1744 ( C = O ), 1593 (C = C aromatic); 1 H-NMR (400 MHz, DMSO- d 6 ) δ (ppm): 10.90 (s, 1H, OH), 7.60 (d, J = 8.40 Hz, 1H, Ar-H), 7.42 (d, J = 10.00 Hz, 2H, Ar-H), 7.39 (d, J = 10 Hz, 2H, Ar-H), 7.36 (s, 1H, Ar-H), 7.26–7.21 (m, 3H, 2Ar-H, pyrazole-H), 7.00 (d, J = 8.00 Hz, 2H, Ar-H), 6.75 (d, J = 8.00 Hz, 2H, Ar-H), 3.74 (s, 3H, OCH 3 ), 2.07 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 160.00, 153.68, 151.87, 147.38, 143.64, 142.89, 139.83, 139.48, 136.84, 134.19, 132.02, 131.01, 129.71, 128.38, 126.38, 123.19, 119.48, 115.55, 109.63, 55.38, 12.45; ESI-MS m/z [M + H] + for C 25 H 21 ClN 3 O 4 calculated: 462.11889, found: 462.11846. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(3,4-dimethoxy phenyl)-1-phenyl-1 H -pyrazole-3-carboxylate (8e) : Yellowish white powder; yield (77%); mp: 115–118°C; IR (ATR) cm − 1 ; 3300 (OH), 1740 ( C = O ), 1593 (C = C aromatic); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.43 (s, 1H, OH), 7.56 (s, 1H, Ar-H), 7.46 (s, 1H, Ar-H), 7.40 (d, J = 6.50 Hz, 1H, Ar-H), 7.25–7.19 (m, 4H, Ar-H), 7.01–6.98 (m, 2H, Ar-H), 7.16 (s, 1H, pyrazole-H), 6.79 (d, J = 8.50, 2H, Ar-H), 3.73 (s, 3H, OCH 3 ), 3.70 (s, 3H, OCH 3 ), 3.65 (s, 3H, OCH 3 ), 2.06 (s, 3H, CH 3 ); 13 C NMR (100MHz, DMSO- d 6 ) δ (ppm): 161.75, 159.58, 153.64, 150.58, 150.19, 149.29, 148.01, 145.05, 144.15, 142.86, 139.50, 130.41, 127.84, 125.76, 121.67, 121.62, 121.41, 119.71, 115.86, 112.90, 109.84, 56.74, 55.85, 54.56, 11.65; ESI-MS m/z [M + H] + for C 27 H 24 N 2 NaO 6 calculated: 488.1816, found: 488.18086. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-phenyl-1-(4-sulfamoyl phenyl)-1 H -pyrazole-3-carboxylate (8f) : Yellowish powder; yield (53%); mp:122–124°C; IR (ATR) cm − 1 ; 3681 (OH), 1744 ( C = O ), 1590 (C = C aromatic), 1165 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 11.17 (s, 1H, OH), 7.86 (d, J = 6.50 Hz, 2H, Ar-H), 7.55 (d, J = 6.5 Hz, 2H, Ar-H), 7.51 (s, 2H, SO 2 NH 2 ),7.40–7.35 (m, 4H, Ar-H), 7.32–7.30 (m, 3H, Ar-H), 7.24 (s, 1H, Ar-H) 7.19 (s, 1H, pyrazole-H), 3.79 (s, 3H, OCH 3 ), 2.06 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 165.53, 159.99, 153.18, 151.12, 145.63, 143.53, 142.52, 141.49, 139.80, 136.74, 130.18, 129.18, 127.77, 126.80, 123.77, 123.30, 118.83, 111.65, 109.60, 55.71, 12.56; ESI-MS m/z [M + Na] + for C 25 H 22 N 4 Na O 6 S calculated: 529.1158, found: 529.1158. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 1-(4-sulfamoylphenyl) -5- p -tolyl-1 H -pyrazole-3-carboxylate (8g) : Yellowish white powder; yield (69%); mp: 195–197°C; IR (ATR) cm − 1 ; 3255(OH), 1740 ( C = O ), 1594 (C = C aromatic), 1164 (SO 2 NH 2 ), 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 11.34 (s, 1H, OH), 7.95 (d, J = 7.50, 1H, Ar-H), 7.86 (d, J = 7.5 Hz, 2H, Ar-H), 7.68 (d, J = 7.50 Hz, 1H, Ar-H), 7.54 (d, J = 8.50 Hz, 2H, Ar-H), 7.52 (s, 2H, SO 2 NH 2 ), 7.40 (s, 1H, Ar-H), 7.27 (s, 1H, pyrazole-H), 7.24–7.19 (m, 4H, Ar-H), 3.78 (s, 3H, OCH 3 ), 2.28 (s, 3H, CH 3 ), 2.16 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 160.42, 153.19, 151.27, 145.80, 144.61, 142.05, 139.88, 139.41, 136.67, 130.21, 129.17, 127.98, 127.33, 126.51, 126.10, 123.30, 118.84, 111.67, 110.12, 56.45, 21.50, 12.56; ESI-MS m/z [M + Na] + for C 26 H 24 N 4 NaO 6 S calculated: 543.1314, found: 543.13044. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylate (8h) :Yellowish white powder; yield (49%), mp: 125–127°C; IR (ATR) cm − 1 ; 3400 (OH), 1742 ( C = O ), 1596 (C = C aromatic), 1165 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 11.27 (s, 1H, OH), 7.87(d, J = 8.50 Hz, 2H, Ar-H), 7.55 (d, J = 9.50 Hz, 2H, Ar-H), 7.51 (s, 2H, SO 2 NH 2 ), 7.40 (s, 1H, Ar-H), 7.24–7.20 (m, 4H, Ar-H), 7.19 (s, 1H, pyrazole-H), 6.95 (d, J = 9.50 Hz, 2H, Ar-H), 3.79 (s, 3H, OCH 3 ), 3.74 (s, 3H, OCH 3 ), 2.16 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 161.75, 159.18, 153.97, 151.07, 145.93, 144.64, 143.75, 142.47, 139.90, 136.44, 130.41, 129.62, 127.78, 123.18, 122.03, 121.41, 118.82, 115.47, 111.11, 56.74, 55.85, 12.14; ESI-MS m/z [M + Na] + for C 26 H 24 N 4 NaO 7 S calculated: 559.1263, found: 559.12740. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-chlorophenyl)-1- (4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylate (8i) : Yellowish brown powder; yield (67%); mp:110–112°C; IR (ATR) cm − 1 ; 3371 (OH), 1743 ( C = O ), 1595 (C = C aromatic), 1161 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.44 (s, 1H, OH), 7.85 (d, J = 10.00 Hz, 2H, Ar-H), 7.76 (d, J = 6.50 Hz, 1H, Ar-H), 7.61 (s, 1H, Ar-H), 7.49 (s, 2H, SO 2 NH 2 ), 7.40–7.33 (m, 4H, 3Ar-H, pyrazole-H), 7.26 (d, J = 10.00 Hz, 2H, Ar-H), 7.19 (d, J = 10.00 Hz, 2H, Ar-H), 3.71 (s, 3H, OCH 3 ), 1.90 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 159.57, 150.82, 144.50, 144.38, 143.30, 141.53, 136.59, 134.61, 132.25,, 129.43, 128.25, 127.46, 126.50, 124.90, 123.26, 120.17, 119.66, 118.84, 109.89, 56.38, 11.76; ESI-MS m/z [M + 1] + for C 25 H 22 ClN 4 O 6 S calculated: 541.0943, found: 541.09486. ( E )-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(3,4-dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxylate (8j) : Yellowish powder; yield (48%); mp: 129–131°C; IR (ATR) cm − 1 ; 3300 (OH), 1731( C = O ), 1595 (C = C aromatic); 1164 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 11.26 (s, 1H, OH), 7.87 (d, J = 9.00 Hz, 2H, Ar-H), 7.85 (d, J = 9.00 Hz, 2H, Ar-H), 7.52 (s, 2H, SO 2 NH 2 ), 7.41 (s, 1H, Ar-H), 7.32 (s, 1H, Ar-H), 7.25–7.24 (m, 1H, Ar-H), 6.93 (d, J = 8.50 Hz, 1H, Ar-H), 6.88 (s, 1H, pyrazole-H ) , 6.81–6.75 (m, 2H, Ar-H), 3.79 (s, 3H, OCH 3 ), 3.72 (s, 3H, OCH 3 ), 3.60 (s, 3H, OCH 3 ), 2.17 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 167.78, 159.18, 152.75, 149.29, 148.50, 145.44, 144.65, 143.36, 141.58, 140.30, 136.44, 132.58, 129.12, 127.83, 126.15, 122.30, 121.40, 119.33, 112.91, 110.73, 109.84, 56.28, 56.06, 55.89, 11.75; ESI-MS m/z [M + Na] + for C 27 H 26 N 4 NaO 8 S calculated: 589.1369, found: 589.13803. 4.1.7. General procedure for synthesis of N -(4-acetylphenyl)-5-(4-subistitutedphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxamide 9a-c. To the suspension of 1,5-diarylpyrazole carboxylic acid derivatives 6f, 6h and 6j (0.001 mol) in 20 mL of benzene, thionyl chloride (2 mL) was added and heated under reflux for 4 h. Evaporation of the solvent was carried out under vacuum to give a residue of the corresponding acyl chloride that was utilized in the following steps without purification. A mixture of acyl chloride in dry DMF, few drops of triethylamine and 4-aminoacetophenone (0.270 g, 0.002 mol) were heated under reflux for 8h. Then, 20 mL of cold distilled water was added, followed by acidification with dil. HCl and extraction twice with ethyl acetate. Purification was performed by column chromatography using chloroform: methanol 98:2 as eluent to afford compounds 9a-c [ 64 ]. N -(4-Acetylphenyl)-5-phenyl-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxamide (9a) : Yellowish brown solid; yield (88%); mp: 81–83°C; IR (ATR) cm − 1 ; 1725 ( CO -CH 3 ), 1675 ( CO NH), 1591 (C = C aromatic), 1161 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.53 (s, 1H, NH), 7.96 (d, J = 9.00 Hz, 2H, Ar-H), 7.86 (d, J = 9.00 Hz, 2H, Ar-H), 7.68–7.62 (m, 3H, Ar-H), 7.57 (d, J = 9.00 Hz, 2H, Ar-H), 7.49 (s, 2H, SO 2 NH 2 ), 7.40 (d, J = 9.00 Hz, 2H, Ar-H), 7.30–7.32 (m, 2H, Ar-H), 7.20 (s, 1H, pyrazole-H), 2.52 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.09, 167.91, 154.18, 145.63, 144.56, 143.96, 142.53, 133.27, 132.23, 131.21, 129.83, 127.84, 125.96, 125.26, 120.60, 119.04, 109.61, 26.95; ESI-MS m/z [M + Na] + for C 24 H 20 N 4 NaO 4 S calculated: 483.1103, found: 483.11090. N -(4-Acetylphenyl)-5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1 H - pyrazole-3-carboxamide (9b) :Yellowish brown solid; yield (66%); mp: 74–76°C; IR (ATR) cm − 1 ; 1715 ( CO -CH 3 ),1681 ( CO NH), 1594 (C = C aromatic), 1160 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 11.18 (s, 1H, NH), 7.87 (d, J = 7.60 Hz, 2H, Ar-H), 7.84 (d, J = 7.6.0 Hz, 2H, Ar-H), 7.80 (d, J = 9.00 Hz, 2H, Ar-H), 7.62 (d, J = 9.00 Hz, 2H, Ar-H), 7.54 (s, 2H, SO 2 NH 2 ), 7.33 (s, 1H, pyrazole-H), 6.98 (d, J = 7.60 Hz, 2H, Ar-H), 6.96 (d, J = 7.60 Hz, 2H, Ar-H), 3.82 (s, 3H, OCH 3 ), 2.47 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 195.96, 167.20, 154.55, 146.06, 144.59, 142.88, 133.27, 132.59, 131.09, 130.12, 129.18, 126.18, 125.30, 123.46, 120.64, 118.98, 113.06, 55.42, 26.61; ESI-MS m/z [M-H] − for C 25 H 21 N 4 O 5 S calculated: 489.1238, found: 489.12547. N -(4-Acetylphenyl)-5-(3,4-dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1 H - pyrazole-3-carboxamide (9c) : Brownish solid; yield (60%); mp: 81–83°C; IR (ATR) cm − 1 ; 1720 ( CO -CH 3 ), 1669 ( CO NH), 1590 (C = C aromatic), 1160 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.55 (s, 1H, NH), 7.95 (s, 1H, Ar-H), 7.92 (d, J = 9.00 Hz, 2H, Ar-H), 7.84 (d, J = 7.50 Hz, 2H, Ar-H), 7.76 (d, J = 8.50 Hz, 1H, Ar-H), 7.72 (d, J = 9.00 Hz, 2H, Ar-H), 7.62–7.56 (m, 4H, 2 Ar-H, SO 2 NH 2 ), 7.37 (s, 1H, pyrazole-H), 7.01 (d, J = 8.50 Hz, 1H, Ar-H), 3.82 (s, 3H, OCH 3 ), 3.80 (s, 3H, OCH 3 ), 2.53 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 197.06, 164.12, 153.21, 152.80, 150.42, 148.06, 145.60, 143.23, 142.71, 132.93, 130.56, 129.86, 129.15, 126.10, 124.03, 123.54, 119.19,116.49, 110.95, 56.76, 55.71, 26.62; ESI-MS m/z [M-H] − for C 26 H 23 N 4 O 6 S calculated: 519.1344, found: 519.13451. 4.1.8. General procedure for synthesis of ( E )- N -(4-(1-(hydroxyimino)ethyl) phenyl)-5-(4-subistituted phenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3- carboxamide 10a-c. A mixture of the appropriate ketone derivatives 9a-c (0.001 mol) and hydroxylamine hydrochloride (0.138 g, 0.002 mol) in 30 mL of absolute ethanol was heated under reflux for 8–12 h and then left to cool to room temperature. The separated solid was filtered off, washed with 10% ammonia solution, then washed with distilled water, dried, and recrystallized from absolute ethanol to afford the target products 10a-c . ( E )- N -(4-(1-(Hydroxyimino)ethyl)phenyl)-5-phenyl-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxamide (10a) :Yellowish powder; yield (55%); mp: 168–170°C ; IR (ATR) cm − 1 ; 3681 (OH), 1680 ( CO NH), 1598 (C = C aromatic), 1162 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.30 (s, 1H, NH), 8.75 (s, 1H, OH), 7.79 (d, J = 8.00 Hz, 2H, Ar-H), 7.63 (d, J = 8.00 Hz, 2H, Ar-H), 7.50–7.56 (m, 3H, Ar-H), 7.47–7.44 (m, 2H, Ar-H), 7.40 (s, 2H, SO 2 NH 2 ), 7.35 (d, J = 10.00 Hz, 2H, Ar-H), 7.29 (s, 1H, pyrazole-H), 7.21 (d, J = 10.00 Hz, 2H, Ar-H), 2.06 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 160.05, 153.24, 151.71, 148.09, 143.55, 141.92, 133.69, 129.88, 129.39, 129.30, 127.44, 126.52, 126.42, 121.34, 120.78, 120.33, 109.58, 12.14; ESI-MS m/z [M + H] + for C 24 H 22 N 5 O 4 S calculated: 476.1387, found: 476.13970. ( E )- N -(4-(1-(Hydroxyimino)ethyl)phenyl)-5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1 H -pyrazole-3-carboxamide (10b) : Yellowish brown powder; yield (51%); mp: 110–112°C; IR (ATR) cm − 1 ; 3350 (OH), 1677 ( CO NH), 1596 (C = C aromatic), 1162 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.31 (s, 1H, NH), 10.02 (s, 1H, OH), 7.84 (d, J = 8.00 Hz, 2H, Ar-H), 7.78 (d, J = 8.50 Hz, 2H, Ar-H), 7.52 (d, J = 8.50 Hz, 2H, Ar-H), 7.46 (d, J = 9.00 Hz, 2H, Ar-H), 7.41 (s, 2H, SO 2 NH 2 ), 7.30 (s, 1H, pyrazole-H), 6.97 (d, J = 8.00 Hz, 2H, Ar-H), 6.91 (d, J = 9.00 Hz, 2H, Ar-H), 3.80 (s, 3H, CH 3 ), 2.07 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 167.18, 155.50, 144.08, 142.51, 133.80, 132.35, 131.38, 130.65, 129.47, 127.11, 126.41, 126.09, 125.97, 123.46, 121.51, 114.33, 111.98, 56.12, 17.34; ESI-MS m/z [M-H] − for C 25 H 22 N 5 O 5 S calculated: 504.1347, found: 504.13773. ( E )-5-(3,4-Dimethoxyphenyl)- N -(4-(1-(hydroxyimino)ethyl)phenyl)-1- (4-sulfamoylphenyl)-1 H -pyrazole-3-carboxamide (10c) :Brownish powder; yield (44%), mp: 105–107°C; IR (ATR) cm − 1 ; 3220 (OH), 1680 ( CO NH), 1591 (C = C aromatic), 1161 (SO 2 NH 2 ); 1 H-NMR (500 MHz, DMSO- d 6 ) δ (ppm): 10.36 (s, 1H, NH), 10.09 (s, 1H, OH), 7.92 (s, 1H, Ar-H), 7.79 (d, J = 7.50 Hz, 1H, Ar-H), 7.63 (d, J = 8.50 Hz, 2H, Ar-H), 7.52 (d, J = 7.50 Hz, 2H, Ar-H), 7.50–7.44 (m, 4H, 2ArH, SO 2 NH 2 ), 7.36 (d, J = 8.50 Hz, 2H, Ar-H), 7.27 (s, 1H, pyrazole-H), 6.99 (d, J = 7.50 Hz, 1H, Ar-H), 3.77 (s, 3H, OCH 3 ), 3.74 (s, 3H, OCH 3 ), 2.08 (s, 3H, CH 3 ); 13 C-NMR (100MHz, DMSO- d 6 ) δ (ppm): 167.78, 159.18, 152.75, 149.29, 148.50, 145.44, 144.65, 143.36, 140.30, 136.44, 132.58, 129.12, 127.83, 126.15, 122.30, 121.40, 119.33, 112.91, 109.84, 56.19, 55.96, 12.14; ESI-MS m/z [M-H] − for C 26 H 24 N 5 O 6 S calculated: 534.14473, found: 534.14307. 4.2. Measurement of nitric oxide release Materials and methods The nitrite calibration curve in addition to the absorbance of the tested compounds were measured on Shimadzu UV-160, UV-Visible spectrophotometer (Shimadzu, Tokyo, Japan). The tested compounds were prepared as solutions in DMF and diluted with the buffer system till a concentration of 100 mM. N - Acetyl cystiene solution was prepared in a concentration of 500 mM in methanol. Griess reagent consists of 0.1% w/v NED solution in water and sulfanilamide solution (1% w/v of sulfanilamide in 5% w/v phosphoric acid). Nitrite standard solution (0.1 M sodium nitrite in water) stock solution was prepared from which a dilute solution of 100 µM nitrite solution was prepared by dilution 1 mL of the stock solution to 1000 mL with phosphate buffer of pH 7.4. From this solution, 4 serials two-fold dilutions were performed to generate different concentrations of the nitrite solution (100.00, 50.00, 25.00 and 12.50 µM) and these concentrations were used for nitrite calibration curve. Preparation of nitrite standard curve Sulfanilamide and NEDD solutions were kept at 25°C, 100 mL of sulfanilamide solution was added to each dilution of the prepared standard nitrite solution. The mixture was left at 25°C for 5–10 minutes protected from light. To this mixture 100 mL of the NEDD solution was added and the mixture was again left for 5-10minutes at 25°C protected from light. The absorbance of the formed purple color was measured within 30 minutes at λ max 546 nm, a blank experiment was performed under the same conditions, the procedure was repeated three times for each dilution of the nitrite and the average absorbance was calculated. A plot of the average absorbance value for each concentration of the nitrite standard solution as a function of “ Y ” against nitrite concentration as a function of “ X “ was constructed to generate a standard nitrite calibration curve at pH 7.4. NO release assay The amount of NO released from the tested compounds 8a-b, 8d-i and 10a-c was measured using the Griess colorimetric method[ 65 ] either in phosphate buffer of pH 7.4 in the presence of N -acetyl cysteine which serves as a source of thiols. The amount of NO released from the tested compounds was measured relative to NO released from standard sodium nitrite solution. Procedure Different solutions of the tested compounds 8a-b, 8d-i and 10a-c in DMF were diluted using phosphate buffer of pH 7.4 till a final concentration of 100 mM (test solutions). To 100 mL of different test solutions, 100 mL of N -acetyl cysteine solution was added and the obtained solution was kept in an incubator at 37°C (treated solutions). The solutions were treated similarly as for nitrite standard solution with Griess reagent components, 100 mL of sulfanilamide solution was added to each tube of the treated solution. The mixture was left at 25°C for 5 − 10 minutes protected from light. To this mixture 100 mL of the NED solution was added and the mixture was again left at 25°C for 5–10 minutes protected from light. The absorbance of the formed purple color, if any, was measured within 30 minutes at λmax 546 nm. A blank experiment was performed under the same conditions, the procedure was repeated three times for each tested compound and the average absorbance was calculated. The corresponding concentration of nitrite was determined by comparison to the nitrite standard calibration curve and the amount of NO released (attributed by the corresponding nitrite concentration) was calculated as percentage of moles of NO released from 1 mole of the tested compounds. 4.3. Biology Materials and methods Evaluation of anticancer activity was performed according to the standard water-soluble tetrazolium-8 (WST-8) assay at the Faculty of Engineering, Yamagata University, Yonezawa, Japan, using an MTP-310 absorbance microplate reader. The EGFR inhibitory assay was carried out at the Faculty of Engineering, Yamagata University, Yonezawa, Japan, according to the protocol enzyme linked immunosorbent assay (ELISA) kits (Douset sandwich ELISA test, recombinant mouse EGFR). The JNK-2 inhibitory assay was performed at the Faculty of Engineering, Yamagata University, Yonezawa, Japan, according to the protocol of enzyme-linked immunosorbent assay (ELISA), the assay was carried out using the JNK-2 kit (Simple Step ELISA, pT183/Y185, Abacam Company, Japan). Apoptosis and cell cycle analysis was performed at Faculty of medicine, Yamagata University, Yamagata, Japan, using (BD FACS melody). Molecular docking was performed at the Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Researchers, RIEKN, Japan, using ICM-Pro 3.8 software (MolSoft L.L.C, USA). 4.3.1. Evaluation of anticancer activity According to the standard water-soluble tetrazolium-8 (WST-8) assay, the current synthesized compounds have been tested for their anticancer activities against different five cancer cell lines; DLD-1, Hela, K562, SUIT-2 and HepG2 and daunorubicin was used as reference drug by the WST-8 assay. The five cells were maintained in a suspension culture, (Dulbecco's modified eagle medium (DMEM) for SUIT-2, Hela and HepG2 or PRIM for K562 and DLD), supplemented with 5% FBS (Fetal Bovine Serum) containing 1% of a penicillin-streptomycin (1:1) mixture. A 100 µL aliquot of cells (10000 cells/mL) was added to a 96 well plate and incubated for 24 h at 37°C in a humidified incubator containing 5% CO 2 in the air. After 24 h, a 10 µL aliquot of test compound (concentrations varying in the range of 10–150 µM) was added to each of the 96 wells and incubated for 24 h. Then A 10 µL WST-8 solution (mixture of WST-8 and 1-methoxy PMS) was added to each well and the incubation continued for 3 h. The visible absorbance at 450 nm and 630 nm as the reference wavelength of each well was quantified using an MTP-310 absorbance microplate reader. Daunorubicin was used as a positive control. The results of cytotoxicity were recorded as growth inhibition percentages and as IC 50 values [ 66 , 67 ]. 4.3.2. EGFR inhibitory assay This assay was carried out according to the protocol for enzyme linked immunosorbent assay (ELISA) kits (Douset sandwich ELISA test, recombinant mouse EGFR) [ 68 , 69 ]. This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for EGFR has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any EGFR present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-conjugated antibody specific for EGFR is added to each well and incubated. Following a wash to remove unbound substances streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of EGFR bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ± 2nm. The concentration of EGFR in the sample is then determined by comparing the O.D of samples to the standard curve. 4.3.3. JNK-2 inhibitory assay JNK-2 inhibitory assay was performed according to the protocol of enzyme-linked immunosorbent assay (ELISA). The assay was carried out using the JNK-2 kit (Simple Step ELISA, pT183/Y185, Abacam Company, Japan).[ 70 , 71 ] for the semi-quantitative measurement of JNK-2 protein in human cell lysate. The SimpleStep ELISA employs an affinity tag labelled capture antibody and a reporter attached detector antibody to immunocapture the sample analyte in solution. This complete complex (capture antibody/analyte/detector antibody) is then immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or controls are added to the wells, followed by the antibody cocktail. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by the addition of stop solution, completing any color change from blue to yellow. The signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, the development of TMB can be recorded kinetically at 600 nm. An antibody cocktail can be prepared by combining an appropriate volume of the capture and detector antibodies immediately prior to assay. To make 3 mL of the antibody cocktail, combine 1.5 mL of capture antibody with 1.5 mL of detector antibody. Mix thoroughly and gently. Control lysate can be prepared from HEK293 cells, cultured in 10% FBS containing medium, then treated with 1µg/mL anisomycin. After preparing all the reagents, samples and control as instructed and following the previously published procedures [ 70 , 71 ], add 50 µL of sample or control to the well, add 50 µL of the antibody cocktail, then incubate at room temperature for 1 hour on a plate shaker set to 400 rpm, aspirate and wash each well three times with 350 µL with wash buffer, add 100 µL TMB substrate to each well and incubate for 15 minutes, then add 100 µL stop solution and measure the absorbance at 450 nm. 4.3.4. Apoptosis analysis For apoptosis induction measurement, seeding of Hela cells in 96-well plates (1×10 4 cell/well) and DMEM medium was added and incubated for 24 h, at the second day the cells were treated with the test compounds (at IC 50 , double and half of IC 50 ) then incubated overnight, on the third day wash each well twice with 100 µL phosphate buffered saline (PBS), add Trypsin 100 µL and incubate plates for 3 to 5 minutes at 37 degrees, then 200 µL of medium was added to separate cells, centrifuge cells for 5 minutes and decant supernatant. Wash twice with 100 µL PBS and add 200µL of Buffer, 5 µL of PI& Annexin and 200 µl of PBS for the test compounds. For control, add 400 µL of PBS. Finally, the cell suspension was observed under the fluorescent microscope or transfer to a round bottom tube for flowcytometric analysis using (BD FACS melody). 4.3.5. Cell cycle analysis To cultured Hela cells in 96-well plates (1×10 4 cell/well), DMEM medium and the IC 50 concentration of the test compounds were added, then incubated for 24 h after the addition of test compound. Transfer supernatant to falcon tube for each plate and wash each well twice with 3 mL PBS, add Trypsin 1 mL and incubate plates for 3 to 5 minutes at 37 degrees, then 3 mL of the medium was added to separate cells, centrifuge cells for 5 minutes and decant supernatant, wash again with 3 mL PBS, and add 10 mL of medium. Cells were immediately fixed in ice cold 70% ethanol overnight at -20°C. In the day of the analysis, cells were washed ×3 with PBS and re-suspended in propidium iodide (PI) for 15 minutes at room temperature, protected from light. Cell-cycle analysis was performed using flowcytometric analysis (BD FACS melody) and data obtained from cell cycle distribution was analyzed using FSC-W (Watson model) to estimate the percentage of cells in G1, S, and G2. 4.3.6. Evaluation of cytotoxicity against PC12 cells According to the standard water-soluble tetrazolium-8 (WST-8) assay, the cytotoxicity of compounds 8g and 8i against the PC12 cell line was evaluated, and daunorubicin was used as the reference drug by WST-8. The PC12 cells were maintained in a suspension culture, (Dulbecco's modified eagle medium (DMEM), supplemented with 5% FBS (Fetal Bovine Serum) containing 1% of a penicillin-streptomycin (1:1) mixture. A 100 µL aliquot of cells (10000 cells/mL) was added to a 96 well plate and incubated for 24 h at 37 ° C in a humidified incubator containing 5% CO 2 in the air. After 24 h, a 10 µL aliquot of test compound (concentrations varying in the range of 10–150 µM) was added to each of the 96 wells and incubated for 24 h. Then A 10 µL WST-8 solution (mixture of WST-8 and 1-methoxy PMS) was added to each well and the incubation continued for 3 h. The visible absorbance at 450 nm and 630 nm as the reference wavelength of each well was quantified using an MTP-310 absorbance microplate reader. Daunorubicin was used as a positive control. The results of cytotoxicity were recorded as growth inhibition percentages and as IC 50 values. 4.3.7. Molecular docking on EGFR and JNK-2 Docking simulation was performed by the Inter-coordinate Mechanics (ICM) using ICM-Pro 3.8 software (MolSoft L.L.C, USA)[ 72 ]. First, the 3D structures of the tested compounds and sorafenib (a reference multi-target kinase inhibitor) [ 73 ] were generated to perform well-suited docking. Then, the enzyme was prepared by adjusting the interface properties, including water molecules deletion, hydrogen atoms optimization and formal charges refinement. In addition, enzyme relaxation was logged to run flexible docking. The ligands binding affinities were calculated by the Gaussian potential based on the ligand electrostatic potential and shape complementarity at the binding site [ 14 , 49 ]. In these studies, the template-docking method was used by selecting pre-defined binding pockets of the study receptors. The structural models of the tested comounds against human EGFR complexed with AZD9291 inhibitor (2.80 ; PDB ID: 4ZAU) were used [74] and crystal structure of human JNK-2 complexed with an indazole inhibitor (2.14 Å; PDB ID: 3E7O) was used for c-Jun N -terminal kinase 2 (JNK-2)[ 13 ]. Declarations Ethics approval and consent to participate Not applicable in this manuscript Consent for publication Not applicable Availability of data and materials All data generated or analysed during this study are included in this published article [and its supplementary information files. Competing interests "The authors declare that they have no competing interests" Funding This article is not funded (Not applicable). Authors' contributions A- Kamal S. Abdelrahman. did the practical chemistry part B- Heba A.Hassan, suggested the idea of work, wrote and reviewed the whole manuscript C- Salah A. Abdel-Aziz, helped in the revision and writing of the introduction part D- Adel A. Marzouk, helped in the revision and writing of the chemistry part E- raef shams, helped in the revision and writing of the docking part F- Keima Osawa, helped in the biological part practical and writing G- Mohamed Abdel-Aziz, suggested the idea of work and helped in revision of the manuscript H- Hiroyuki Konno, helped in the practical chemistry part Acknowledgement We are grateful to Dr. Hideyuki Miyatake of RIKEN for using ICM software. References Gallorini, M.; Cataldi, A.; di Giacomo, V. Cyclin-dependent kinase modulators and cancer therapy. BioDrugs 2012 , 26 , 377-391. Kibria, G.; Hatakeyama, H.; Harashima, H. Cancer multidrug resistance: mechanisms involved and strategies for circumvention using a drug delivery system. Archives of pharmacal research 2014 , 37 , 4-15. Abdelgawad, M. A.; Bakr, R. B.; Alkhoja, O. A.; Mohamed, W. R. Design, synthesis and antitumor activity of novel pyrazolo [3, 4-d] pyrimidine derivatives as EGFR-TK inhibitors. Bioorganic chemistry 2016 , 66 , 88-96. Mitsudomi, T.; Yatabe, Y. Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. The FEBS journal 2010 , 277 , 301-308. Rego, R.; Foster, N.; Smyrk, T.; Le, M.; O'connell, M.; Sargent, D.; Windschitl, H.; Sinicrope, F. Prognostic effect of activated EGFR expression in human colon carcinomas: comparison with EGFR status. British journal of cancer 2010 , 102 , 165-172. Cao, C.; Lu, S.; Sowa, A.; Kivlin, R.; Amaral, A.; Chu, W.; Yang, H.; Di, W.; Wan, Y. Priming with EGFR tyrosine kinase inhibitor and EGF sensitizes ovarian cancer cells to respond to chemotherapeutical drugs. Cancer letters 2008 , 266 , 249-262. Bridges, A. The rationale and strategy used to develop a series of highly potent, irreversible, inhibitors of the epidermal growth factor receptor family of tyrosine kinases. Current medicinal chemistry 1999 , 6 , 825-844. Iida, K.; Nakayama, K.; Rahman, M.; Rahman, M.; Ishikawa, M.; Katagiri, A.; Yeasmin, S.; Otsuki, Y.; Kobayashi, H.; Nakayama, S. EGFR gene amplification is related to adverse clinical outcomes in cervical squamous cell carcinoma, making the EGFR pathway a novel therapeutic target. British journal of cancer 2011 , 105 , 420-427. Huang, P.; Xu, X.; Wang, L.; Zhu, B.; Wang, X.; Xia, J. The role of EGF‐EGFR signalling pathway in hepatocellular carcinoma inflammatory microenvironment. Journal of cellular and molecular medicine 2014 , 18 , 218-230. Allen, H.; Hsuan, J.; Clark, S.; Maziarz, R.; Waterfield, M.; Flavell, R.; Haley, J. Expression of epidermal-growth-factor receptor in the K562 cell line by transfection. Altered receptor biochemistry. Biochemical journal 1990 , 271 , 785-790. Mathew, M. P.; Tan, E.; Saeui, C. T.; Bovonratwet, P.; Sklar, S.; Bhattacharya, R.; Yarema, K. J. Metabolic flux-driven sialylation alters internalization, recycling, and drug sensitivity of the epidermal growth factor receptor (EGFR) in SW1990 pancreatic cancer cells. Oncotarget 2016 , 7 , 66491. Butterworth, S.; Cross, D. A.; Finlay, M. R. V.; Ward, R. A.; Waring, M. J. The structure-guided discovery of osimertinib: the first US FDA approved mutant selective inhibitor of EGFR T790M. Medchemcomm 2017 , 8 , 820-822. Shaw, D.; Wang, S. M.; Villaseñor, A. G.; Tsing, S.; Walter, D.; Browner, M. F.; Barnett, J.; Kuglstatter, A. The crystal structure of JNK2 reveals conformational flexibility in the MAP kinase insert and indicates its involvement in the regulation of catalytic activity. Journal of molecular biology 2008 , 383 , 885-893. Kuglstatter, A.; Ghate, M.; Tsing, S.; Villaseñor, A. G.; Shaw, D.; Barnett, J. W.; Browner, M. F. X-ray crystal structure of JNK2 complexed with the p38α inhibitor BIRB796: insights into the rational design of DFG-out binding MAP kinase inhibitors. Bioorganic & medicinal chemistry letters 2010 , 20 , 5217-5220. Abdelrahman, K. S.; Hassan, H. A.; Abdel-Aziz, S. A.; Marzouk, A. A.; Narumi, A.; Konno, H.; Abdel-Aziz, M. JNK signaling as a target for anticancer therapy. Pharmacological Reports 2021 , 1-30. Ozgur, E.; Kayhan, H.; Kismali, G.; Senturk, F.; Sensoz, M.; Ozturk, G. G.; Sel, T. Effects of radiofrequency radiation on colorectal cancer cell proliferation and inflammation. Turkish Journal of Biochemistry 2021 , 46 , 525-532. Yue, Y.; Yan, L.; Yan-Ping, G.; Li, Z.; Lu-Bo, G. GL-V9 reverses adriamycin resistance in hepatocellular carcinoma cells by affecting JNK2-related autophagy. Chinese journal of natural medicines 2020 , 18 , 491-499. Tian, X.; Traub, B.; Shi, J.; Huber, N.; Schreiner, S.; Chen, G.; Zhou, S.; Henne-Bruns, D.; Knippschild, U.; Kornmann, M. c-Jun N-terminal kinase 2 suppresses pancreatic cancer growth and invasion and is opposed by c-Jun N-terminal kinase 1. Cancer Gene Therapy 2022 , 29 , 73-86. Merkerova, M.; Bruchova, H.; Brdicka, R. JNK2 and p38 MAPK over-expressions do not represent key events in chronic myeloid leukemia transformation. Neoplasma 2007 , 54 , 503-510. Lin, C. S.; Lin, C. L.; Ying, T. H.; Chiou, H. L.; Hung, C. H.; Liao, W. S.; Hsieh, Y. H.; Kao, S. H. β‐Mangostin inhibits the metastatic power of cervical cancer cells attributing to suppression of JNK2/AP‐1/Snail cascade. Journal of Cellular Physiology 2020 , 235 , 8446-8460. Kardosh, A.; Wang, W.; Uddin, J.; Petasis, N. A.; Hofman, F. M.; Chen, T. C.; Schonthal, A. H. Dimethyl-celecoxib (DMC), a derivative of celecoxib that lacks cyclooxygenase-2-inhibitory function, potently mimics the anti-tumor effects of celecoxib on Burkitt’s lymphoma in vitro and in vivo. Cancer biology & therapy 2005 , 4 , 571-582. Sharma, V.; Bhatia, P.; Alam, O.; Naim, M. J.; Nawaz, F.; Sheikh, A. A.; Jha, M. Recent advancement in the discovery and development of COX-2 inhibitors: Insight into biological activities and SAR studies (2008–2019). Bioorganic chemistry 2019 , 89 , 103007. Chandrakantha, B.; Isloor, A. M.; Shetty, P.; Isloor, S.; Malladi, S.; Fun, H. K. Synthesis, characterization and antimicrobial activity of novel ethyl 1-(N-substituted)-5-phenyl-1 H-pyrazole-4-carboxylate derivatives. Medicinal Chemistry Research 2012 , 21 , 2702-2708. Manvar, D.; Pelliccia, S.; La Regina, G.; Famiglini, V.; Coluccia, A.; Ruggieri, A.; Anticoli, S.; Lee, J.-C.; Basu, A.; Cevik, O. New 1-phenyl-5-(1H-pyrrol-1-yl)-1H-pyrazole-3-carboxamides inhibit hepatitis C virus replication via suppression of cyclooxygenase-2. European journal of medicinal chemistry 2015 , 90 , 497-506. Pathak, R. B.; Chovatia, P.; Parekh, H. Synthesis, antitubercular and antimicrobial evaluation of 3-(4-chlorophenyl)-4-substituted pyrazole derivatives. Bioorganic & medicinal chemistry letters 2012 , 22 , 5129-5133. Yuan, J.-W.; Wang, S.-F.; Luo, Z.-L.; Qiu, H.-Y.; Wang, P.-F.; Zhang, X.; Yang, Y.-A.; Yin, Y.; Zhang, F.; Zhu, H.-L. Synthesis and biological evaluation of compounds which contain pyrazole, thiazole and naphthalene ring as antitumor agents. Bioorganic & medicinal chemistry letters 2014 , 24 , 2324-2328. Yu, L.; Wu, W. K. K.; Li, Z. J.; Liu, Q. C.; Li, H. T.; Wu, Y. C.; Cho, C. H. Enhancement of doxorubicin cytotoxicity on human esophageal squamous cell carcinoma cells by indomethacin and 4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (SC236) via inhibiting P-glycoprotein activity. Molecular pharmacology 2009 , 75 , 1364-1373. Zhang, L.; Peterson, T. E.; Lu, V. M.; Parney, I. F.; Daniels, D. J. Antitumor activity of novel pyrazole-based small molecular inhibitors of the STAT3 pathway in patient derived high grade glioma cells. Plos one 2019 , 14 , e0220569. Sharp, S. Y.; Boxall, K.; Rowlands, M.; Prodromou, C.; Roe, S. M.; Maloney, A.; Powers, M.; Clarke, P. A.; Box, G.; Sanderson, S. Correction: In vitro Biological Characterization of a Novel, Synthetic Diaryl Pyrazole Resorcinol Class of Heat Shock Protein 90 Inhibitors. Cancer research 2019 , 79 , 287-287. Abuo-Rahma, G. E.-D. A.; Abdel-Aziz, M.; Beshr, E. A.; Ali, T. F. 1, 2, 4-Triazole/oxime hybrids as new strategy for nitric oxide donors: Synthesis, anti-inflammatory, ulceroginicity and antiproliferative activities. European journal of medicinal chemistry 2014 , 71 , 185-198. Shen, S.; Xu, N.; Klamer, G.; Ko, K.-H.; Khoo, M.; Ma, D.; Moore, J.; O'Brien, T. A.; Dolnikov, A. Small-molecule inhibitor of glycogen synthase kinase 3β 6-bromoindirubin-3-oxime inhibits hematopoietic regeneration in stem cell recipient mice. Stem Cells and Development 2015 , 24 , 724-736. Zhang, X.; Castanotto, D.; Nam, S.; Horne, D.; Stein, C. 6BIO enhances oligonucleotide activity in cells: a potential combinatorial anti-androgen receptor therapy in prostate cancer cells. Molecular Therapy 2017 , 25 , 79-91. Xiong, B.; Chen, S.; Zhu, P.; Huang, M.; Gao, W.; Zhu, R.; Qian, J.; Peng, Y.; Zhang, Y.; Dai, H. Design, synthesis, and biological evaluation of novel thiazolyl substituted bis-pyrazole oxime derivatives with potent antitumor activities by selectively inducing apoptosis and ROS in cancer cells. Medicinal Chemistry 2019 , 15 , 743-754. Hong, S.; Shin, Y.; Jung, M.; Ha, M. W.; Park, Y.; Lee, Y.-J.; Shin, J.; Oh, K. B.; Lee, S. K.; Park, H.-g. Efficient synthesis and biological activity of Psammaplin A and its analogues as antitumor agents. European Journal of Medicinal Chemistry 2015 , 96 , 218-230. Bednarczyk-Cwynar, B.; Zaprutko, L. Recent advances in synthesis and biological activity of triterpenic acylated oximes. Phytochemistry Reviews 2015 , 14 , 203-231. Schepetkin, I. A.; Khlebnikov, A. I.; Potapov, A. S.; Kovrizhina, A. R.; Matveevskaya, V. V.; Belyanin, M. L.; Atochin, D. N.; Zanoza, S. O.; Gaidarzhy, N. M.; Lyakhov, S. A. Synthesis, biological evaluation, and molecular modeling of 11H-indeno [1, 2-b] quinoxalin-11-one derivatives and tryptanthrin-6-oxime as c-Jun N-terminal kinase inhibitors. European journal of medicinal chemistry 2019 , 161 , 179-191. Schepetkin, I. A.; Kirpotina, L. N.; Khlebnikov, A. I.; Hanks, T. S.; Kochetkova, I.; Pascual, D. W.; Jutila, M. A.; Quinn, M. T. Identification and characterization of a novel class of c-Jun N-terminal kinase inhibitors. Molecular pharmacology 2012 , 81 , 832-845. Lu, L.; Sha, S.; Wang, K.; Zhang, Y.-H.; Liu, Y.-D.; Ju, G.-D.; Wang, B.; Zhu, H.-L. Discovery of chromeno [4, 3-c] pyrazol-4 (2H)-one containing carbonyl or oxime derivatives as potential, selective inhibitors PI3Kα. Chemical and Pharmaceutical Bulletin 2016 , c16-00388. Schepetkin, I. A.; Plotnikov, M. B.; Khlebnikov, A. I.; Plotnikova, T. M.; Quinn, M. T. Oximes: Novel therapeutics with anticancer and anti-inflammatory potential. Biomolecules 2021 , 11 , 777. Soliman, R. Preparation and antidiabetic activity of some sulfonylurea derivatives of 3, 5-disubstituted pyrazoles. Journal of medicinal chemistry 1979 , 22 , 321-325. Aghazadeh Tabrizi, M.; Baraldi, P. G.; Baraldi, S.; Ruggiero, E.; De Stefano, L.; Rizzolio, F.; Di Cesare Mannelli, L.; Ghelardini, C.; Chicca, A.; Lapillo, M. Discovery of 1, 5-diphenylpyrazole-3-carboxamide derivatives as potent, reversible, and selective monoacylglycerol lipase (MAGL) inhibitors. Journal of medicinal chemistry 2018 , 61 , 1340-1354. Shin, U. c.; Choi, J. S.; Beak, Y. J.; Lee, M. w.; Kim, H. S.; Choi, D. W.; Kim, D. G.; Kim, S. W. Development of a 68Ga‐labelled PET tracer for carbonic anhydrase IX‐overexpressed tumors using the artificial sweetener saccharin. Journal of Labelled Compounds and Radiopharmaceuticals 2020 . Gonzalez, R. M.; Seurynck-Servoss, S. L.; Crowley, S. A.; Brown, M.; Omenn, G. S.; Hayes, D. F.; Zangar, R. C. Development and validation of sandwich ELISA microarrays with minimal assay interference. Journal of proteome research 2008 , 7 , 2406-2414. El-Kashef, D. H.; El-Sheakh, A. R. Hepatoprotective effect of celecoxib against tamoxifen-induced liver injury via inhibiting ASK-1/JNK pathway in female rats. Life sciences 2019 , 231 , 116573. Tsutsumi, R.; Ito, H.; Hiramitsu, T.; Nishitani, K.; Akiyoshi, M.; Kitaori, T.; Yasuda, T.; Nakamura, T. Celecoxib inhibits production of MMP and NO via down-regulation of NF-κB and JNK in a PGE2 independent manner in human articular chondrocytes. Rheumatology international 2008 , 28 , 727-736. Maayah, Z. H.; Levasseur, J.; Piragasam, R. S.; Abdelhamid, G.; Dyck, J. R.; Fahlman, R. P.; Siraki, A. G.; El-Kadi, A. O. 2-Methoxyestradiol protects against pressure overload-induced left ventricular hypertrophy. Scientific reports 2018 , 8 , 1-15. Pozarowski, P.; Grabarek, J.; Darzynkiewicz, Z. Flow cytometry of apoptosis. Current protocols in cell biology 2003 , 21 , 18.18. 11-18.18. 33. Yosaatmadja, Y.; Silva, S.; Dickson, J. M.; Patterson, A. V.; Smaill, J. B.; Flanagan, J. U.; McKeage, M. J.; Squire, C. J. Binding mode of the breakthrough inhibitor AZD9291 to epidermal growth factor receptor revealed. Journal of structural biology 2015 , 192 , 539-544. Jousserandot, A.; Boucher, J.-L.; Henry, Y.; Niklaus, B.; Clement, B.; Mansuy, D. Microsomal cytochrome P450 dependent oxidation of N-hydroxyguanidines, amidoximes, and ketoximes: mechanism of the oxidative cleavage of their CN (OH) bond with formation of nitrogen oxides. Biochemistry 1998 , 37 , 17179-17191. Radwan, A. A.; Ghorab, M. M.; Alsaid, M. S.; Alanazi, F. K. Novel ethyl 1, 5-disubstituted-1H-pyrazole-3-carboxylates as a new class of antimicrobial agents. Acta Pharmaceutica 2014 , 64 , 335-344. Li, Z.; Wang, Z.-C.; Li, X.; Abbas, M.; Wu, S.-Y.; Ren, S.-Z.; Liu, Q.-X.; Liu, Y.; Chen, P.-W.; Duan, Y.-T. Design, synthesis and evaluation of novel diaryl-1, 5-diazoles derivatives bearing morpholine as potent dual COX-2/5-LOX inhibitors and antitumor agents. European journal of medicinal chemistry 2019 , 169 , 168-184. Penning, T. D.; Talley, J. J.; Bertenshaw, S. R.; Carter, J. S.; Collins, P. W.; Docter, S.; Graneto, M. J.; Lee, L. F.; Malecha, J. W.; Miyashiro, J. M. Synthesis and biological evaluation of the 1, 5-diarylpyrazole class of cyclooxygenase-2 inhibitors: identification of 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1 H-pyrazol-1-yl] benzenesulfonamide (SC-58635, celecoxib). Journal of medicinal chemistry 1997 , 40 , 1347-1365. Batchu, H.; Bhattacharyya, S.; Kant, R.; Batra, S. Palladium-catalyzed chelation-assisted regioselective oxidative dehydrogenative homocoupling/ortho-hydroxylation in N-phenylpyrazoles. The Journal of organic chemistry 2015 , 80 , 7360-7374. Murray, W. V.; Wachter, M. P. A simple regioselective synthesis of ethyl 1, 5‐diarylpyrazole‐3‐carboxylates. Journal of heterocyclic chemistry 1989 , 26 , 1389-1392. Yamali, C.; Gul, H. I.; Ece, A.; Bua, S.; Angeli, A.; Sakagami, H.; Sahin, E.; Supuran, C. T. Synthesis, biological evaluation and in silico modelling studies of 1, 3, 5-trisubstituted pyrazoles carrying benzenesulfonamide as potential anticancer agents and selective cancer-associated hCA IX isoenzyme inhibitors. Bioorganic chemistry 2019 , 92 , 103222. Rogez-Florent, T.; Meignan, S.; Foulon, C.; Six, P.; Gros, A.; Bal-Mahieu, C.; Supuran, C. T.; Scozzafava, A.; Frédérick, R.; Masereel, B. New selective carbonic anhydrase IX inhibitors: synthesis and pharmacological evaluation of diarylpyrazole-benzenesulfonamides. Bioorganic & medicinal chemistry 2013 , 21 , 1451-1464. Harrison, C. 18 months sliced off Celebrex's patent protection. Nature reviews. Drug discovery 2014 , 13 , 326. Lu, X.-Y.; Wang, Z.-C.; Ren, S.-Z.; Shen, F.-Q.; Man, R.-J.; Zhu, H.-L. Coumarin sulfonamides derivatives as potent and selective COX-2 inhibitors with efficacy in suppressing cancer proliferation and metastasis. Bioorganic & medicinal chemistry letters 2016 , 26 , 3491-3498. Raiford, L. C.; Hill, E. L. Effect of Constitution on the Rearrangement of the Phenylhydrazones of Some Unsymmetrically Substituted Dibenzalacetones1. Journal of the American Chemical Society 1934 , 56 , 174-176. Hwang, S. H.; Wagner, K. M.; Morisseau, C.; Liu, J.-Y.; Dong, H.; Wecksler, A. T.; Hammock, B. D. Synthesis and structure− activity relationship studies of urea-containing pyrazoles as dual inhibitors of cyclooxygenase-2 and soluble epoxide hydrolase. Journal of medicinal chemistry 2011 , 54 , 3037-3050. Uddin, M. J.; Crews, B. C.; Ghebreselasie, K.; Marnett, L. J. Design, synthesis, and structure–activity relationship studies of fluorescent inhibitors of cycloxygenase-2 as targeted optical imaging agents. Bioconjugate chemistry 2013 , 24 , 712-723. Chen, L. W.; Wang, P. F.; Tang, D. J.; Tao, X. X.; Man, R. J.; Qiu, H. Y.; Wang, Z. C.; Xu, C.; Zhu, H. L. Metronidazole containing pyrazole derivatives potently inhibit tyrosyl‐tRNA synthetase: design, synthesis, and biological evaluation. Chemical biology & drug design 2016 , 88 , 592-598. Abdelazeem, A. H.; El-Din, A. G. S.; Abdel-Fattah, M. M.; Amin, N. H.; El-Moghazy, S. M.; El-Saadi, M. T. Discovery of novel urea-diarylpyrazole hybrids as dual COX-2/sEH inhibitors with improved anti-inflammatory activity and highly reduced cardiovascular risks. European Journal of Medicinal Chemistry 2020 , 205 , 112662. Guevara, I.; Iwanejko, J.; Dembińska-Kieć, A.; Pankiewicz, J.; Wanat, A.; Anna, P.; Goła̧bek, I.; Bartuś, S.; Malczewska-Malec, M.; Szczudlik, A. Determination of nitrite/nitrate in human biological material by the simple Griess reaction. Clinica Chimica Acta 1998 , 274 , 177-188. Ishiyama, M.; Miyazono, Y.; Sasamoto, K.; Ohkura, Y.; Ueno, K. A highly water-soluble disulfonated tetrazolium salt as a chromogenic indicator for NADH as well as cell viability. Talanta 1997 , 44 , 1299-1305. Hotsumi, M.; Tajiri, M.; Nikaido, Y.; Sato, T.; Makabe, K.; Konno, H. Design, synthesis, and evaluation of a water soluble C5-monoketone type curcumin analogue as a potent amyloid β aggregation inhibitor. Bioorganic & medicinal chemistry letters 2019 , 29 , 2157-2161. Gangemi, S.; Franchina, T.; Minciullo, P. L.; Profita, M.; Zanghì, M.; David, A.; Kennez, I.; Adamo, V. IL‐33/IL‐31 axis: A new pathological mechanisms for EGFR tyrosine kinase inhibitors‐associated skin toxicity. Journal of cellular biochemistry 2013 , 114 , 2673-2676. Masago, K.; Fujita, S.; Hatachi, Y.; Fukuhara, A.; Sakuma, K.; Ichikawa, M.; Kim, Y. H.; Mio, T.; Mishima, M. Clinical significance of pretreatment serum amphiregulin and transforming growth factor‐α, and an epidermal growth factor receptor somatic mutation in patients with advanced non‐squamous, non‐small cell lung cancer. Cancer science 2008 , 99 , 2295-2301. Gil-Martínez, A. L.; Cuenca, L.; Estrada, C.; Sánchez-Rodrigo, C.; Fernández-Villalba, E.; Herrero, M. T. Unexpected exacerbation of neuroinflammatory response after a combined therapy in old Parkinsonian mice. Frontiers in cellular neuroscience 2018 , 12 , 451. Cenariu, D.; Fischer-Fodor, E.; Țigu, A. B.; Bunea, A.; Virág, P.; Perde-Schrepler, M.; Toma, V.-A.; Mocan, A.; Berindan-Neagoe, I.; Pintea, A. Zeaxanthin-Rich Extract from Superfood Lycium barbarum Selectively Modulates the Cellular Adhesion and MAPK Signaling in Melanoma versus Normal Skin Cells In Vitro. Molecules 2021 , 26 , 333. Neves, M. A.; Totrov, M.; Abagyan, R. Docking and scoring with ICM: the benchmarking results and strategies for improvement. Journal of computer-aided molecular design 2012 , 26 , 675-686. An, J.; Totrov, M.; Abagyan, R. Pocketome via comprehensive identification and classification of ligand binding envelopes. Molecular & Cellular Proteomics 2005 , 4 , 752-761. Hynes, N. E.; Lane, H. A. ERBB receptors and cancer: the complexity of targeted inhibitors. Nature Reviews Cancer 2005 , 5 , 341-354. Additional Declarations No competing interests reported. Supplementary Files Oximesupplementrydata1.docx Scheme1.png Scheme 1. Synthesis of compounds 7a-j and 8a-j. Reagent and conditions ; a) diethyl oxalate, sodium ethoxide, absolute ethanol,4-6h; b) sodium nitrite, conc HCl, ice bath stirring, sodium sulfite, sodium hydroxide; c) sodium acetate, absolute ethanol, reflux for 2-3h; d) sodium hydroxide reflux for 3hr, HCl; e) EDC, HOBT, dry DMF, 4-hydroxy-3-methoxy acetophenone, DIPEA, stirring for 12h; f) hydroxylamine hydrochloride, absolute ethanol, reflux for 12h. Scheme2.png Scheme 2. Synthesis of compounds 9a-c and 10a-c. Reagent and conditions ; g) thionyl chloride, benzene, reflux for 4h; h) 4-aminoacetophenone, triethylamine, DMF, reflux for 8h; i) hydroxylamine hydrochloride, absolute ethanol, reflux for 12h. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2772431","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":190157339,"identity":"27a4f526-8ac0-43d7-89e4-33b72d916f52","order_by":0,"name":"Kamal S. Abdelrahman","email":"","orcid":"","institution":"Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al-Azhar University, Assiut Branch, Assiut 71524, Egypt","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Kamal","middleName":"S.","lastName":"Abdelrahman","suffix":""},{"id":190157343,"identity":"770ab8fe-18a0-438d-8882-0d897edee23f","order_by":1,"name":"Heba A. Hassan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7klEQVRIiWNgGAWjYBACAwYeBgbGBhCT+QCIhLCJ1MKWQLIWHgPitJiz9x78wLjDzp5/ds/XzTwMNrIbDjAffoFPi2XPuWQJxjPJiTPunN12m4chzXjDAbY0C7wOu5FjIMHYxpzAcCMXpOVw4oYDPGYGeLXcf2P8g7Gt3l7+Rs4zoJb/QC383/BrucFjBrTlMOOGGzlsQC0HQLYwP8Cr5UxemkVi2/HEjTfSzG7OMUg2nnmYzQyfDgaD42cP3/jYVm0vdyP52Y03FXayfcebH3/AqwcEEhAmADEzA5sEQS3ogJmwLaNgFIyCUTCSAACTQVBdGBO4AAAAAABJRU5ErkJggg==","orcid":"","institution":"Department of Medicinal Chemistry Faculty of Pharmacy, Minia University, Minia 61519, Egypt","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Heba","middleName":"A.","lastName":"Hassan","suffix":""},{"id":190157347,"identity":"02b0d988-182b-47bd-bc42-4c3b425dad91","order_by":2,"name":"Salah A. Abdel-Aziz","email":"","orcid":"","institution":"Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al-Azhar University, Assiut Branch, Assiut 71524, Egypt","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Salah","middleName":"A.","lastName":"Abdel-Aziz","suffix":""},{"id":190157349,"identity":"a3ff5f40-5de7-48cf-81e1-374d637a1a04","order_by":3,"name":"Adel A. Marzouk","email":"","orcid":"","institution":"Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al-Azhar University, Assiut Branch, Assiut 71524, Egypt","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Adel","middleName":"A.","lastName":"Marzouk","suffix":""},{"id":190157350,"identity":"e708581e-8676-4fe7-9861-1b6fd7e162c2","order_by":4,"name":"Raef shams","email":"","orcid":"","institution":"Emergent Bioengineering Materials Research Team, RIKEN Centre for Emergent Matter Science, RIKEN, Wako, Saitama 351-0198, Japan","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Raef","middleName":"","lastName":"shams","suffix":""},{"id":190157354,"identity":"823f081c-1dce-436b-b21a-7f260c03d306","order_by":5,"name":"Keima Oswa","email":"","orcid":"","institution":"Graduate School of Science and Engineering, Yamagata University, Yonezawa, Yamagata 992-8510, Japan.","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Keima","middleName":"","lastName":"Oswa","suffix":""},{"id":190157357,"identity":"50bfd874-ad5c-4c32-93cd-184c95d108de","order_by":6,"name":"Mohamed Abdel-Aziz","email":"","orcid":"","institution":"Department of Medicinal Chemistry Faculty of Pharmacy, Minia University, Minia 61519, Egypt","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"","lastName":"Abdel-Aziz","suffix":""},{"id":190157359,"identity":"9b201d9b-6e01-471a-bdec-d8842f5acefe","order_by":7,"name":"Hiroyuki Konno","email":"","orcid":"","institution":"Graduate School of Science and Engineering, Yamagata University, Yonezawa, Yamagata 992-8510, Japan.","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hiroyuki","middleName":"","lastName":"Konno","suffix":""}],"badges":[],"createdAt":"2023-04-03 14:59:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2772431/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2772431/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":35589283,"identity":"e7e63ddc-0a5b-4de8-a318-5fdeeae7f22f","added_by":"auto","created_at":"2023-04-11 14:39:21","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":60106,"visible":true,"origin":"","legend":"\u003cp\u003eStructure of diarylpyrazole derivatives with anticancer activity.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/b58810ccaa352290cbf93c8e.png"},{"id":35589284,"identity":"1ec3ea7f-eca2-46e5-b9c8-e587f35896f0","added_by":"auto","created_at":"2023-04-11 14:39:21","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":45808,"visible":true,"origin":"","legend":"\u003cp\u003eStructure of Oxime derivatives with anticancer activity.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/6ed9a736f8c1b1fdce961ee6.png"},{"id":35590786,"identity":"dab664de-5aae-449e-92aa-f130ac365658","added_by":"auto","created_at":"2023-04-11 14:47:21","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":101049,"visible":true,"origin":"","legend":"\u003cp\u003eGeneral structure of scaffold \u003cstrong\u003eA\u003c/strong\u003e and\u003cstrong\u003e B\u003c/strong\u003e.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/5fcd25d6e7905aa6f0d5fb08.png"},{"id":35591253,"identity":"105b62e9-339b-41ef-9f7a-cef58ab011bd","added_by":"auto","created_at":"2023-04-11 14:55:21","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":241257,"visible":true,"origin":"","legend":"\u003cp\u003eFACS analysis using PI-stained Hela cell line after treatment of compounds \u003cstrong\u003e8g, 8i \u003c/strong\u003eand\u003cstrong\u003eDMSO\u003c/strong\u003e as control.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/c4f45cbfb2c4f0a567d7f14a.png"},{"id":35592600,"identity":"6dea45d2-6120-47cf-8d13-c549ba303693","added_by":"auto","created_at":"2023-04-11 15:03:22","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":980463,"visible":true,"origin":"","legend":"\u003cp\u003eMicroscopic examination of apoptosis and necrosis of compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003eusing annexin-v and PI on Hela cell line.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/ea6605fc0fa760cca3e4625a.png"},{"id":35592597,"identity":"661b08ed-3304-4acd-a236-8c0edc76bf5c","added_by":"auto","created_at":"2023-04-11 15:03:21","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":83602,"visible":true,"origin":"","legend":"\u003cp\u003ePercentages of apoptosis and necrosis of compounds \u003cstrong\u003e8g \u003c/strong\u003eand \u003cstrong\u003e8i\u003c/strong\u003e on Hela cell.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/f3e360346b8b1651d500ac5f.png"},{"id":35591255,"identity":"d9d9660f-b2bf-4362-b12d-1fa3ce79a836","added_by":"auto","created_at":"2023-04-11 14:55:21","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":328478,"visible":true,"origin":"","legend":"\u003cp\u003eRibbon representation of EGFR showing the binding pocket with a grey skin mesh (PDB ID: 4ZAU).\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/86f86b15c7783b243d898ca3.png"},{"id":35589289,"identity":"cdea10d6-0290-4fce-8090-ef7dbfcf3e33","added_by":"auto","created_at":"2023-04-11 14:39:21","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":811316,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of sorafenib at the EGFR binding pocket\u003c/strong\u003e. \u003cstrong\u003ea\u003c/strong\u003e, Binding of sorafenib (blue) to EGFR (light green) showing the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb\u003c/strong\u003e, \u003cstrong\u003eThe binding pocket's corresponding 2D representation of sorafenib.\u003c/strong\u003eHydrophobic interactions are highlighted in light green, while hydrogen bonding is highlighted in light blue (shown as a dashed arrow).\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/b88a7e83cfd2752002ce362c.png"},{"id":35591258,"identity":"2bd66a84-f479-4bee-a098-7698f53572d1","added_by":"auto","created_at":"2023-04-11 14:55:22","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":832622,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of 8d at the EGFR binding pocket\u003c/strong\u003e. \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003e8d \u003c/strong\u003e(yellow) to EGFR (light green) showing the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb,\u003c/strong\u003e \u003cstrong\u003ethe binding pocket's corresponding 2D representation of 8d. \u003c/strong\u003eHydrophobic interactions are highlighted in light green, while hydrogen bonding is highlighted in light blue (shown as a dashed arrow).\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/0e9fdcd36f64a7a9fa3f9c90.png"},{"id":35591260,"identity":"4f8b3838-80b3-4470-b43f-6ce2cd6bc053","added_by":"auto","created_at":"2023-04-11 14:55:22","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":861999,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of 8g at the EGFR binding pocket\u003c/strong\u003e. \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003e8g\u003c/strong\u003e (orchid) to EGFR (light green) shows the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb, the binding pocket's corresponding 2D representation of 8g. \u003c/strong\u003eHydrophobic interactions are highlighted in light green, while hydrogen bonding is highlighted in light blue (shown as a dashed arrow).\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/726deccce060499cfa512289.png"},{"id":35589294,"identity":"ce359a52-2127-40f5-b2a7-79fd64deea62","added_by":"auto","created_at":"2023-04-11 14:39:22","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":666195,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of 8i at the EGFR binding pocket.\u003c/strong\u003e \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003e8i \u003c/strong\u003e(magenta) to EGFR (light green) showing the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb\u003c/strong\u003e, \u003cstrong\u003ethe binding pocket's corresponding 2D representation of 8i. \u003c/strong\u003eHydrophobic interactions are highlighted in light green, while hydrogen bonding is highlighted in light blue (shown as a dashed arrow).\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/06d79496cfdfb6b59d000c50.png"},{"id":35590792,"identity":"24c27d95-c9f5-4381-a8ef-62329f227956","added_by":"auto","created_at":"2023-04-11 14:47:22","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":905864,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of 10c at the EGFR binding pocket\u003c/strong\u003e. \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003e10c\u003c/strong\u003e (khaki) to EGFR (light green) showing the interactive residues. Inter- and intra-molecular hydrogen bonds are shown as red dots. \u003cstrong\u003eb\u003c/strong\u003e, \u003cstrong\u003ethe binding pocket's corresponding 2D representation of 10c. \u003c/strong\u003eHydrophobic interactions are highlighted in light green, while hydrogen bonding is highlighted in light blue (shown as a dashed arrow).\u003c/p\u003e","description":"","filename":"13.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/bad52c49b04f66f046f7ecd9.png"},{"id":35592601,"identity":"1c21c9c7-a16a-454a-a10d-de60db29a2ab","added_by":"auto","created_at":"2023-04-11 15:03:22","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":270339,"visible":true,"origin":"","legend":"\u003cp\u003eRibbon representation of JNK-2 showing the binding pocket with a grey skin mesh (PDB ID: 3E7O) [13].\u003c/p\u003e","description":"","filename":"14.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/733813c164e6be921c707bdd.png"},{"id":35592858,"identity":"6bff4ba9-9b84-482a-9c4e-26c0671014c6","added_by":"auto","created_at":"2023-04-11 15:11:21","extension":"png","order_by":15,"title":"Figure 15","display":"","copyAsset":false,"role":"figure","size":704266,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of sorafenib at the JNK-2 binding pocket.\u003c/strong\u003e \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003esorafenib\u003c/strong\u003e (blue) to JNK-2 (sky blue) showing the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb\u003c/strong\u003e, The corresponding 2D representation of \u003cstrong\u003esorafenib\u003c/strong\u003e at the binding pocket of JNK-2. Light green highlight are for hydrophobic interactions, light blue highlight is for hydrogen bonding (shown as a dashed arrow) and grey highlight is for Van der Waals forces.\u003c/p\u003e","description":"","filename":"15.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/b94b25de10aebf75aa00b602.png"},{"id":35589300,"identity":"695ffe9a-adbe-43cc-888b-f42adf748a4d","added_by":"auto","created_at":"2023-04-11 14:39:22","extension":"png","order_by":16,"title":"Figure 16","display":"","copyAsset":false,"role":"figure","size":531166,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of compound 8i at the JNK-2 binding pocket.\u003c/strong\u003e \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003e8i \u003c/strong\u003e(magenta) to JNK-2 (sky blue) showing the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb\u003c/strong\u003e \u003cstrong\u003ethe corresponding 2D representation of 8i in the JNK-2 binding pocket\u003c/strong\u003e, Light green highlights are for hydrophobic interactions, light blue highlights are for hydrogen bonding (shown as a dashed arrow) and gray highlights are for Van der Waals forces.\u003c/p\u003e","description":"","filename":"16.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/90de514ecbec78df85318dc6.png"},{"id":35590797,"identity":"9e16ca54-f592-4a5f-ae01-5d15af12c71a","added_by":"auto","created_at":"2023-04-11 14:47:22","extension":"png","order_by":17,"title":"Figure 17","display":"","copyAsset":false,"role":"figure","size":658979,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDocking pose of compound 8d at the JNK-2 binding pocket\u003c/strong\u003e. \u003cstrong\u003ea\u003c/strong\u003e, Binding of \u003cstrong\u003e8d \u003c/strong\u003e(Pink) to JNK-2 (sky blue) showing the interactive residues. Hydrogen bonds are shown as red dots. \u003cstrong\u003eb\u003c/strong\u003e, the corresponding 2D representation of \u003cstrong\u003e8d\u003c/strong\u003e at the binding pocket of JNK-2. Light green highlights are for hydrophobic interactions. Light green highlights are for hydrophobic interactions, light blue highlights are for hydrogen bonding (shown as a dashed arrow) and grey highlights are for Van der Waals forces.\u003c/p\u003e","description":"","filename":"17.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/f6569e64cd11b952ad2c70e0.png"},{"id":37668250,"identity":"9669dfa8-fe79-4135-bc9b-ffdcad0fdee5","added_by":"auto","created_at":"2023-05-30 14:53:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":9597931,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/6bb690fc-c1f9-46dc-b2a1-9d898992531f.pdf"},{"id":35589303,"identity":"49e7e479-1817-4450-b95c-7c9600b4b416","added_by":"auto","created_at":"2023-04-11 14:39:22","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":20140938,"visible":true,"origin":"","legend":"","description":"","filename":"Oximesupplementrydata1.docx","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/55d4d95199318a4de2f6f94c.docx"},{"id":35589292,"identity":"f2eef075-73b3-40b0-aaf4-ca6f99583ae7","added_by":"auto","created_at":"2023-04-11 14:39:22","extension":"png","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":99416,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScheme 1.\u003c/strong\u003e Synthesis of compounds\u003cstrong\u003e 7a-j \u003c/strong\u003eand \u003cstrong\u003e8a-j\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cu\u003eReagent and conditions\u003c/u\u003e; a) diethyl oxalate, sodium ethoxide, absolute ethanol,4-6h; b) sodium nitrite, conc HCl, ice bath stirring, sodium sulfite, sodium hydroxide; c) sodium acetate, absolute ethanol, reflux for 2-3h; d) sodium hydroxide reflux for 3hr, HCl; e) EDC, HOBT, dry DMF, 4-hydroxy-3-methoxy acetophenone, DIPEA, stirring for 12h; f) hydroxylamine hydrochloride, absolute ethanol, reflux for 12h.\u003c/p\u003e","description":"","filename":"Scheme1.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/0be1875d40836b0b4d6c5fe6.png"},{"id":35590783,"identity":"40b97eb2-3f00-4d0b-810e-203a6e8f855d","added_by":"auto","created_at":"2023-04-11 14:47:21","extension":"png","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":72613,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScheme 2. \u003c/strong\u003eSynthesis of compounds\u003cstrong\u003e 9a-c \u003c/strong\u003eand\u003cstrong\u003e 10a-c.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cu\u003eReagent and conditions\u003c/u\u003e; g) thionyl chloride, benzene, reflux for 4h; h) 4-aminoacetophenone, triethylamine, DMF, reflux for 8h; i) hydroxylamine hydrochloride, absolute ethanol, reflux for 12h.\u003c/p\u003e","description":"","filename":"Scheme2.png","url":"https://assets-eu.researchsquare.com/files/rs-2772431/v1/dcf18eb7c506cbc3f00ff71a.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Synthesis, molecular docking and evaluation of 1,5-diarylpyrazole/oxime hybrids targeting EGFR and JNK-2 as antiproliferative agents","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCancer is the second-leading cause of death throughout the world. Hence, the incidence rate of cancer mortality is becoming increasingly important on a global scale [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Chemotherapy, which uses drugs that target cell division and angiogenesis or that induces cancer cell death \u003cem\u003evia\u003c/em\u003e various signaling pathways, is one of the cancer treatment strategies. However, because of chemotherapy side effects and the development of drug resistance in cancer cells, there is an urgent need for the design, synthesis, and development of effective and safe chemotherapy[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eEGFR is a tyrosine kinase receptor that plays an important role in cellular signaling activities including cell growth, division, differentiation, metabolism, adhesion, and death [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Four tyrosine kinase-related receptors (EGFR, HER2, HER3, and HER4) have been classified into the HER family. Deregulation of HER family signals enhances proliferation, invasion, metastasis, angiogenesis, and cancer cell survival [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. EGFR receptors are over-expressed in different human tumors such as breast, ovarian, prostate, colon, renal, pancreatic, hepatocellular carcinoma, cervical cancer, non-small cell lung cancer (NSCLS) and leukemia [\u003cspan additionalcitationids=\"CR5 CR6 CR7 CR8 CR9 CR10\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Thus, EGFR inhibition has been developed as one of the most efficient strategies for cancer therapy. Several small molecules targeting EGFR are now clinically available such as gefitinib, erlotinib, lapatinib and dacomitinib [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eJNK-2 is a member of the MAP kinase family involved in signaling pathways that has been implicated in several diseases like cancer and inflammatory diseases [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Due to the important key roles of JNK-2 in cancer progression through control of proliferation, differentiation, survival and migration, JNK-2 becomes an appealing oncogenic target for cancer therapy due to its high expression in a variety of cancers, including colorectal adenocarcinoma, cervical cancer, pancreatic cancer, hepatocellular carcinoma, and leukemia.[\u003cspan additionalcitationids=\"CR15 CR16 CR17 CR18 CR19\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. JNK signaling is apparently involved in cancer development and progression as in lymphoma cancer cells through protection it from apoptosis by decreasing ROS accumulation. Also, JNK regulates micro RNA-92a and glucose regulating protein 78 (GRP78) in human pancreatic cancer, which promotes cell proliferation and survival. In hepatocellular carcinoma (HCC) JNK pathway is responsible for its development and progression and becomes the target for therapeutic treatment of HCC. Otherwise, blocking of the JNK pathway leading to inhibition of proliferation human B lymphoma cell due to downregulation of early growth response gen-1(Egr-1) protein. Moreover, the relation between the JNK pathway and other pathways like kappa B(NF- KB) and p38, which are acting together for the regulation of cell proliferation and survival, Also, there is a close relation between JNK and immune evasion regulatory factors such as transforming growth factor-β (TGF-β) and interferon-γ (IFN-γ) mediate cell survival. In addition, JNK can promote cancer cells survival by autophagy to counteract apoptosis. To date, the majority of JNK inhibitors target the highly conserved ATP- binding site, while number of this inhibitors were proven \u003cem\u003ein vivo\u003c/em\u003e in animal model, but not applied therapeutically until now due to lack of its specificity and its side effect, in addition, to increase in the concentration of ATP decrease its efficacy[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePyrazoles are an important class of heterocyclic compounds and exhibit broad range of biological activities including anticancer [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], anti-inflammatory [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], antimicrobial [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], antiviral [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] and antitubercular activities [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Biological evaluation revealed that some pyrazole-containing compounds, such as compounds \u003cb\u003eI\u003c/b\u003e and \u003cb\u003eII\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) exhibited antiproliferative activity against the human cervical cancer cell line Hela by inhibiting cell migration and potent EGFR tyrosine kinase inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e values of 0.07 and 0.06 \u0026micro;M, respectively, in comparison to the positive control erlotinib (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.03 \u0026micro;M) [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Moreover, the selective COX-2 inhibitor compound \u003cb\u003eSC-236\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) showed antitumor activity through blocking of tumor promotor-induced activator protein-1 (AP1) activation as result of suppression of JNK expression and used to treat hepatocellular cancer in conjunction with doxorubicin [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Furthermore, compound \u003cb\u003eSC74102\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) displayed JNK-2 inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e of 1.35 \u0026micro; mol/l as well as its p38α inhibitory activity. Additionally, diarypyrazoles have been reported to have STAT3 inhibitory activity as in compound \u003cb\u003eMNS1-Leu V\u003c/b\u003e [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] and heat shock protein inhibitory activity as in compound \u003cb\u003eCCT072453 VI\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eOximes became with high interest in pharmaceutical chemistry as well. The oxime moiety can hydrogen bind with amino acid residues in the active site of many enzymes and is easy to coordinate with metal ions. Thus, the oxime moiety can improve the binding of the whole molecule to its binding site. Oximes can also release nitric oxide free radicals, which have a cytostatic and cytotoxic effect on cancer cells. It can prevent cancer cells from spreading and help macrophages to kill cancer cells. Several targets have been reported to combine NO with cancer therapy, including either the synergistic effect between anticancer drugs and nitric oxide, increasing the flow of anticancer therapy by NO to intracellular compartments, or increasing the efficiency of cytostatic therapy and overcome of resistance to anticancer agents.[\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. On the other hand, the introduction of an oxime group into an appropriate chemical backbone is a reasonable approach for the preparation of cytotoxic agents, and many oxime derivatives have been reported to have therapeutic activity for cancer[\u003cspan additionalcitationids=\"CR33\" citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Also, the introduction of oxime moiety in some natural compounds such as \u003cb\u003epsammaplin A\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) analog was responsible for high anticancer activity [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Triterpene-derived acylated oximes have demonstrated cytotoxic or antiproliferative action against numerous cancer cell lines [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] and several indirubin oxime showed anticancer activity better than natural alkaloid \u003cb\u003eindirubin\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Furthermore, the oxime derivative of natural alkaloid \u003cb\u003etryptanthrin\u003c/b\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) showed JNK1/2/3 inhibition [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Finally. Oximes have been employed in the development of several kinase inhibitors, including those for JNK [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], phosphorylase kinase (PhK), and phosphatidyl inositol 3-kinase (PI3K)[\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Indirubin oximes, for example, have a high affinity for binding to the ATP-binding site of protein kinases involved in the development of tumors, such as cyclin-dependent kinases (CDK), glycogen synthase kinase 3 (GSK) 1, vascular endothelial growth factor receptor 2 (VEGFR-2), c-Src, and casein kinase 2(CK2). Many of these kinases could serve as molecular targets for drugs that combat cancer. [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBased on the information mentioned above and in continuing of our efforts to find small compounds with potential anticancer activity, the aim of this work was to construct a hybrid series of 1,5-diarylpyrazole derivatives (Scaffold \u003cb\u003eA\u003c/b\u003e and \u003cb\u003eB\u003c/b\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) that target EGFR and JNK-2 and contain oxime as a NO release moiety to enhance anti-cancer activity. To have the crucial pharmacophoric characteristics of EGFR/JNK-2 inhibitors, Scaffolds \u003cb\u003eA\u003c/b\u003e and \u003cb\u003eB\u003c/b\u003e were designed using the ester (Scaffold A) or amide moiety (Scaffold B), as well as oxime moiety to produce hydrogen bonding connections. Moreover, vicinal 1,5-diarylpyrazole seems to be more flexible in occupying both enzymes allosteric hydrophobic sites. Different substitutions (electron donating and withdrawing groups) were used in order to study their SAR. Additionally, the distinction between scaffolds \u003cb\u003eA\u003c/b\u003e and \u003cb\u003eB\u003c/b\u003e guarantees that the optimal pharmacophore with the best replacement for enzyme binding is preserved. This hybridization was carried out to produce a synergistic impact, increase anticancer effectiveness, and/or lessen adverse effects, if any.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"2. Results And Discussion","content":"\u003cdiv class=\"Section2\" id=\"Sec3\"\u003e\n \u003ch2\u003e2.1. Chemistry\u003c/h2\u003e\n \u003cp\u003eClaisen condensation of different substituted acetophenone \u003cstrong\u003e1a-e\u003c/strong\u003e with diethyl oxalate in the presence of sodium ethoxide to gives 1,3-dicarbonyl compounds (\u0026beta;-diketoester) gave compounds \u003cstrong\u003e2a-e\u003c/strong\u003e in a good yield. 4-Hydrazinylbenzenesulfonamide hydrochloride \u003cstrong\u003e4b\u003c/strong\u003e was synthesized by diazotization of sulfanilamide with sodium nitrite and hydrochloric acid followed by reduction with sodium sulfite in the presence of sodium hydroxide and hydrochloric acid [\u003cspan class=\"CitationRef\"\u003e41\u003c/span\u003e]. 1,5-Diarylpyarzole carboxylate derivatives \u003cstrong\u003e5a-j\u003c/strong\u003e were synthesized by condensation of 1,3-dicarbonyl compounds (\u0026beta;-diketoester) \u003cstrong\u003e3a-e\u003c/strong\u003e with phenyl hydrazine \u003cstrong\u003e4a\u003c/strong\u003e directly or in the presence of sodium acetate as in compound \u003cstrong\u003e4b.\u003c/strong\u003e Hydrolysis of 1,5-diarylpyrazole ester \u003cstrong\u003e5a-j\u003c/strong\u003e derivatives with alcoholic potassium hydroxide yielded 1,5-diarylpyrazole carboxylic acid derivatives \u003cstrong\u003e6a-j\u003c/strong\u003e [\u003cspan class=\"CitationRef\"\u003e42\u003c/span\u003e]. Compounds \u003cstrong\u003e7a-j\u003c/strong\u003e were synthesized according to Steglich esterification by coupling of 1,5-diarylpyarzole carboxylic acid derivatives \u003cstrong\u003e6a-j\u003c/strong\u003e with 4-hydroxy-3-methoxy acetophenone using EDC as coupling agent and HOBt as additives in presence of DIPEA as base (Scheme \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The structures of the synthesized compounds \u003cstrong\u003e7a-j\u003c/strong\u003e were confirmed by IR, \u003csup\u003e1\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR and HRMS (ESI) spectroscopy. The IR spectra showed significant stretching bands at 1658\u0026ndash;1746 cm\u003csup\u003e-1\u003c/sup\u003e related to carbonyl of ester group (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003eO-Ph) and at 1162\u0026ndash;1164 cm\u003csup\u003e-1\u003c/sup\u003e for compounds \u003cstrong\u003e7f-j\u003c/strong\u003e related to (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) group. The \u003csup\u003e1\u003c/sup\u003eH-NMR spectra of compounds \u003cstrong\u003e7a-j\u003c/strong\u003e showed two common singlet peaks at \u0026delta; 3.71\u0026ndash;3.85 ppm related to methoxy group of acetophenone and at \u0026delta; 2.51\u0026ndash;2.75 ppm attributed to (CO-\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCH\u003c/span\u003e\u003csub\u003e\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003e3\u003c/span\u003e\u003c/sub\u003e). The \u003csup\u003e13\u003c/sup\u003eC-NMR spectra of compounds \u003cstrong\u003e7a-j\u003c/strong\u003e showed significant signals related to carbonyl carbon of ketone (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e) appeared at \u0026delta; 197.11\u0026ndash;197.80 ppm. Peak at \u0026delta; 159.99-167.77 ppm was related to carbonyl carbon of ester (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003eO-Ph), at \u0026delta; 54.96\u0026ndash;57.14 ppm attributed to carbon of methoxy group that attached to acetophenone moiety and at \u0026delta; 26.65\u0026ndash;28.85 ppm attributed to carbon of methyl group (CO-\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCH\u003c/span\u003e\u003csub\u003e\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003e3\u003c/span\u003e\u003c/sub\u003e). HRMS (ESI) data for compounds \u003cstrong\u003e7a-j\u003c/strong\u003e further confirmed their assigned structure. The m/z value of molecular ion peak [M\u0026thinsp;+\u0026thinsp;1]\u003csup\u003e+\u003c/sup\u003e or [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e was close to calculated one in all target compounds.\u003c/p\u003e\n \u003cp\u003eThe target oxime derivatives \u003cstrong\u003e8a-j\u003c/strong\u003e were prepared by refluxing a mixture of ketone intermediates \u003cstrong\u003e7a-j\u003c/strong\u003e and hydroxylamine hydrochloride in absolute ethanol. The chemical structure of the prepared compounds was elucidated by IR, \u003csup\u003e1\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR and HRMS spectroscopy. The IR spectra of compounds \u003cstrong\u003e8a-j\u003c/strong\u003e were characterized by the appearance of intense broad bands at 3139\u0026ndash;3681 cm\u003csup\u003e-1\u003c/sup\u003e related to OH group, in addition to (COO-Ph) that exhibited stretching vibration at 1717\u0026ndash;1756 cm\u003csup\u003e-1\u003c/sup\u003e. A characteristic feature of the \u003csup\u003e1\u003c/sup\u003eH-NMR spectra for oximes \u003cstrong\u003e8a-j\u003c/strong\u003e is the appearance of downfield singlets in the range \u0026delta; 10.36\u0026ndash;11.34 ppm related to the hydroxyl group. The resonances of CH\u003csub\u003e3\u003c/sub\u003e protons were observed in the expected regions at \u0026delta; 1.90\u0026ndash;2.28 ppm and appeared to be more upfield shifted than the CH\u003csub\u003e3\u003c/sub\u003e protons of the corresponding ketones by 0.40\u0026ndash;0.60 ppm due to the low electronegativity of N atom of the oxime relative to O atom of the ketone. Also, all the aromatic protons appeared in their expected chemical shift. One of the characteristic features of \u003csup\u003e13\u003c/sup\u003eC-NMR spectra of compounds \u003cstrong\u003e8a-j\u003c/strong\u003e is the disappearance of ketonic carbonyl due to its conversation to ketoxime group (C\u0026thinsp;=\u0026thinsp;N-OH), which appeared at \u0026delta; 150.82-159.99 ppm. Also, the methyl group attached to ketoxime appeared at \u0026delta; 11.65\u0026ndash;14.71 ppm. HRMS (ESI) data for compounds \u003cstrong\u003e8a-j\u003c/strong\u003e further confirmed their assigned structure. The m/z value of molecular ion peak [M\u0026thinsp;+\u0026thinsp;1]\u003csup\u003e+\u003c/sup\u003e or [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e was close to calculated one in all cases.\u003c/p\u003e\n \u003cp\u003eCompounds \u003cstrong\u003e9a-c\u003c/strong\u003e were synthesized by activating 1,5-diarypyrazole carboxylic acid derivatives \u003cstrong\u003e6f\u003c/strong\u003e, \u003cstrong\u003e6h\u003c/strong\u003e and \u003cstrong\u003e6j\u003c/strong\u003e with thionyl chloride in benzene to obtain acyl chloride derivatives, which were coupled with 4-aminoacetophenone by heating in dry DMF in presence of triethylamine as base (\u003cstrong\u003eScheme 2\u003c/strong\u003e). The structure of the synthesized compounds \u003cstrong\u003e9a-c\u003c/strong\u003e was confirmed by IR, \u003csup\u003e1\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR and HRMS (ESI) spectroscopy. The IR spectra showed significant stretching bands at 1669\u0026ndash;1681 cm\u003csup\u003e-1\u003c/sup\u003e assigned to (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003eNH) and at 1160\u0026ndash;1161 cm\u003csup\u003e-1\u003c/sup\u003e related to (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e). In the \u003csup\u003e1\u003c/sup\u003eH-NMR spectra for compounds \u003cstrong\u003e9a-c\u003c/strong\u003e two singlet peaks are common and appeared at \u0026delta; 10.53\u0026ndash;11.18 ppm related to the amidic (NH) proton and at \u0026delta; 2.47\u0026ndash;2.53 ppm related to (CO-\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCH\u003c/span\u003e\u003csub\u003e\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003e3\u003c/span\u003e\u003c/sub\u003e). The \u003csup\u003e13\u003c/sup\u003eC-NMR spectra of compounds \u003cstrong\u003e9a-c\u003c/strong\u003e showed significant signals related to carbonyl carbon of ketone (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e) appeared at \u0026delta; 195.96-197.09 ppm, peak at \u0026delta; 164.12-167.91 ppm related to carbonyl carbon of amide (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003eNH) and at \u0026delta; 26.61\u0026ndash;26.95 ppm related to carbon of methyl (CO-\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCH\u003c/span\u003e\u003csub\u003e\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003e3\u003c/span\u003e\u003c/sub\u003e). HRMS (ESI) data for compounds \u003cstrong\u003e9a-c\u003c/strong\u003e confirmed their assigned structure. The m/z value of molecular ion peak [M-1]\u003csup\u003e-\u003c/sup\u003e or [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e was close to the calculated one in all cases .\u003c/p\u003e\n \u003cp\u003eOxime derivatives \u003cstrong\u003e10a-c\u003c/strong\u003e were synthesized by refluxing a mixture of ketone intermediates \u003cstrong\u003e9a-c\u003c/strong\u003e and hydroxylamine hydrochloride in absolute ethanol. The chemical structure of the prepared compounds was elucidated by IR, \u003csup\u003e1\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR and HRMS spectroscopy. The IR spectra of compounds \u003cstrong\u003e10a-c\u003c/strong\u003e was characterized by the appearance of intense broad bands at 3220\u0026ndash;3681 cm\u003csup\u003e-1\u003c/sup\u003e related to OH and NH groups, in addition to (\u003cspan class=\"Underline\" name=\"Emphasis\" type=\"Underline\"\u003eCO\u003c/span\u003e-NH) and (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) groups that exhibited stretching vibration at 1677\u0026ndash;1680 cm\u003csup\u003e-1\u003c/sup\u003e and 1161\u0026ndash;1162 cm\u003csup\u003e-1\u003c/sup\u003e, respectively. A characteristic feature of the \u003csup\u003e1\u003c/sup\u003eH-NMR spectra for oximes \u003cstrong\u003e10a-c\u003c/strong\u003e was the appearance of downfield singlets in the range \u0026delta; 8.75\u0026ndash;10.9 ppm related to the hydroxyl group. The resonances of NH and CH\u003csub\u003e3\u003c/sub\u003e protons were observed in the expected regions at \u0026delta; 10.30-10.36 ppm and \u0026delta; 2.06\u0026ndash;2.08 ppm, respectively. The CH\u003csub\u003e3\u003c/sub\u003e protons appeared to be more upfield shifted than the CH\u003csub\u003e3\u003c/sub\u003e protons of the corresponding ketones by 0.40\u0026ndash;0.45 ppm due to the low electronegativity of the N atom of the oxime relative to O atom of the ketone. One of the characteristic features of \u003csup\u003e13\u003c/sup\u003eC-NMR spectra of compounds \u003cstrong\u003e10a-c\u003c/strong\u003e is the disappearance of ketonic carbonyl due to its conversation to ketoxime group (C\u0026thinsp;=\u0026thinsp;N-OH), which appeared at \u0026delta; 153.24-159.18 ppm. Also, the methyl group attached to ketoxime appeared at \u0026delta; 12.14\u0026ndash;17.34 ppm. The HRMS (ESI) data for compounds \u003cstrong\u003e10a-c\u003c/strong\u003e further confirmed their assigned structure. The m/z value of the molecular ion peak [M-1]\u003csup\u003e-\u003c/sup\u003e or [M\u0026thinsp;+\u0026thinsp;1]\u003csup\u003e+\u003c/sup\u003e was close to calculated one in all target compounds.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec4\"\u003e\n \u003ch2\u003e2.2 Biology\u003c/h2\u003e\n \u003cdiv class=\"Section3\" id=\"Sec5\"\u003e\n \u003ch2\u003e2.2.1. \u003cem\u003eIn vitro\u003c/em\u003e antiproliferative screening activities.\u003c/h2\u003e\n \u003cp\u003eCompounds \u003cstrong\u003e7a-j, 8a-j, 9a-c\u003c/strong\u003e, and \u003cstrong\u003e10a-c\u003c/strong\u003e were evaluated for \u003cem\u003ein vitro\u003c/em\u003e anticancer activities against different five cancer cell lines namely, human colorectal adenocarcinoma cell lines DLD-1, human cervical cancer cell line Hela, human pancreatic cancer cell line SUIT-2, human myelogenous leukemia cell line K562 and human hepatocellular carcinoma cell line HepG2 using WST-8 assay at concentration of 100 \u0026micro;M to investigate the growth inhibition percent (GI%) of each compound using \u003cstrong\u003edaunorubicin\u003c/strong\u003e as reference drug [\u003cspan class=\"CitationRef\"\u003e43\u003c/span\u003e] (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u0026nbsp;\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAntiproliferative activity of compounds \u003cstrong\u003e7a-j\u003c/strong\u003e, \u003cstrong\u003e8a-j\u003c/strong\u003e, \u003cstrong\u003e9a-c, 10a-c\u003c/strong\u003e and \u003cstrong\u003edaunorubicin\u003c/strong\u003e against DLD-1, Hela, K562, SUIT-2 and HepG2 cell lines at 100 \u0026micro;M using WST-8 assay.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eCompound no.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eGrowth inhibition (GI%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDLD-1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHela\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eK562\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSUIT-2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHepG2\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e7a\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e101\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e81\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e83\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e70\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e103\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e85\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e52\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e48\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e68\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e72\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e104\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e104\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e59.40\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e73\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e101\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e14\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7f\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e18.65\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e16\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e24\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e75\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e96.70\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e77\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e92\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e66\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e73\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e46.00\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e101\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e101\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e101\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e63\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7j\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e81\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e25.70\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e33\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e94\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e77\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e92.00\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e60\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e99\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e102\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e107\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e78\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e67\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e61\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e51\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e72.00\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e48\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e95\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e77\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e98\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e29.20\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e77\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e93\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e83\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e99\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8f\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e72.90\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e107\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e76\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e95\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e80\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e85.80\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e78\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e87\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e96\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e93\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e52.30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e78\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e73\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e63\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e34.00\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e97\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e98\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e90\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8j\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e9.60\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e53\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e34\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e79\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e60\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e9a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e60.70\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e66\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e60\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e36\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e9b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e42.15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e45\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e19\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e9c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e16.80\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e57\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e42\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e99.00\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e84\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e88\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e72\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e72.50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e84\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e92.40\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e75\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e87\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e103\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e52\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eDaunorubicin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e82.45\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e92\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003eFrom the screening results in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, compounds \u003cstrong\u003e7a-j\u003c/strong\u003e and \u003cstrong\u003e9a-c\u003c/strong\u003e that are 1,5-diarylpyrazole acetophenone derivatives displayed considerable cytotoxicity towards the pancreatic cell line SUIT-2 with GI% ranging from 68\u0026ndash;104% for compounds \u003cstrong\u003e7a-j\u003c/strong\u003e and 42\u0026ndash;60% for compounds \u003cstrong\u003e9a-c\u003c/strong\u003e. Compounds \u003cstrong\u003e7b\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H) demonstrated moderate to high cytotoxicity against five cancer cell lines with GI% ranging from 46\u0026ndash;103%, while compound \u003cstrong\u003e7d\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-Cl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H) established an excellent cytotoxicity towards DLD-1, Hela, and SUIT-2 with GI% of 72%, 104%, and 104%, respectively, and moderate cytotoxicity against K562 cell line with GI% 50. Furthermore, compound \u003cstrong\u003e7i\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-Cl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) displayed superior antiproliferative activity against Hela, K562 and SUIT-2 cell lines with GI% of 101%,101% and 101% and moderate activity against HepG2 cell line with GI% of 63%. Importantly, all oxime derivatives \u003cstrong\u003e8a-j\u003c/strong\u003e and \u003cstrong\u003e10a-c\u003c/strong\u003e exhibited marked antiproliferative activity in comparison to ketone derivatives \u003cstrong\u003e9a-c\u003c/strong\u003e and \u003cstrong\u003e11a-c\u003c/strong\u003e as a result of the role of oxime moiety in cytotoxicity. Compounds \u003cstrong\u003e8b-j\u003c/strong\u003e demonstrated a significant cytotoxicity against SUIT-2 and HepG2 cell lines with GI% ranging from 51\u0026ndash;107%. Compounds \u003cstrong\u003e8b\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H), \u003cstrong\u003e8f\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) and \u003cstrong\u003e8g\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) exhibited broadness cytotoxicity in five cancer cell lines with GI% ranging from 60\u0026ndash;107%. Moreover, compounds \u003cstrong\u003e8a-i\u003c/strong\u003e exhibited high antiproliferative activity against leukemia cell line K562 with GI% ranging from 67\u0026ndash;99%. Compound \u003cstrong\u003e8e\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3,4-OCH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H) displayed remarked cytotoxicity in Hela, K562, SUIT-2 and HepG2 with GI% of 77%, 93%, 83% and 99% respectively, while compound \u003cstrong\u003e8i\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-Cl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) exhibited high antiproliferative activity against Hela, K562, SUIT-2 and HepG2 with GI% of 100%, 97%, 98% and 90% respectively. Furthermore, compounds \u003cstrong\u003e10a-c\u003c/strong\u003e showed moderate to high cytotoxicity in pancreatic cancer cell line SUIT-2 with GI% of 88%, 50% and 103% respectively, only compound \u003cstrong\u003e10c\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u0026thinsp;=\u0026thinsp;3,4-OCH\u003csub\u003e3\u003c/sub\u003e) demonstrated antiproliferative activity in the five cancer cell lines with GI% ranging from 52\u0026ndash;103%. So as a conclusion on the SAR study of those compounds as antiproliferative agents, oxime moiety potentiated the anticancer activity and electron donating groups on R\u003csup\u003e1\u003c/sup\u003e played an important role on the activity. Moreover, the sulfamoyl moiety on R\u003csup\u003e2\u003c/sup\u003e seems to be of potential role in the activity of the prepared compounds.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec6\"\u003e\n \u003ch2\u003e2.2.2 \u003cem\u003eIn vitro\u003c/em\u003e cytotoxicity measurements (IC\u003csub\u003e50\u003c/sub\u003e) against five cancer cell lines.\u003c/h2\u003e\n \u003cp\u003eFor further investigation, compounds with committed antiproliferative activity against five cancer cell lines (DLD-1, Hela, K562, SUIT-2 and HepG2) at 100 \u0026micro;M were selected to measure growth inhibition percentage using WST-8 assay at different six concentrations of 1, 10, 20, 50, 80, 100 \u0026micro;M for calculating their IC\u003csub\u003e50\u003c/sub\u003e using the daunorubicin as reference. All selected compounds and daunorubicin were recorded as the minimum concentration required to inhibit half cell growth (IC\u003csub\u003e50\u003c/sub\u003e) and results were listed in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab2\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003e\u003cem\u003eIn vitro\u003c/em\u003e antiproliferative activity of the most active compounds expressed as IC\u003csub\u003e50\u003c/sub\u003e values using WST-8 assay against DLD-1, Hela, K562, SUIT-2 and HepG2 cancer cell lines. The results recorded as IC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;M) using \u003cstrong\u003edaunorubicin\u003c/strong\u003e as reference.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eCompound no.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;M)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDLD-1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHela\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eK562\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSUIT-2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHepG2\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e7b\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNT\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e81\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e43\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eND\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e93\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e92\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e32\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e27\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e35.7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e14.4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e57\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e23.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e20\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8f\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e15.6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e22.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e32.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7.6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e19\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e12.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e74\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e21\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e26\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eND\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e71\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e62\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eND\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e26\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e16\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eND\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e36\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eND\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e29\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eDaunorubicin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.097\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e13.30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003ea: ND means not determined, b: NT means not tested\u003c/p\u003e\n \u003cp\u003eThe target oxime derivatives exhibited promising antiproliferative activity against five cancer cell line as listed in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e more than the corresponding ketone, such as compounds \u003cstrong\u003e8b\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H), \u003cstrong\u003e8d\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-Cl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H), \u003cstrong\u003e8g\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), \u003cstrong\u003e10a\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u0026thinsp;=\u0026thinsp;H), and \u003cstrong\u003e10b\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e- OCH\u003csub\u003e3\u003c/sub\u003e) that showed remarkable cytotoxicity against DLD-1 cell line with IC\u003csub\u003e50\u003c/sub\u003e of 10, 14.4, 32.30, 26 and 36 \u0026micro;M, respectively in comparison to daunorubicin (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;30 \u0026micro;M). Also, compounds \u003cstrong\u003e8g\u003c/strong\u003e, \u003cstrong\u003e8i\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) and \u003cstrong\u003e10c\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3,4-di-OCH\u003csub\u003e3\u003c/sub\u003e) demonstrated high antiproliferative activity against Hela cell line with IC\u003csub\u003e50\u003c/sub\u003e of 8, 13, and 5 \u0026micro;M respectively, while compounds \u003cstrong\u003e8b, 8d, 8e\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3,4-OCH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H), \u003cstrong\u003e8f\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) and \u003cstrong\u003e8h\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-OCH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) showed a moderate antiproliferative activity with IC\u003csub\u003e50\u003c/sub\u003e of 32, 57, 22, 22 and 74 \u0026micro;M in comparison with daunorubicin. Moreover, compounds \u003cstrong\u003e8b\u003c/strong\u003e, \u003cstrong\u003e8d\u003c/strong\u003e, \u003cstrong\u003e8f\u003c/strong\u003e, \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e10a\u003c/strong\u003e established excellent anticancer activity in comparison to daunorubicin (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;13 \u0026micro;M) against the human myelogenous leukemia cell line K562 with IC\u003csub\u003e50\u003c/sub\u003e of 13, 9, 15.6, 7.6 and 16 \u0026micro;M. Compounds \u003cstrong\u003e8a\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H), \u003cstrong\u003e8e\u003c/strong\u003e, \u003cstrong\u003e8h\u003c/strong\u003e, and \u003cstrong\u003e10c\u003c/strong\u003e exhibited good anticancer activity at the same cell line with IC\u003csub\u003e50\u003c/sub\u003e of 22, 20, 21 and 29 \u0026micro;M. Furthermore, compounds \u003cstrong\u003e8b\u003c/strong\u003e, \u003cstrong\u003e8g\u003c/strong\u003e, \u003cstrong\u003e8h\u003c/strong\u003e and \u003cstrong\u003e10c\u003c/strong\u003e showed a significant anticancer activity against the human pancreatic cancer cell line SUIT-2 with IC\u003csub\u003e50\u003c/sub\u003e of 27, 19, 26 and 13 \u0026micro;M, respectively. Compounds \u003cstrong\u003e8e\u003c/strong\u003e and \u003cstrong\u003e8g\u003c/strong\u003e demonstrated excellent antiproliferative activity better than daunorubicin (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;22 \u0026micro;M) against the hepatocellular carcinoma cell line HepG2 with IC\u003csub\u003e50\u003c/sub\u003e of 4.7 and 12.3 \u0026micro;M, respectively. In addition, compounds \u003cstrong\u003e8d\u003c/strong\u003e and \u003cstrong\u003e8f\u003c/strong\u003e showed equal anticancer activity to daunorubicin at the same cell line with IC\u003csub\u003e50\u003c/sub\u003e of 23.3 and 22.3 \u0026micro;M, respectively. Unlike to anticancer activity of ketone derivatives \u003cstrong\u003e7a-j\u003c/strong\u003e, compounds \u003cstrong\u003e7b\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H) and \u003cstrong\u003e7g\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) displayed excellent antiproliferative activity in comparison with daunorubicin against human colorectal adenocarcinoma DLD-1 with IC\u003csub\u003e50\u003c/sub\u003e of 13 \u0026micro;M. Also, compound \u003cstrong\u003e7d\u003c/strong\u003e established remarkable anticancer activity towards the human cervical cancer cell line Hela with IC\u003csub\u003e50\u003c/sub\u003e of 15 \u0026micro;M.\u003c/p\u003e\n \u003cp\u003eSubstituents on the terminal phenyl ring of the 1,5-diarylpyrazole part, showed a significant effect on the biological profile of anticancer activity. Compounds \u003cstrong\u003e8b\u003c/strong\u003e, \u003cstrong\u003e8f, 8g, 8h\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e, which are considered the most potent anticancer oxime derivatives showed substituents on \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H, \u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ep\u003c/em\u003e-Cl, but when \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;OCH\u003csub\u003e3\u003c/sub\u003e, moderate anticancer activity was observed as in compound \u003cstrong\u003e8h\u003c/strong\u003e, which indicated the presence of a lipophilic group (Cl, CH\u003csub\u003e3\u003c/sub\u003e) improving anticancer activity Also, the sulfamoyl group at the \u003cem\u003epara\u003c/em\u003e position of the phenyl ring of the diarylpyrazole part is essential for the broadness of anticancer activities such as compounds \u003cstrong\u003e8g\u003c/strong\u003e, \u003cstrong\u003e8i\u003c/strong\u003e, \u003cstrong\u003e8h\u003c/strong\u003e and compound \u003cstrong\u003e10c\u003c/strong\u003e as results of hydrogen bonding formation on the active site. Meanwhile, compound \u003cstrong\u003e10c\u003c/strong\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3,4-di OCH\u003csub\u003e3\u003c/sub\u003e established good anticancer activity with (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;5\u0026ndash;29 \u0026micro;M) and the remaining compounds in scaffold B showed weak anticancer activity. The difference between the hydrophilic and hydrophobic substitutions in scaffold A, as well as the presence of the methoxy group in the 4-hydroxy-3-methoxyl acetophenone carrying oxime moiety, led to scaffold A superior anticancer efficacy compared to scaffold B. The anticancer properties of 3,4-di-OCH3-containing compounds, however, were goo in both scaffolds.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec7\"\u003e\n \u003ch2\u003e2.2.3. Evaluation of EGFR inhibitory activity\u003c/h2\u003e\n \u003cp\u003eBeing over-expressed in a variety of human cancers and connected to cancer proliferation, angiogenesis, and metastasis, EGFR has received substantial study and clinical validation as a target for cancer treatment. Most of these medications are designed to bind to the ATP active site of EGFR-TK. The structural study of previously reported instances of tyrosine kinase anticancer medications served as the basis for the introduction of such drugs [\u003cspan class=\"CitationRef\"\u003e4\u003c/span\u003e]. Herein, the human EGFR-TK Elisa kit assay was performed to evaluate the \u003cem\u003ein vitro\u003c/em\u003e EGFR-inhibitory potency of the more active anticancer compounds \u003cstrong\u003e8b\u003c/strong\u003e, \u003cstrong\u003e8d\u003c/strong\u003e, \u003cstrong\u003e8g\u003c/strong\u003e, \u003cstrong\u003e8i\u003c/strong\u003e and \u003cstrong\u003e10c\u003c/strong\u003e using the multi-target kinase inhibitor drug sorafenib as a reference using quantitative sandwich enzyme immunoassay technology [\u003cspan class=\"CitationRef\"\u003e44\u003c/span\u003e]. The test investigated the potential of the test compounds to bind to EGFR, resulting in suppression of epidermal growth factor from binding to EGFR that led to inhibition of receptor dimerization and tyrosine autophosphorylation and suppression of cancer cell proliferation. Screening results of EGFR inhibitory activity of tested compounds and sorafenib expressed as IC\u003csub\u003e50\u003c/sub\u003e in \u0026micro;M and recorded in Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. Results showed that compound \u003cstrong\u003e8g\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep-\u003c/em\u003eCH\u003csub\u003e3\u003c/sub\u003e, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), \u003cstrong\u003e8i\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep-\u003c/em\u003eCl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) and \u003cstrong\u003e10c (R\u003c/strong\u003e\u0026thinsp;=\u0026thinsp;3,4-di OCH\u003csub\u003e3\u003c/sub\u003e) exhibited moderate EGFR inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e of 18, 21 and 12 \u0026micro;M, respectively. On the other hand, compound \u003cstrong\u003e8d\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep-\u003c/em\u003eCl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H) displayed successful EGFR inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e of 8 \u0026micro;M in comparison to positive control drug sorafenib (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;3.5 \u0026micro;M). These findings revealed that lipophilic substitutions on vicinal 1,5- diarylpyarzole, together with the oxime moiety as in compound \u003cstrong\u003e8d\u003c/strong\u003e, have good fitting and binding on EGFR, making it an effective EGFR inhibitor.\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab3\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eEGFR-TK inhibitory activity of target compounds represented as IC\u003csub\u003e50\u003c/sub\u003e using sorafenib as reference.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCompound no\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;M)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8b\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026gt;\u0026thinsp;1000\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8d\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e21\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e12\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003esorafenib\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec8\"\u003e\n \u003ch2\u003e2.2.4. Evaluation of JNK-2 inhibitory activity\u003c/h2\u003e\n \u003cp\u003eJNK-2 is a member of the MAP kinase family involved in signaling pathways, which has been implicated in several diseases like cancer and inflammatory diseases [\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e]. So, this family is widely considered for targeting by small molecule therapeutics [\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e]. JNK-2 inhibitors are classified to two types: Type I inhibitors target the DFG-in conformation (open conformation), while type II inhibitors target the DFG-out conformation (closed conformation) [\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e14\u003c/span\u003e]. Based on the vital role of JNK-2 in human malignancies as a result of its contribution to multiple cancer-related pathways and the reported role of celecoxib as JNK inhibitor, we examined the JNK-2 inhibitory activity of compounds \u003cstrong\u003e8d\u003c/strong\u003e, \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e using the multitarget kinase drug sorafenib as reference [\u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e]. The JNK-2 inhibitory activity of the selected compounds was investigated \u003cem\u003ein vitro\u003c/em\u003e using a simple step ELISA kit for quantitative measurement of JNK-2 (pT138/Y185) protein in human cells, which investigated the possible binding of tested compounds in the ATP binding site of JNK-2, leading to inhibition of substrate binding on JNK-2 enzyme and inhibition of the JNK-2 pathway that could explain the antiproliferative activity of these compounds [\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e]. Screening results of JNK-2 inhibitory activity of tested compounds and sorafenib expressed as IC\u003csub\u003e50\u003c/sub\u003e in \u0026micro;M and recorded in Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e. Results showed that the oxime derivative \u003cstrong\u003e8i\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-Cl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e) is a potent inhibitor of JNK-2 with an IC\u003csub\u003e50\u003c/sub\u003e of 1 \u0026micro;M same as the activity of the multi-target kinase sorafenib. These results indicated that the anticancer activity of compound \u003cstrong\u003e8i\u003c/strong\u003e was due to the dual inhibition of EGFR and JNK-2. Also, compound \u003cstrong\u003e8d\u003c/strong\u003e (\u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ep\u003c/em\u003e-Cl, \u003cstrong\u003eR\u003c/strong\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H) showed moderate JNK-2-inhibiting activity with an IC\u003csub\u003e50\u003c/sub\u003e of 49 \u0026micro;M, which indicated its anticancer activity because of dual inhibition of EGFR and JNK-2. It was clear from these data that the oxime moiety and \u003cem\u003ep\u003c/em\u003e-Cl substitution improved the anticancer activity of two compounds in addition to the sulfamoyl moiety that makes compound \u003cstrong\u003e8i\u003c/strong\u003e more potent as JNK-2 inhibitor.\u0026nbsp;\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab4\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eJNK-2 inhibitory activity of target compounds represented as IC\u003csub\u003e50\u003c/sub\u003e using sorafenib as reference.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCompound no\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;M)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8d\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8g\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026gt;\u0026thinsp;200\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSorafenib\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec9\"\u003e\n \u003ch2\u003e2.2.5. Cell cycle analysis and apoptosis detection\u003c/h2\u003e\n \u003cdiv class=\"Section4\" id=\"Sec10\"\u003e\n \u003ch2\u003e2.2.5.1. Cell cycle analysis\u003c/h2\u003e\n \u003cp\u003eThis analysis was applied to investigate the effects of tested compounds on cell cycle distribution and on cell-death associated DNA fragmentation. The Hela cell line was analyzed flow cytometrically after propidium iodide (PI) staining following treatment of compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e. As shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, compound \u003cstrong\u003e8g\u003c/strong\u003e established G2/M arrest of Hela cancer cells indicating cell death and DNA fragmentation and the percentage of Hela cells at G2 phase increased from 7.3% (DMSO treated Hela cell) to 16.5% after 24h and 20.5% after 48h. While compound \u003cstrong\u003e8i\u003c/strong\u003e showed a combined S phase and G2 phase arrest and the percentage of Hela cells at S phase increased from 8.47% (control) to 59.4%, while the percentage of Hela cells at G2 phase raised from 7.3% (control) to 18.2%.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section4\" id=\"Sec11\"\u003e\n \u003ch2\u003e2.2.5.2. Apoptosis assay\u003c/h2\u003e\n \u003cp\u003eFor further investigation of the anticancer activity of compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e, studies of apoptotic changes after treatment of Hela cells with these inhibitors were examined using fluorescent microscope and flowcytometry. Moreover, to discriminate between apoptosis and necrosis after treatment of Hela cell with two inhibitors \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e at different concentrations, a fluorescent microscope was used to distinguish between apoptosis and necrosis based on their characteristic difference in morphology using annexin V and PI [\u003cspan class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e\n \u003cp\u003eAs shown in Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e, compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e established remarkable apoptosis at low concentrations that was marked in green because of the binding of annexin-v with phosphatidylserine. This was exposed in the cell membrane of Hela cells after treatment with two the inhibitors due to plasma membrane sprouting and chromatin concentration. Prolonged incubation of a Hela cells with two the inhibitors directed cells to necrosis, which was marked in red because of PI staining of necrotic cells as a result of losing their dye-excluding ability owing to loss of plasma membrane integrity and dissolution of nuclear chromatin [\u003cspan class=\"CitationRef\"\u003e49\u003c/span\u003e].\u003c/p\u003e\n \u003cp\u003eSubsequently, flowcytometric analysis of the effect of compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e on Hela cell apoptosis at concentration double the IC\u003csub\u003e50\u003c/sub\u003e of each compound for 24h using Annexin v/PI were investigated and apoptotic marker changes for each compound on Hela cells were analyzed in comparison to control untreatable Hela cells. As appeared in Figs. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e the parentage of apoptotic Hela cells was increased significantly after treatment with compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e from 1.82% for control untreatable Hela cells to 34.7% for compound \u003cstrong\u003e8g\u003c/strong\u003e and 17.3% for compound \u003cstrong\u003e8i\u003c/strong\u003e. These results attributed that those apoptotic cells were increased after treatment of these compounds as result of antiproliferative activity not due to cytotoxicity.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec12\"\u003e\n \u003ch2\u003e2.2.6. Evaluation of cytotoxicity towards the normal cell line PC12\u003c/h2\u003e\n \u003cp\u003eEvaluation of normal cell viability is fundamental to analyze the efficacy of target compounds and is often used in conjunction with cytotoxicity tests to help understand how target compounds toxicity affects normal cell health.\u003c/p\u003e\n \u003cp\u003eTo investigate the selectivity of the target compounds towards cancer cells, the cytotoxicity of compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e was measured against the normal cell line PC12 (rat adrenal-derived pheochromocytoma cells) using a WST-8 assay at six concentrations 1, 10, 20, 50, 80, 100 \u0026micro;M to calculate CC\u003csub\u003e50\u003c/sub\u003e in comparison to \u003cstrong\u003edaunorubicin\u003c/strong\u003e as reference drug as recorded in Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e. The results listed in Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e showed that no cytotoxicity was observed with compound \u003cstrong\u003e8i\u003c/strong\u003e against PC12 cell line with CC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;\u0026gt;\u0026thinsp;100 \u0026micro;M. While compound \u003cstrong\u003e8g\u003c/strong\u003e showed a moderate cytotoxicity with CC\u003csub\u003e50\u003c/sub\u003e of 16.1 \u0026micro;M in comparison to daunorubicin, which established high cytotoxicity towards PC12 cell line with percentage growth inhibition of 113% at 100\u0026micro;M, 107% at 50\u0026micro;M and 81% at 1\u0026micro;M. These results indicated that these target compounds had no or little cytotoxicity against the normal cell line and demonstrated selectivity towards cancer cells.\u0026nbsp;\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab5\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCytotoxicity of compounds \u003cstrong\u003e8g, 8i\u003c/strong\u003e, and \u003cstrong\u003edaunorubicin\u003c/strong\u003e against the PC12 cell line represented as CC\u003csub\u003e50\u003c/sub\u003e.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCompound no\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;M)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8g\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e16.1\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8i\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026gt;\u0026thinsp;100\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eDaunorubicin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec13\"\u003e\n \u003ch2\u003e2.3. Measurement of nitric oxide release\u003c/h2\u003e\n \u003cp\u003eGriess colorimetric method was used for the indirect determination of NO, which includes spectrophotometry measurements of the stable decomposition products NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. This method requires that NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e first reduced to NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and then NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e is determined by the Griess reaction that includes two steps, the first step is a diazotization reaction where the NO-derived nitrogen agent, dinitrogen trioxide (N\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e) resulting from the spontaneous oxidation of NO with sulfanilamide to produce the diazonium ion. The second step is the coupling of diazonium with \u003cem\u003eN\u003c/em\u003e-(1-napthyl)ethylenediamine dihydrochloride (NEDD) to form a strongly absorbed azo colorimetric product at \u0026lambda;\u003csub\u003emax\u003c/sub\u003e 546 nm. In order to evaluate thiol-induced NO generation from the appropriate compounds including NO-donating oximes \u003cstrong\u003e8a-b, 8d-i\u003c/strong\u003e and \u003cstrong\u003e10a-c\u003c/strong\u003e, they were incubated in aqueous phosphate buffer of pH 7.4 in the presence of excess \u003cem\u003eN\u003c/em\u003e-acetylcysteine, which serves as a source of thiols that are essential for release of NO from oximes [\u003cspan class=\"CitationRef\"\u003e50\u003c/span\u003e]. The signal intensity of the dye is proportional to the amount of NO released. To quantify the amount of NO released, a standard curve was made by measuring the change in absorbance of various concentration of standard sodium nitrite solutions treated by the same way. The results expressed as amount of NO released (mol/mol) and listed in Table \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e. The obtained results indicated that the NO-donating oximes \u003cstrong\u003e8a-b, 8d-i\u003c/strong\u003e and \u003cstrong\u003e10a-c\u003c/strong\u003e achieved maximum amount of NO released after 2 hours. \u0026nbsp;\u003c/p\u003e\n \u003ctable border=\"1\" id=\"Tab6\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eThe amount of NO released from compounds \u003cstrong\u003e8a-b, 8d-i\u003c/strong\u003e and \u003cstrong\u003e10a-c\u003c/strong\u003e in phosphate buffer of pH\u0026thinsp;=\u0026thinsp;7.4.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eCompound\u003c/p\u003e\n \u003cp\u003eNo.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eAmount of NO released (mol/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e1h\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e2h\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e3h\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e4h\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8a\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0.095\u0026thinsp;\u0026plusmn;\u0026thinsp;0.036\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0.121\u0026thinsp;\u0026plusmn;\u0026thinsp;0.047\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0.108\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0.056\u0026thinsp;\u0026plusmn;\u0026thinsp;0.022\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.102\u0026thinsp;\u0026plusmn;\u0026thinsp;0.039\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.159\u0026thinsp;\u0026plusmn;\u0026thinsp;0.019\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.105\u0026thinsp;\u0026plusmn;\u0026thinsp;0.025\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.095\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.096\u0026thinsp;\u0026plusmn;\u0026thinsp;0.036\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.106\u0026thinsp;\u0026plusmn;\u0026thinsp;0.040\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.101\u0026thinsp;\u0026plusmn;\u0026thinsp;0.038\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.079\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.064\u0026thinsp;\u0026plusmn;\u0026thinsp;0.022\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.120\u0026thinsp;\u0026plusmn;\u0026thinsp;0.046\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.081\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.048\u0026thinsp;\u0026plusmn;\u0026thinsp;0.018\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8f\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.096\u0026thinsp;\u0026plusmn;\u0026thinsp;0.036\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.116\u0026thinsp;\u0026plusmn;\u0026thinsp;0.045\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.079\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.048\u0026thinsp;\u0026plusmn;\u0026thinsp;0.018\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.083\u0026thinsp;\u0026plusmn;\u0026thinsp;0.035\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.154\u0026thinsp;\u0026plusmn;\u0026thinsp;0.044\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.141\u0026thinsp;\u0026plusmn;\u0026thinsp;0.039\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.089\u0026thinsp;\u0026plusmn;\u0026thinsp;0.019\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.099\u0026thinsp;\u0026plusmn;\u0026thinsp;0.029\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.158\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.104\u0026thinsp;\u0026plusmn;\u0026thinsp;0.033\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.094\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0022\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.105\u0026thinsp;\u0026plusmn;\u0026thinsp;0.029\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.115\u0026thinsp;\u0026plusmn;\u0026thinsp;0.018\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.109\u0026thinsp;\u0026plusmn;\u0026thinsp;0.046\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.087\u0026thinsp;\u0026plusmn;\u0026thinsp;0.035\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.054\u0026thinsp;\u0026plusmn;\u0026thinsp;0.023\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.108\u0026thinsp;\u0026plusmn;\u0026thinsp;0.043\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.069\u0026thinsp;\u0026plusmn;\u0026thinsp;0.024\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.036\u0026thinsp;\u0026plusmn;\u0026thinsp;0.012\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.095\u0026thinsp;\u0026plusmn;\u0026thinsp;0.035\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.114\u0026thinsp;\u0026plusmn;\u0026thinsp;0.046\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.079\u0026thinsp;\u0026plusmn;\u0026thinsp;0.031\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.031\u0026thinsp;\u0026plusmn;\u0026thinsp;0.018\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.075\u0026thinsp;\u0026plusmn;\u0026thinsp;0.036\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.151\u0026thinsp;\u0026plusmn;\u0026thinsp;0.049\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.114\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.081\u0026thinsp;\u0026plusmn;\u0026thinsp;0.020\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003eThe data recorded in Table \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e indicated that the NO-donating oximes \u003cstrong\u003e8a, 8b, 8d, 8e, 8f, 8g, 8h, 8i\u003c/strong\u003e and \u003cstrong\u003e10a-c\u003c/strong\u003e achieved maximum amount of NO release after 2 hours. At the first hour the amount of nitric oxide released is increased in compounds \u003cstrong\u003e8i\u003c/strong\u003e, \u003cstrong\u003e8b\u003c/strong\u003e and \u003cstrong\u003e8h\u003c/strong\u003e, while maximum release of nitric oxide occurred in compounds \u003cstrong\u003e8b\u003c/strong\u003e and \u003cstrong\u003e8h\u003c/strong\u003e was at 2 hours. Then, the amount of nitic oxide released was declined, which may explain the biological important of oxime moiety as source of nitic oxide release in comparison to ketone intermediates \u003cstrong\u003e7a-j\u003c/strong\u003e and \u003cstrong\u003e9a-c\u003c/strong\u003e as shown in anticancer activity of oxime derivatives.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec14\"\u003e\n \u003ch2\u003e2.4. Docking\u003c/h2\u003e\n \u003cdiv class=\"Section3\" id=\"Sec15\"\u003e\n \u003ch2\u003e2.4.1. \u003cem\u003eIn silico\u003c/em\u003e molecular docking study into EGFR\u003c/h2\u003e\n \u003cp\u003eFor mechanistic investigation of antiproliferative activity of compounds \u003cstrong\u003e8d, 8g, 8i\u003c/strong\u003e and \u003cstrong\u003e10c\u003c/strong\u003e, the \u003cem\u003ein silico\u003c/em\u003e simulation studies targeting EGFR tyrosine kinase domain (Fig. \u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e using sorafenib, a multitarget kinase inhibitor drug, as reference was done. For the Epidermal Growth Factor Receptor (EGFR), the structural models of the selected ligands were built against the human EGFR complexed with AZD9291 inhibitor (2.80 \u0026Aring;; PDB ID: 4ZAU)[\u003cspan class=\"CitationRef\"\u003e49\u003c/span\u003e]. Since the docking simulation scored the ligands based on the structural compatibility and the electrostatic potential, the ligands differentially bound the active site of EGFR at different affinities, as illustrated in Table \u003cspan class=\"InternalRef\"\u003e7.\u003c/span\u003e \u0026nbsp;\u003c/p\u003e\n \u003ctable border=\"1\" id=\"Tab8\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCalculated binding properties of test compounds and sorafenib against EGFR.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTest compounds\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eBinding score\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHydrogen\u003c/p\u003e\n \u003cp\u003ebonding energy\u003c/p\u003e\n \u003cp\u003e(Kcal/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHydrophobic\u003c/p\u003e\n \u003cp\u003einteraction energy\u003c/p\u003e\n \u003cp\u003e(Kcal/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSorafenib\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-38.13\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-2.52\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-7.12\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8d\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-32.33\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-1.19\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-7.97\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-16.46\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2.25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-7.62\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-12.98\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2.47\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-7.50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e10c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-23.74\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-3.36\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-7.35\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003cp\u003eAs recorded in Table \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e, the predicted binding affinities, in turn, come with the evaluated inhibitory values of the EGFR enzymatic activity (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Sorafenib and oxime ligands (\u003cstrong\u003e8d, 8g, 8i\u003c/strong\u003e and \u003cstrong\u003e10c\u003c/strong\u003e) bound the active site by hydrogen bonding and hydrophobic interactions with respect to sorafenib, which had the highest affinity to the active site. Importantly, the amine group of M793 residue acts as a hydrogen donor that forms hydrogen bonding with the carbonyl groups of the ligands and sorafenib. Like sorafenib, compound \u003cstrong\u003e8d\u003c/strong\u003e was shown to occupy the active site with a binding affinity higher than the other compounds due to its flexibility to fit the active site \u003cstrong\u003e(\u003c/strong\u003eFigs. 9 and \u003cspan class=\"InternalRef\"\u003e10\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e. The sandwiching effect was achieved by the hydrophobic contacts formed by the non-polar residues L718, V726, A743 and L792. Compound \u003cstrong\u003e10c\u003c/strong\u003e showed a relatively similar binding score but without Van der Waals forces \u003cstrong\u003e(\u003c/strong\u003eFig. \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e. Compounds \u003cstrong\u003e8g\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e showed the same affinity levels to bind the active site. \u003cstrong\u003e(\u003c/strong\u003eFigs. \u003cspan class=\"InternalRef\"\u003e11\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec16\"\u003e\n \u003ch2\u003e\u003cstrong\u003e2.3.2\u003c/strong\u003e. \u003cspan class=\"BoldItalic\" name=\"Emphasis\" type=\"BoldItalic\"\u003eIn silico\u003c/span\u003e \u003cstrong\u003emolecular docking study on JNK-2\u003c/strong\u003e\u003c/h2\u003e\n \u003cp\u003eFor further mechanistic investigation of the antiproliferative activity, compounds \u003cstrong\u003e8d\u003c/strong\u003e and \u003cstrong\u003e8i\u003c/strong\u003e were investigated for their \u003cem\u003ein silico\u003c/em\u003e simulation studies targeting the JNK-2 binding pocket (Fig. \u003cspan class=\"InternalRef\"\u003e14\u003c/span\u003e) using sorafenib as reference drug. JNK-2 inhibitors are classified to two types: Type I inhibitors target the DFG-in conformation (open conformation), while type II inhibitors target the DFG-out conformation (closed conformation) [\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e14\u003c/span\u003e]. Based on open confirmation, we docked the selected compounds against a pre-defined open conformation of JNK-2 (Fig. \u003cspan class=\"InternalRef\"\u003e14\u003c/span\u003e) [\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e]. Sorafenib is a common MAP kinase inhibitor, and it was shown to have a higher binding affinity to JNK-2 active site compared to the other ligands (Table \u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e). Sorafenib extends at the active site from the hinge region (L110 and M111) to the DFG conformation (D169), which makes it structurally compatible to fit the active site (Fig. \u003cspan class=\"InternalRef\"\u003e14\u003c/span\u003e). Sorafenib binds to the ATP site of JNK-2 and forms two hydrogen bonds: one with M111 of the hinge and the other with K55 of the \u003cem\u003eN\u003c/em\u003e-terminal b3-strand. In addition, the binding of sorafenib is supported by Van der Waals forces with the hinge residues E109 and Q117 as well as hydrophobic interactions. Like sorafenib, compound \u003cstrong\u003e8i\u003c/strong\u003e is bound to the active site by the same machinery, forming hydrogen bonds with M111 and K55 (\u003cstrong\u003eFigs.\u0026nbsp;15 and 16\u003c/strong\u003e). However, the weaker affinity of compound \u003cstrong\u003e8i\u003c/strong\u003e than that of sorafenib comes from the lesser extent of compound \u003cstrong\u003e8i\u003c/strong\u003e to the DFG conformation, resulting in weaker hydrophobic interactions. Furthermore, the sulfonamide group of compounds \u003cstrong\u003e8i\u003c/strong\u003e was shown to be extended outside the binding pocket near the hinge region, describing the group\u0026apos;s lower interactivity. Compound \u003cstrong\u003e8i\u003c/strong\u003e binding is supported by Van der Waals forces with the hinge region (E109, N114 and Q117). However, the binding energy is lower than that of sorafenib. Compound \u003cstrong\u003e8i\u003c/strong\u003e binds to the hinge region by only one hydrogen bond with M111 supported by Van der Waals force with Q117 (\u003cstrong\u003eFig.\u0026nbsp;16)\u003c/strong\u003e. On the other hand, compound \u003cstrong\u003e8d\u003c/strong\u003e hydrophobically binds with the hinge region with extension to the \u003cem\u003eN\u003c/em\u003e-terminal b3-strand, where it forms a hydrogen bond with K55 and a Van der Waals contact with E109 (\u003cstrong\u003eFig.\u0026nbsp;17\u003c/strong\u003e). The structural incompatibility of compound \u003cstrong\u003e8d\u003c/strong\u003e results in a weaker binding score, which in turn, results in weaker binding energy and consequently, a weaker inhibitory effect.\u0026nbsp;\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab9\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCalculated binding properties of test compounds andsorafenib against JNK-2.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTest compounds\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eBinding score\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHydrogen\u003c/p\u003e\n \u003cp\u003ebonding energy\u003c/p\u003e\n \u003cp\u003e(Kcal/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHydrophobic\u003c/p\u003e\n \u003cp\u003einteraction energy\u003c/p\u003e\n \u003cp\u003e(Kcal/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVan der Waals\u003c/p\u003e\n \u003cp\u003einteraction energy\u003c/p\u003e\n \u003cp\u003e(Kcal/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSorafenib\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-21.48\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-2.72\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-8.46\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-35.60\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8d\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-9.16\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-0.99\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-7.97\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e-33.70\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e8i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-12.00\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-3.19\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-7.66\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e-32.59\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Conclusion","content":"\u003cp\u003eA series of 1,5-diarylpyrzole derivatives targeting EGFR and JNK-2 were developed and synthesized and biologically evaluated for their anticancer activity against a panel of five cancer cell lines, namely human colorectal adenocarcinoma cell line DLD-1, human cervical cancer cell line Hela, human leukemia cell line K562, human pancreatic cell line SUIT-2 and human hepatocellular carcinoma cell line HepG2. Oxime derivative compounds \u003cb\u003e8a-j\u003c/b\u003e and \u003cb\u003e10a-c\u003c/b\u003e showed better anticancer activity than their corresponding ketones. Regarding substituents on the terminal phenyl ring of the 1,5-diarylpyrazole part, they showed a significant effect on the biological profile of anticancer activity especially when \u003cb\u003eR\u003c/b\u003e\u003csup\u003e\u003cb\u003e1\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;H, \u003cem\u003ep\u003c/em\u003e-CH\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ep\u003c/em\u003e-Cl, such as compounds \u003cb\u003e8b\u003c/b\u003e, \u003cb\u003e8f, 8g, 8h\u003c/b\u003e and \u003cb\u003e8i\u003c/b\u003e, which are considered the most potent anticancer oxime derivatives. Also, oxime derivatives \u003cb\u003e8g\u003c/b\u003e, \u003cb\u003e8i\u003c/b\u003e and \u003cb\u003e10c\u003c/b\u003e exhibited moderate EGFR inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e of 18, 21 and 12 \u0026micro;M respectively, while compound \u003cb\u003e8d\u003c/b\u003e displayed good EGFR inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e of 8 \u0026micro;M. Moreover, compound \u003cb\u003e8i\u003c/b\u003e showed potent JNK-2 inhibitory activity with IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.00 \u0026micro;M, similar to the positive reference drug sorafenib. Selectivity of compounds \u003cb\u003e8i\u003c/b\u003e and \u003cb\u003e8g\u003c/b\u003e towards cancer cells rather than normal cells was evaluated and compound \u003cb\u003e8i\u003c/b\u003e observed no cytotoxicity against the PC12 cell line with CC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;\u0026gt;\u0026thinsp;100 \u0026micro;M, while compound \u003cb\u003e8g\u003c/b\u003e showed moderate cytotoxicity with CC\u003csub\u003e50\u003c/sub\u003e of 16.1 \u0026micro;M in comparison to reference drug daunorubicin. Furthermore, compound \u003cb\u003e8g\u003c/b\u003e exhibited cell cycle arrest at the G2/M phase in the cell cycle analysis of the Hela cell line, while compound \u003cb\u003e8i\u003c/b\u003e showed combined S phase and G2 phase arrest. Additionally, Hela cell apoptotic changes after treatments of compounds \u003cb\u003e8g\u003c/b\u003e and \u003cb\u003e8i\u003c/b\u003e were investigated by fluorescent microscope and flowcytometry as results of their antiproliferative activity. Lastly, \u003cem\u003ein silico\u003c/em\u003e molecular docking studies showed a good fitting of compounds \u003cb\u003e8d\u003c/b\u003e, \u003cb\u003e8g\u003c/b\u003e, \u003cb\u003e8i\u003c/b\u003e and \u003cb\u003e10c\u003c/b\u003e at the EGFR binding site with binding free energies in the range of -12.98 to 32.30 kcl/mol, while compounds \u003cb\u003e8d\u003c/b\u003e and \u003cb\u003e8i\u003c/b\u003e showed binding free energies in range of -9.16 to -12.00 kcl/mol at the JNK-2 binding site.\u003c/p\u003e"},{"header":"4. Experimental","content":"\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e4.1. Chemistry\u003c/h2\u003e \u003cp\u003e \u003cb\u003eMaterial and equipment\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eAll chemicals used for the preparation of the target compounds are of analytical grade and can be used without further purification. Solvents were purified and freshly distilled before use according to the standard procedures.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eReaction progress was monitored by thin layer chromatography (Merck Silica gel 60 F254) on glass plates and visualized with a UV lamp (254 nm).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eColumn chromatography was performed using spherical, neutral silica gel of diameter 40\u0026ndash;100 \u0026micro;m (Kanto chemical co. Inc., Tokyo, Japan).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eMelting points were recorded at a ATM-02 (AS ONE, Tokyo, Japan).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eIR spectra were recorded at FT/IR-Spectrum Two (PerkinElmer, MA, USA) at the Faculty of Engineering, Yamagata University, Yonezawa, Japan.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eH-NMR (400 or 500 MHz) and \u003csup\u003e13\u003c/sup\u003eC-NMR (100 or 125 MHz) spectra were recorded on either a JNM-ECX400 or JNM-ECX500 (JEOL, Tokyo, Japan) in Faculty of Engineering, Yamagata University, Yonezawa, Japan. Chemical shifts are reported in ppm relative to tetramethylsilane (0 ppm), chloroform (7.26 ppm: 1H, 77.1 ppm: 13C) and dimethyl sulfoxide (2.50 ppm: 1H, 39.6 ppm: 13C). Coupling constant (\u003cem\u003eJ\u003c/em\u003e) is measured in hertz (Hz). Multiplicity was designated as: s, singlet; d, doublet; t, triplet; q, quartet; p, pentet; dd, doublet of doublet; and m for multiplet.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eMass spectra (ESI-MS) were carried out using the AccuTOF JMS-T100LC (JEOL, Tokyo, Japan) at the Faculty of Engineering, Yamagata University, Yonezawa, Japan.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cdiv id=\"Sec20\" class=\"Section3\"\u003e \u003ch2\u003e4.1.1. General procedure for the synthesis of ethyl 4-(substituted phenyl)-2-hydroxy-4-oxobut-2-enoates (2a-e).\u003c/h2\u003e \u003cp\u003eA mixture of diethyl oxalate (2.92 g, 0.02 mol) and substituted acetophenone derivatives (0.01 mol) in ethanol (50 mL) was added to previously prepared sodium ethoxide (sodium, 0.46 g, 0.02 mol, ethanol 100 mL) at 50\u0026deg;C. The reaction mixture was heated under reflux for 2\u0026ndash;3 h. After cooling, the solvent was removed, and the residue was taken up in water (200 mL) and acidified with concentrated HCl (1 mL). The aqueous mixture was extracted with ethyl acetate (3x150 mL). The combined extracts were washed with brine (100 mL), dried (MgSO\u003csub\u003e4\u003c/sub\u003e), and concentrated. The obtained solid was recrystallized from methanol to give compounds \u003cb\u003e2a-e\u003c/b\u003e and the produced compounds used in the next step without further purification [\u003cspan additionalcitationids=\"CR52\" citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section3\"\u003e \u003ch2\u003e4.1.2. General procedure for synthesis of 4-hydrazinylbenzenesulfonamide hydrochloride 4b.\u003c/h2\u003e \u003cp\u003eA cold, stirred mixture of sulfanilamide (3.42 g, 0.02 mol), hydrochloric acid (10 mL) and crushed ice (200 g) was diazotized by the dropwise addition of sodium nitrite (1.4 g, 0.02 mol) in water (25 mL) over 30 min. The cold diazonium salt solution thus formed was rapidly added to a well-cooled solution of sodium sulfite (2.52 g) and sodium hydroxide (0.800g) in water (50 mL) with vigorous stirring, and the resulting mixture was left in the ice bath for 15 min, followed by acidification with 10 mL HCl and concentrated. The precipitated 4-hydrazineylbenzenesulfonamide hydrochloride \u003cb\u003e4b\u003c/b\u003e was collected and dried: white crystals; mp :225\u0026deg;C (lit. mp: 225\u0026deg;C); yield 3.9 g (88%) [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section3\"\u003e \u003ch2\u003e4.1.3. General procedure for the synthesis of ethyl 1,5 diarypyarzole-3-carboxylate (5a-j).\u003c/h2\u003e \u003cp\u003eA mixture of diketoesters \u003cb\u003e2a-c\u003c/b\u003e (0.01mole) and phenylhydrazine \u003cb\u003e4a\u003c/b\u003e (0.01mole) was dissolved in suitable amount of absolute ethanol (40 mL) and refluxed for 5 h to afford compounds \u003cb\u003e5a-c\u003c/b\u003e. A mixture of diketoesters \u003cb\u003e2d-f\u003c/b\u003e and 4-hydrazinylbenzenesulfonamide hydrochloride \u003cb\u003e4b\u003c/b\u003e was refluxed in absolute ethanol for 5 h in the presence of sodium acetate (0.02 mole) to afford compounds \u003cb\u003e5d-f.\u003c/b\u003e The reaction process was monitored by TLC using chloroform: methanol (98: 2) solvent system for compounds \u003cb\u003e5a-c\u003c/b\u003e and chloroform: methanol (95: 5) solvent system for compounds \u003cb\u003e5d-f\u003c/b\u003e. The content of reaction mixture was evaporated under vacuum and the crude product was purified by column chromatography [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 1,5-diphenyl-1H-pyrazole-3-carboxylate (5a)\u003c/b\u003e: Reddish brown solid; yield (75%); mp: 85\u0026ndash;87\u0026deg;C (lit. 86\u0026deg;C)[\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 1-phenyl-5-(p-tolyl)-1H-pyrazole-3-carboxylate (5b)\u003c/b\u003e: Reddish solid; yield (80%), mp: 87\u0026ndash;88\u0026deg;C (lit. 84\u0026ndash;86\u0026deg;C)[\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-(4-methoxyphenyl)-1-phenyl-1H-pyrazole-3-carboxylate(5c)\u003c/b\u003e: Reddish brown solid; yield (81%); mp: 97\u0026ndash;99\u0026deg;C (lit. 97\u0026deg;C)[\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-(4-chlorophenyl)-1-phenyl-1H-pyrazole-3-carboxylate (5d)\u003c/b\u003e: Reddish brown solid; yield (89%); mp: 92\u0026ndash;94\u0026deg;C (lit. 95\u0026ndash;97\u0026deg;C)[\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] .\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-(3,4-dimethoxyphenyl)-1-phenyl-1H-pyrazole-3-carboxylate (5e)\u003c/b\u003e: Brownish solid, yield (63%); mp: 174\u0026ndash;176\u0026deg;C (lit. 177\u0026deg;C)[\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-phenyl-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (5f)\u003c/b\u003e: Reddish powder; yield (66%), mp: 192\u0026ndash;194\u0026deg;C (lit. 192)[\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 1-(4-sulfamoylphenyl)-5-(p-tolyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (5g)\u003c/b\u003e: Reddish brown; yield (75%), mp: 227\u0026ndash;228\u0026deg;C (lit. 227\u0026deg;C)[\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (5h)\u003c/b\u003e: Reddish brown powder; yield (71%), mp: 207\u0026ndash;209\u0026deg;C (lit. 205\u0026ndash;207\u0026deg;C)[\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-(4-chlorophenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (5i)\u003c/b\u003e: Reddish brown powder; yield (80%), mp: 107\u0026ndash;109\u0026deg;C (lit. 108\u0026deg;C)[\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eEthyl 5-(3,4-dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1H-pyrazole-3-carboxylate (5j)\u003c/b\u003e: Reddish brown powder; yield (64%); mp: 214\u0026ndash;215\u0026deg;C[\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003e4.1.4. General procedure for the synthesis of 1,5-diarypyrazole carboxylic acids 6a-j.\u003c/h2\u003e \u003cp\u003eA mixture of methanolic solution of compounds \u003cb\u003e5a-j\u003c/b\u003e (4 mmol), potassium hydroxide (KOH, 20%, 10 mL) was stirred at 60\u0026deg;C for 4 h. After cooling, the mixture solution was poured into water and acidified with hydrochloric acid solution (1 M) to pH\u0026thinsp;=\u0026thinsp;3. The aqueous mixture was extracted with ethyl acetate (3x50 mL) and the aqueous layer was discarded. The combined organic extracts were dried with anhydrous MgSO\u003csub\u003e4\u003c/sub\u003e. The organic solvent was evaporated under vacuum to obtain solid products \u003cb\u003e6a-j\u003c/b\u003e [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e1,5-Diphenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6a)\u003c/b\u003e: Brown powder; yield (84%); mp: 180\u0026ndash;182\u0026deg;C (lit. 182\u0026ndash;183)[\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e1-Phenyl-5-(p-tolyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6b)\u003c/b\u003e: Reddish powder; yield (87%); mp: 171\u0026ndash;172\u0026deg;C[\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-(4-Methoxyphenyl)-1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6c)\u003c/b\u003e: Reddish brown powder; yield (79%); mp: 192\u0026ndash;195\u0026deg;C (lit. 196\u0026ndash;197\u0026deg;C)[\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-(4-Chlorophenyl)-1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6d)\u003c/b\u003e: Yellowish brown powder; yield (78%); mp: \u0026gt; 300\u0026deg;C[\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-(3,4-Dimethoxyphenyl)-1-phenyl-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003e1H\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6e)\u003c/b\u003e: Brown powder; yield (84%); mp: 213\u0026ndash;214\u0026deg;C; \u003csup\u003e1\u003c/sup\u003eH-NMR (400 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) δ (ppm): 7.94 7.52 (m, 5H, Ar-H), 7.39 (s, 1H, pyrazole-H), 6.94\u0026ndash;6.68 (m, 3H, Ar-H), 3.79 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.77 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 163.36, 160.36, 145.81, 144.09, 142.55, 130.71, 128.76, 127.45, 126.47, 125.40, 122.05, 120.40, 115.26, 110.45, 56.23, 56.12; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e calculated: 325.1183, found: 325.256.\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-Phenyl-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylica acid(6f)\u003c/b\u003e: Yellowish brown powder; yield (78%), mp: 184\u0026ndash;186\u0026deg;C (lit. 188\u0026deg;C)[\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e1-(4-Sulfamoylphenyl)-5-(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003ep\u003c/span\u003e \u003cb\u003e-tolyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6g)\u003c/b\u003e: Reddish brown powder; yield (88%); mp: 194\u0026ndash;195\u0026deg;C[\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-(4-Methoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6h)\u003c/b\u003e: Brownish powder; yield (73%), mp: 197\u0026ndash;198\u0026deg;C[\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-(4-Chlorophenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6i)\u003c/b\u003e: Yellowish brown powder; yield (84%); mp: 212\u0026ndash;214\u0026deg;C[\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e5-(3,4-Dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylic acid (6j)\u003c/b\u003e: Reddish brown powder; yield (77%); mp: 206\u0026ndash;208\u0026deg;C; \u003csup\u003e1\u003c/sup\u003eH-NMR (400 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) δ (ppm): 10.76 (s, 1H, OH), 7.99 (s, 1H, Ar-H), 7.91(d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.79 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.65\u0026ndash;7.60 (m, 4H, 2Ar-H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.43 (s, 1H, pyrazole-H), 3.89 (s, 6H, 2 OCH\u003csub\u003e3\u003c/sub\u003e);\u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 163.36, 159.98, 147.36, 145.23, 141.21, 131.47, 130.17, 128.82, 126.87, 125.54, 122.87, 119.99, 115.14, 107.53, 56.38, 56.21; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS calculated: 404.0911, found: 404.0271.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section3\"\u003e \u003ch2\u003e4.1.5. General procedure for synthesis of 4-acetyl-2-methoxyphenyl 5-(4-subistituted-phenyl) 1-(4-substituted-phenyl)-1\u003cem\u003eH\u003c/em\u003e-pyrazole-3-carboxylate 7a-j.\u003c/h2\u003e \u003cp\u003eA mixture of pyrazole carboxylic acid derivatives \u003cb\u003e6a-j\u003c/b\u003e (0.001 mol), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (0.384 g, 0.002 mol), 1-hydroxybenzotriazole (HOBt) (0.306 g, 0.002 mol), were stirred in dry DMF (5 mL) for 30 min, then \u003cem\u003eN,N-\u003c/em\u003ediisopropylethylamine (DIPEA) (0.258 g, 0.002 mol) and 4-hydroxy-3-methoxyacetophenone (0.002 mol) were added to the mixture and stirred for 12 h. 20 mL Distilled water was added followed by acidification with dil. HCl. Extraction twice with ethyl acetate and purification were performed by using column chromatography with chloroform as eluent for compounds \u003cb\u003e7a-e\u003c/b\u003e and chloroform: methanol 98:2 for compounds \u003cb\u003e7f-j.\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl 1,5-diphenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (7a)\u003c/b\u003e: Yellowish brown solid; yield (75%); mp: 98\u0026ndash;100\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1748 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1725 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCo-\u003c/span\u003eCH\u003csub\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e3\u003c/span\u003e\u003c/sub\u003e), 1574 (C\u0026thinsp;=\u0026thinsp;C); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) δ (ppm): 7.65 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 7.46 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.43\u0026ndash;7.39 (m, 3H, Ar-H), 7.38 (s, 1H, Ar-H), 7.34\u0026ndash;7.32 (m, 3H, Ar-H), 7.30 (s, 1H, pyrazole-H), 7.28\u0026ndash;7.27 (m, 2H, Ar-H), 6.84 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 3.75 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.57 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.35, 160.07, 151.86, 147.61, 146.33, 145.04, 143.48, 142.86, 141.18, 139.68, 136.44, 129.91, 129.45, 129.11, 126.55, 123.97, 122.69, 114.98, 111.62, 56.74, 27.17; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eNaO\u003csub\u003e4\u003c/sub\u003e calculated: 435.13208, found: 435.13095.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-1-phenyl 5-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003ep\u003c/span\u003e \u003cb\u003e-tolyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (7b)\u003c/b\u003e: Yellowish solid; yield (81%); mp: 110\u0026ndash;112\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1743 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u003c/span\u003eOO-Ph), 1710 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1575 (C\u0026thinsp;=\u0026thinsp;C); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ (ppm): 7.65 (s, 1H, Ar-H), 7.53\u0026ndash;7.50 (m, 6H, Ar-H), 7.36\u0026ndash;7.35 (m, 2H, Ar-H), 7.15 (s, 1H, pyrazole-H), 6.92\u0026ndash;6.94 (m, 3H, Ar-H), 3.88 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.62 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e), 2.33 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, CDCl\u003csub\u003e3\u003c/sub\u003e ) δ (ppm): 197.40, 159.67, 150.10, 146.64, 143.95, 142.51, 139.46, 138.90, 136.03, 130.16, 129.85, 128.80, 125.35, 124.00, 122.97, 121.94, 111.97, 109.93, 56.48, 26.65, 21.49; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eNaO\u003csub\u003e4\u003c/sub\u003e calculated: 449.1477, found: 449.14830.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl \u0026minus;\u0026thinsp;5-(4-methoxyphenyl) 1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e3-carboxylate (7c)\u003c/b\u003e: Yellowish brown solid; yield (85%); mp: 69\u0026ndash;71\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1743 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1725 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1577 (C\u0026thinsp;=\u0026thinsp;C); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 7.64 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.00 Hz, 1H, Ar-H), 7.46 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.42 (s, 1H, Ar-H), 7.38 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.00 Hz, 1H, Ar-H), 7.34 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 2H, Ar-H), 7.22 (s, 1H, pyrazole-H), 7.17 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 6.88\u0026ndash;6.85 (m, 3H, Ar-H), 3.84 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.79 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.58 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.73, 159.99, 151.77, 148.69, 145.56, 143.22, 142.20, 139.78, 136.74, 130.95, 130.16, 129.55, 125.76, 124.02, 122.28, 116.09, 115.07, 111.96, 110.36, 57.17, 55.74, 26.65; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eNaO\u003csub\u003e5\u003c/sub\u003e calculated: 465.1426, found: 465.14282.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-5-(4-chlorophenyl) 1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole- ---3-carboxylate (7d)\u003c/b\u003e: Yellowish brown solid; yield (73%); mp: 85\u0026ndash;87\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1739 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1728 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO-\u003c/span\u003eCH\u003csub\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e3\u003c/span\u003e\u003c/sub\u003e), 1575 (C\u0026thinsp;=\u0026thinsp;C); \u003csup\u003e1\u003c/sup\u003eH-NMR (400 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ (ppm): 7.60 (s, 1H, Ar-H), 7.56 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.00 Hz, 1H, Ar-H), 7.50\u0026ndash;7.46 (m, 4H, Ar-H), 7.32\u0026ndash;7.35 (m, 2H, Ar-H), 7.17\u0026ndash;7.15 (m, 2H, Ar-H), 7.14 (s, 1H, pyrazole-H), 6.90 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.50 Hz, 2H, Ar-H), 3.88 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.51 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ (ppm): 197.11, 160.00, 150.43, 147.01, 146.31, 143.53, 143.23, 138.32, 135.31, 133.65, 129.23, 128.88, 127.17, 126.08, 125.74, 123.01, 113.97, 111.69, 109.80, 56.14, 27.26; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eClN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e calculated: 447.1106, found: 447.10859.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-5-(3,4-dimethoxyphenyl) 1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (7e)\u003c/b\u003e: Reddish yellow solid; yield (71%); mp: 74\u0026ndash;76\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1740 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1715 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1589 (C\u0026thinsp;=\u0026thinsp;C); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 7.65 (s, 1H, Ar-H), 7.63 (s, 1H, Ar-H), 7.56 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 7.46 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.38\u0026ndash;7.41 (m, 3H, Ar-H), 7.30 (s, 1H, pyrazole-H), 6.97 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 6.85\u0026ndash;6.83 (m, 2H, Ar-H), 3.84 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.76 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.71 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.59 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-d6) δ (ppm): 197.38, 160.00, 153.54, 151.71, 149.03, 147.99, 147.03, 142.92, 140.14, 136.74, 130.54, 130.34, 129.55, 127.07, 123.73, 122.65, 121.95, 115.07, 112.67, 112.01, 110.57, 57.15, 56.48, 55.44, 26.95; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e25\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e calculated: 473.1707, found: 473.17144.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-5-phenyl 1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e-carboxylate (7f)\u003c/b\u003e: Yellowish solid; yield (65%); mp: 69\u0026ndash;72\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1735 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1724 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e3\u003c/span\u003e\u003c/sub\u003e), 1589 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1162 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 7.91 (s, 1H, Ar-H), 7.87 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.80 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 1H, Ar-H), 7.68\u0026ndash;7.64 (m, 3H, Ar-H), 7.55 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H), 7.53 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.41\u0026ndash;7.39 (m, 2H, Ar-H), 7.35 (s, 1H, pyrazole-H), 7.31 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 1H, Ar-H), 3.85 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.57 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.75, 167.77, 159.18, 151.90, 144.65, 144.14, 142.86, 141.18, 135.65, 132.33,129.48, 129.40, 127.04, 125.27, 124.50, 123.58, 122.00, 112.02, 111.12, 56.74, 28.85; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e21\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003e Na O\u003csub\u003e6\u003c/sub\u003eS calculated: 514.1049, found: 514.10449.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-1-(4-sulfamoylphenyl) 5-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003ep\u003c/span\u003e \u003cb\u003e-tolyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (7g)\u003c/b\u003e: Yellowish solid; yield (78%), mp: 83\u0026ndash;85\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1746 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCOO-Ph\u003c/span\u003e), 1718 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1595 (C\u0026thinsp;=\u0026thinsp;C), 1162 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 7.95 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 7.86 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.69\u0026ndash;7.63 (m, 2H, Ar-H), 7.55 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.52 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.39 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.29 (s, 1H, pyrazole-H), 7.19 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 3.84 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.60 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e), 2.28 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.74, 163.64, 151.71, 146.50, 144.25, 143.93, 142.21, 139.63, 130.19, 129.12, 128.82, 128.45, 127.77, 126.14, 125.04, 123.75, 122.31, 112.64, 110.31, 56.44, 27.25, 21.47; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e23\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eNaO\u003csub\u003e6\u003c/sub\u003eS calculated: 528.1205, found: 528.12058.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl 5-(4-methoxyphenyl) 1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003epyrazole-3-carboxylate (7h)\u003c/b\u003e:Yellowish solid; yield (55%), mp: 80\u0026ndash;83\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1750 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1720 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1585 (C\u0026thinsp;=\u0026thinsp;C), 1164 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 7.86 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.66 (s, 1H, Ar-H), 7.64 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 2H, Ar-H), 7.55 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.51 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.39 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 1H, Ar-H), 7.13 (s, 1H, pyrazole-H), 7.26\u0026ndash;7.23 (m, 3H, Ar-H), 3.84 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.74 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.60 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.35, 160.08, 151.86, 148.90, 144.65, 143.35, 141.58, 136.44, 130.40, 129.61, 127.43, 126.55, 123.98, 123.58, 121.01, 115.47, 113.69, 111.12, 57.53, 55.85, 26.68; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e23\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003e Na O\u003csub\u003e7\u003c/sub\u003eS calculated: 544.1154, found: 544.11530.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-5-(4-chlorophenyl) 1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003epyrazole-3-carboxylate (7i)\u003c/b\u003e: Yellowish brown solid; yield (76%); mp: 71\u0026ndash;74\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1744 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1726 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1591 (C\u0026thinsp;=\u0026thinsp;C), 1162 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm):, 7.89 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.86 (s, 1H, Ar-H), 7.80 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 1H, Ar-H), 7.65 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 1H, Ar-H), 7.57 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.52 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.47 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.38 (s, 1H, pyrazole-H), 7.33 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 3.84 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.69 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.80, 162.85, 159.53, 144.43, 143.25, 143.15, 141.69, 136.56, 132.33, 129.46, 127.90, 127.43, 126.56, 126.09, 123.83, 122.31, 120.09, 112.31, 111.90, 57.14, 27.14; ESI-MS m/z [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eClN\u003csub\u003e3\u003c/sub\u003eNaO\u003csub\u003e6\u003c/sub\u003eS calculated: 524.0689, found: 524.06882.\u003c/p\u003e \u003cp\u003e \u003cb\u003e4-Acetyl-2-methoxyphenyl-5-(3,4-dimethoxyphenyl) 1-(4-sulfamoylphenyl)\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (7j)\u003c/b\u003e: Yellowish brown solid; yield (52%); mp: 75\u0026ndash;78\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1749(\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eO-Ph), 1727 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1579 (C\u0026thinsp;=\u0026thinsp;C), 1162 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) δ (ppm): 8.05 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.96 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 7.80 (s, 1H, Ar-H), 7.74 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.69 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.56 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 7.50 (s, 1H, Ar-H), 7.10 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 7.05 (s, 1H, pyrazole-H), 6.95 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 3.99 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.89 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.75 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.75 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.75, 163.93, 151.47, 149.29, 148.89, 145.80, 144.68, 143.33, 142.86, 142.08, 136.04, 129.13, 127.41, 126.55, 125.27, 123.58, 122.29, 121.00, 112.90, 111.12, 109.84, 57.13, 56.74, 54.96, 27.17; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e25\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eNaO\u003csub\u003e8\u003c/sub\u003eS calculated: 574.1260, found: 574.12616.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003e4.1.6. General procedure for synthesis of (\u003cem\u003eE\u003c/em\u003e)-4-(1-(hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-substituted phenyl)-1-(4-substituted phenyl)-1\u003cem\u003eH\u003c/em\u003e-pyrazole-3-carboxylate 8a-j.\u003c/h2\u003e \u003cp\u003eA mixture of the appropriate ketone derivatives \u003cb\u003e7a-j\u003c/b\u003e (0.001 mol) and hydroxylamine hydrochloride (0.138 g, 0.002 mol) in 30 mL of absolute ethanol was heated under reflux for 8\u0026ndash;12 h and then left to cool to room temperature. The separated solid was filtered off, washed with 10% ammonia solution, then with distilled water, dried, and crystallized from absolute ethanol, affording the target products \u003cb\u003e8a-j\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)2-methoxyphenyl 1,5-diphenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8a)\u003c/b\u003e: Yellowish brown solid; yield (67%); mp: 167\u0026ndash;170\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3191 (OH), 1756 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1594 (C\u0026thinsp;=\u0026thinsp;C aromatic); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 11.30 (s, 1H, OH), 7.47 (s, 1H, Ar-H), 7.41\u0026ndash;7.37 (m, 4H, Ar-H), 7.33\u0026ndash;7.30 (m, 3H, Ar-H), 7.28\u0026ndash;7.23 (m, 3H, Ar-H), 7.20 (s, 1H, pyrazole-H), 7.00 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 1H, Ar-H), 6.78 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 1H, Ar-H), 3.73 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.06 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 160.07, 152.75, 148.50, 148.01, 144.05, 142.67, 139.72, 136.71, 132.58, 130.41, 129.62, 128.72, 127.84, 126.55,, 123.37, 119.35, 118.92, 116.26, 109.46, 56.24, 12.14; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e21\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eNaO\u003csub\u003e4\u003c/sub\u003e calculated: 450.1430, found: 450.14238.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 1-phenyl-5-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003ep\u003c/span\u003e \u003cb\u003e-tolyl-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8b)\u003c/b\u003e: Yellowish green powder; yield (75%); mp: 123\u0026ndash;125\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3138 (OH), 1736 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1593 (C\u0026thinsp;=\u0026thinsp;C aromatic); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-d6) δ (ppm): 10.36 (s, 1H, OH), 7.40\u0026ndash;7.35 (m, 3H, Ar-H), 7.29 (s, 1H, Ar-H), 7.15\u0026ndash;7.05 (m, 5H, 4 Ar-H, pyrazole-H), 6.96 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 2H, Ar-H), 6.75 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 2H, Ar-H), 3.74 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.21 (s, 3H, Ph-\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCH\u003c/span\u003e\u003csub\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e3\u003c/span\u003e\u003c/sub\u003e), 2.02 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) δ (ppm): 160.73, 153.20, 152.18, 151.13, 147.97, 147.97, 142.01, 139.43, 136.85, 129.81, 129.06, 128.64, 126.07, 124.01, 119.19, 116.09, 115.42, 111.97, 108.89, 56.78, 21.85, 11.83; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e calculated: 442.1761, found: 442.17438.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-methoxyphenyl) 1\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8c)\u003c/b\u003e: Yellowish brown powder; yield (75%); mp: 132\u0026ndash;135\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3400 (OH), 1739(\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1590 (C\u0026thinsp;=\u0026thinsp;C aromatic); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) δ (ppm): 10.43 (s, 1H, OH), 7.47 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 7.34 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.25 (s, 1H, Ar-H), 7.18 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.11\u0026ndash;7.08 (m, 2H, Ar-H), 7.04 (s, 1H, pyrazole-H), 7.95 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 6.80\u0026ndash;6.77 (m, 3H, Ar-H), 3.68 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.62 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.02 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 162.15, 159.58, 150.58, 147.22, 145.43, 143.76, 142.47, 140.80, 136.88, 130.90, 129.61, 127.83, 126.55, 123.18, 121.16, 120.61, 115.87, 114.57, 109.43, 56.74, 54.96, 14.71; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e calculated: 458.17160, found: 458.17154.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-chlorophenyl)-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8d)\u003c/b\u003e: Yellowish brown powder; yield (65%); mp: 106\u0026ndash;109\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3228 (OH), 1744 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1593 (C\u0026thinsp;=\u0026thinsp;C aromatic); \u003csup\u003e1\u003c/sup\u003eH-NMR (400 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.90 (s, 1H, OH), 7.60 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.40 Hz, 1H, Ar-H), 7.42 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.39 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10 Hz, 2H, Ar-H), 7.36 (s, 1H, Ar-H), 7.26\u0026ndash;7.21 (m, 3H, 2Ar-H, pyrazole-H), 7.00 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 6.75 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 3.74 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.07 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 160.00, 153.68, 151.87, 147.38, 143.64, 142.89, 139.83, 139.48, 136.84, 134.19, 132.02, 131.01, 129.71, 128.38, 126.38, 123.19, 119.48, 115.55, 109.63, 55.38, 12.45; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e21\u003c/sub\u003eClN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e calculated: 462.11889, found: 462.11846.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(3,4-dimethoxy phenyl)-1-phenyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8e)\u003c/b\u003e: Yellowish white powder; yield (77%); mp: 115\u0026ndash;118\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3300 (OH), 1740 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1593 (C\u0026thinsp;=\u0026thinsp;C aromatic); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.43 (s, 1H, OH), 7.56 (s, 1H, Ar-H), 7.46 (s, 1H, Ar-H), 7.40 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.50 Hz, 1H, Ar-H), 7.25\u0026ndash;7.19 (m, 4H, Ar-H), 7.01\u0026ndash;6.98 (m, 2H, Ar-H), 7.16 (s, 1H, pyrazole-H), 6.79 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50, 2H, Ar-H), 3.73 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.70 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.65 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.06 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 161.75, 159.58, 153.64, 150.58, 150.19, 149.29, 148.01, 145.05, 144.15, 142.86, 139.50, 130.41, 127.84, 125.76, 121.67, 121.62, 121.41, 119.71, 115.86, 112.90, 109.84, 56.74, 55.85, 54.56, 11.65; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eNaO\u003csub\u003e6\u003c/sub\u003e calculated: 488.1816, found: 488.18086.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-phenyl-1-(4-sulfamoyl phenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8f)\u003c/b\u003e: Yellowish powder; yield (53%); mp:122\u0026ndash;124\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3681 (OH), 1744 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1590 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1165 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 11.17 (s, 1H, OH), 7.86 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.50 Hz, 2H, Ar-H), 7.55 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.5 Hz, 2H, Ar-H), 7.51 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e),7.40\u0026ndash;7.35 (m, 4H, Ar-H), 7.32\u0026ndash;7.30 (m, 3H, Ar-H), 7.24 (s, 1H, Ar-H) 7.19 (s, 1H, pyrazole-H), 3.79 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.06 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 165.53, 159.99, 153.18, 151.12, 145.63, 143.53, 142.52, 141.49, 139.80, 136.74, 130.18, 129.18, 127.77, 126.80, 123.77, 123.30, 118.83, 111.65, 109.60, 55.71, 12.56; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003efor C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003e Na O\u003csub\u003e6\u003c/sub\u003eS calculated: 529.1158, found: 529.1158.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 1-(4-sulfamoylphenyl)\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e-5-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003ep\u003c/span\u003e \u003cb\u003e-tolyl-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8g)\u003c/b\u003e: Yellowish white powder; yield (69%); mp: 195\u0026ndash;197\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3255(OH), 1740 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1594 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1164 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 11.34 (s, 1H, OH), 7.95 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50, 1H, Ar-H), 7.86 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5 Hz, 2H, Ar-H), 7.68 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 7.54 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.52 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.40 (s, 1H, Ar-H), 7.27 (s, 1H, pyrazole-H), 7.24\u0026ndash;7.19 (m, 4H, Ar-H), 3.78 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.28 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e), 2.16 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 160.42, 153.19, 151.27, 145.80, 144.61, 142.05, 139.88, 139.41, 136.67, 130.21, 129.17, 127.98, 127.33, 126.51, 126.10, 123.30, 118.84, 111.67, 110.12, 56.45, 21.50, 12.56; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eNaO\u003csub\u003e6\u003c/sub\u003eS calculated: 543.1314, found: 543.13044.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8h)\u003c/b\u003e:Yellowish white powder; yield (49%), mp: 125\u0026ndash;127\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3400 (OH), 1742 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1596 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1165 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 11.27 (s, 1H, OH), 7.87(d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.55 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.50 Hz, 2H, Ar-H), 7.51 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.40 (s, 1H, Ar-H), 7.24\u0026ndash;7.20 (m, 4H, Ar-H), 7.19 (s, 1H, pyrazole-H), 6.95 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.50 Hz, 2H, Ar-H), 3.79 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.74 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.16 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 161.75, 159.18, 153.97, 151.07, 145.93, 144.64, 143.75, 142.47, 139.90, 136.44, 130.41, 129.62, 127.78, 123.18, 122.03, 121.41, 118.82, 115.47, 111.11, 56.74, 55.85, 12.14; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eNaO\u003csub\u003e7\u003c/sub\u003eS calculated: 559.1263, found: 559.12740.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(4-chlorophenyl)-1-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8i)\u003c/b\u003e: Yellowish brown powder; yield (67%); mp:110\u0026ndash;112\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3371 (OH), 1743 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1595 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1161 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.44 (s, 1H, OH), 7.85 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.76 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.50 Hz, 1H, Ar-H), 7.61 (s, 1H, Ar-H), 7.49 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.40\u0026ndash;7.33 (m, 4H, 3Ar-H, pyrazole-H), 7.26 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.19 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 3.71 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 1.90 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 159.57, 150.82, 144.50, 144.38, 143.30, 141.53, 136.59, 134.61, 132.25,, 129.43, 128.25, 127.46, 126.50, 124.90, 123.26, 120.17, 119.66, 118.84, 109.89, 56.38, 11.76; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;1]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eClN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS calculated: 541.0943, found: 541.09486.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-4-(1-(Hydroxyimino)ethyl)-2-methoxyphenyl 5-(3,4-dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxylate (8j)\u003c/b\u003e: Yellowish powder; yield (48%); mp: 129\u0026ndash;131\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3300 (OH), 1731(\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eC\u0026thinsp;=\u0026thinsp;O\u003c/span\u003e), 1595 (C\u0026thinsp;=\u0026thinsp;C aromatic); 1164 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 11.26 (s, 1H, OH), 7.87 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.85 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.52 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.41 (s, 1H, Ar-H), 7.32 (s, 1H, Ar-H), 7.25\u0026ndash;7.24 (m, 1H, Ar-H), 6.93 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 6.88 (s, 1H, pyrazole-H\u003cb\u003e)\u003c/b\u003e, 6.81\u0026ndash;6.75 (m, 2H, Ar-H), 3.79 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.72 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.60 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.17 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 167.78, 159.18, 152.75, 149.29, 148.50, 145.44, 144.65, 143.36, 141.58, 140.30, 136.44, 132.58, 129.12, 127.83, 126.15, 122.30, 121.40, 119.33, 112.91, 110.73, 109.84, 56.28, 56.06, 55.89, 11.75; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e26\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eNaO\u003csub\u003e8\u003c/sub\u003eS calculated: 589.1369, found: 589.13803.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003e4.1.7. General procedure for synthesis of \u003cem\u003eN\u003c/em\u003e-(4-acetylphenyl)-5-(4-subistitutedphenyl)-1-(4-sulfamoylphenyl)-1\u003cem\u003eH\u003c/em\u003e-pyrazole-3-carboxamide 9a-c.\u003c/h2\u003e \u003cp\u003eTo the suspension of 1,5-diarylpyrazole carboxylic acid derivatives \u003cb\u003e6f, 6h\u003c/b\u003e and \u003cb\u003e6j\u003c/b\u003e (0.001 mol) in 20 mL of benzene, thionyl chloride (2 mL) was added and heated under reflux for 4 h. Evaporation of the solvent was carried out under vacuum to give a residue of the corresponding acyl chloride that was utilized in the following steps without purification. A mixture of acyl chloride in dry DMF, few drops of triethylamine and 4-aminoacetophenone (0.270 g, 0.002 mol) were heated under reflux for 8h. Then, 20 mL of cold distilled water was added, followed by acidification with dil. HCl and extraction twice with ethyl acetate. Purification was performed by column chromatography using chloroform: methanol 98:2 as eluent to afford compounds \u003cb\u003e9a-c\u003c/b\u003e [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eN\u003c/span\u003e \u003cb\u003e-(4-Acetylphenyl)-5-phenyl-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxamide (9a)\u003c/b\u003e: Yellowish brown solid; yield (88%); mp: 81\u0026ndash;83\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1725 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1675 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eNH), 1591 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1161 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.53 (s, 1H, NH), 7.96 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.86 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.68\u0026ndash;7.62 (m, 3H, Ar-H), 7.57 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.49 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.40 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.30\u0026ndash;7.32 (m, 2H, Ar-H), 7.20 (s, 1H, pyrazole-H), 2.52 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.09, 167.91, 154.18, 145.63, 144.56, 143.96, 142.53, 133.27, 132.23, 131.21, 129.83, 127.84, 125.96, 125.26, 120.60, 119.04, 109.61, 26.95; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e24\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003e NaO\u003csub\u003e4\u003c/sub\u003eS calculated: 483.1103, found: 483.11090.\u003c/p\u003e \u003cp\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eN\u003c/span\u003e \u003cb\u003e-(4-Acetylphenyl)-5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003epyrazole-3-carboxamide (9b)\u003c/b\u003e:Yellowish brown solid; yield (66%); mp: 74\u0026ndash;76\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1715 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e),1681 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eNH), 1594 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1160 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 11.18 (s, 1H, NH), 7.87 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.60 Hz, 2H, Ar-H), 7.84 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.6.0 Hz, 2H, Ar-H), 7.80 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.62 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.54 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.33 (s, 1H, pyrazole-H), 6.98 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.60 Hz, 2H, Ar-H), 6.96 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.60 Hz, 2H, Ar-H), 3.82 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.47 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 195.96, 167.20, 154.55, 146.06, 144.59, 142.88, 133.27, 132.59, 131.09, 130.12, 129.18, 126.18, 125.30, 123.46, 120.64, 118.98, 113.06, 55.42, 26.61; ESI-MS m/z [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e21\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003eS calculated: 489.1238, found: 489.12547.\u003c/p\u003e \u003cp\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eN\u003c/span\u003e \u003cb\u003e-(4-Acetylphenyl)-5-(3,4-dimethoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003epyrazole-3-carboxamide (9c)\u003c/b\u003e: Brownish solid; yield (60%); mp: 81\u0026ndash;83\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 1720 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003e-CH\u003csub\u003e3\u003c/sub\u003e), 1669 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eNH), 1590 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1160 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.55 (s, 1H, NH), 7.95 (s, 1H, Ar-H), 7.92 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.84 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 2H, Ar-H), 7.76 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 7.72 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.62\u0026ndash;7.56 (m, 4H, 2 Ar-H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.37 (s, 1H, pyrazole-H), 7.01 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 1H, Ar-H), 3.82 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.80 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.53 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 197.06, 164.12, 153.21, 152.80, 150.42, 148.06, 145.60, 143.23, 142.71, 132.93, 130.56, 129.86, 129.15, 126.10, 124.03, 123.54, 119.19,116.49, 110.95, 56.76, 55.71, 26.62; ESI-MS m/z [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e23\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS calculated: 519.1344, found: 519.13451.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003e4.1.8. General procedure for synthesis of (\u003cem\u003eE\u003c/em\u003e)-\u003cem\u003eN\u003c/em\u003e-(4-(1-(hydroxyimino)ethyl)\u003c/h2\u003e \u003cp\u003e \u003cb\u003ephenyl)-5-(4-subistituted phenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ecarboxamide 10a-c.\u003c/b\u003e \u003c/p\u003e \u003cp\u003eA mixture of the appropriate ketone derivatives \u003cb\u003e9a-c\u003c/b\u003e (0.001 mol) and hydroxylamine hydrochloride (0.138 g, 0.002 mol) in 30 mL of absolute ethanol was heated under reflux for 8\u0026ndash;12 h and then left to cool to room temperature. The separated solid was filtered off, washed with 10% ammonia solution, then washed with distilled water, dried, and recrystallized from absolute ethanol to afford the target products \u003cb\u003e10a-c\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eN\u003c/span\u003e \u003cb\u003e-(4-(1-(Hydroxyimino)ethyl)phenyl)-5-phenyl-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxamide (10a)\u003c/b\u003e:Yellowish powder; yield (55%); mp: 168\u0026ndash;170\u0026deg;C ; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3681 (OH), 1680 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eNH), 1598 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1162 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.30 (s, 1H, NH), 8.75 (s, 1H, OH), 7.79 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.63 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.50\u0026ndash;7.56 (m, 3H, Ar-H), 7.47\u0026ndash;7.44 (m, 2H, Ar-H), 7.40 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.35 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 7.29 (s, 1H, pyrazole-H), 7.21 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.00 Hz, 2H, Ar-H), 2.06 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 160.05, 153.24, 151.71, 148.09, 143.55, 141.92, 133.69, 129.88, 129.39, 129.30, 127.44, 126.52, 126.42, 121.34, 120.78, 120.33, 109.58, 12.14; ESI-MS m/z [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e for C\u003csub\u003e24\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eS calculated: 476.1387, found: 476.13970.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eN\u003c/span\u003e \u003cb\u003e-(4-(1-(Hydroxyimino)ethyl)phenyl)-5-(4-methoxyphenyl)-1-(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxamide (10b)\u003c/b\u003e: Yellowish brown powder; yield (51%); mp: 110\u0026ndash;112\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3350 (OH), 1677 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eNH), 1596 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1162 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.31 (s, 1H, NH), 10.02 (s, 1H, OH), 7.84 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 7.78 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.52 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.46 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 7.41 (s, 2H, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.30 (s, 1H, pyrazole-H), 6.97 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.00 Hz, 2H, Ar-H), 6.91 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.00 Hz, 2H, Ar-H), 3.80 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e), 2.07 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 167.18, 155.50, 144.08, 142.51, 133.80, 132.35, 131.38, 130.65, 129.47, 127.11, 126.41, 126.09, 125.97, 123.46, 121.51, 114.33, 111.98, 56.12, 17.34; ESI-MS m/z [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e for C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003eS calculated: 504.1347, found: 504.13773.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eE\u003c/span\u003e \u003cb\u003e)-5-(3,4-Dimethoxyphenyl)-\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eN\u003c/span\u003e \u003cb\u003e-(4-(1-(hydroxyimino)ethyl)phenyl)-1-\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e(4-sulfamoylphenyl)-1\u003c/b\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eH\u003c/span\u003e \u003cb\u003e-pyrazole-3-carboxamide (10c)\u003c/b\u003e:Brownish powder; yield (44%), mp: 105\u0026ndash;107\u0026deg;C; IR (ATR) cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 3220 (OH), 1680 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCO\u003c/span\u003eNH), 1591 (C\u0026thinsp;=\u0026thinsp;C aromatic), 1161 (SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e); \u003csup\u003e1\u003c/sup\u003eH-NMR (500 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 10.36 (s, 1H, NH), 10.09 (s, 1H, OH), 7.92 (s, 1H, Ar-H), 7.79 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 7.63 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.52 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 2H, Ar-H), 7.50\u0026ndash;7.44 (m, 4H, 2ArH, SO\u003csub\u003e2\u003c/sub\u003eNH\u003csub\u003e2\u003c/sub\u003e), 7.36 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.50 Hz, 2H, Ar-H), 7.27 (s, 1H, pyrazole-H), 6.99 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.50 Hz, 1H, Ar-H), 3.77 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 3.74 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e), 2.08 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e); \u003csup\u003e13\u003c/sup\u003eC-NMR (100MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e6\u003c/sub\u003e) δ (ppm): 167.78, 159.18, 152.75, 149.29, 148.50, 145.44, 144.65, 143.36, 140.30, 136.44, 132.58, 129.12, 127.83, 126.15, 122.30, 121.40, 119.33, 112.91, 109.84, 56.19, 55.96, 12.14; ESI-MS m/z [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e for C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS calculated: 534.14473, found: 534.14307.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003e4.2. Measurement of nitric oxide release\u003c/h2\u003e \u003cp\u003e \u003cb\u003eMaterials and methods\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eThe nitrite calibration curve in addition to the absorbance of the tested compounds were measured on Shimadzu UV-160, UV-Visible spectrophotometer (Shimadzu, Tokyo, Japan).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eThe tested compounds were prepared as solutions in DMF and diluted with the buffer system till a concentration of 100 mM.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cem\u003eN\u003c/em\u003e- Acetyl cystiene solution was prepared in a concentration of 500 mM in methanol.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eGriess reagent consists of 0.1% w/v NED solution in water and sulfanilamide solution (1% w/v of sulfanilamide in 5% w/v phosphoric acid).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eNitrite standard solution (0.1 M sodium nitrite in water) stock solution was prepared from which a dilute solution of 100 \u0026micro;M nitrite solution was prepared by dilution 1 mL of the stock solution to 1000 mL with phosphate buffer of pH 7.4. From this solution, 4 serials two-fold dilutions were performed to generate different concentrations of the nitrite solution (100.00, 50.00, 25.00 and 12.50 \u0026micro;M) and these concentrations were used for nitrite calibration curve.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ePreparation of nitrite standard curve\u003c/b\u003e \u003c/p\u003e \u003cp\u003eSulfanilamide and NEDD solutions were kept at 25\u0026deg;C, 100 mL of sulfanilamide solution was added to each dilution of the prepared standard nitrite solution. The mixture was left at 25\u0026deg;C for 5\u0026ndash;10 minutes protected from light. To this mixture 100 mL of the NEDD solution was added and the mixture was again left for 5-10minutes at 25\u0026deg;C protected from light. The absorbance of the formed purple color was measured within 30 minutes at λ\u003csub\u003emax\u003c/sub\u003e 546 nm, a blank experiment was performed under the same conditions, the procedure was repeated three times for each dilution of the nitrite and the average absorbance was calculated. A plot of the average absorbance value for each concentration of the nitrite standard solution as a function of \u003csup\u003e\u0026ldquo;\u003c/sup\u003eY\u003csup\u003e\u0026rdquo;\u003c/sup\u003e against nitrite concentration as a function of \u003csup\u003e\u0026ldquo;\u003c/sup\u003eX \u003csup\u003e\u0026ldquo;\u003c/sup\u003ewas constructed to generate a standard nitrite calibration curve at pH 7.4.\u003c/p\u003e \u003cp\u003e \u003cb\u003eNO release assay\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe amount of NO released from the tested compounds \u003cb\u003e8a-b, 8d-i\u003c/b\u003e and \u003cb\u003e10a-c\u003c/b\u003e was measured using the Griess colorimetric method[\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e] either in phosphate buffer of pH 7.4 in the presence of \u003cem\u003eN\u003c/em\u003e-acetyl cysteine which serves as a source of thiols. The amount of NO released from the tested compounds was measured relative to NO released from standard sodium nitrite solution.\u003c/p\u003e \u003cp\u003e \u003cb\u003eProcedure\u003c/b\u003e \u003c/p\u003e \u003cp\u003eDifferent solutions of the tested compounds \u003cb\u003e8a-b, 8d-i\u003c/b\u003e and \u003cb\u003e10a-c\u003c/b\u003e in DMF were diluted using phosphate buffer of pH 7.4 till a final concentration of 100 mM (test solutions). To 100 mL of different test solutions, 100 mL of \u003cem\u003eN\u003c/em\u003e-acetyl cysteine solution was added and the obtained solution was kept in an incubator at 37\u0026deg;C (treated solutions). The solutions were treated similarly as for nitrite standard solution with Griess reagent components, 100 mL of sulfanilamide solution was added to each tube of the treated solution.\u003c/p\u003e \u003cp\u003eThe mixture was left at 25\u0026deg;C for 5 \u0026minus;\u0026thinsp;10 minutes protected from light. To this mixture 100 mL of the NED solution was added and the mixture was again left at 25\u0026deg;C for 5\u0026ndash;10 minutes protected from light. The absorbance of the formed purple color, if any, was measured within 30 minutes at λmax 546 nm. A blank experiment was performed under the same conditions, the procedure was repeated three times for each tested compound and the average absorbance was calculated. The corresponding concentration of nitrite was determined by comparison to the nitrite standard calibration curve and the amount of NO released (attributed by the corresponding nitrite concentration) was calculated as percentage of moles of NO released from 1 mole of the tested compounds.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section2\"\u003e \u003ch2\u003e4.3. Biology\u003c/h2\u003e \u003cp\u003e \u003cb\u003eMaterials and methods\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eEvaluation of anticancer activity was performed according to the standard water-soluble tetrazolium-8 (WST-8) assay at the Faculty of Engineering, Yamagata University, Yonezawa, Japan, using an MTP-310 absorbance microplate reader.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eThe EGFR inhibitory assay was carried out at the Faculty of Engineering, Yamagata University, Yonezawa, Japan, according to the protocol enzyme linked immunosorbent assay (ELISA) kits (Douset sandwich ELISA test, recombinant mouse EGFR).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eThe JNK-2 inhibitory assay was performed at the Faculty of Engineering, Yamagata University, Yonezawa, Japan, according to the protocol of enzyme-linked immunosorbent assay (ELISA), the assay was carried out using the JNK-2 kit (Simple Step ELISA, pT183/Y185, Abacam Company, Japan).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eApoptosis and cell cycle analysis was performed at Faculty of medicine, Yamagata University, Yamagata, Japan, using (BD FACS melody).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eMolecular docking was performed at the Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Researchers, RIEKN, Japan, using ICM-Pro 3.8 software (MolSoft L.L.C, USA).\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cdiv id=\"Sec30\" class=\"Section3\"\u003e \u003ch2\u003e4.3.1. Evaluation of anticancer activity\u003c/h2\u003e \u003cp\u003eAccording to the standard water-soluble tetrazolium-8 (WST-8) assay, the current synthesized compounds have been tested for their anticancer activities against different five cancer cell lines; DLD-1, Hela, K562, SUIT-2 and HepG2 and daunorubicin was used as reference drug by the WST-8 assay. The five cells were maintained in a suspension culture, (Dulbecco's modified eagle medium (DMEM) for SUIT-2, Hela and HepG2 or PRIM for K562 and DLD), supplemented with 5% FBS (Fetal Bovine Serum) containing 1% of a penicillin-streptomycin (1:1) mixture. A 100 \u0026micro;L aliquot of cells (10000 cells/mL) was added to a 96 well plate and incubated for 24 h at 37\u0026deg;C in a humidified incubator containing 5% CO\u003csub\u003e2\u003c/sub\u003e in the air. After 24 h, a 10 \u0026micro;L aliquot of test compound (concentrations varying in the range of 10\u0026ndash;150 \u0026micro;M) was added to each of the 96 wells and incubated for 24 h. Then A 10 \u0026micro;L WST-8 solution (mixture of WST-8 and 1-methoxy PMS) was added to each well and the incubation continued for 3 h. The visible absorbance at 450 nm and 630 nm as the reference wavelength of each well was quantified using an MTP-310 absorbance microplate reader. Daunorubicin was used as a positive control. The results of cytotoxicity were recorded as growth inhibition percentages and as IC\u003csub\u003e50\u003c/sub\u003e values [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec31\" class=\"Section3\"\u003e \u003ch2\u003e4.3.2. EGFR inhibitory assay\u003c/h2\u003e \u003cp\u003eThis assay was carried out according to the protocol for enzyme linked immunosorbent assay (ELISA) kits (Douset sandwich ELISA test, recombinant mouse EGFR) [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e, \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e]. This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for EGFR has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any EGFR present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-conjugated antibody specific for EGFR is added to each well and incubated. Following a wash to remove unbound substances streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of EGFR bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm\u0026thinsp;\u0026plusmn;\u0026thinsp;2nm. The concentration of EGFR in the sample is then determined by comparing the O.D of samples to the standard curve.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec32\" class=\"Section3\"\u003e \u003ch2\u003e4.3.3. JNK-2 inhibitory assay\u003c/h2\u003e \u003cp\u003eJNK-2 inhibitory assay was performed according to the protocol of enzyme-linked immunosorbent assay (ELISA). The assay was carried out using the JNK-2 kit (Simple Step ELISA, pT183/Y185, Abacam Company, Japan).[\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e] for the semi-quantitative measurement of JNK-2 protein in human cell lysate. The SimpleStep ELISA employs an affinity tag labelled capture antibody and a reporter attached detector antibody to immunocapture the sample analyte in solution. This complete complex (capture antibody/analyte/detector antibody) is then immobilized \u003cem\u003evia\u003c/em\u003e immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or controls are added to the wells, followed by the antibody cocktail. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by the addition of stop solution, completing any color change from blue to yellow. The signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, the development of TMB can be recorded kinetically at 600 nm.\u003c/p\u003e \u003cp\u003eAn antibody cocktail can be prepared by combining an appropriate volume of the capture and detector antibodies immediately prior to assay. To make 3 mL of the antibody cocktail, combine 1.5 mL of capture antibody with 1.5 mL of detector antibody. Mix thoroughly and gently. Control lysate can be prepared from HEK293 cells, cultured in 10% FBS containing medium, then treated with 1\u0026micro;g/mL anisomycin. After preparing all the reagents, samples and control as instructed and following the previously published procedures [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e], add 50 \u0026micro;L of sample or control to the well, add 50 \u0026micro;L of the antibody cocktail, then incubate at room temperature for 1 hour on a plate shaker set to 400 rpm, aspirate and wash each well three times with 350 \u0026micro;L with wash buffer, add 100 \u0026micro;L TMB substrate to each well and incubate for 15 minutes, then add 100 \u0026micro;L stop solution and measure the absorbance at 450 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec33\" class=\"Section3\"\u003e \u003ch2\u003e4.3.4. Apoptosis analysis\u003c/h2\u003e \u003cp\u003eFor apoptosis induction measurement, seeding of Hela cells in 96-well plates (1\u0026times;10\u003csup\u003e4\u003c/sup\u003e cell/well) and DMEM medium was added and incubated for 24 h, at the second day the cells were treated with the test compounds (at IC\u003csub\u003e50\u003c/sub\u003e, double and half of IC\u003csub\u003e50\u003c/sub\u003e) then incubated overnight, on the third day wash each well twice with 100 \u0026micro;L phosphate buffered saline (PBS), add Trypsin 100 \u0026micro;L and incubate plates for 3 to 5 minutes at 37 degrees, then 200 \u0026micro;L of medium was added to separate cells, centrifuge cells for 5 minutes and decant supernatant. Wash twice with 100 \u0026micro;L PBS and add 200\u0026micro;L of Buffer, 5 \u0026micro;L of PI\u0026amp; Annexin and 200 \u0026micro;l of PBS for the test compounds. For control, add 400 \u0026micro;L of PBS. Finally, the cell suspension was observed under the fluorescent microscope or transfer to a round bottom tube for flowcytometric analysis using (BD FACS melody).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec34\" class=\"Section3\"\u003e \u003ch2\u003e4.3.5. Cell cycle analysis\u003c/h2\u003e \u003cp\u003eTo cultured Hela cells in 96-well plates (1\u0026times;10\u003csup\u003e4\u003c/sup\u003e cell/well), DMEM medium and the IC\u003csub\u003e50\u003c/sub\u003e concentration of the test compounds were added, then incubated for 24 h after the addition of test compound. Transfer supernatant to falcon tube for each plate and wash each well twice with 3 mL PBS, add Trypsin 1 mL and incubate plates for 3 to 5 minutes at 37 degrees, then 3 mL of the medium was added to separate cells, centrifuge cells for 5 minutes and decant supernatant, wash again with 3 mL PBS, and add 10 mL of medium. Cells were immediately fixed in ice cold 70% ethanol overnight at -20\u0026deg;C. In the day of the analysis, cells were washed \u0026times;3 with PBS and re-suspended in propidium iodide (PI) for 15 minutes at room temperature, protected from light. Cell-cycle analysis was performed using flowcytometric analysis (BD FACS melody) and data obtained from cell cycle distribution was analyzed using FSC-W (Watson model) to estimate the percentage of cells in G1, S, and G2.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec35\" class=\"Section3\"\u003e \u003ch2\u003e4.3.6. Evaluation of cytotoxicity against PC12 cells\u003c/h2\u003e \u003cp\u003eAccording to the standard water-soluble tetrazolium-8 (WST-8) assay, the cytotoxicity of compounds \u003cb\u003e8g\u003c/b\u003e and \u003cb\u003e8i\u003c/b\u003e against the PC12 cell line was evaluated, and daunorubicin was used as the reference drug by WST-8. The PC12 cells were maintained in a suspension culture, (Dulbecco's modified eagle medium (DMEM), supplemented with 5% FBS (Fetal Bovine Serum) containing 1% of a penicillin-streptomycin (1:1) mixture. A 100 \u0026micro;L aliquot of cells (10000 cells/mL) was added to a 96 well plate and incubated for 24 h at 37\u003csup\u003e\u0026deg;\u003c/sup\u003eC in a humidified incubator containing 5% CO\u003csub\u003e2\u003c/sub\u003e in the air. After 24 h, a 10 \u0026micro;L aliquot of test compound (concentrations varying in the range of 10\u0026ndash;150 \u0026micro;M) was added to each of the 96 wells and incubated for 24 h. Then A 10 \u0026micro;L WST-8 solution (mixture of WST-8 and 1-methoxy PMS) was added to each well and the incubation continued for 3 h. The visible absorbance at 450 nm and 630 nm as the reference wavelength of each well was quantified using an MTP-310 absorbance microplate reader. Daunorubicin was used as a positive control. The results of cytotoxicity were recorded as growth inhibition percentages and as IC\u003csub\u003e50\u003c/sub\u003e values.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec36\" class=\"Section3\"\u003e \u003ch2\u003e4.3.7. Molecular docking on EGFR and JNK-2\u003c/h2\u003e \u003cp\u003eDocking simulation was performed by the Inter-coordinate Mechanics (ICM) using ICM-Pro 3.8 software (MolSoft L.L.C, USA)[\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. First, the 3D structures of the tested compounds and sorafenib (a reference multi-target kinase inhibitor) [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e] were generated to perform well-suited docking. Then, the enzyme was prepared by adjusting the interface properties, including water molecules deletion, hydrogen atoms optimization and formal charges refinement. In addition, enzyme relaxation was logged to run flexible docking. The ligands binding affinities were calculated by the Gaussian potential based on the ligand electrostatic potential and shape complementarity at the binding site [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. In these studies, the template-docking method was used by selecting pre-defined binding pockets of the study receptors. The structural models of the tested comounds against human EGFR complexed with AZD9291 inhibitor (2.80 ; PDB ID: 4ZAU) were used [74] and crystal structure of human JNK-2 complexed with an indazole inhibitor (2.14 \u0026Aring;; PDB ID: 3E7O) was used for c-Jun \u003cem\u003eN\u003c/em\u003e-terminal kinase 2 (JNK-2)[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable in this manuscript\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analysed during this study are included in this published article [and its supplementary information files.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026quot;The authors declare that they have no competing interests\u0026quot;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis article is not funded (Not applicable).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA- Kamal S. Abdelrahman. did the practical chemistry part\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eB- Heba A.Hassan, suggested the idea of work, wrote and reviewed the whole manuscript\u003c/p\u003e\n\u003cp\u003eC- Salah A. Abdel-Aziz, helped in the revision and writing of the introduction part\u003c/p\u003e\n\u003cp\u003eD- Adel A. Marzouk, helped in the revision and writing of the chemistry part\u003c/p\u003e\n\u003cp\u003eE- raef shams, helped in the revision and writing of the docking part\u003c/p\u003e\n\u003cp\u003eF- Keima Osawa, helped in the biological part practical and writing\u003c/p\u003e\n\u003cp\u003eG- Mohamed Abdel-Aziz, suggested the idea of work and helped in revision of the manuscript\u003c/p\u003e\n\u003cp\u003eH- Hiroyuki Konno, helped in the practical chemistry part\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe are grateful to Dr. Hideyuki Miyatake of RIKEN for using ICM software.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eGallorini, M.; Cataldi, A.; di Giacomo, V. Cyclin-dependent kinase modulators and cancer therapy. \u003cem\u003eBioDrugs\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e26\u003c/em\u003e, 377-391.\u003c/li\u003e\n\u003cli\u003eKibria, G.; Hatakeyama, H.; Harashima, H. Cancer multidrug resistance: mechanisms involved and strategies for circumvention using a drug delivery system. \u003cem\u003eArchives of pharmacal research\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e37\u003c/em\u003e, 4-15.\u003c/li\u003e\n\u003cli\u003eAbdelgawad, M. A.; Bakr, R. B.; Alkhoja, O. A.; Mohamed, W. R. Design, synthesis and antitumor activity of novel pyrazolo [3, 4-d] pyrimidine derivatives as EGFR-TK inhibitors. \u003cem\u003eBioorganic chemistry\u003c/em\u003e \u003cstrong\u003e2016\u003c/strong\u003e, \u003cem\u003e66\u003c/em\u003e, 88-96.\u003c/li\u003e\n\u003cli\u003eMitsudomi, T.; Yatabe, Y. Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. \u003cem\u003eThe FEBS journal\u003c/em\u003e \u003cstrong\u003e2010\u003c/strong\u003e, \u003cem\u003e277\u003c/em\u003e, 301-308.\u003c/li\u003e\n\u003cli\u003eRego, R.; Foster, N.; Smyrk, T.; Le, M.; O\u0026apos;connell, M.; Sargent, D.; Windschitl, H.; Sinicrope, F. Prognostic effect of activated EGFR expression in human colon carcinomas: comparison with EGFR status. \u003cem\u003eBritish journal of cancer\u003c/em\u003e \u003cstrong\u003e2010\u003c/strong\u003e, \u003cem\u003e102\u003c/em\u003e, 165-172.\u003c/li\u003e\n\u003cli\u003eCao, C.; Lu, S.; Sowa, A.; Kivlin, R.; Amaral, A.; Chu, W.; Yang, H.; Di, W.; Wan, Y. Priming with EGFR tyrosine kinase inhibitor and EGF sensitizes ovarian cancer cells to respond to chemotherapeutical drugs. \u003cem\u003eCancer letters\u003c/em\u003e \u003cstrong\u003e2008\u003c/strong\u003e, \u003cem\u003e266\u003c/em\u003e, 249-262.\u003c/li\u003e\n\u003cli\u003eBridges, A. The rationale and strategy used to develop a series of highly potent, irreversible, inhibitors of the epidermal growth factor receptor family of tyrosine kinases. \u003cem\u003eCurrent medicinal chemistry\u003c/em\u003e \u003cstrong\u003e1999\u003c/strong\u003e, \u003cem\u003e6\u003c/em\u003e, 825-844.\u003c/li\u003e\n\u003cli\u003eIida, K.; Nakayama, K.; Rahman, M.; Rahman, M.; Ishikawa, M.; Katagiri, A.; Yeasmin, S.; Otsuki, Y.; Kobayashi, H.; Nakayama, S. EGFR gene amplification is related to adverse clinical outcomes in cervical squamous cell carcinoma, making the EGFR pathway a novel therapeutic target. \u003cem\u003eBritish journal of cancer\u003c/em\u003e \u003cstrong\u003e2011\u003c/strong\u003e, \u003cem\u003e105\u003c/em\u003e, 420-427.\u003c/li\u003e\n\u003cli\u003eHuang, P.; Xu, X.; Wang, L.; Zhu, B.; Wang, X.; Xia, J. The role of EGF‐EGFR signalling pathway in hepatocellular carcinoma inflammatory microenvironment. \u003cem\u003eJournal of cellular and molecular medicine\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e18\u003c/em\u003e, 218-230.\u003c/li\u003e\n\u003cli\u003eAllen, H.; Hsuan, J.; Clark, S.; Maziarz, R.; Waterfield, M.; Flavell, R.; Haley, J. Expression of epidermal-growth-factor receptor in the K562 cell line by transfection. Altered receptor biochemistry. \u003cem\u003eBiochemical journal\u003c/em\u003e \u003cstrong\u003e1990\u003c/strong\u003e, \u003cem\u003e271\u003c/em\u003e, 785-790.\u003c/li\u003e\n\u003cli\u003eMathew, M. P.; Tan, E.; Saeui, C. T.; Bovonratwet, P.; Sklar, S.; Bhattacharya, R.; Yarema, K. J. Metabolic flux-driven sialylation alters internalization, recycling, and drug sensitivity of the epidermal growth factor receptor (EGFR) in SW1990 pancreatic cancer cells. \u003cem\u003eOncotarget\u003c/em\u003e \u003cstrong\u003e2016\u003c/strong\u003e, \u003cem\u003e7\u003c/em\u003e, 66491.\u003c/li\u003e\n\u003cli\u003eButterworth, S.; Cross, D. A.; Finlay, M. R. V.; Ward, R. A.; Waring, M. J. The structure-guided discovery of osimertinib: the first US FDA approved mutant selective inhibitor of EGFR T790M. \u003cem\u003eMedchemcomm\u003c/em\u003e \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e8\u003c/em\u003e, 820-822.\u003c/li\u003e\n\u003cli\u003eShaw, D.; Wang, S. M.; Villase\u0026ntilde;or, A. G.; Tsing, S.; Walter, D.; Browner, M. F.; Barnett, J.; Kuglstatter, A. The crystal structure of JNK2 reveals conformational flexibility in the MAP kinase insert and indicates its involvement in the regulation of catalytic activity. \u003cem\u003eJournal of molecular biology\u003c/em\u003e \u003cstrong\u003e2008\u003c/strong\u003e, \u003cem\u003e383\u003c/em\u003e, 885-893.\u003c/li\u003e\n\u003cli\u003eKuglstatter, A.; Ghate, M.; Tsing, S.; Villase\u0026ntilde;or, A. G.; Shaw, D.; Barnett, J. W.; Browner, M. F. X-ray crystal structure of JNK2 complexed with the p38\u0026alpha; inhibitor BIRB796: insights into the rational design of DFG-out binding MAP kinase inhibitors. \u003cem\u003eBioorganic \u0026amp; medicinal chemistry letters\u003c/em\u003e \u003cstrong\u003e2010\u003c/strong\u003e, \u003cem\u003e20\u003c/em\u003e, 5217-5220.\u003c/li\u003e\n\u003cli\u003eAbdelrahman, K. S.; Hassan, H. A.; Abdel-Aziz, S. A.; Marzouk, A. A.; Narumi, A.; Konno, H.; Abdel-Aziz, M. JNK signaling as a target for anticancer therapy. \u003cem\u003ePharmacological Reports\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, 1-30.\u003c/li\u003e\n\u003cli\u003eOzgur, E.; Kayhan, H.; Kismali, G.; Senturk, F.; Sensoz, M.; Ozturk, G. G.; Sel, T. Effects of radiofrequency radiation on colorectal cancer cell proliferation and inflammation. \u003cem\u003eTurkish Journal of Biochemistry\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e46\u003c/em\u003e, 525-532.\u003c/li\u003e\n\u003cli\u003eYue, Y.; Yan, L.; Yan-Ping, G.; Li, Z.; Lu-Bo, G. GL-V9 reverses adriamycin resistance in hepatocellular carcinoma cells by affecting JNK2-related autophagy. \u003cem\u003eChinese journal of natural medicines\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e18\u003c/em\u003e, 491-499.\u003c/li\u003e\n\u003cli\u003eTian, X.; Traub, B.; Shi, J.; Huber, N.; Schreiner, S.; Chen, G.; Zhou, S.; Henne-Bruns, D.; Knippschild, U.; Kornmann, M. c-Jun N-terminal kinase 2 suppresses pancreatic cancer growth and invasion and is opposed by c-Jun N-terminal kinase 1. \u003cem\u003eCancer Gene Therapy\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e29\u003c/em\u003e, 73-86.\u003c/li\u003e\n\u003cli\u003eMerkerova, M.; Bruchova, H.; Brdicka, R. JNK2 and p38 MAPK over-expressions do not represent key events in chronic myeloid leukemia transformation. \u003cem\u003eNeoplasma\u003c/em\u003e \u003cstrong\u003e2007\u003c/strong\u003e, \u003cem\u003e54\u003c/em\u003e, 503-510.\u003c/li\u003e\n\u003cli\u003eLin, C. S.; Lin, C. L.; Ying, T. H.; Chiou, H. L.; Hung, C. H.; Liao, W. S.; Hsieh, Y. H.; Kao, S. H. \u0026beta;‐Mangostin inhibits the metastatic power of cervical cancer cells attributing to suppression of JNK2/AP‐1/Snail cascade. \u003cem\u003eJournal of Cellular Physiology\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e235\u003c/em\u003e, 8446-8460.\u003c/li\u003e\n\u003cli\u003eKardosh, A.; Wang, W.; Uddin, J.; Petasis, N. A.; Hofman, F. M.; Chen, T. C.; Schonthal, A. H. Dimethyl-celecoxib (DMC), a derivative of celecoxib that lacks cyclooxygenase-2-inhibitory function, potently mimics the anti-tumor effects of celecoxib on Burkitt\u0026rsquo;s lymphoma in vitro and in vivo. \u003cem\u003eCancer biology \u0026amp; therapy\u003c/em\u003e \u003cstrong\u003e2005\u003c/strong\u003e, \u003cem\u003e4\u003c/em\u003e, 571-582.\u003c/li\u003e\n\u003cli\u003eSharma, V.; Bhatia, P.; Alam, O.; Naim, M. J.; Nawaz, F.; Sheikh, A. A.; Jha, M. Recent advancement in the discovery and development of COX-2 inhibitors: Insight into biological activities and SAR studies (2008\u0026ndash;2019). \u003cem\u003eBioorganic chemistry\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e89\u003c/em\u003e, 103007.\u003c/li\u003e\n\u003cli\u003eChandrakantha, B.; Isloor, A. M.; Shetty, P.; Isloor, S.; Malladi, S.; Fun, H. K. Synthesis, characterization and antimicrobial activity of novel ethyl 1-(N-substituted)-5-phenyl-1 H-pyrazole-4-carboxylate derivatives. \u003cem\u003eMedicinal Chemistry Research\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e21\u003c/em\u003e, 2702-2708.\u003c/li\u003e\n\u003cli\u003eManvar, D.; Pelliccia, S.; La Regina, G.; Famiglini, V.; Coluccia, A.; Ruggieri, A.; Anticoli, S.; Lee, J.-C.; Basu, A.; Cevik, O. New 1-phenyl-5-(1H-pyrrol-1-yl)-1H-pyrazole-3-carboxamides inhibit hepatitis C virus replication via suppression of cyclooxygenase-2. \u003cem\u003eEuropean journal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e90\u003c/em\u003e, 497-506.\u003c/li\u003e\n\u003cli\u003ePathak, R. B.; Chovatia, P.; Parekh, H. Synthesis, antitubercular and antimicrobial evaluation of 3-(4-chlorophenyl)-4-substituted pyrazole derivatives. \u003cem\u003eBioorganic \u0026amp; medicinal chemistry letters\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e22\u003c/em\u003e, 5129-5133.\u003c/li\u003e\n\u003cli\u003eYuan, J.-W.; Wang, S.-F.; Luo, Z.-L.; Qiu, H.-Y.; Wang, P.-F.; Zhang, X.; Yang, Y.-A.; Yin, Y.; Zhang, F.; Zhu, H.-L. Synthesis and biological evaluation of compounds which contain pyrazole, thiazole and naphthalene ring as antitumor agents. \u003cem\u003eBioorganic \u0026amp; medicinal chemistry letters\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e24\u003c/em\u003e, 2324-2328.\u003c/li\u003e\n\u003cli\u003eYu, L.; Wu, W. K. K.; Li, Z. J.; Liu, Q. C.; Li, H. T.; Wu, Y. C.; Cho, C. H. Enhancement of doxorubicin cytotoxicity on human esophageal squamous cell carcinoma cells by indomethacin and 4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (SC236) via inhibiting P-glycoprotein activity. \u003cem\u003eMolecular pharmacology\u003c/em\u003e \u003cstrong\u003e2009\u003c/strong\u003e, \u003cem\u003e75\u003c/em\u003e, 1364-1373.\u003c/li\u003e\n\u003cli\u003eZhang, L.; Peterson, T. E.; Lu, V. M.; Parney, I. F.; Daniels, D. J. Antitumor activity of novel pyrazole-based small molecular inhibitors of the STAT3 pathway in patient derived high grade glioma cells. \u003cem\u003ePlos one\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e14\u003c/em\u003e, e0220569.\u003c/li\u003e\n\u003cli\u003eSharp, S. Y.; Boxall, K.; Rowlands, M.; Prodromou, C.; Roe, S. M.; Maloney, A.; Powers, M.; Clarke, P. A.; Box, G.; Sanderson, S. Correction: In vitro Biological Characterization of a Novel, Synthetic Diaryl Pyrazole Resorcinol Class of Heat Shock Protein 90 Inhibitors. \u003cem\u003eCancer research\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e79\u003c/em\u003e, 287-287.\u003c/li\u003e\n\u003cli\u003eAbuo-Rahma, G. E.-D. A.; Abdel-Aziz, M.; Beshr, E. A.; Ali, T. F. 1, 2, 4-Triazole/oxime hybrids as new strategy for nitric oxide donors: Synthesis, anti-inflammatory, ulceroginicity and antiproliferative activities. \u003cem\u003eEuropean journal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e71\u003c/em\u003e, 185-198.\u003c/li\u003e\n\u003cli\u003eShen, S.; Xu, N.; Klamer, G.; Ko, K.-H.; Khoo, M.; Ma, D.; Moore, J.; O\u0026apos;Brien, T. A.; Dolnikov, A. Small-molecule inhibitor of glycogen synthase kinase 3\u0026beta; 6-bromoindirubin-3-oxime inhibits hematopoietic regeneration in stem cell recipient mice. \u003cem\u003eStem Cells and Development\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e24\u003c/em\u003e, 724-736.\u003c/li\u003e\n\u003cli\u003eZhang, X.; Castanotto, D.; Nam, S.; Horne, D.; Stein, C. 6BIO enhances oligonucleotide activity in cells: a potential combinatorial anti-androgen receptor therapy in prostate cancer cells. \u003cem\u003eMolecular Therapy\u003c/em\u003e \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e25\u003c/em\u003e, 79-91.\u003c/li\u003e\n\u003cli\u003eXiong, B.; Chen, S.; Zhu, P.; Huang, M.; Gao, W.; Zhu, R.; Qian, J.; Peng, Y.; Zhang, Y.; Dai, H. Design, synthesis, and biological evaluation of novel thiazolyl substituted bis-pyrazole oxime derivatives with potent antitumor activities by selectively inducing apoptosis and ROS in cancer cells. \u003cem\u003eMedicinal Chemistry\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e15\u003c/em\u003e, 743-754.\u003c/li\u003e\n\u003cli\u003eHong, S.; Shin, Y.; Jung, M.; Ha, M. W.; Park, Y.; Lee, Y.-J.; Shin, J.; Oh, K. B.; Lee, S. K.; Park, H.-g. Efficient synthesis and biological activity of Psammaplin A and its analogues as antitumor agents. \u003cem\u003eEuropean Journal of Medicinal Chemistry\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e96\u003c/em\u003e, 218-230.\u003c/li\u003e\n\u003cli\u003eBednarczyk-Cwynar, B.; Zaprutko, L. Recent advances in synthesis and biological activity of triterpenic acylated oximes. \u003cem\u003ePhytochemistry Reviews\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e14\u003c/em\u003e, 203-231.\u003c/li\u003e\n\u003cli\u003eSchepetkin, I. A.; Khlebnikov, A. I.; Potapov, A. S.; Kovrizhina, A. R.; Matveevskaya, V. V.; Belyanin, M. L.; Atochin, D. N.; Zanoza, S. O.; Gaidarzhy, N. M.; Lyakhov, S. A. Synthesis, biological evaluation, and molecular modeling of 11H-indeno [1, 2-b] quinoxalin-11-one derivatives and tryptanthrin-6-oxime as c-Jun N-terminal kinase inhibitors. \u003cem\u003eEuropean journal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e161\u003c/em\u003e, 179-191.\u003c/li\u003e\n\u003cli\u003eSchepetkin, I. A.; Kirpotina, L. N.; Khlebnikov, A. I.; Hanks, T. S.; Kochetkova, I.; Pascual, D. W.; Jutila, M. A.; Quinn, M. T. Identification and characterization of a novel class of c-Jun N-terminal kinase inhibitors. \u003cem\u003eMolecular pharmacology\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e81\u003c/em\u003e, 832-845.\u003c/li\u003e\n\u003cli\u003eLu, L.; Sha, S.; Wang, K.; Zhang, Y.-H.; Liu, Y.-D.; Ju, G.-D.; Wang, B.; Zhu, H.-L. Discovery of chromeno [4, 3-c] pyrazol-4 (2H)-one containing carbonyl or oxime derivatives as potential, selective inhibitors PI3K\u0026alpha;. \u003cem\u003eChemical and Pharmaceutical Bulletin\u003c/em\u003e \u003cstrong\u003e2016\u003c/strong\u003e, c16-00388.\u003c/li\u003e\n\u003cli\u003eSchepetkin, I. A.; Plotnikov, M. B.; Khlebnikov, A. I.; Plotnikova, T. M.; Quinn, M. T. Oximes: Novel therapeutics with anticancer and anti-inflammatory potential. \u003cem\u003eBiomolecules\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e11\u003c/em\u003e, 777.\u003c/li\u003e\n\u003cli\u003eSoliman, R. Preparation and antidiabetic activity of some sulfonylurea derivatives of 3, 5-disubstituted pyrazoles. \u003cem\u003eJournal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e1979\u003c/strong\u003e, \u003cem\u003e22\u003c/em\u003e, 321-325.\u003c/li\u003e\n\u003cli\u003eAghazadeh Tabrizi, M.; Baraldi, P. G.; Baraldi, S.; Ruggiero, E.; De Stefano, L.; Rizzolio, F.; Di Cesare Mannelli, L.; Ghelardini, C.; Chicca, A.; Lapillo, M. Discovery of 1, 5-diphenylpyrazole-3-carboxamide derivatives as potent, reversible, and selective monoacylglycerol lipase (MAGL) inhibitors. \u003cem\u003eJournal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e61\u003c/em\u003e, 1340-1354.\u003c/li\u003e\n\u003cli\u003eShin, U. c.; Choi, J. S.; Beak, Y. J.; Lee, M. w.; Kim, H. S.; Choi, D. W.; Kim, D. G.; Kim, S. W. Development of a 68Ga‐labelled PET tracer for carbonic anhydrase IX‐overexpressed tumors using the artificial sweetener saccharin. \u003cem\u003eJournal of Labelled Compounds and Radiopharmaceuticals\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e.\u003c/li\u003e\n\u003cli\u003eGonzalez, R. M.; Seurynck-Servoss, S. L.; Crowley, S. A.; Brown, M.; Omenn, G. S.; Hayes, D. F.; Zangar, R. C. Development and validation of sandwich ELISA microarrays with minimal assay interference. \u003cem\u003eJournal of proteome research\u003c/em\u003e \u003cstrong\u003e2008\u003c/strong\u003e, \u003cem\u003e7\u003c/em\u003e, 2406-2414.\u003c/li\u003e\n\u003cli\u003eEl-Kashef, D. H.; El-Sheakh, A. R. Hepatoprotective effect of celecoxib against tamoxifen-induced liver injury via inhibiting ASK-1/JNK pathway in female rats. \u003cem\u003eLife sciences\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e231\u003c/em\u003e, 116573.\u003c/li\u003e\n\u003cli\u003eTsutsumi, R.; Ito, H.; Hiramitsu, T.; Nishitani, K.; Akiyoshi, M.; Kitaori, T.; Yasuda, T.; Nakamura, T. Celecoxib inhibits production of MMP and NO via down-regulation of NF-\u0026kappa;B and JNK in a PGE2 independent manner in human articular chondrocytes. \u003cem\u003eRheumatology international\u003c/em\u003e \u003cstrong\u003e2008\u003c/strong\u003e, \u003cem\u003e28\u003c/em\u003e, 727-736.\u003c/li\u003e\n\u003cli\u003eMaayah, Z. H.; Levasseur, J.; Piragasam, R. S.; Abdelhamid, G.; Dyck, J. R.; Fahlman, R. P.; Siraki, A. G.; El-Kadi, A. O. 2-Methoxyestradiol protects against pressure overload-induced left ventricular hypertrophy. \u003cem\u003eScientific reports\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e8\u003c/em\u003e, 1-15.\u003c/li\u003e\n\u003cli\u003ePozarowski, P.; Grabarek, J.; Darzynkiewicz, Z. Flow cytometry of apoptosis. \u003cem\u003eCurrent protocols in cell biology\u003c/em\u003e \u003cstrong\u003e2003\u003c/strong\u003e, \u003cem\u003e21\u003c/em\u003e, 18.18. 11-18.18. 33.\u003c/li\u003e\n\u003cli\u003eYosaatmadja, Y.; Silva, S.; Dickson, J. M.; Patterson, A. V.; Smaill, J. B.; Flanagan, J. U.; McKeage, M. J.; Squire, C. J. Binding mode of the breakthrough inhibitor AZD9291 to epidermal growth factor receptor revealed. \u003cem\u003eJournal of structural biology\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e192\u003c/em\u003e, 539-544.\u003c/li\u003e\n\u003cli\u003eJousserandot, A.; Boucher, J.-L.; Henry, Y.; Niklaus, B.; Clement, B.; Mansuy, D. Microsomal cytochrome P450 dependent oxidation of N-hydroxyguanidines, amidoximes, and ketoximes: mechanism of the oxidative cleavage of their CN (OH) bond with formation of nitrogen oxides. \u003cem\u003eBiochemistry\u003c/em\u003e \u003cstrong\u003e1998\u003c/strong\u003e, \u003cem\u003e37\u003c/em\u003e, 17179-17191.\u003c/li\u003e\n\u003cli\u003eRadwan, A. A.; Ghorab, M. M.; Alsaid, M. S.; Alanazi, F. K. Novel ethyl 1, 5-disubstituted-1H-pyrazole-3-carboxylates as a new class of antimicrobial agents. \u003cem\u003eActa Pharmaceutica\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e64\u003c/em\u003e, 335-344.\u003c/li\u003e\n\u003cli\u003eLi, Z.; Wang, Z.-C.; Li, X.; Abbas, M.; Wu, S.-Y.; Ren, S.-Z.; Liu, Q.-X.; Liu, Y.; Chen, P.-W.; Duan, Y.-T. Design, synthesis and evaluation of novel diaryl-1, 5-diazoles derivatives bearing morpholine as potent dual COX-2/5-LOX inhibitors and antitumor agents. \u003cem\u003eEuropean journal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e169\u003c/em\u003e, 168-184.\u003c/li\u003e\n\u003cli\u003ePenning, T. D.; Talley, J. J.; Bertenshaw, S. R.; Carter, J. S.; Collins, P. W.; Docter, S.; Graneto, M. J.; Lee, L. F.; Malecha, J. W.; Miyashiro, J. M. Synthesis and biological evaluation of the 1, 5-diarylpyrazole class of cyclooxygenase-2 inhibitors: identification of 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1 H-pyrazol-1-yl] benzenesulfonamide (SC-58635, celecoxib). \u003cem\u003eJournal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e1997\u003c/strong\u003e, \u003cem\u003e40\u003c/em\u003e, 1347-1365.\u003c/li\u003e\n\u003cli\u003eBatchu, H.; Bhattacharyya, S.; Kant, R.; Batra, S. Palladium-catalyzed chelation-assisted regioselective oxidative dehydrogenative homocoupling/ortho-hydroxylation in N-phenylpyrazoles. \u003cem\u003eThe Journal of organic chemistry\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e80\u003c/em\u003e, 7360-7374.\u003c/li\u003e\n\u003cli\u003eMurray, W. V.; Wachter, M. P. A simple regioselective synthesis of ethyl 1, 5‐diarylpyrazole‐3‐carboxylates. \u003cem\u003eJournal of heterocyclic chemistry\u003c/em\u003e \u003cstrong\u003e1989\u003c/strong\u003e, \u003cem\u003e26\u003c/em\u003e, 1389-1392.\u003c/li\u003e\n\u003cli\u003eYamali, C.; Gul, H. I.; Ece, A.; Bua, S.; Angeli, A.; Sakagami, H.; Sahin, E.; Supuran, C. T. Synthesis, biological evaluation and in silico modelling studies of 1, 3, 5-trisubstituted pyrazoles carrying benzenesulfonamide as potential anticancer agents and selective cancer-associated hCA IX isoenzyme inhibitors. \u003cem\u003eBioorganic chemistry\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e92\u003c/em\u003e, 103222.\u003c/li\u003e\n\u003cli\u003eRogez-Florent, T.; Meignan, S.; Foulon, C.; Six, P.; Gros, A.; Bal-Mahieu, C.; Supuran, C. T.; Scozzafava, A.; Fr\u0026eacute;d\u0026eacute;rick, R.; Masereel, B. New selective carbonic anhydrase IX inhibitors: synthesis and pharmacological evaluation of diarylpyrazole-benzenesulfonamides. \u003cem\u003eBioorganic \u0026amp; medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2013\u003c/strong\u003e, \u003cem\u003e21\u003c/em\u003e, 1451-1464.\u003c/li\u003e\n\u003cli\u003eHarrison, C. 18 months sliced off Celebrex\u0026apos;s patent protection. \u003cem\u003eNature reviews. Drug discovery\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e13\u003c/em\u003e, 326.\u003c/li\u003e\n\u003cli\u003eLu, X.-Y.; Wang, Z.-C.; Ren, S.-Z.; Shen, F.-Q.; Man, R.-J.; Zhu, H.-L. Coumarin sulfonamides derivatives as potent and selective COX-2 inhibitors with efficacy in suppressing cancer proliferation and metastasis. \u003cem\u003eBioorganic \u0026amp; medicinal chemistry letters\u003c/em\u003e \u003cstrong\u003e2016\u003c/strong\u003e, \u003cem\u003e26\u003c/em\u003e, 3491-3498.\u003c/li\u003e\n\u003cli\u003eRaiford, L. C.; Hill, E. L. Effect of Constitution on the Rearrangement of the Phenylhydrazones of Some Unsymmetrically Substituted Dibenzalacetones1. \u003cem\u003eJournal of the American Chemical Society\u003c/em\u003e \u003cstrong\u003e1934\u003c/strong\u003e, \u003cem\u003e56\u003c/em\u003e, 174-176.\u003c/li\u003e\n\u003cli\u003eHwang, S. H.; Wagner, K. M.; Morisseau, C.; Liu, J.-Y.; Dong, H.; Wecksler, A. T.; Hammock, B. D. Synthesis and structure\u0026minus; activity relationship studies of urea-containing pyrazoles as dual inhibitors of cyclooxygenase-2 and soluble epoxide hydrolase. \u003cem\u003eJournal of medicinal chemistry\u003c/em\u003e \u003cstrong\u003e2011\u003c/strong\u003e, \u003cem\u003e54\u003c/em\u003e, 3037-3050.\u003c/li\u003e\n\u003cli\u003eUddin, M. J.; Crews, B. C.; Ghebreselasie, K.; Marnett, L. J. Design, synthesis, and structure\u0026ndash;activity relationship studies of fluorescent inhibitors of cycloxygenase-2 as targeted optical imaging agents. \u003cem\u003eBioconjugate chemistry\u003c/em\u003e \u003cstrong\u003e2013\u003c/strong\u003e, \u003cem\u003e24\u003c/em\u003e, 712-723.\u003c/li\u003e\n\u003cli\u003eChen, L. W.; Wang, P. F.; Tang, D. J.; Tao, X. X.; Man, R. J.; Qiu, H. Y.; Wang, Z. C.; Xu, C.; Zhu, H. L. Metronidazole containing pyrazole derivatives potently inhibit tyrosyl‐tRNA synthetase: design, synthesis, and biological evaluation. \u003cem\u003eChemical biology \u0026amp; drug design\u003c/em\u003e \u003cstrong\u003e2016\u003c/strong\u003e, \u003cem\u003e88\u003c/em\u003e, 592-598.\u003c/li\u003e\n\u003cli\u003eAbdelazeem, A. H.; El-Din, A. G. S.; Abdel-Fattah, M. M.; Amin, N. H.; El-Moghazy, S. M.; El-Saadi, M. T. Discovery of novel urea-diarylpyrazole hybrids as dual COX-2/sEH inhibitors with improved anti-inflammatory activity and highly reduced cardiovascular risks. \u003cem\u003eEuropean Journal of Medicinal Chemistry\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e205\u003c/em\u003e, 112662.\u003c/li\u003e\n\u003cli\u003eGuevara, I.; Iwanejko, J.; Dembińska-Kieć, A.; Pankiewicz, J.; Wanat, A.; Anna, P.; Goła̧bek, I.; Bartuś, S.; Malczewska-Malec, M.; Szczudlik, A. Determination of nitrite/nitrate in human biological material by the simple Griess reaction. \u003cem\u003eClinica Chimica Acta\u003c/em\u003e \u003cstrong\u003e1998\u003c/strong\u003e, \u003cem\u003e274\u003c/em\u003e, 177-188.\u003c/li\u003e\n\u003cli\u003eIshiyama, M.; Miyazono, Y.; Sasamoto, K.; Ohkura, Y.; Ueno, K. A highly water-soluble disulfonated tetrazolium salt as a chromogenic indicator for NADH as well as cell viability. \u003cem\u003eTalanta\u003c/em\u003e \u003cstrong\u003e1997\u003c/strong\u003e, \u003cem\u003e44\u003c/em\u003e, 1299-1305.\u003c/li\u003e\n\u003cli\u003eHotsumi, M.; Tajiri, M.; Nikaido, Y.; Sato, T.; Makabe, K.; Konno, H. Design, synthesis, and evaluation of a water soluble C5-monoketone type curcumin analogue as a potent amyloid \u0026beta; aggregation inhibitor. \u003cem\u003eBioorganic \u0026amp; medicinal chemistry letters\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e29\u003c/em\u003e, 2157-2161.\u003c/li\u003e\n\u003cli\u003eGangemi, S.; Franchina, T.; Minciullo, P. L.; Profita, M.; Zangh\u0026igrave;, M.; David, A.; Kennez, I.; Adamo, V. IL‐33/IL‐31 axis: A new pathological mechanisms for EGFR tyrosine kinase inhibitors‐associated skin toxicity. \u003cem\u003eJournal of cellular biochemistry\u003c/em\u003e \u003cstrong\u003e2013\u003c/strong\u003e, \u003cem\u003e114\u003c/em\u003e, 2673-2676.\u003c/li\u003e\n\u003cli\u003eMasago, K.; Fujita, S.; Hatachi, Y.; Fukuhara, A.; Sakuma, K.; Ichikawa, M.; Kim, Y. H.; Mio, T.; Mishima, M. Clinical significance of pretreatment serum amphiregulin and transforming growth factor‐\u0026alpha;, and an epidermal growth factor receptor somatic mutation in patients with advanced non‐squamous, non‐small cell lung cancer. \u003cem\u003eCancer science\u003c/em\u003e \u003cstrong\u003e2008\u003c/strong\u003e, \u003cem\u003e99\u003c/em\u003e, 2295-2301.\u003c/li\u003e\n\u003cli\u003eGil-Mart\u0026iacute;nez, A. L.; Cuenca, L.; Estrada, C.; S\u0026aacute;nchez-Rodrigo, C.; Fern\u0026aacute;ndez-Villalba, E.; Herrero, M. T. Unexpected exacerbation of neuroinflammatory response after a combined therapy in old Parkinsonian mice. \u003cem\u003eFrontiers in cellular neuroscience\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e12\u003c/em\u003e, 451.\u003c/li\u003e\n\u003cli\u003eCenariu, D.; Fischer-Fodor, E.; Țigu, A. B.; Bunea, A.; Vir\u0026aacute;g, P.; Perde-Schrepler, M.; Toma, V.-A.; Mocan, A.; Berindan-Neagoe, I.; Pintea, A. Zeaxanthin-Rich Extract from Superfood Lycium barbarum Selectively Modulates the Cellular Adhesion and MAPK Signaling in Melanoma versus Normal Skin Cells In Vitro. \u003cem\u003eMolecules\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e26\u003c/em\u003e, 333.\u003c/li\u003e\n\u003cli\u003eNeves, M. A.; Totrov, M.; Abagyan, R. Docking and scoring with ICM: the benchmarking results and strategies for improvement. \u003cem\u003eJournal of computer-aided molecular design\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e26\u003c/em\u003e, 675-686.\u003c/li\u003e\n\u003cli\u003eAn, J.; Totrov, M.; Abagyan, R. Pocketome via comprehensive identification and classification of ligand binding envelopes. \u003cem\u003eMolecular \u0026amp; Cellular Proteomics\u003c/em\u003e \u003cstrong\u003e2005\u003c/strong\u003e, \u003cem\u003e4\u003c/em\u003e, 752-761.\u003c/li\u003e\n\u003cli\u003eHynes, N. E.; Lane, H. A. ERBB receptors and cancer: the complexity of targeted inhibitors. \u003cem\u003eNature Reviews Cancer\u003c/em\u003e \u003cstrong\u003e2005\u003c/strong\u003e, \u003cem\u003e5\u003c/em\u003e, 341-354.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Pyrazole, EGFR, JNK-2, Docking, Anti-proliferative ","lastPublishedDoi":"10.21203/rs.3.rs-2772431/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2772431/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA series of new 1,5-diarylpyrazole oxime hybrid derivatives (\u003cstrong\u003escaffold A\u003c/strong\u003e and \u003cstrong\u003eB\u003c/strong\u003e) were designed, synthesized, and their structures were examined for purity with different spectroscopic techniques. All the synthesized compounds (\u003cstrong\u003e7a-j\u003c/strong\u003e), (\u003cstrong\u003e8a-j\u003c/strong\u003e), (\u003cstrong\u003e9a-c\u003c/strong\u003e) and (\u003cstrong\u003e10a-c\u003c/strong\u003e) were biologically evaluated for their \u003cem\u003ein vitro\u003c/em\u003e cytotoxicity against a panel of five cancer cell lines known to express EGFR and JNK-2, namely human colorectal adenocarcinoma cell line DLD-1, human cervical cancer cell line Hela, human leukemia cell line K562, human pancreatic cell line SUIT-2 and human hepatocellular carcinoma cell line HepG2. The oxime containing compounds (\u003cstrong\u003e8a-j\u003c/strong\u003e) and (\u003cstrong\u003e10a-c\u003c/strong\u003e) were more active as antiproliferative agents than their non-oxime congeners (\u003cstrong\u003e7a-j\u003c/strong\u003e) and (\u003cstrong\u003e9a-c\u003c/strong\u003e). Compounds \u003cstrong\u003e8d, 8g, 8i,\u003c/strong\u003e and \u003cstrong\u003e10c\u003c/strong\u003e inhibited EGFR with IC\u003csub\u003e50\u003c/sub\u003e values ranging from 8 to 21 µM when compared to sorafenib. Compound \u003cstrong\u003e8i\u003c/strong\u003e inhibited JNK-2 as effectively as sorafenib, with an IC\u003csub\u003e50\u003c/sub\u003e of 1.00 µM. Furthermore, compound \u003cstrong\u003e8g \u003c/strong\u003eshowed cell cycle arrest at the G2/M phase in the Hela cell line cell cycle analysis, whereas compound \u003cstrong\u003e8i\u003c/strong\u003e showed combined S phase and G2 phase arrest. Docking studies revealed that oxime derivatives fit well at the EGFR binding site, with binding free energies ranging from -12.98 to 32.30 kcl/mol for compounds \u003cstrong\u003e8d, 8g, 8i,\u003c/strong\u003e and \u003cstrong\u003e10c\u003c/strong\u003e, while compounds \u003cstrong\u003e8d \u003c/strong\u003eand \u003cstrong\u003e8i\u003c/strong\u003e had binding free energies ranging from -9.16 to -12.00 kcl/mol at the JNK-2 binding site.\u003c/p\u003e","manuscriptTitle":"Synthesis, molecular docking and evaluation of 1,5-diarylpyrazole/oxime hybrids targeting EGFR and JNK-2 as antiproliferative agents","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-04-11 14:39:16","doi":"10.21203/rs.3.rs-2772431/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"23aee33a-662b-47c3-8688-d14637eac256","owner":[],"postedDate":"April 11th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-05-30T14:45:19+00:00","versionOfRecord":[],"versionCreatedAt":"2023-04-11 14:39:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2772431","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2772431","identity":"rs-2772431","version":["v1"]},"buildId":"pf3fE39SIOqb-0xH_OWvX","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}