Rational Design and Synthesis of Matrine Containing Coumarin Derivatives as Hsp90 (NTD&CTD) Isoform selective Inhibitors for the Treatment of Lung Carcinoma

Rational Design and Synthesis of Matrine Containing Coumarin Derivatives as Hsp90 (NTD&CTD) Isoform selective Inhibitors for the Treatment of Lung Carcinoma | 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 Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Rational Design and Synthesis of Matrine Containing Coumarin Derivatives as Hsp90 (NTD&CTD) Isoform selective Inhibitors for the Treatment of Lung Carcinoma Jamal A.H Kowah, Chenxi Guan, Meiyan Jiang, Ruobing Gao, Yufang Li, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4632508/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 Matrine serves as the molecular backbone, targeting the Hsp90 protein N-terminal domain (NTD) and C-terminal domain (CTD), both highly expressed in lung tumor cells. In this study, Matrine Contains Coumarins derivatives were designed and synthesized based on our previously reported compound C . Employing primary structure-activity relationships and docking analysis, a series of derivatives were biologically assessed for their antiproliferative effects against three cancer cell lines: A549, HepG-2, and HeLa cells. Based on the bioactivity results, derivative 5a emerged as the most potent, significantly enhancing antiproliferation against A549, HepG-2, and HeLa cells, with IC 50 values of 7.35 ± 0.097, 7.727 ± 0.10, and 8.02 ± 0.065 µM, respectively. Subsequent mechanistic investigations confirmed 5a 's ability to inhibit A549 cell proliferation and suppress colony formation and migration. In in vivo studies utilizing a xenograft mouse model inoculated with A549 cells in female Balb/c nude mice, compound 5a displayed superior antitumor activity compared to reference compounds 5-Fluorouracil and Matrine. Notably, the tumor growth inhibition (TGI) values for 5a , 5-Fluorouracil, and Matrine were 72.4%, 64.3%, and 46.8%, respectively. Physical sciences/Chemistry/Biochemistry Physical sciences/Chemistry/Biosynthesis Biological sciences/Biochemistry Biological sciences/Cell biology Biological sciences/Chemical biology Biological sciences/Drug discovery Biological sciences/Evolution Biological sciences/Molecular biology Physical sciences/Chemistry Matrine-based Coumarin Anticancer Agents Hsp90 Inhibitors Lung Carcinoma Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 10 Figure 11 Figure 12 1. Introduction Cancer represents a significant global health risk, despite the remarkable strides made in the field of medical science for cancer treatment. Consequently, substantial health concerns persist among individuals affected by this devastating disease[ 1 ]. The incidence and mortality of cancer have continued to rise over time [ 2 ]. Chemotherapy remains an essential component in the clinical treatment of solid tumors. However, various challenges, including drug resistance and unfavorable side effects, limit the widespread application of chemotherapy. It is vitally necessary to develop new anticancer medicines that are more effective and less toxic to healthy cells. Type II programmed cell death or autophagy is new and vital for preserving cellular homeostasis via lysosome-dependent degradation of cytoplasmic proteins and organelles. Recent research has demonstrated that activating autophagic cell death has a significant effect on cancer treatment [ 3 ] and autophagic cell death has become more and more widely used in the field of tumor treatment [ 4 ]. Long identified as an anticancer target, Hsp90 is an ATP-dependent chaperone necessary for various protein assembly and folding processes. Recently, scientists discovered that Hsp90 plays a significant role in inflammation, neurodegenerative disease and viral infection. Developing drugs that inhibit this chaperone might help treat these previously untreatable conditions. Numerous client proteins implicated in cancer pathogenesis have been identified several of which possess carcinogenic properties. Notable examples include the tumor protein 53 (p53), the human epidermal growth factor receptor 2 (HER2/neu), signal transduction proteins Braf and Akt, and cell cycle regulators CDK4/6. These proteins play pivotal roles in critical cellular processes associated with cancer development, progression, and therapeutic response. [ 5 , 6 ]. A promising target for anticancer medicines has been identified as Hsp90 due to its involvement in crucial cancer activities [ 7 ]. Hsp90 is essential for the development of tumors and the expression of such an enzyme is significantly correlated with tumor malignancy and a poor prognosis. Hsp90 inhibitors affect more than 30 oncoproteins directly impacting several oncogenic processes [ 8 ]. As a result of Hsp90's prominence, it has become an attractive drug target for treating multiple diseases [ 9 ]. Several studies have discovered that it can be used as a potential strategy for cancer treatment [ 10 ]. According to various interaction sites, Hsp90 inhibitors are generally categorized as N-terminal, C-terminal and middle-domain inhibitors. The main clinical downside of Hsp90 N-terminal domain (NTD) inhibitors is the heat shock response (HSR). This pro-survival mechanism is activated by N-terminal Hsp90 inhibitors leading to increased Hsp70 transcription [ 11 ]. Due to NTD inhibitors' disadvantages, CTD and MD inhibitors were developed. The issues posed by NTD inhibitors motivated the development of C-terminal domain (CTD) and middle domain (MD) inhibitors. The discovery of the Hsp90 CTD ATP-binding site has led to the discovery of specific CTD inhibitors as potent anticancer therapeutic options including Novobiocin, a natural drug that contains coumarin [ 12 , 13 ] and Gambogic acid was also a selective inhibitor that binds to Hsp90 middle domain Fig. 1 . [ 14 ]. Twenty Hsp90 inhibitors have enrolled in clinical studies since this time [ 15 ]. Therefore, the imperative to create new Hsp90 inhibitors is still very important [ 16 ]. Traditional Chinese medicine is a rich natural product library containing several natural medicines with pharmacological effects. Matrine, a primary constituent found in leguminous plants like Sophora flavescens and Sophora alopecuroides, has demonstrated clinical potential. Studies have indicated that the combination of matrine with other anticancer drugs notably enhances patients' immune capacity and treatment efficacy [ 17 ]. Currently, the scientific literature on the synthesis of matrine derivatives is significant, indicating that matrine has various pharmacological properties and clinical potential. Moreover, matrine has several advantages over traditional Chinese medicine injections in terms of quality control as a one-component drug [ 18 ]. Synthesis of anticancer drugs such as matrine-containing coumarins derivatives is very attractive[ 19 ]. Previously, our group focused on matrine derivatives bearing benzo-α-pyrone structures scaffold, a privileged structure with a broad pharmacological activity that exerted potent anticancer activity by inhibiting lung cancer cell proliferation in vitro and in vivo with no apparent side effects [ 20 ]. This study builds upon the findings of our previous investigations, as we continue our research efforts in designing and synthesizing novel matrine-containing coumarin derivatives with a specific focus on targeting Hsp90 inhibitors for the treatment of Lung Carcinoma. The primary objective of our current work is to enhance the antiproliferative efficacy of these compounds against cancer cells. To achieve this, we have implemented strategic structural modifications to the matrine scaffold, aiming to optimize their pharmacological properties and bolster the potency of these Hsp90 inhibitors in combating cancer, as illustrated in Fig. 2 . The importance of coumarin has led to many efforts to develop new synthetic procedures. There is a wide range of applications of coumarins in biology, medicine, and polymer science [ 21 ]. Modern and recent medicine uses several coumarins as anticancer activities[ 22 ]. Organic and medicinal chemists have synthesized bioactive natural products containing coumarin heterocyclic nucleus. A variety of methods can be used to synthesize coumarins including the Perkin reaction [ 23 ], Knoevenagel condensation[ 24 ], Pechmann condensation [ 25 ], Wittig reaction, Baylis Hillman reaction, Claisen rearrangement, Vilsmeier-Haack and Suzuki cross‐coupling reaction[ 26 ]. The medicinal properties of coumarins have been studied by several authors[ 27 ]. In our quest to develop Hsp90 inhibitory autophagy activators with heightened anticancer potential, we undertook a series of modifications on matrine-based coumarin compounds. These alterations encompassed substituting the 12-N atom with various benzoyl halides, acyl halides, or sulfonyl halides, alongside exploring variations in the carboxyl group at the C-11 position. To evaluate the cytotoxicity of the synthesized compounds, we conducted the 3-(4,5-dimethylthiazol-2-diphenyltetrazolium) bromide (MTT) assay using three distinct cancer cell lines: human cervical carcinoma (HeLa), human hepatocellular carcinoma (HepG-2), and human lung adenocarcinoma (A549) cells. Additionally, we carried out structure-activity relationship (SAR) analysis, providing valuable insights into the correlation between compound structure and anticancer activity, as illustrated in Fig. 3 [ 28 ]. To deepen our comprehension of the most effective compound's anticancer mechanism, we explored its potential as an autophagy-inducing agent. We further evaluated the compound's impact on colony formation, cellular migration, and apoptosis, as well as investigated in vivo antitumor efficacy and toxicity. Moreover, docking studies were conducted to assess the design of the target compounds (a, b, and c). Importantly, to the best of our knowledge [ 29 ]. 2. Matrine Coumarin Derivatives Design of an Hsp90(NTD&CTD) Isoform Selective Inhibitor. Employing matrine as the foundational scaffold, our investigation was centered on targeting the N-terminal domain (NTD) and C-terminal domain (CTD) of the Hsp90 protein. Informed by insights derived from the two-dimensional crystal structures of the Hsp90 protein NTD (PDB code: 3T0Z) and CTD (PDB code: 2CG9), recognized for their upregulated expression in tumor cells, a specific array of matrine inhibitors was meticulously crafted to interact with both the NTD and CTD domains. This deliberate strategy resulted in the development of a group of matrine compounds intricately designed to engage with and modulate both the NTD and CTD regions of the Hsp90 protein. 3. Results and discussion 3.1. Chemistry As a part of this investigation, we synthesized different series of new compounds containing other various moieties in one structure. To obtain primary groups important for Hsp90 inhibitory activity target compounds, we first designed three matrine-based coumarin derivatives as shown in Fig. 2 . The matrinic acid derivatives were efficiently synthesized according to the protocol outlined in Scheme 1 . The detailed chemical structures of compounds are organized in Tables 1 , 2 , and 3 . The experimental section describes a novel target compound synthesis using commercially available matrine as the initial material. The primary synthetic pathway involved the hydrolysis of matrine 1 with aqueous KOH to obtain intermediate 2 , which was then subjected to esterification through the addition of SOCl 2 and an alcohol reaction, resulting in intermediate 3 . Subsequent substitution reactions with acyl, sulfonyl, or benzoyl halides and anhydrous potassium carbonate yielded intermediate 4 , further hydrolyzed to produce intermediate 5 with a 90% yield [ 30 ]. Reduction reactions were instrumental in transforming functional groups, with selective reduction of the carbonyl group yielding allyl alcohol compounds 6 , 9 , and 10 using reducing LiAlH 4 agent. The drop-wise addition of LiAlH 4 to the esters significantly enhanced product yield, as validated by literature reports [ 31 ]. Additionally, matrinic acid 2 was processed to yield protected 12 N-Fmoc Matrinic acid 7 and 12 N-Boc Matrinic acid 8 with a higher yield of 85% [ 32 ] as depicted in Scheme 1 . In another synthesis pathway, 3-aminocoumarins 14 were synthesized through Dakin's procedure from N-acetylglycine 11 and various salicylaldehyde derivatives 12. Hydrolysis of 3-acetamidocoumarins 13 was conducted using H 2 SO 4 , with 70% H 2 SO 4 exclusively yielding 3-aminocoumarin within 15 minutes [ 33 , 34 ]. Coumarin-3-carboxylic acids 16 were efficiently synthesized through Knoevenagel condensation between salicylaldehyde derivatives 12 and Meldrum's acid 15 utilizing sodium azide or potassium carbonate as catalysts [ 35 , 36 ] as depicted in Scheme 2 . The synthesis of target compounds ( a ) involved the reaction of matrinic acid ( 5, 7, 8 ) with 3-aminocoumarins 14 in the presence of EDC, HOBt, and NMM, resulting in targets ( 1a-21a ) [ 37 , 38 ]. Furthermore, DCC esterification emerged as a pivotal methodology in the esterification reactions of Coumarin-3-carboxylic acids 16 and matrine alcohols ( 6, 9, 10) . The addition of a small quantity of 4-methylaminopyridine (DMAP) as a catalyst in the reaction system accelerated the reaction, enhancing the conversion rate and yielding compound ( 1b-8b ) effectively [ 30 ]. Additionally, the synthesis of compound ( c ) from matrinic acid ( 5, 7, 8 ) and 2-hydroxybenzaldehyde derivatives using anhydrous potassium acetate and acetic anhydride yielded target compounds ( 1c-16c ) with high yields of 60–70% as depicted in Scheme 3 [ 39 ]. 3.2. Biological evaluation and SAR study 3.2.1. In vitro biological activity In the context of in vitro cell growth activities, all synthesized matrine-based coumarin derivatives ( a, b , and c ) were evaluated against three human cancer cell lines, including HeLa (human cervical cancer cell line), A549 (Human lung cancer cells) and HepG-2 (Human Hepatoma Cells) serving as subjects for the MTT assay, with matrine employed as the positive control. The IC 50 values of target matrine derivatives ( a , b and c ) were illustrated in Tables 1 , 2 and 3 . According to Table 1 , the parent matrine displayed unsatisfactory growth activities across these three cancer cell lines, all target compounds exhibited moderate to potent growth activities against the tested cell lines. In an effort to identify more potent inhibitors of Hsp90 a series of matrine-based coumarin derivatives were synthesized and initial structure-activity relationships (SARs) were established. The SARs observed between matrine-based coumarin compounds and their Hsp90 inhibitory activity is summarized in Fig. 3 . The SAR investigation involved modifications inspired by previous studies [ 30 , 40 ] particularly the opening of the D-ring from matrine. Chemical modifications of the nitrogen atoms at the 12-N position were substituted with benzoyl halides, acyl halides, or sulfonyl halides and the focus shifted to variations of the carboxyl group at the C-11 attachment aiming to enhance the activity of matrine [ 41 , 42 ]. Significantly, among these derivatives, compounds 5a and 4c , modified by 4-tert-butylbenzenesulfonyl chloride at the 12-N atom and carboxyl group by 3-aminocoumarins for compound 5a , and 2-hydroxy-4-methylbenzaldehyde for compound 4c , exhibited the most potent cytotoxicity with IC 50 values of A549, HepG-2, and HeLa (7.35 ± 0.097, 7.727 ± 0.10, 8.02 ± 0.065 µM) and (10.75 ± 0.88, 12.66 ± 1.36, 14.62 ± 2.3), respectively. Additionally, compound 1b , modified by 4-bromobenzoyl chloride at the 12-N atom and carboxyl group by coumarin-3-carboxylic acids, also demonstrated significant cytotoxicity with IC 50 values of A549, HepG-2, and HeLa (8.94 ± 0.58, 10.65 ± 0.07, 11.22 ± 1.25) respectively. Among the compounds tested derivative 5a exhibited the highest potency. Considering these observations, the antiproliferative efficacy of the three newly designed matrine-based coumarin derivatives on the aforementioned cell lines prompted further investigation of their cytotoxicity profile against additional cancer types. Furthermore, the most effective compound, 5a was evaluated for its cytotoxic effect on normal lung epithelial cells (BEAS-2B). As presented in Table 5S, 5a exhibited a substantially higher IC 50 value against normal lung epithelial cells (BEAS-2B) indicating its exceptional selectivity between A549 (human lung cancer cells) and normal lung epithelial cells (BEAS-2B). Given its potent efficacy and increased selectivity for cancer cells compound 5a was selected for further investigation of its mechanism of growth inhibition in cancer cells. 3.2.1.1. Cell viability of compound 5a against A549 cell lines In the MTT assay, the A549 cells were studied to determine the potential anticancer mechanism of compound 5a . To evaluate its effectiveness, different concentrations of drug 5a (0, 1, 2, 4, 8, 16, and 32 µM) were administered to the A549 cells for durations of 24, 48, and 72 h. The results are depicted in Fig. 4 and Table 4S. demonstrate that the proliferation of human lung cancer cells A549 was considerably diminished in a dose and time-dependent manner by 5a . 3.2.1.2. Antiproliferative activity of compound 5a on the normal cell line BEAS-2B and A549 The objective of using drugs to cure malignant tumors is to destroy the cancer cells without causing harm or toxicity to the healthy cells or overall health. Consequently, the effect of 5a on the survival of normal lung epithelial cells (BEAS-2B) and lung cancer cells (A549) was assessed by the MTT assay at various concentrations (0, 1, 2, 4, 8, 16 and 32 µM) after a 48 h treatment. As illustrated in Fig. 5 and Table 5S. the findings establish that 5a is comparatively less harmful to normal cells in contrast to lung cancer cells. 3.2.1.3. Effect of compound 5a on cancer cell colony formation of A549 cell lines Cancer cell colony formation was widely used to estimate neoplastic transformation. After being exposed to 5a at various concentrations for 48 h human lung cancer cell A549 exhibited evidence of cell cloning. According to Fig. 6 , the number of cell clones in A549 cells was significantly reduced after treatment with 5a at various concentrations (0, 2, 4, 8, 16, and 32 µM) compared to the negative control and the inhibitory effect grew more assertive with the increase in concentration. 3.2.1.4. Antimigration activity of compound 5a on A549 cell lines Several matrine inhibitors demonstrated anti-angiogenesis activities against tumor endothelium and angiogenesis is crucial for the progression development and metastasis of human cancer. Therefore, utilizing human lung cancer A549 cells as a model, we evaluated 5a at various concentrations (0, 2, 4 and 8 µM) for 24 and 48 h to see how it affected endothelial cells. We also observed cell migration and calculated the scratch area which is a standard method to confirm anti-angiogenesis effects. We first evaluated 5a ability to inhibit A549 migration which is a crucial stage in the development of new blood vessels as shown in Fig. 7 and Table 6S. The study's findings illustrated that within 48 hours, untreated cells repopulated the initially scraped area. Conversely, compound 5a distinctly and dose-dependently inhibited the migration of A549 cells, as indicated by the results [ 3 ]. 3.2.1.5. Cell apoptosis assay of 5a on A549. We first evaluated its anticancer activities to determine whether derivative 5a inhibitory effects on the A549 cell lines resulted from apoptosis by DAPI staining and apoptosis morphological analysis. Derivative 5a caused apoptosis after 48 h of incubation with it at various concentrations (0, 1, 2, 4, 6 and 8 µM) in DAPI staining, as demonstrated by the micronuclei of proliferating A549 cells. 5a caused a significant amount of dense blue staining and the development of apoptotic bodies as the nuclear membrane was ruptured, indicating that 5a had an anticancer activity on A549 cells as illustrated in Fig. 8 A. The specific cytotoxic effects of A549 cells were also evaluated utilizing flow cytometry for the apoptotic analysis method and double stained with annexin V-FITC/PI. As illustrated in Fig. 8 B and Table 7S. we evaluated the apoptosis induced by 5a in A549 and compared it with the standard. As a result, different concentrations of 5a (0, 2, 4 and 8 µM) were used to analyze human lung cancer A549 cells for apoptosis using Annexin V-FITC. The fluorescence of Annexin V labeled with FITC can be detected by flow cytometry when it binds to necrotic cells; PI (propidium iodide) is a nucleic acid dye and normal cells and early apoptosis can be detected. PI cannot penetrate the cell membrane in normal cells and early apoptosis. However, it can penetrate the membrane of necrotic and apoptotic cells to form a red color. Annexin V-FITC and PI can be used to detect early apoptosis, late apoptosis, and necrosis. As a result, flow cytometry can be used to quantitatively determine the apoptosis rate of A549 cells. Based on the flow cytometry results shown in Fig. 8 B, 5 a can induce early and late apoptosis in A549 cells. With the increase of dose, the proportion of A549 cell apoptosis induced by 5a low medium and high dose groups was 6.89% ±1.12%, 8.645 ± 1.12%, and 58.79 ± 7.35%, which were significantly different from the control group 4.244 ± 0.80% (P < 0.001) (* p < 0.05, ** p < 0.01, *** p < 0.001). 3.2.1.6. The effect of compound 5a on the apoptosis-related client proteins Hsp90, Bcl-2 and Bax. Human lung cancer cells A549 were treated with 5a (0, 2, 4 and 8 µM) for 48 h and apoptosis-related proteins were determined to understand the effects of various concentrations of drugs on related proteins. The client protein Hsp90, Bcl-2 and Bax family members play a crucial role in the process of cell apoptosis, which can be divided into two main categories. The changes in the expression levels of Hsp90, Bcl-2 and Bax in the A549 cells were detected by Western blot. As shown in Fig. 9 and Table 8S. the experiment confirmed significant changes in the expression levels. The expression of Bcl-2 and Hsp90 proteins that inhibit apoptosis decreased and the presentation of Bax a protein that promotes apoptosis increased. 3.2.2. In Vivo antiproliferative activity 3.2.2.1. Acute Toxicity Study of Compound 5a According to the Sub-Acute toxicity assay results, compound 5a did not cause death in mice and the body weight of the treatment group gradually increased while remaining identical to that of the vehicle, Matrine and 5-Fluorouracil groups as positive control Fig. 10 A and Table 9S. In addition, HE staining revealed that mice receiving 40 mg/kg of the drug did not exhibit overt toxicity in their hearts, spleens, kidneys, lungs, livers and tumors Fig. 10 B. Overall, molecule 5a had reasonable pharmacokinetic parameters and was safe for oral administration in vivo. 3.2.2.2. Compound 5a inhibited A549 tumor growth in vivo To evaluate compound 5a antitumor activity in vivo we established the A549 cell inoculated xenograft mouse model in which female Balb/c nude mice were divided into four groups: vehicle, 5-Fluorouracil, Matrine and compound 5a (40 mg/kg intraperitoneal QD). In each group, mice were given an oral treatment after three days. Following 28 days of continuous treatment as shown in Fig. 11 A, B compound 5a exhibited more potent antitumor activity than 5-Fluorouracil and Matrine as the dosage increased its antitumor activity improved. The tumor growth inhibition (TGI) value of 5a , Matrine and 5-Fluorouracil was 72.4%, 64.3% and 46.8% respectively Fig. 11 C. In contrast the body weight of the 5a -treated group did not differ from that of the vehicle, 5-Fluorouracil and Matrine groups further demonstrating the good safety profile of compound 5a in vivo. The relationship curves of tumor volumes at various times of treatments over 28 days after administration as shown in Fig. 11 D. 4. Molecular Docking In order to comprehensively assess the potency of Matrine Contains Coumarins derivatives and to provide guidance for further Structure-Activity Relationship (SAR) studies, [ 29 ] conducted molecular docking of the target compounds, elucidating their action mechanisms on the N-terminal domain (NTD) and C-terminal domain (CTD) action targets of the Hsp90 protein.[ 29 ]. Molecular docking of various designed compounds with corresponding action targets is established to establish the basis of various inhibitor molecules [ 40 , 41 ]. Demonstrates compound docking into the Hsp90 ATP-binding site of NTD (PDB ID: 3T0Z) the poses of compounds 5a into the active site of protein were generated based on the scores. As seen in Fig. 12 A; B the resulting docking model with minimum relative binding energy is -10.5 kcal mol − 1 indicates that compound 5a has interaction with NTD action targets of Hsp90. The interaction between compound 5a and protein was formed through hydrogen bonds from GLY137; GLY135; ASN51 and LYS112 in protein with the carbonyl group of the amide bond in compound 5a as well as CH-π interaction and also the interaction between compound 5a and protein was formed through N-H donor bond from GLY135. The binding model of compound 5a into the Hsp90 ATP-binding site of NTD revealed several molecular interactions thought to be responsible for the observed affinity: pi–anion interaction between the benzene ring of coumarin and ASP 54 and other interactions, including Alkyl MET 98; LEU 107; ALA 55 and Van der Waals. As seen in Fig. 12 C; D compound 5a docking into the Hsp90 ATP-binding active sites of CTD (PDB ID: 2CG9) formed interactions with the key amino acid residues, the resulting docking model with minimum relative binding energy is -10.2 kcal mol − 1 indicates that compound 5a has interaction with CTD action targets of Hsp90. The interaction between compound 5a and protein was formed through hydrogen bonds from LEU454; GLU453; ASN446 and SER419 in protein with carbonyl group of the amide bond in compound 5a as well as CH-π interaction, and interaction also was formed through N-H donor bond from GLU415. The binding model of compound 5a into Hsp90 ATP-binding active sites revealed several molecular interactions thought to be responsible for the observed affinity: including carbon-hydrogen bond -OCH 3 from ASP503 and Van der Waals. This result suggests that 4-tert-Butylbenzenesulfonyl introduced to the nitrogen atom at the 12-position and 3-amino-5-methoxy-coumarin through the carboxyl group might be beneficial for anticancer activity. 5. Conclusion In summary, Matrine Contains Coumarins derivatives were designed, synthesized and evaluated for their cytotoxic activity against three cancer cell lines (A549, HepG-2 and HeLa cells) as Hsp90 inhibitors for the treatment of lung carcinoma. SAR studies indicated that compound 5a with 4-tert-butylbenzenesulfonyl chloride at the 12-N atom and carboxyl group by 3-amino-5-methyl-2H-chromen-2-one, had the best anti-cancer activity. Based on the bioactivity results derivative 5a was the most potent compound exhibiting excellent antiproliferative activity against A549, HepG-2 and Hela cells with IC 50 values of 7.35 ± 0.097, 7.727 ± 0.10 and 8.02 ± 0.065 µM respectively. Additionally, derivative 5a demonstrated the most potent activity against A549 cells and was non-toxic to normal lung epithelial cells BEAS-2B. Further mechanism investigations confirmed that 5a inhibited A549 cell proliferation, and suppressed colony formation and migration. In vivo, experiments showed that 5a possessed different mechanisms against A549 cancer cells and exhibited more potent antitumor activity than Matrine and 5-Fluorouracil with a tumor growth inhibition (TGI) value of 72.4% compared to 64.3% and 46.8% respectively. Based on the in vitro and in vivo experiments, compound 5a showed promise as an antitumor agent and further studies including pharmacokinetic profiles and prophase tests are encouraged. Molecular docking analysis revealed that the interactions between 5a functionalities and residues were crucial for binding compounds to proteins. Enhancing these interactions could positively impact the inhibitory activity of the Hsp90 ATP-binding at both CTD and NTD sites. This work provides useful information for further structural modifications of these compounds and the synthesis of new, potent antitumor agents. 6. Experimental section 6.1. General Chemistry Reagents and solvents were purchased from commercial sources and used without further purification. Reactions were monitored by thin-layer chromatography (TLC) using precoated silica gel plates (silica gel GF/UV 254), and spots were visualized under UV light (254 nm). The products were purified by flash column chromatography equipped with commercial silica gel (300–400 mesh). All the compounds were characterized with 1 H-NMR, 13 C-NMR and MS. 1 H and 13 C nuclear magnetic resonance (NMR) data were recorded on Bruker Avance 500 spectrometers operating at 1 H and 13 C frequencies of 500 and 126 MHz respectively, in the indicated solvents (DMSO-d6 or CDCl 3 , TMS as internal standard). Chemical shifts (δ) are in ppm relative to the residual solvent signal (DMSO-d6 with 2.48 and 39.52 ppm and CDCl 3 with 7.26 and 77.16 ppm for 1 H and 13 C, respectively). As an internal standard, chemical shifts were given in ppm (d) relative to SiMe4. Coupling constants (J) were in hertz (Hz) and signals were designated as follows: s, singlet; d, doublet; t, triplet; m, multiple; br, broad singlet, etc. Mass spectra were obtained from a ThermoFisher LCQ Fleet (ESI). Melting points were determined in open capillary tubes on the X-4 melting point apparatus without correction. Synthesis of intermediate 1–14 is shown in the supplementary information file. 6.1.1. Synthesis of compound (1a-20a) Matrinic acid derivatives (5,7,8) (2.10 mmol), HOBt (2.10 mmol) and EDCI (2.10 mmol) were dissolved in DCM (10 mL). After 10 min of stirring NMM (3.15 mmol) and 3-aminocoumarins 12 (3.15 mmol) were added and stirred for 12 h. and monitored the reaction by TLC. Their action mixture was poured into water and extracted with EtOAc. The organic layer was washed with saturated bicarbonate solution followed by brine solution. It was dried under anhydrous Na 2 SO 4 and the solvent was evaporated. The gained residue was purified by flash column chromatography on silica gel with CH 2 Cl 2 /CH 3 OH as the eluent to afford the title compounds 1a-21a [ 44 ]. tert-butyl1-(4-((6-chloro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (1a) Yield 64%; Mp 185–188°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.29 (s, 1H, NH), 8.02 (d, J = 1.4 Hz, 1H, CH), 7.99 (s, 1H, ArH), 7.39 (dd, J = 7.5, 1.4 Hz,1H, ArH), 7.30 (d, J = 7.5 Hz, 1H, ArH), 4.29 (dd, J = 12.4, 7.7 Hz, 1H, CH), 3.21 (dd, J = 12.4, 7.7 Hz, 1H), 2.80 (dt, J = 10.8, 4.0 Hz, 1H), 2.51 (dt, J = 12.3, 5.5 Hz, 2H), 2.41 (dt, J = 12.4, 5.4 Hz, 2H), 2.30–2.23 (m, 1H), 2.19–2.09 (m, 1H), 2.04 (dd, J = 5.0 Hz, 3H), 1.92–1.84 (m, 2H), 1.83–1.78 (m, 1H), 1.62 (td, J = 7.8, 4.1 Hz, 3H), 1.57–1.49 (m, 2H), 1.47–1.43 (m, 2H), 1.42 (s, 9H, C-(CH 3 ) 3 ), 1.40–1.28 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 157.65, 155.50, 149.07, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 81.20, 67.56, 54.76, 53.21, 46.17, 40.85, 37.70, 36.88, 30.89, 28.31, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 29 H 38 ClN 3 O 5 , [M + H] + m/z: 543.2500, found 544.2949. Anal: C, 64.02; H, 7.04; Cl, 6.52; N, 7.72; O, 14.70. 4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(6-nitro-2-oxo-2H-chromen-3-yl)butanamide (2a) Yield 60%; Light yellow oil. 1 H NMR (500 MHz, CDCl 3 ) δ 9.33 (s, 1H, NH), 8.58 (d, J = 2.9 Hz,1H, ArH), 8.21 (dd, J = 14.9, 3.0 Hz, 1H, ArH), 8.05 (s, 1H, ArH), 7.71–7.50 (m, 5H, Ar-5H), 3.47 (dd, J = 24.7, 13.7 Hz, 1H), 2.91–2.81 (m, 2H), 2.51 (dt, J = 24.8, 10.9 Hz, 2H), 2.41 (dt, J = 24.8, 10.8 Hz, 2H), 2.04 (td, J = 16.2, 1.8 Hz, 2H), 1.96–1.82 (m, 2H), 1.82–1.69 (m, 3H),1.63 (t, J = 21.8 Hz, 1H), 1.57–1.49 (m, 2H), 1.50–1.43 (m, 4H), 1.44–1.35 (m, 2H), 1.32 (s, 9H, C-(CH 3 ) 3 ), 1.32–1.26 (m, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 155.50, 154.56, 154.40, 144.74, 133.92, 128.79, 128.32, 126.05, 125.20, 124.51, 121.90, 117.69, 113.24, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 34 H 42 N 4 O 7 S, [M + H] + m/z: 650.2774, found 650.2900. Anal: C, 62.75; H, 6.51; N, 8.61; O, 17.21; S, 4.93. tert-butyl1-(4-((6-nitro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (3a) Yield 49%; brown oil. 1 H NMR (500 MHz, CDCl 3 ) δ 9.27 (s, 1H, NH), 8.59 (d, J = 2.9 Hz, 1H, ArH), 8.24 (dd, J = 15.0, 2.9 Hz, 1H, ArH), 8.04 (s, 1H, CH), 7.62 (d, J = 15.0 Hz, 1H, ArH), 4.37 (dd, J = 24.8, 14.7 Hz,1H, CH), 3.04 (dd, J = 24.8, 14.6 Hz, 1H, CH), 2.51 (dt, J = 24.8, 10.9 Hz, 3H), 2.41 (dt, J = 24.8, 10.8 Hz, 2H), 2.07 (dd, J = 15.6, 2.7 Hz, 3H), 1.95 (dt, J = 16.2, 5.8 Hz, 1H), 1.86–1.76 (m, 2H), 1.77–1.68 (m, 2H), 1.67–156 (m, 3H), 1.55–1.49 (m, 2H), 1.48–1.43 (m, 2H), 1.42 (s, 9H, C-(CH 3 ) 3 ), 1.39–1.27 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 157.65, 155.50, 154.56, 144.74, 126.05, 125.20, 124.51, 121.90, 117.69, 113.24, 81.20, 67.56, 54.76, 53.21, 46.17, 40.85, 37.70, 36.88, 30.89, 28.31, 26.28, 23.44, 23.25, 22.61 (s). HRMS (ESI): Calcd. C 29 H 38 N 4 O 7 , [M + H] + m/z: 554.2740, found 555.2471. Anal: C, 62.80; H, 6.91; N, 10.10; O, 20.19. 4-(2-cinnamoyldecahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(5-methoxy-2-oxo-2H-chromen-3-yl)butanamide (4a) Yield 57%; Mp 189–192°C. 1 H NMR (500 MHz, DMSO) δ 9.22 (s, 1H, NH), 7.97 (s, 1H, CH), 7.92 (dt, J = 4.7, 2.1 Hz, 1H, ArH), 7.90 (dd, J = 3.1, 1.5 Hz, 1H, ArH), 7.59 (t, J = 13.7 Hz, 2H), 7.50 (ddd, J = 17.6, 6.3, 3.2 Hz, 1H), 7.40 (s, 1H, ArH), 7.36 (dd, J = 20.0 Hz, 1H, ArH), 7.32 (s, 1H, CH), 7.23 (dd, J = 13.6, 4.4 Hz, 1H, ArH), 7.03 (d, J = 30.2 Hz, 1H, CH), 3.83 (s, 3H, Ar-OCH 3 ), 3.61 (dd, J = 24.8, 14.6 Hz, 1H), 3.05 (dd, J = 24.8, 14.5 Hz, 1H), 2.63 (dt, J = 22.2, 8.2 Hz, 1H), 2.57–2.48 (m, 2H), 2.47–2.37 (m, 2H), 2.34–2.22 (m, 1H), 2.21–2.11 (m, 1H), 2.10–2.06 (m, 1H), 2.04 (dd, J = 16.3, 12.4 Hz, 1H), 1.90–1.71 (m, 3H), 1.66–1.55 (m, 3H), 1.55–1.47 (m, 2H), 1.46–1.40 (m, 2H), 1.38–1.25 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 171.53, 163.97, 155.06, 145.89, 139.85, 138.48, 136.15, 128.70 (d, J = 16.4 Hz), 128.05, 124.68, 124.14, 123.52, 119.86, 119.20, 115.18, 114.05, 67.56, 58.77, 56.83, 53.21, 46.87, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 34 H 39 N 3 O 5 , [M + H] + m/z: 569.2890, found 569.2653. Anal: C, 71.68; H, 6.90; N, 7.38; O, 14.04. 4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(5-methoxy-2-oxo-2H-chromen-3-yl)butanamide (5a )Yield 59%; Mp 195–196°C. 1H NMR (500 MHz, CDCl 3 ) δ 9.21 (s, 1H, NH), 7.93 (s, 1H, CH), 7.65–7.58 (m, 4H, ArH), 7.42–7.31 (m, 2H, ArH), 7.04 (dd, J = 15.0 Hz, 1H, Ar-1H), 3.83 (s, 3H, Ar-OCH 3 ), 3.82 (dd, J = 24.9, 13.7 Hz, 1H, CH), 3.37 (dd, J = 24.8, 13.7 Hz, 1H), 3.17 (dt, J = 22.0, 8.0 Hz, 1H), 2.51 (dt, J = 23.2, 12.4 Hz, 2H), 2.41 (dt, J = 14.0, 10.8 Hz, 2H), 2.05 (dd, J = 15.7, 5.4 Hz, 2H),1.85–1.70 (m, 4H), 1.60 (d, J = 21.8 Hz, 1H), 1.54–1.48 (m, 3H), 1.47–1.42 (m, 3H), 1.41–1.35 (m, 2H), 1.33 (s, 9H), 1.28 (dd, J = 11.7, 4.1 Hz, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 161.08, 155.50, 154.40, 150.91, 133.92, 128.79, 128.32, 127.48, 124.19, 116.58, 114.16, 111.15, 102.14, 67.56, 59.57, 56.08, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 35 H 45 N 3 O 6 S, [M + H] + m/z: 635.3029, found 635.2773. Anal: C, 66.12; H, 7.13; N, 6.61; O, 15.10; S, 5.04. 4-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide (6a) Yield 55%; Mp 175–178°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.25 (s, 1H, NH), 8.12 (dd, J = 7.5, 5.2 Hz, 2H, Ar-2H), 7.95 (s, 1H, CH), 7.68 (dd, J = 7.5, 1.4 Hz,1H, ArH), 7.55 (td, J = 7.5, 1.5 Hz, 1H, ArH), 7.44 (dd, J = 8.6, 6.9 Hz, 2H, Ar-2H), 7.29 (dd, J = 7.0, 5.6 Hz, 2H, Ar-2H), 3.36 (dd, J = 12.4, 7.3 Hz, 1H), 2.99 (dd, J = 12.4, 7.3 Hz, 1H), 2.68 (q, J = 11.0, 4.1 Hz, 1H), 2.51 (dt, J = 12.3, 5.4 Hz, 2H), 2.41 (dt, J = 12.4, 5.4 Hz, 2H), 2.34–2.23 (m, 1H), 2.21–2.10 (m, 1H), 2.04 (dd, J = 5.0 Hz, 3H), 1.87–1.80 (m, 2H), 1.80–1.72 (m, 1H), 1.72–1.60 (m, 3H), 1.57–1.49 (m, 2H), 1.48–1.38 (m, 2H), 1.38–1.27 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 173.57, 153.33, 149.07, 133.61, 131.63, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 115.98, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 31 H 34 FN 3 O 4 , [M + H] + m/z: 531.2533, found 532.2344. Analysis: C, 70.04; H, 6.45; F, 3.57; N, 7.90; O, 12.04. 4-(decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide (7a) Yield 50%; Mp 190–192°C. 1 H NMR (500 MHz, DMSO) δ 9.25 (s, 1H, NH), 7.96 (s, 1H, CH), 7.68 (dd, J = 14.3, 2.8 Hz, 1H, ArH), 7.61–7.50 (m, 1H),1H, ArH), 7.38–7.30 (m, 2H, Ar-2H), 3.01 (dd, J = 24.8, 14.9 Hz, 1H), 2.51 (dt, J = 12.3, 5.5 Hz, 2H), 2.46 (d, J = 9.3 Hz, 1H), 2.41 (dt, J = 12.4, 5.4 Hz, 2H), 2.17–2.07 (m, 2H), 2.04 (dd, J = 6.0, 4.6 Hz, 2H), 1.86–1.66 (m, 5H), 1.61–1.48 (m, 2H), 1.51–1.45 (m, 4H), 1.45–1.29 (m, 4H). 13 C NMR (126 MHz, DMSO) δ 171.53, 155.50, 149.07, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 68.36, 58.08, 53.21, 47.89, 41.90, 37.70, 36.11, 32.24, 28.36, 26.36, 23.44, 23.25, 21.56. HRMS (ESI): Calcd. C 24 H 31 N 3 O 3 , [M + H] + m/z: 409.2365, found 409.7183. Anal: C, 70.39; H, 7.63; N, 10.26; O, 11.72. 4-(2-(4-cyanobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide (8a) Yield 49%; Light yellow oil. 1 H NMR (500 MHz, DMSO) δ 9.25 (s, 1H, NH), 8.19 (dd, J = 15.4, 2.9 Hz, 2H, Ar-2H), 8.09 (dd, J = 15.4, 2.9 Hz, 2H, Ar-2H), 7.95 (s, 1H, Ar-CH), 7.68 (dd, J = 14.5, 2.7 Hz, 1H, Ar-H), 7.55 (dd, J = 14.6, 12.2 Hz, 1H, Ar-H), 7.36 (dt, J = 14.0, 3.1 Hz, 2H, Ar-H), 3.34 (dd, J = 24.8, 14.4 Hz, 1H), 3.00 (dd, J = 24.7, 14.3 Hz, 1H), 2.69 (dt, J = 22.0, 8.1 Hz, 1H), 2.51 (dt, J = 24.8, 10.8 Hz, 2H), 2.41 (dt, J = 24.8, 10.8 Hz, 2H), 2.34–2.14 (m, 2H), 2.10–1.97, 1.88–1.72 (m, 3H), 1.72–1.57 (m, 3H), 1.59–1.44 (m, 3H), 1.44–1.22 (m, 3H). 13 C NMR (126 MHz, DMSO) δ 171.53, 170.91, 155.50, 149.07, 139.16, 132.92, 129.44, 128.88, 125.69, 125.46, 124.19, 123.65, 118.94, 117.56, 114.76, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61 HRMS (ESI): Calcd. C 32 H 34 N 4 O 4 , [M + H] + m/z: 538.2580, found 538.6747. Anal: C, 71.35; H, 6.36; N, 10.40; O, 11.88. 4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(6-methyl-2-oxo-2H-chromen-3-yl)butanamide (9a) Yield 56%; Mp 190–193°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.28 (s, 1H, NH), 7.96 (s, 1H, CH), 7.66–7.54 (m, 4H, Ar-4H), 7.52 (d, J = 2.9 Hz, 1H, Ar-H), 7.28 (d, J = 14.9 Hz, 1H, Ar-H), 7.19 (dd, J = 14.9, 2.8 Hz, 1H, Ar-H), 3.31 (dd, J = 24.9, 13.7 Hz, 1H, CH), 2.91 (dd, J = 24.8, 13.7 Hz, 1H), 2.60–2.51 (m, 1H), 2.51–2.41 (m, 2H), 2.41 (s, 3H, Ar-CH 3 ), 2.38 (dd, J = 17.7, 7.0 Hz, 1H), 2.19–2.13 (m, 3H), 2.04 (t, J = 10.6 Hz, 3H), 1.87–1.78 (m, 1H), 1.75–1.66 (m, 3H), 1.63– 158 (m, 3H), 1.57–1.47 (m, 2H), 1.47–1.41 (m, 1H), 1.40–1.35 (m, 1H), 1.32 (s, 9H, C-(CH 3 ) 3 ), 1.29 (dd, J = 13.1, 8.9 Hz, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 155.50, 154.40, 148.30, 134.77, 133.92, 128.79, 128.32, 127.63, 125.71, 124.51, 121.57, 117.43, 113.24, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61, 21.23. HRMS (ESI): Calcd. C 35 H 45 N 3 O 5 S, [M + H] + m/z: 619.3080, found 620.3302. Anal: C, 67.82; H, 7.32; N, 6.78; O, 12.91; S, 5.17. N-(6-chloro-2-oxo-2H-chromen-3-yl)-4-(2-(3-chlorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butanamide (10a) Yield 47%; Mp 188–190°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.25 (s, 1H, NH), 8.02 (d, J = 2.9 Hz, 1H, ArH), 8.01 (s, 1H, CH), 7.87 (t, J = 2.9 Hz,1H, ArH), 7.85 (dt, J = 14.7, 3.2 Hz, 1H, ArH), 7.69 (dt, J = 15.0, 3.1 Hz, 1H, ArH), 7.58 (t, J = 14.8 Hz, 1H, ArH), 7.37 (dd, J = 15.0, 2.9 Hz, 1H, ArH), 7.30 (d, J = 14.9 Hz, 1H, ArH), 3.42 (dd, J = 24.8, 14.4 Hz, 1H, CH), 3.10–2.86 (m, 2H), 2.51 (dt, J = 24.7, 10.8 Hz, 2H), 2.41 (dt, J = 24.8, 10.6 Hz, 2H), 2.24–2.10 (m, 1H), 2.04 (q, J = 12.9 Hz, 3H), 2.00–1.85 (m, 1H), 1.76 (t, J = 7.1 Hz, 3H), 1.65 (ddd, J = 8.3 Hz, 3H), 1.62–1.54 (m, 2H), 1.45–1.39 (m, 1H), 1.37–1.26 (m, 3H). 13 C NMR (126 MHz, CDCl3) δ 174.53, 169.06, 155.50, 148.29, 137.27, 134.68, 132.01, 130.13, 129.96, 129.61, 126.77, 126.37, 124.51, 123.35, 122.38, 118.80, 113.24, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 31 H 33 Cl 2 N 3 O 4 , [M + H] + m/z: 581.1848, found 581.2474. Anal: C, 67.94; H, 6.25; Cl, 6.47; N, 7.67; O, 11.68. 4-(2-(3-chlorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide (11a) Yield 45%; Mp 178–180°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.25 (s, 1H, NH), 7.95 (s, 1H, CH), 7.86 (dt, J = 12.7, 3.0 Hz, 1H, ArH), 7.82 (t, J = 3.2 Hz, 1H, ArH), 7.70 (dd, J = 7.2, 3.1 Hz, 1H, ArH), 7.67 (dd, J = 6.6, 3.4 Hz, 1H, ArH), 7.59 (d, J = 14.8 Hz, 1H, ArH), 7.54 (dd, J = 14.3, 2.9 Hz, 1H, ArH), 7.34 (ddd, J = 18.0, 8.2, 2.0 Hz, 2H, Ar-2H), 3.38 (dd, J = 24.8, 14.4 Hz, 1H, CH), 2.98 (dd, J = 24.8, 14.3 Hz, 1H), 2.68 (dt, J = 22.0, 8.2 Hz, 1H), 2.51 (dt, J = 24.8, 10.9 Hz, 2H), 2.41 (dt, J = 24.8, 10.7 Hz, 2H), 2.35–2.12 (m, 2H), 2.11–1.98 (m, 3H), 1.88–1.73 (m, 3H), 1.71–1.61 (m, 3H), 1.60–1.53 (m, 2H), 1.52–1.41 (m, 2H), 1.42–1.27 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 169.06, 155.50, 149.07, 137.27, 134.68, 130.13, 129.96 (s), 129.61, 128.88, 126.37, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 31 H 34 ClN 3 O 4 , [M + H] + m/z: 547.2238, found 548.2740. Anal: C, 67.94; H, 6.25; Cl, 6.47; N, 7.67; O, 11.68. 4-(2-(3-bromobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide (12a) Yield 39%; Mp 187–188°C. 1 H NMR (500 MHz, DMSO) δ 9.25 (s, 1H, NH), 8.14 (t, J = 3.0 Hz, 1H, ArH), 7.95 (s, 1H, CH), 7.89 (dt, J = 14.8, 2.9 Hz, 1H, ArH), 7.79 (dt, J = 14.8, 3.0 Hz, 1H, ArH), 7.68 (dd, J = 14.3, 2.8 Hz, 1H, ArH), 7.57 (dd, J = 13.4, 2.4 Hz, 1H, ArH), 7.50 (t, J = 14.9 Hz, 1H, ArH) 7.37–7.30 (m, 2H, Ar-2H), 3.35 (dd, J = 24.7, 14.5 Hz, 1H, CH), 2.96 (dd, J = 24.8, 14.4 Hz, 1H), 2.66 (dt, J = 22.0, 8.1 Hz, 1H), 2.51 (dt, J = 24.8, 10.9 Hz, 2H), 2.41 (dt, J = 24.8, 10.7 Hz, 2H), 2.28–2.11 (m, 2H), 2.08 (dd, J = 7.1, 2.7 Hz, 1H), 2.03 (dd, J = 16.1, 12.2 Hz, 2H), 1.88–1.73 (m, 3H), 1.71–1.61 (m, 3H), 1.60–1.53 (m, 2H), 1.52–1.41 (m, 2H), 1.42–1.27 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 171.53, 169.06, 155.50, 149.07, 136.35, 133.80, 131.78, 130.77, 128.88, 126.18, 125.69, 125.46, 124.19, 123.65, 123.31, 117.56, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 31 H 34 BrN 3 O 4 , [M + H] + m/z: 591.1733, found 592.2801. Anal: C, 62.84; H, 5.78; Br, 13.49; N, 7.09; O, 10.80. N-(2-oxo-2H-chromen-3-yl)-4-(2-(phenylsulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butanamide (13a) Yield 51%; Mp 199–201°C. 1 H NMR (500 MHz, DMSO) δ 9.33 (s, 1H, NH), 7.92 (s, 1H, CH), 7.89 (dd, J = 5.8, 2.5 Hz, 1H, ArH), 7.68 (dd, J = 14.3, 2.8 Hz, 1H, ArH), 7.64 (dd, J = 9.3, 3.2 Hz, 3H, Ar-3H), 7.59–7.52 (m, 1H, ArH), 7.38–7.30 (m, 2H, Ar-2H), 3.33 (dd, J = 24.9, 13.8 Hz, 1H, CH), 2.93 (dd, J = 24.9, 13.7 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 2H), 2.49 (dt, J = 20.3, 6.2 Hz, 2H), 2.41 (dt, J = 24.8, 20.9, 10.8 Hz, 2H), 2.20–2.09 (m, 1H), 2.05 (t, J = 10.6 Hz, 2H), 1.87–1.77 (m, 4H), 1.65 (d, J = 21.6 Hz, 2H), 1.52–1.47 (m, 2H), 1.46–1.42 (m, 2H), 1.41–1.35 (m, 2H), 1.36–1.27 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 171.53, 155.50, 149.07, 138.88, 134.48, 129.76, 129.05, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 30 H 35 N 3 O 5 S, [M + H] + m/z: 549.2297, found 549.2999. Anal: C, 65.55; H, 6.42; N, 7.64; O, 14.55; S, 5.83. 4-(2-benzoyldecahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide (14a) Yield 61%; Mp 196–199°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.25, (s, 1H, NH), 7.97 (d, J = 3.3 Hz, 1H, ArH), 7.96 (s, 1H, Ar-1H), 7.93(dd, J = 3.3, 1.7 Hz,1H, CH ), 7.68 (dd, J = 14.6, 3.1 Hz, 1H, ArH), 7.60 (ddd, J = 8.2, 7.3, 4.0 Hz, 1H, ArH), 7.57–7.55 (m, 1H, Ar-1H), 7.52 (dd, J = 6.9, 2.7 Hz, 1H, Ar-1H), 7.38–7.30 (m, 2H), 3.44 (dd, J = 24.8, 14.5 Hz, 1H, CH), 3.08–2.94 (m, 2H), 2.51 (dt, J = 24.7, 10.8 Hz, 2H), 2.41 (dt, J = 24.8, 10.6 Hz, 2H), 2.14 (ddd, J = 17.0, 11.7, 1.5 Hz, 1H), 2.05 (t, J = 12.2 Hz, 2H), 2.00–1.88 (m, 3H), 1.80–1.71 (m, 3H), 1.69–1.63 (m, 2H), 1.58 (dd, J = 11.9, 8.7 Hz, 2H), 1.50 (dd, J = 20.9, 10.2 Hz, 1H), 1.45–1.40 (m, 1H), 1.39–1.28 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 170.91, 155.50, 149.07, 135.71, 130.18, 128.88, 128.26, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 31 H 35 N 3 O 4 , [M + H] + m/z: 513.2628, found 513.1701. Anal: C, 72.49; H, 6.87; N, 8.18; O, 12.46. 4-(2-(4-cyanobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(6-nitro-2-oxo-2H-chromen-3-yl)butanamide (15a) Yield 53%; Mp 198–200°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.14 (s, 1H, NH), 8.58 (d, J = 3.1 Hz, 1H, ArH), 8.25 (d, J = 2.9 Hz, 1H, ArH), 8.21 (dd, J = 6.1, 2.7 Hz, 1H, ArH), 8.18–8.17 (m, 1H), 8.09 (dd, J = 15.5, 2.8 Hz, 2H, ArH), 8.01 (s, 1H, CH), 7.62 (d, J = 15.0 Hz, 1H, ArH), 3.66 (dd, J = 24.9, 11.4 Hz, 1H), 3.50 (dt, J = 16.8, 13.6 Hz, 1H), 2.51 (dt, J = 2.0 Hz, 2H), 2.48 (d, J = 10.9 Hz, 1H), 2.41 (dt, J = 24.8, 20.9, 10.8 Hz, 2H), 2.17–1.96 (m, 4H), 1.86 (s, 1H), 1.78–1.67 (m, 1H), 1.68–1.59 (m, 4H), 1.61–1.51 (m, 2H), 1.50–1.42 (m, 2H), 1.41–1.33 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 170.91, 155.50, 154.56, 144.74, 139.16, 132.92, 129.44, 126.05, 125.20, 124.51, 121.90, 118.94, 117.69, 114.76, 113.24, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 32 H 33 N 5 O 6 , [M + H] + m/z: 583.2431, found 584.2520. Anal: C, 65.85; H, 5.70; N, 12.00; O, 16.45. 4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)-N-(6-chloro-2-oxo-2H-chromen-3-yl)butanamide (16a) Yield 49%; Mp 195–198°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.28 (s, 1H, NH), 8.00 (d, J = 2.9 Hz,, 1H, ArH), 7.98 (s, 1H, CH), 7.67–7.57 (m, 4H, Ar-4H), 7.38 (dd, J = 14.8, 2.9 Hz, 1H, ArH), 7.28 (d, J = 15.0 Hz, 1H, ArH), 3.32 (dd, J = 24.8, 13.8 Hz, 1H, CH), 2.91 (dd, J = 24.8, 13.7 Hz, 1H), 2.51 (dt, J = 24.8, 10.9 Hz, 2H), 2.49 (t, J = 10.8 Hz, 1H), 2.41 (dt, J = 24.8, 10.7 Hz, 2H), 2.18–2.09 (m, 1H), 2.05 (td, J = 10.7, 0.6 Hz, 2H), 1.87–1.76 (m, 2H), 1.76–1.66 (m, 2H), 1.60 (d, J = 2.1 Hz, 1H), 1.56–1.50 (m, 2H), 1.56–1.50 (m, 2H), 1.49–1.45 (m, 3H), 1.41–1.34 (m, 3H), 1.33 (s, 9H, C-(CH 3 ) 3 ), 1.31–1.23 (m, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 155.50, 154.40, 148.29, 133.92, 132.01, 128.79, 128.32, 126.77, 124.51, 123.35, 122.38, 118.80, 113.24, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 34 H 42 ClN 3 O 5 S, [M + H] + m/z: 639.2534, found 640.3938. Anal: C, 63.78; H, 6.61; Cl, 5.54; N, 6.56; O, 12.49; S, 5.01. N-(6-chloro-2-oxo-2H-chromen-3-yl)-4-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butanamide (17a) Yield 44%; Mp 187–189°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.28 (s, 1H, NH), 8.12 (dd, J = 15.0, 9.9 Hz, 2H, Ar-2H), 8.02 (d, J = 2.9 Hz, 1H, ArH), 8.00 (s, 1H, CH), 7.40 (q, J = 3.2 Hz, 1H, ArH), 7.36 (dd, J = 8.9, 6.0 Hz, 2H, Ar-2H), 7.29 (d, J = 14.9 Hz, 1H, ArH), 3.40 (dd, J = 24.7, 14.5 Hz, 1H, CH), 3.11–2.94 (m, 2H), 2.51 (dt, J = 24.7, 10.7 Hz, 2H), 2.41 (dt, J = 24.8, 10.6 Hz, 2H), 2.19–2.10 (m, 1H), 2.05 (dd, J = 15.8, 9.1 Hz, 3H), 2.00–1.86 (m, 1H), 1.81–1.71 (m, 3H), 1.72–1.62 (m, 3H), 1.62–1.46 (m, 3H), 1.46–1.39 (m, 2H), 1.39–1.29 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 173.30, 154.16, 148.29, 133.61, 132.01, 131.63, 126.77, 124.51, 123.35, 122.38, 118.80, 115.98, 113.24, 67.56, 58.99 (s), 53.21 (s), 47.57 (s), 40.85 (s), 37.70 (s), 36.88 (s), 30.89 (s), 28.28 (s), 26.28 (s), 23.44 (s), 23.25 (s), 22.61 (s). HRMS (ESI): Calcd. C 31 H 33 ClFN 3 O 4 , [M + H] + m/z: 565.2144, found 566.2206. Anal: C, 65.78; H, 5.88; Cl, 6.26; F, 3.36; N, 7.42; O, 11.31. 4-(decahydro-1H,4H-pyrido[3,2,1-ij][, ]naphthyridin-1-yl)-N-(6-methyl-2-oxo-2H-chromen-3-yl)butanamide (18a) Yield 39%; Mp 200–202°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.24 (s, 1H, NH), 7.97 (s, 1H, CH), 7.52 (d, J = 2.9 Hz, 1H, ArH), 7.31 (d, J = 15.0 Hz, 1H, ArH), 7.22 (dd, J = 15.0, 2.9 Hz, 1H, ArH), 3.01 (dd, J = 24.8, 14.9 Hz, 1H, CH), 2.51 (dt, J = 24.7, 10.7 Hz, 2H), 2.42 (s, 3H, CH 3 ), 2.41 (dt, J = 24.8, 10.6 Hz, 2H), 2.11 (ddd, J = 15.9, 13.8, 8.0 Hz, 2H), 2.03 (dd, J = 16.4, 9.6 Hz, 2H), 1.82–1.68 (m, 5H), 1.56–1.49 (m, 2H), 1.46 (t, J = 5.5 Hz, 3H), 1.43–1.40 (m, 2H), 1.39–1.31 (m, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 155.50, 148.30, 134.77, 127.63, 125.71, 124.51, 121.57, 117.43, 113.24, 68.36, 58.08, 53.21, 47.89, 41.90, 37.70, 36.11, 32.24, 28.36, 26.36, 23.44, 23.25, 21.56, 21.23. HRMS (ESI): Calcd. C 25 H 33 N 3 O 3 , [M + H] + m/z: 423.2522, found 423.3215. Anal: C, 70.89; H, 7.85; N, 9.92; O, 11.33. tert-butyl 1-(4-oxo-4-((2-oxo-2H-chromen-3-yl)amino)butyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (19a) Yield 47%; Light yellow oil. 1 H NMR (500 MHz, CDCl 3 ) δ 9.30 (s, 1H, NH), 7.96 (s, 1H, CH), 7.68 (dd, J = 14.6, 3.1 Hz, 1H, ArH), 7.59–7.52 (m, 1H, ArH), 7.34 (td, J = 14.9, 7.6 Hz, J = 10.0 Hz, 2H, Ar-2H), 4.30 (dd, J = 24.8, 15.2 Hz, 1H), 3.21 (dd, J = 24.8, 15.2 Hz, 1H), 2.83 (dt, J = 21.6, 8.0 Hz, 1H), 2.57–2.46 (m, 2H), 2.46–2.35 (m, 2H), 2.24–2.10 (m, 2H), 2.10–1.98 (m, 3H), 1.93–1.78 (m, 3H), 1.68–1.59 (m, 3H), 1.60–1.50 (m, 2H), 1.51–1.43 (m, 2H), 1.42 (s, 9H), 1.40–1.25 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 157.65, 155.50, 149.07, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 81.20, 67.56, 54.76, 53.21, 46.17, 40.85, 37.70, 36.88, 30.89, 28.31, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 29 H 39 N 3 O 5 , [M + H] + m/z: 509.2890, found 509.8866. Anal: C, 68.35; H, 7.71; N, 8.25; O, 15.70. (9H-fluoren-9-yl)methyl1-(4-((5-chloro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (20a) Yield 52%; Mp 196–199°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.29 (s, 1H, NH), 8.27 (s, 1H, CH), 7.90 (dd, J = 14.7, 3.2 Hz, 2H, Ar-2H), 7.57 (t, J = 15.0 Hz, 1H, ArH), 7.41 (dd, J = 14.6, 3.4 Hz, 2H, Ar-2H), 7.34 (td, J = 14.9, 3.4 Hz, 2H, Ar-2H),7.26 (dd, J = 7.1, 3.1 Hz, 1H, ArH), 7.24 (d, J = 3.1 Hz, 2H, Ar-2H), 7.17 (dd, J = 14.9, 3.0 Hz, 1H, ArH), 5.31 (d, J = 11.8 Hz, 2H, CH2), 4.46 (dd, J = 24.8, 15.3 Hz, 1H, CH), 3.99 (t, J = 11.8 Hz, 1H), 3.37 (dd, J = 24.8, 15.4 Hz, 1H), 2.82 (dt, J = 21.6, 8.1 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 2H), 2.41 (dt, J = 50.0 Hz, 2H), 2.28–2.13 (m, 2H), 2.07–1.98 (m, 3H), 1.94–1.79 (m, 3H), 1.71–1.56 (m, 3H), 1.55–1.53 (m, 2H), 1.55–1.39 (m, 3H), 1.38–1.24 (m, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53 (s), 157.88 (s), 155.50 (s), 149.92 (s), 143.41 (s), 139.81 (s), 128.10 (s), 127.80 (s), 127.52 (s), 126.50–126.07 (m), 125.07 (s), 120.89 (s), 117.47 (s), 116.92 (s), 116.65 (s), 67.56 (s), 67.18 (s), 54.76 (s), 53.21 (s), 48.95 (s), 46.17 (s), 40.85 (s), 37.70 (s), 36.88 (s), 30.89 (s), 28.28 (s), 26.28 (s), 23.44 (s), 23.25 (s), 22.61 (s). HRMS (ESI): Calcd. C 39 H 40 ClN 3 O 5 , [M + H] + m/z: 665.2656, found 666.4602. Anal: C, 70.31; H, 6.05; Cl, 5.32; N, 6.31; O, 12.01. (9H-fluoren-9-yl)methyl1-(4-((5-fluoro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (21a) Yield 56%; Mp 195–198°C. 1 H NMR (500 MHz, CDCl 3 ) δ 9.25 (s, 1H, NH), 8.06 (s, 1H, CH), 7.90 (dd, J = 14.7, 3.2 Hz, 2H, ArH), 7.61 (td, J = 15.0, 10.0 Hz, 1H, ArH), 7.41 (dt, J = 12.1, 6.0 Hz, 2H, ArH), 7.33 (dd, J = 14.8, 3.4 Hz, 2H, ArH), 7.24 (td, J = 14.8, 3.2 Hz, 2H, ArH), 7.13 (dd, J = 15.0, 3.0 Hz, 1H, ArH), 6.91 (ddd, J = 15.9, 14.9, 2.9 Hz, 1H, ArH), 5.31 (d, J = 11.8 Hz, 2H, CH 2 ), 4.46 (dd, J = 24.8, 15.4 Hz, 1H, CH), 3.99 (t, J = 11.8 Hz, 1H, CH), 3.37 (dd, J = 24.8, 15.3 Hz, 1H), 2.82 (dt, J = 21.4, 8.0 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 2H), 2.41 (dt, J = 50.0 Hz, 2H), 2.24–2.10 (m, 2H), 2.10–2.00 (m, 3H), 1.94–1.78 (m, 3H), 1.72–1.57 (m, 3H), 1.57–1.49 (m, 2H), 1.48–1.42 (m, 2H), 1.40–1.23 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 174.53, 157.88, 155.50, 153.38, 149.70, 143.41, 139.81, 127.52, 126.18, 125.07, 120.89, 116.42, 115.03, 112.00, 110.97, 67.56, 67.18, 54.76, 53.21, 48.95, 46.17, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C 39 H 40 FN 3 O 5 , [M + H] + m/z: 649.2952, found 650.7940. Anal: C, 72.09; H, 6.21; F, 2.92; N, 6.47; O, 12.31. 6.1.2. Synthesis of compound (1b-7b) The solution of Matrinic alcohol derivatives (6,9,10) (1.0 eq) and coumarin-3-carboxylic acids derivatives 14 ( 2.0 eq) in 10 mL of anhydrous dichloromethane (DCM) was cooled to 0°C. DCC (77 mg, 1.5 eq) and pyridine (0.168 mL 2.0 eq) were added to this, and a catalytic amount of DMAP (0.1 eq). The reaction mixture was stirred overnight at room temperature diluted with DCM and washed with water and brine solution. The dried organic layer on column chromatography yielded the desired product [ 40 ]. 4-(2-(4-bromobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl2-oxo-2H-chromene-3-carboxylate (1b) Yield 58%; Light yellow oil. 1 H NMR (500 MHz, CDCl 3 ) δ 8.55 (s, 1H, CH), 7.96 (d, J = 2.5 Hz, 2H, Ar-2H), 7.93 (d, J = 2.3 Hz, 2H), 7.69 (d, J = 2.5 Hz, 1H), 7.67 (dd, J = 4.2, 2.4 Hz, 1H, ArH), 7.65 (dd, J = 3.8, 3.2 Hz, ArH, 1H), 7.39–7.28 (m, 3H, Ar-3H), 3.97 (t, J = 9.9 Hz, 2H), 3.36 (dt, J = 21.6, 9.2 Hz, 1H), 3.18 (dd, J = 24.7, 14.7 Hz, 1H), 2.65 (dd, J = 24.7, 14.7 Hz, 1H), 2.51 (dt, J = 24.7, 10.8 Hz, 2H), 2.41 (dt, J = 24.7, 10.7 Hz, 2H), 2.16 (t, J = 58.8 Hz, 1H), 2.08–1.99 (m, 1H), 1.84–1.70 (m, 2H), 1.71–1.67 (m, 3H), 1.64 (d, J = 8.3 Hz, 2H), 1.62–1.54 (m, 2H), 1.43 (ddd, J = 19.2 Hz, 2H), 1.35 (dt, J = 23.5, 7.5 Hz, 2H), 1.30–1.19 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 170.91, 163.31, 160.42, 152.43, 136.97, 132.91, 131.51, 131.20, 130.81, 128.55, 125.32, 124.45, 122.09, 120.12, 117.43, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C 32 H 35 BrN 2 O 5 , [M + H] + m/z: 606.1729, found 606.3915. Anal: C, 63.26; H, 5.81; Br, 13.15; N, 4.61; O, 13.17. 4-(2-(2-naphthoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl2-oxo-2H-chromene-3-carboxylate (2b) Yield 57%; Light yellow oil 1 H NMR (500 MHz, CDCl 3 ) δ 8.55 (s, 1H, CH), 8.40 (t, J = 1.4 Hz, 1H, ArH), 8.15 (dd, J = 7.5, 1.4 Hz, 1H, ArH), 7.90 (ddt, J = 6.7, 4.9, 1.5 Hz, 2H, Ar-2H), 7.83 (dd, J = 7.5, 1.6 Hz, 1H, ArH), 7.68 (dd, J = 7.5, 1.4 Hz, 1H, ArH), 7.64 (dd, J = 7.4, 1.5 Hz, 1H, ArH), 7.60 (td, J = 7.4, 1.6 Hz, 1H, ArH), 7.56 (ddd, J = 9.0, 8.4, 1.6 Hz, 1H, ArH), 7.36 (dd, J = 6.4, 1.2 Hz, 1H, ArH), 7.34 (dd, J = 10.0 Hz, 1H, ArH), 3.97 (t, J = 7.5 Hz, 2H), 3.49 (dd, J = 12.3, 7.4 Hz, 1H), 3.02 (dd, J = 12.5, 7.3 Hz, 1H), 2.72 (dt, J = 11.0, 4.1 Hz, 1H), 2.51 (dt, J = 12.3, 5.4 Hz, 2H), 2.41 (dt, J = 12.4, 5.4 Hz, 2H), 2.27–2.16 (m, 1H), 2.04 (t, J = 11.2 Hz, 1H), 1.96 (ddt, J = 12.0, 8.2, 5.9 Hz, 1H), 1.80–1.71 (m, 2H), 1.66 (ddd, J = 11.8, 7.2, 3.5 Hz, 2H), 1.63–1.59 (m, 2H), 1.53 (dq, J = 11.1, 5.6 Hz, 2H), 1.43 (tt, J = 10.2, 5.2 Hz, 2H), 1.39–1.30 (m, 2H), 1.25 (p, J = 7.8 Hz, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 169.44, 163.31, 160.42, 152.43, 134.48, 134.11, 132.91, 131.20, 129.46, 128.60, 127.71, 127.31, 126.64, 125.32, 122.09, 120.12, 118.49, 117.43, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C 36 H 38 N 2 O 5 , [M + H] + m/z: 578.2781, found 579.1513. Anal: C, 74.72; H, 6.62; N, 4.84; O, 13.82. 4-(2-(2-methylbenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl-5-fluoro-2-oxo-2H-chromene-3-carboxylate (3b) Yield 49%; Mp 201–203°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.55 (s, 1H, CH), 7.76 (dd, J = 14.9, 2.9 Hz, 1H, ArH), 7.61 (td, J = 15.0, 10.0 Hz, 1H, ArH), 7.34 (td, J = 14.8, 2.9 Hz, 1H, ArH), 7.24 (dd, J = 15.0, 3.3 Hz, 1H, ArH), 7.11 (ddd, J = 18.0, 10.6, 3.1 Hz, Ar-2H), 6.94 (ddd, J = 15.8, 15.2, 3.0 Hz, 1H, ArH), 3.97 (t, J = 15.0 Hz, 2H), 3.33 (dd, J = 24.8, 14.9 Hz, 1H), 2.87 (dd, J = 24.8, 14.8 Hz, 1H), 2.69 (dt, J = 21.4, 8.2 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 2H), 2.41 (dt, J = 50.0 Hz, 2H), 2.22 (s, 3H, Ar-CH 3 ), 2.16–2.02 (m, 2H), 2.00–1.84 (m, 2H), 1.81–1.71 (m, 1H), 1.73–1.66 (m, 4H), 1.66–1.60 (m, 2H), 1.48–1.35 (m, 2H), 1.33–1.27 (m, 2H), 1.26–1.18 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 171.46, 164.39, 160.30, 153.40, 138.49, 136.46, 130.96, 130.26, 130.06, 128.72, 125.57, 124.80, 115.39, 110.48, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25, 20.26. HRMS (ESI): Calcd. C 33 H 37 FN 2 O 5 , [M + H] + m/z: 560.2687, found 560.2420. Anal: C, 70.69; H, 6.65; F, 3.39; N, 5.00; O, 14.27. 4-(2-(phenylsulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl-5-chloro-2-oxo-2H-chromene-3-carboxylate (4b) Yield 59%; Mp 199–201°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.52 (s, 1H, CH), 7.85 (dd, J = 6.6, 2.8 Hz, 2H, ArH), 7.63–7.59 (m, 3H, Ar-3H), 7.55 (dd, J = 15.5, 14.3 Hz, 1H, ArH), 7.21 (d, J = 1.7 Hz, 2H, Ar-2H), 3.96 (t, J = 14.8 Hz, 2H), 3.76 (dd, J = 24.8, 14.6 Hz, 1H), 3.33 (dd, J = 24.8, 14.6 Hz, 1H), 2.96 (dt, J = 21.8, 12.5 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 3H), 2.41 (dt, J = 50.0 Hz, 3H), 1.85–1.72 (m, 2H), 1.65 (d, J = 22.3 Hz, 1H), 1.59–1.51 (m, 4H), 1.50–1.42 (m, 4H), 1.43–1.35 (m, 2H), 1.34–1.19 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 163.31, 160.42, 152.74, 138.88, 138.69, 134.48, 131.17, 129.74, 129.05, 126.27, 125.07, 117.40, 116.61, 67.56, 66.85, 59.57, 53.21, 48.99, 41.06, 36.19, 31.85, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C 31 H 35 ClN 2 O 6 S, [M + H] + m/z: 598.1904, found 599.1418. Anal: C, 62.15; H, 5.89; Cl, 5.92; N, 4.68; O, 16.02; S, 5.35. 4-(2-(2-methylbenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl-2-oxo-2H-chromene-3-carboxylate (5b) Yield 67%; Mp 200–202°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.53 (s, 1H, CH), 7.74 (dd, J = 14.9, 3.0 Hz, 1H, ArH), 7.66 (dd, J = 14.3, 2.8 Hz, 1H, ArH), 7.57–7.50 (m, 1H, ArH), 7.38–7.31 (m, 2H, Ar-2H), 7.29 (t, J = 2.8 Hz, 1H), 7.22 (dd, J = 14.9, 3.4 Hz, 1H, ArH), 7.09 (td, J = 14.7, 3.3 Hz, 1H, ArH), 3.96 (t, J = 14.9 Hz, 2H), 3.35 (dd, J = 24.7, 14.7 Hz,1H), 2.83 (dd, J = 24.7, 14.7 Hz, 1H), 2.62 (dt, J = 21.8, 8.0 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 2H), 2.41 (dt, J = 50.0 Hz, 2H), 2.21 (s, 3H, Ar-CH 3 ), 2.17–2.01 (m, 2H), 2.01–1.88 (m, 1H), 1.79–1.70 (m, 1H), 1.69–1.60 (m, 5H), 1.58–1.49 (m, 2H), 1.49–1.42 (m, 2H), 1.41–1.29 (m, 2H), 1.30–1.21 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 171.46, 163.31, 160.42, 152.43, 136.46, 132.91, 131.20, 130.96, 130.26, 128.72, 128.55, 125.57, 125.32, 122.09, 120.12, 117.43, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25, 20.26. HRMS (ESI): Calcd. C 33 H 38 N 2 O 5 , [M + H] + m/z: 542.2781, found 542.2996. Anal: C, 73.04; H, 7.06; N, 5.16; O, 14.74. 4-(2-(2-methylbenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl-6-methyl-2-oxo-2H-chromene-3-carboxylate (6b) Yield 48%; Mp 200–203°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.55 (s, 1H, CH), 7.76 (dd, J = 14.9, 2.9 Hz, 1H, ArH), 7.52 (d, J = 2.7 Hz, 1H, ArH), 7.36 (dd, J = 15.0, 2.5 Hz, 2H, ArH), 7.31 (dd, J = 9.0, 6.0 Hz, 1H, ArH),, 7.24 (dd, J = 15.0, 3.3 Hz, 1H, ArH), 7.11 (td, J = 14.7, 3.3 Hz, 1H, ArH), 3.97 (t, J = 14.9 Hz, 2H), 3.35 (dd, J = 24.8, 14.8 Hz, 1H), 2.84 (dd, J = 24.8, 14.8 Hz, 1H), 2.63 (dt, J = 21.8, 8.0 Hz, 1H), 2.52–2.44 (m, 3H), 2.42 (s, 3H, Ar-CH 3 ), 2.39 (dd, J = 17.8, 7.0 Hz, 1H), 2.22 (s, 3H, Ar-CH 3 ), 2.13–2.02 (m, 2H), 2.01–1.89 (m, 1H), 1.79–1.71 (m, 3H), 1.70–1.63 (m, 4H), 1.48–1.42 (m, 3H), 1.41–1.37 (m, 2H), 1.29–1.21 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 171.46, 163.31, 160.42, 151.23, 136.46, 135.61, 131.32, 130.89, 130.26, 128.72, 128.18, 125.57, 123.00, 120.49, 117.16, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25, 21.23, 20.26. HRMS (ESI): Calcd. C 34 H 40 N 2 O 5 , [M + H] + m/z: 556.2937, found 557.4050. Anal: C, 73.36; H, 7.24; N, 5.03; O, 14.37. 4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)butyl-5-bromo-2-oxo-2H-chromene-3-carboxylate (7b) Yield 51%; Mp 195–198°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.55 (s, 1H, CH), 7.67–7.60 (m, 4H, Ar-4H), 7.52 (t, J = 14.8 Hz, 1H, ArH), 7.37 (dd, J = 15.0, 3.3 Hz, 1H, ArH), 7.30 (dd, J = 14.7, 3.2 Hz, 1H, ArH), 3.97 (t, J = 14.9 Hz, 2H), 3.88 (dd, J = 24.1, 14.0 Hz, 1H), 3.33 (dd, J = 24.4, 14.3 Hz, 1H), 3.05 (dt, J = 21.4, 13.6 Hz, 1H), 2.51 (dt, J = 50.0 Hz, 2H), 2.41 (dt, J = 50.0 Hz, 2H), 1.87–1.79 (m, 2H), 1.69–1.60 (m, 4H), 1.54–1.49 (m, 3H), 1.47–1.41 (m, 4H), 1.41–1.34 (m, 2H), 1.33 (s, 9H, C-(CH 3 ) 3 ), 1.32–1.18 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 163.31, 160.42, 154.40, 151.68, 133.92, 133.24, 131.14, 128.86, 128.32, 123.66, 118.81, 118.30, 117.05, 67.56, 66.85, 59.57, 53.21, 48.99, 41.06, 36.19, 34.58, 31.85, 31.36, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C 35 H 43 BrN 2 O 6 S, [M + H] + m/z: 698.2025, found 699.4138. Anal: C, 60.08; H, 6.19; Br, 11.42; N, 4.00; O, 13.72; S, 4.58. 6.1.3. Synthesis of compound (1c-11c) A solution containing anhydrous potassium acetate (CH 3 CO 2 K, 2.94 mmol) the conveniently substituted matrinic acid derivatives ( 5,7,8 ) (1.67 mmol) and the corresponding 2-hydroxybenzaldehyde (1.67 mmol) in acetic anhydride (Ac 2 O, 10 mL) was refluxed (138°C) for 16 h. The reaction mixture was cooled, neutralized with 10% aqueous sodium bicarbonate (NaHCO 3 ), and extracted (3 × 30 mL) with ethyl acetate (EtOAc). The organic layers were combined and washed with distilled water dried with anhydrous sodium sulfate (Na 2 SO 4 ) and evaporated under reduced pressure. The product was purified by recrystallization in ethanol (EtOH) and dried to afford the desired compound [ 39 ]. 3-(2-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-6-nitro-2H-chromen-2-one (1c) Yield 53%; Mp 173–174°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.76 (d, J = 2.9 Hz, 1H), 8.28 (dd, J = 15.0, 2.9 Hz, 1H, ArH), 8.22 (s, 1H, ArH), 8.12 (dd, J = 14.9, 10.0 Hz, 2H, Ar-2H), 7.62 (d, J = 15.0 Hz, 1H, ArH), 7.42–7.32 (m, 2H, Ar-2H), 3.72 (td, J = 13.4, 12.0 Hz, 1H), 3.49 (dd, J = 24.7, 14.3 Hz, 1H), 2.64 (td, J = 11.6, 2.0 Hz, 2H), 2.51 (dt, J = 24.8, 15.4, 12.6 Hz, 3H), 2.41 (dt, J = 24.8, 20.9, 10.0 Hz, 2H), 2.22–2.10 (m, 1H), 2.04 (t, J = 21.7 Hz, 1H), 1.91–1.75 (m, 4H), 1.72 (dt, J = 13.4, 7.2 Hz, 2H), 1.53 (dd, J = 12.8, 11.4 Hz, 2H), 1.50–1.38 (m, 2H), 1.39–1.20 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 167.32, 160.95, 156.78, 145.89, 144.19, 133.61, 131.63, 127.46, 127.16, 126.86, 122.40, 117.90, 115.98, 67.56, 58.76, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 29 H 30 FN 3 O 5 , [M + H] + m/z: 519.2169, found 519.3663. Anal: C, 67.04; H, 5.82; F, 3.66; N, 8.09; O, 15.40. 3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-6-nitro-2H-chromen-2-one (2c) Yield 59%; Mp 175–177°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.56 (s, 1H, CH), 8.28 (dd, J = 14.9, 3.0 Hz, 1H, ArH), 7.68–7.59 (m, 6H, Ar-6H), 3.62 (dd, J = 24.8, 14.7 Hz, 1H), 3.20 (dd, J = 24.8, 14.7 Hz, 1H), 3.09 (dt, J = 21.4, 12.4 Hz, 1H), 2.63–2.55 (m, 1H), 2.51 (dt, J = 24.8, 15.4, 12.6 Hz, 2H), 2.41 (dt, J = 24.8, 20.9, 10.0 Hz, 2H), 2.39–2.32 (m, 1H), 2.26 (ddd, J = 12.5, 1.9 Hz, 2H), 1.90–1.72 (m, 2H), 1.63 (d, J = 22.3 Hz, 1H), 1.59–1.52 (m, 4H), 1.46–1.41 (m, 1H), 1.39–1.34 (m, 2H), 1.33 (s, 9H, C-(CH 3 ) 3 ), 1.31–1.21 (m, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 160.95, 156.78, 154.40, 145.89, 144.19, 133.92, 128.79, 128.32, 127.46, 127.16, 126.86, 122.40, 117.90, 67.56, 57.82, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 32 H 39 N 3 O 6 S, [M + H] + m/z: 593.2560, found 593.1752. Anal: C, 64.73; H, 6.62; N, 7.08; O, 16.17; S, 5.40. 3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij][ 1 , 6 ]naphthyridin-1-yl)ethyl)-8-methoxy-2H-chromen-2-one( 3c )Yield 45%; Mp 170–173°C. 1H NMR (500 MHz, CDCl3) δ 7.65 (t, J = 10.6 Hz, 4H), 7.42 (d, J = 15.0 Hz, 1H), 7.36 (dd, J = 15.9, 5.3 Hz, 1H), 7.25 (s, 1H), 7.23 (dd, J = 12.8, 3.6 Hz, 1H), 3.83 (s, 3H, Ar-OCH3), 3.79 (d, J = 14.5 Hz, 1H), 3.42 (dd, J = 24.8, 14.6 Hz, 1H), 3.15 (dt, J = 21.6, 12.3 Hz, 1H), 2.51 (dt, J = 24.8, 15.4, 12.6 Hz, 2H), 2.41 (dt, J = 24.8, 20.9, 10.0 Hz, 2H), 2.11 (td, J = 14.2, 0.6 Hz, 2H), 1.86–1.71 (m, 2H), 1.68–1.62 (m, 2H), 1.61 (t, J = 22.5 Hz, 1H), 1.48 (dd, J = 13.7, 8.7 Hz, 6H), 1.39–1.34 (m, 2H), 1.33 (s, 9H, C-(CH 3 ) 3 ). 13 C NMR (126 MHz, CDCl 3 ) δ 160.48, 154.40, 145.53, 144.99, 143.50, 133.92, 128.79, 128.32, 127.13, 125.21, 122.84, 121.33, 115.54, 67.56, 57.82, 56.83, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 33 H 42 N 2 O 5 S, [M + H] + m/z: 578.2814, found 578.3625. Anal: C, 68.48; H, 7.31; N, 4.84; O, 13.82; S, 5.54. 3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-6-methyl-2H-chromen-2-one (4c) Yield 59%; Mp 180–183°C. 1 H NMR (500 MHz, CDCl 3 ) δ 7.80 (s, 1H, Ar-H), 7.63 (t, J = 15.0 Hz, 4H, Ar-4H), 7.52 (d, J = 2.5 Hz, 1H, Ar-H), 7.29 (t, J = 16.6 Hz, 2H, Ar-2H), 3.36 (dd, J = 24.9, 13.9 Hz, 1H), 2.97 (dd, J = 24.8, 13.9 Hz, 1H), 2.69 (t, J = 8.0 Hz, 1H), 2.69–2.61 (m, 2H), 2.59–2.51 (m, 1H), 2.51 (dt, J = 24.8, 15.4, 12.6 Hz, 2H), 2.42 (s, 3H, Ar-CH 3 ), 2.41 (dt, J = 24.8, 20.9, 10.0 Hz, 2H), 2.14–2.02 (m, 1H), 1.92–1.74 (m, 2H), 1.70–1.62 (m, 2H), 1.74–1.63 (m, 4H), 1.48–1.42 (m, 2H), 1.41–1.34 (m, 3H), 1.33 (s, 9H, C-(CH 3 ) 3 ). 13 C NMR (126 MHz, CDCl 3 ) δ 160.95, 154.40, 150.53, 145.89, 135.05, 133.92, 129.82, 128.79, 128.32, 127.46, 127.13, 122.25, 116.94, 67.56, 57.82, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25, 21.23. HRMS (ESI): Calcd. C 33 H 42 N 2 O 4 S, [M + H] + m/z: 562.2865, found 563.5492. Anal: C, 70.43; H, 7.52; N, 4.98; O, 11.37; S, 5.70. 3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-2H-chromen-2-one (5c) Yield 57%; Mp 176–178°C. 1 H NMR (500 MHz, CDCl 3 ) δ 7.68 (s, 1H, ArH), 7.63 (t, J = 4.4 Hz, 3H, Ar-3H), 7.61–7.54 (m, 1H, ArH), 7.53 (tdt, J = 24.0 Hz, 2H, Ar-2H), 7.35 (ttd, J = 10.0 Hz, 2H, Ar-2H), 3.83 (dd, J = 24.9, 14.6 Hz, 1H), 3.69 (dd, J = 24.9, 14.5 Hz, 1H), 3.23 (dt, J = 21.6, 12.5 Hz, 1H), 2.51 (dt, J = 24.7, 10.8 Hz, 1H), 2.41 (dt, J = 10.0, 7.5 Hz, 1H), 2.39 (dt, J = 10.0, 7.5 Hz, 2H), 2.10 (td, J = 10.7, 1.8 Hz, 2H), 1.87–1.71 (m, 2H), 1.63 (t, J = 22.3 Hz, 1H), 1.54 (ddd, J = 10.8, 6.1, 3.1 Hz, 4H), 1.43 (ddd, J = 11.7, 7.1, 3.2 Hz, 2H), 1.43–1.36 (m, 2H), 1.33 (s, 9H, C-(CH 3 ) 3 ), 1.30–1.16 (m, 1H). 13 C NMR (126 MHz, CDCl 3 ) δ 160.95, 154.40, 151.59, 146.28, 133.92, 131.84, 128.79, 128.32, 127.52, 126.97, 124.91, 120.68, 117.32, 67.56, 57.82, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 32 H 40 N 2 O 4 S, [M + H] + m/z: 548.2709, found 549.2999. Anal: C, 70.04; H, 7.35; N, 5.11; O, 11.66; S, 5.84. tert-butyl-1-(2-(6-methyl-2-oxo-2H-chromen-3-yl)ethyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (6c) Yield 55%; Light brown viscous oil. 1 H NMR (500 MHz, CDCl 3 ) δ 7.91 (s, 1H, ArH), 7.52 (d, J = 2.5 Hz, 1H, ArH), 7.31 (d, J = 15.0 Hz, 1H, ArH), 7.27 (dd, J = 19.3 Hz, 1H, ArH), 4.32 (dd, J = 24.8, 15.1 Hz, 1H), 3.24 (dd, J = 24.7, 15.2 Hz, 1H), 2.87 (dt, J = 32.5 Hz, 3H), 2.80 (td, J = 15.6, 2.0 Hz, 2H), 2.51 (dt, J = 24.7, 10.8 Hz, 1H), 2.42 (s, 3H, Ar-CH 3 ), 2.41 (dt, J = 10.0, 7.5 Hz, 1H), 2.26–2.11 (m, 2H), 2.04 (t, J = 21.8 Hz,1H), 1.92–1.74 (m, 1H), 1.74–1.63 (m, 3H), 1.61–1.49 (m, 2H), 1.49–1.43 (m, 2H), 1.42 (s, 9H, C-(CH 3 ) 3 ), 1.39–1.28 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 160.95, 157.65, 150.53, 145.89, 135.05, 129.82, 127.46, 127.13, 122.25, 116.94, 81.20, 67.56, 54.22, 53.21, 46.17, 40.85, 36.88, 29.51, 28.31, 27.54, 26.28, 23.44, 23.25, 21.23. HRMS (ESI): Calcd. C 28 H 38 N 2 O 4 , [M + H] + m/z: 466.2832, found 466.2039. Anal: C, 72.07; H, 8.21; N, 6.00; O, 13.71. (9H-fluoren-9-yl)methyl 1-(2-(2-oxo-2H-chromen-3-yl)ethyl)octahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridine-2(3H)-carboxylate (7c) Yield 49%; Mp 166–169°C. 1 H NMR (500 MHz, CDCl 3 ) δ 7.90 (dd, J = 14.7, 3.2 Hz, 2H, Ar-2H), 7.77 (s, 1H, CH), 7.68 (dd, J = 14.6, 3.1 Hz, 1H, ArH), 7.56 (ddd, J = 18.5, 10.7, 3.2 Hz, 3H, Ar-3H), 7.37 (d, J = 3.2 Hz, 1H, ArH), 7.33 (dd, J = 14.8, 3.3 Hz, 1H, ArH), 7.24 (td, J = 14.9, 3.3 Hz, 2H, Ar-2H), 5.33 (d, J = 11.4 Hz, 2H, CH 2 ), 4.15 (dd, J = 24.8, 16.2 Hz, 1H), 4.05 (t, J = 11.4 Hz, 1H), 3.56 (q, J = 12.3 Hz, 1H), 3.44 (dd, J = 24.8, 16.1 Hz, 1H), 2.71 (td, J = 15.8, 1.9 Hz, 2H), 2.51 (dt, J = 24.7, 10.9 Hz, 2H), 2.41 (dt, J = 24.7, 10.9 Hz, 2H), 2.28–2.12 (m, 1H), 2.04 (t, J = 6.3 Hz, 2H), 2.13–1.79 (m, 2H), 1.68 (td, J = 15.7, 12.2 Hz, 2H), 1.58–1.47 (m, 4H), 1.46–1.33 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 160.95, 157.88, 151.59, 146.28, 143.41, 139.81, 131.84, 127.52, 126.97, 126.18, 125.07, 124.91, 120.89, 120.68, 117.32, 67.56, 67.18, 54.22, 53.21, 48.95, 46.17, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 37 H 38 N 2 O 4 [M + H] + m/z: 574.2832, found 574.2695. Anal: C, 77.33; H, 6.66; N, 4.87; O, 11.14. 3-(2-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-2H-chromen-2-one (8c) Yield 54%; Mp 181–183°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.17–8.08 (m, 2H, Ar-2H), 7.73 (s, 1H, CH), 7.68 (dd, J = 14.7, 3.0 Hz, 1H, ArH), 7.55 (dtd, J = 11.8 Hz, 1H, ArH), 7.42–7.28 (m, 2H, Ar-2H), 3.39 (dd, J = 24.8, 14.4 Hz, 1H), 3.02 (dd, J = 24.8, 14.4 Hz, 1H), 2.75 (dt, J = 22.0, 8.2 Hz, 1H), 2.68 (td, J = 15.6, 1.9 Hz, 1H), 2.51 (dt, J = 24.7, 10.8 Hz, 2H), 2.41 (dt, J = 24.7, 10.6 Hz, 2H), 2.25–2.09 (m, 2H), 2.04 (t, J = 22.0 Hz, 1H), 1.81–1.58 (m, 4H), 1.60–1.48 (m, 2H), 1.48–1.42 (m, 2H), 1.41– 1.31(m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 170.91, 162.34, 151.59, 146.28, 133.61, 131.84, 131.63, 127.52, 126.97, 124.91, 120.68, 117.32, 115.98, 67.56, 58.76, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 29 H 31 FN 2 O 3 [M + H] + m/z: 474.2319, found 475.3319. Analysis: C, 73.40; H, 6.58; F, 4.00; N, 5.90; O, 10.11. 3-(2-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-8-methoxy-2H-chromen-2-one (9c) Yield 49%; Mp 173–176°C. 1 H NMR (500 MHz, CDCl 3 ) δ 8.12 (dd, J = 14.9, 10.0 Hz, 2H, Ar-2H ), 7.74 (s, 1H, CH), 7.40 (dd, J = 4.0, 1.8 Hz, 1H, ArH), 7.37 (dd, J = 4.5, 1.7 Hz, 2H, Ar-2H), 7.34 (t, J = 3.1 Hz, 1H, ArH ), 7.23 (dd, J = 13.6, 4.4 Hz, 1H, ArH), 3.83 (s, 3H, Ar-OCH 3 ), 3.39 (dd, J = 24.8, 14.4 Hz, 1H), 3.01 (dd, J = 24.8, 14.4 Hz, 1H), 2.75 (dt, J = 22.0, 8.2 Hz, 1H), 2.67 (ddd, J = 15.6, 1.9 Hz, 2H), 2.51 (dt, J = 24.7, 10.8 Hz, 2H), 2.41 (dt, J = 24.7, 10.7 Hz, 2H), 2.25–2.09 (m, 2H), 2.04 (t, J = 22.0 Hz, 1H), 1.83–1.72 (m, 2H), 1.69 (dt, J = 7.7, 4.7 Hz, 2H), 1.64–1.51 (m, 2H), 1.48–1.38 (m, 2H), 1.37–1.29 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 172.31, 164.23, 163.33, 160.25, 145.53, 144.99, 143.50, 133.61, 131.63, 127.13, 125.21, 122.84, 121.33, 115.98, 115.54, 67.56, 58.76, 56.83, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 30 H 33 FN 2 O 4 , [M + H] + m/z: 504.2424, found 504.2653. Anal: C, 71.41; H, 6.59; F, 3.77; N, 5.55; O, 12.68. tert-butyl-1-(2-(6-methoxy-2-oxo-2H-chromen-3-yl)ethyl)decahydro-1H-benzo[de]isoquinoline-2(3H)-carboxylate( 10c ) Yield 53%; Light yellow oil; 1 H NMR (500 MHz, CDCl 3 ) δ 7.92 (s, 1H, CH), 7.85 (d, J = 15.0 Hz, 1H, ArH), 7.23 (dd, J = 15.0, 2.9 Hz, 1H, ArH), 6.98 (d, J = 2.9 Hz, 1H, ArH), 4.32 (dd, J = 24.8, 15.2 Hz, 1H), 3.77 (s, 3H, Ar-OCH 3 ), 3.24 (dd, J = 24.8, 15.1 Hz, 1H), 2.90–2.83 (m, 1H), 2.80 (td, J = 15.4, 2.0 Hz, 2H), 2.51 (td, J = 23.2, 12.5 Hz, 2H), 2.41 (td, J = 23.2, 12.5 Hz, 2H), 2.22–2.09 (m, 2H), 2.04 (t, J = 21.8 Hz, 1H), 1.89–1.72 (m, 1H), 1.72–1.62 (m, 3H), 1.60–1.50 (m, 2H), 1.49–1.44 (m, 2H), 1.42 (s, 9H, C-(CH 3 ) 3 ), 1.39–1.27 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 160.95, 157.65, 156.09, 146.95, 145.89, 127.46, 122.22, 117.51, 116.26, 109.66, 81.20, 67.56, 56.08, 54.22, 53.21, 46.17, 40.85, 36.88, 29.51, 28.31, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 28 H 38 N 2 O 5 , [M + H] + m/z: 482.2781, found 483.2837. Anal: C, 69.68; H, 7.94; N, 5.80; O, 16.58. 3-(2-(2-(3-chlorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij] [ 1 , 6 ] naphthyridin-1-yl)ethyl)-2H-chromen-2-on (11c) Yield 59%; Mp 189–192°C. 1 H NMR (500 MHz, CDCl 3 ) δ 7.82 (dt, J = 12.5, 3.0 Hz, 1H, ArH), 7.78 (t, J = 3.2 Hz, 1H, ArH), 7.70 (s, 1H, ArH), 7.67 (dd, J = 7.2, 3.2 Hz, 1H, ArH), 7.64 (dd, J = 7.2, 3.2 Hz, 1H, Ar-1H), 7.56 (dd, J = 6.6, 3.4 Hz, 1H, CH), 7.50 (dd, J = 10.0 Hz, 1H, Ar-1H), 7.34 (d, J = 14.8 Hz, 1H), 7.29 (td, J = 1.9 Hz, 1H), 3.40 (dd, J = 24.7, 14.4 Hz,, 1H), 2.99 (dd, J = 24.7, 14.5 Hz, 1H), 2.73 (td, J = 13.8, 8.2 Hz, 1H), 2.66 (ddd, J = 15.5, 1.8 Hz, 2H), 2.51 (td, J = 23.2, 12.5 Hz, 2H), 2.41 (td, J = 23.2, 12.5 Hz, 2H), 2.25–2.10 (m, 2H), 2.03 (t, J = 22.0 Hz, 1H), 1.84–1.67 (m, 4H), 1.53 (ddd, J = 20.0, 11.2, 6.8 Hz, 2H), 1.48–1.41 (m, 2H), 1.40–1.27 (m, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 169.06, 160.95, 151.59, 146.28, 137.27, 134.68, 131.84, 130.13, 129.96, 129.61, 127.52, 126.97, 126.37, 124.91, 120.68, 117.32, 67.56, 58.76, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C 29 H 31 ClN 2 O 3 [M + H] + m/z: 490.2023, found 491.2933. Anal: C, 70.94; H, 6.36; Cl, 7.22; N, 5.71; O, 9.77. 6.2. Biological evaluation assay 6.2.1. Cell culture A549 (Human lung cancer cells), HepG-2 (Human Hepatoma Cells), and HeLa (human cervical cancer cell line) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI-1640 (Solarbio) medium containing 10% Fetal Bovine Serum (FBS, GIBCO), 100U/ml Penicillin, and 100 mg/ml Streptomycin at 37°C under a 5% CO 2 humidified atmosphere. 6.2.2. Antiproliferative activity A549, HeLa and HepG-2 cells were subjected to an MTT assay to evaluate the investigated compounds' antiproliferative activity. In a CO 2 incubator at 37°C cells were cultured in RPMI-1640 or DMEM complete medium with 10% fetal bovine serum. 96-well plates with a cell density of 2 * 10 3 cells per well were employed to plate exponentially growing cells which were then incubated for 24 h at 37°C to evaluate for attachment. The test compound was dissolved in DMSO. Different concentrations of compounds (100, 50, 25, 12.5, and 6.25µM, respectively) were used to treat the cells for 48 h. Each group consisted of three replicate wells, and subsequent incubation of the plate was conducted at a temperature of 37 ◦C for a duration of 48 hours. MTT solution was added to each well and incubated at 37 ◦C for 4 h. Absorbance of each well was recorded at a wavelength of 570 nm with the multi-function microplate reader. Finally, the data are expressed as means of three independent experiments ± standard deviation (SD). the growth inhibition data represented as IC 50 values were calculated through the prism statistical package [ 45 , 46 ]. 6.2.3. Cell cycle assays A549 and BEAS-2B cell lines were added to a 6-well plate at a final concentration of 2 * 10 5 cells per well and various concentrations of 5a (0, 1, 2, 4, 8, 16, and 32 µM) were subsequently applied. After 48 h the wells were detached utilizing a trypsin/EDTA solution fixed using 70% ice-cold ethanol for 12 h and subsequently exposed to a 50 µL RNase (50 g/ml) treatment for an hour at 37°C. After that, 250 µl 50 g/ml PI was added and the cells were incubated at 4°C for 4 h. Cell Quest software (BD) and Flow Jo 4.8 were used to evaluate the effects on the cell cycle using a FACS Calibur (BD). In this manner, the cell cycle phase of 2 * 10 4 cells was examined. 6.2.4. Colony formation assay A549 cells were seeded in 6-well plates with a density of 1 * 10 3 cells/well and incubated for 48 h before the medium was shifted to a fresh medium containing 5a at concentrations of (0, 2, 4, 8, 16 and 32 µM). The cells were treated for 7 days before being washed twice with PBS, fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and imaged under a microscope (Nikon). After being dissolved in alcohol the violet crystals were read by a microplate reader (BioTek, USA) at an absorbance of 595 nm. GraphPad Prism 7.0 was utilized to analyze the readers. 6.2.5. Cell migration assay A549 cells were cultured on a six-well plate allowing space to adhere to other cells. With a 200 µL pipette tip, a wound was created by drawing a line through the middle of each well. Following three PBS washes to remove floating cells the wells were grown in DMEM with 2% FBS as an addition. After adding various doses of 5a (0, 2, 4 and 8 µM) to the wells the plate was incubated for 48 h. Following the removal of the medium three distinct areas were chosen randomly for imaging using a microscope (Nikon, Japan) (×100 magnification). The following formula was used to determine the scratch area healing rate: Scratch area healing rate is calculated as follows: [initial scratch area - scratch area at a specific time point initial scratch area 100%]. 6.2.6. 4,6-Diamidino-2-phenylindole (DAPI) staining analysis. Cells were seeded in 12-well plates, and exposed to 5a at a preset concentration (0, 1, 2, 4, 6 and 8) for 48 h and then the nutrient supernatant was discarded. The uncovered cells were fixed in 4% paraformaldehyde for 15 min and washed with PBS twice. Finally, the cells were stained in DAPI dye liquor (Southern Biotech Company, USA) at the final concentration of 1 mg/L for 15 min and washed in PBS once again. The samples were observed and photographed by fluorescence microscope (FM) (Olympus BX51, Olympus, Japan). 6.2.7. Annexin V-FITC/PI apoptosis assay For apoptosis determination, 3 * 10 5 cells/well were seeded in a six-well plate and treated with various concentrations of 5a (0, 2, 4 and 8 µM). After 24 h, the medium was removed and the cells were washed once with PBS, followed by detachment using trypsin/EDTA solution. The cells were stained with annexin V‐FITC/PI solution, and viability was measured on a FACS Calibur (BD) using Cell Quest software (BD) and analyzed using Flow Jo (Flow Jo LLC) as described before. 6.2.8. Western blot analysis To examine the effect of 5a on the protein expression level in A549 cells the various concentrations (0, 2, 4 and 8 µM) of 5a -treated cells were harvested and lysed in sodium dodecyl sulfate (SDS) buffer for 30 min at 4°C. The supernatant obtained by centrifugation at 12000 g for 20 min was used to determine protein concentration by the bicinchoninic acid assay. Proteins (20–50 µg) were separated by SDS polyacrylamide gel electrophoresis in 8%-12% gels and transferred onto polyvinylidene difluoride membranes (Millipore) at a low temperature. After blocking the membrane for 1 h at room temperature in 5% skim milk in Tris‐buffered saline (TBS) containing 0.1% of Tween‐20 (TBST), they were incubated with an appropriate primary antibody (1:1000) at 4°C for 12 h. The membranes were then washed with TBST and probed with secondary antibodies (1:1000) for 1 h at room temperature. After washing protein expression was measured using an enhanced chemiluminescence kit. 6.3. Animal Experiments 6.3.1 Ethics Approval This study was conducted by the guidelines of the Institutional Animal Care and Use Committee (IACUC). The protocol was approved by Animal Experimental Ethical Committee of Guangxi University under the No: Gxu-2024-258. 6.3.2. Acute-Toxicity Assessment The Balb/c nude mice were divided randomly (six mice for each group) into one treatment group and the control group. Compound 5a (40 mg/kg, 10% DMSO/10% Tween-80/80% normal saline) and Vehicle (10% DMSO/10% Tween 80/80% normal saline) were administered one day. Subsequently, mice were observed for 28 days and the Mice were placed in a chamber gradually filled with CO 2 gas until unconsciousness and death occurred and the weight was recorded daily. 6.3.3. A549 Xenograft Tumor Model Prior to implantation, A549 cell suspension was collected at a concentration of 1 * 10 7 cells/ml, in PBS and 0.1 ml of each cell was inoculated subcutaneously in the right armpit of the Balb/c nude mice. The diameter of transplanted tumors in mice was measured with a vernier caliper, and when the tumors grew to 100 mm 3 the animals were randomly divided into 6 groups. At the same time, the mice in each group started to be administered. The method of measuring the tumor diameter was used to observe the anti-tumor effect of the test samples dynamically. Immediately after the end of the experiment, the mice were sacrificed and the tumor mass was surgically removed and weighed. Body weight and tumor size were measured every 3 days for 28 days after treatment. The formula for calculating tumor volume (TV) is as follows: TV = 1/2×a×b 2 where a and b represent length and width, respectively [ 47 ]. 6.3.4. Euthanasia study The experimental mice were introduced into a controlled environment within a CO 2 anesthesia chamber. The flow of CO 2 agent was regulated to gradually induce unconsciousness in the mice. Upon achieving unconsciousness, the concentration of CO 2 was elevated to 100%. During this phase, observations were made to confirm the absence of finger pinching reflex and muscle tone, indicative of complete unconsciousness. Following verification, ventilation was sustained for an additional 2 minutes to ensure the cessation of vital signs, thereby confirming the animals' death. 6.3.5. Histological examination The hearts, spleens, kidneys, lungs, livers, and tumor tissues were isolated to observe the integrity and injury in different groups by HE staining. Briefly, after ischemia and reperfusion, mice hearts, spleens, kidneys, lungs, livers, and tumors were isolated and the tissues were fixed in 4% paraformaldehyde overnight at room temperature, and then dehydrated by passing through gradient concentrations of ethanol (80% for 2 h, 90% for 2 h, 95% overnight, 100% for 0.5, 0.5 and 1 h) at room temperature, followed by embedding in paraffin wax. The paraffin‑embedded samples were then sectioned at 4 µm for staining with Mayer's Hematoxylin (H8070, Solarbio Life Sciences, Beijing, China) for 10 min and then by 0.5% aqueous eosin (DH0050, Leigene Biotech, Beijing, China) for 3 min at room temperature. With this method, the nucleus and other acidic structures are stained blue, while the cytoplasm is stained red. Images were acquired using a light microscope at x400 magnification [ 48 ]. 6.4. Molecular docking studies The CTD (PDB ID: 3T0Z) and NTD (PDB ID: 2CG9) atomic coordinates have been retrieved from the RCSB PDB (protein data bank) database [ 49 ] charge assignment, solution measurements, and fragment volumes to the protein have been performed using Autodock Vina PyRx and Discovery Studio 2019 Client for docking analysis. The ligand's 2D structure was drawn and analyzed using ChemDraw, standardized, and converted into PDBQT format using the PyRx Virtual Screening Tool. To show the binding modes 2D, 3D and ligand interaction we carried out using the Discovery Studio 2019 Client. Abbreviations Hsp90 Heat Shock Protein 90 NTD N-terminal domain MD Middle Domain CTD C-Terminal Domain DMAP Dimethylaminopyridine; EDCI 1-ethyl-3-(3-(dimethylamino)propyl) carbodiimide hydrochloride; DCC N, N-dicyclohexylcarbodiimide; DCM Dichloromethane STD Saturation Transfer Difference HSR Heat Shock Response NMR Nuclear Magnetic Resonance DMSO Dimethyl Sulfoxide; IC 50 Half Maximal Inhibitory Concentration H&E Hematoxylin and Eosin Declarations Author Contribution W.L. Supervision, Funding acquisition, Writing - review & editing, Project administration.K.J. Conceptualization, Methodology, Formal analysis, Writing - original draft, Visualization. G.CH. Conceptualization, Methodology, Formal analysis, Visualization.J.M, G.R, L.Y, H.K, Q.G and Y.S Investigation, Data curation, review & editing.L.X. Software, Validation, Resources, Writing - review & editing.Each author contributed significantly to the study's conception, design, data analysis, and interpretation. All authors have read and approved the final manuscript. Acknowledgement This work was supported by the local funding project for scientific and technological development under the guidance of the central government (GuiKe ZY21195012) and Guangxi Innovation-Driven Development Project (GuiKe AA18242040). Data Availability The manuscript and supplementary information file contain the data. The original raw data can be obtained by contacting the Corresponding Author. References “About WHO.” https://www.who.int/about/ (accessed Mar. 10, 2023). WHO, “IARC Research – IARC,” IARC GLOBOCAN cancer incidence and mortality rates , 2018. https://www.iarc.who.int/research-home/ (accessed Dec. 08, 2022). Y. Xu et al. , “Novel matrinic acid derivatives bearing 2-anilinothiazole structure for non-small cell lung cancer treatment with improved Hsp90 targeting effect.,” Drug Dev. Res. , no. June, pp. 1–21, 2022, doi: 10.1002/ddr.21974 . M. Li, X. She, Y. Ou, J. Liu, Z. Yuan, and Q. shi Zhao, “Design, synthesis and biological evaluation of a new class of Hsp90 inhibitors vibsanin C derivatives,” Eur. J. Med. Chem., vol. 244, no. July, p. 114844, 2022, doi: 10.1016/j.ejmech.2022.114844 . C. Prodromou and L. Pearl, “Structure and Functional Relationships of Hsp90,” Curr. Cancer Drug Targets, vol. 3, no. 5, pp. 301–323, 2005, doi: 10.2174/1568009033481877 . A. Röhl, J. Rohrberg, and J. Buchner, “The chaperone Hsp90: Changing partners for demanding clients,” Trends Biochem. Sci., vol. 38, no. 5, pp. 253–262, 2013, doi: 10.1016/j.tibs.2013.02.003 . S. Shantanam and MUELLER, “HHS Public Access,” Physiol. Behav., vol. 176, no. 1, pp. 139–148, 2018, doi: 10.1038/nrc2887.Targeting . I. Ny, “Spring 2009 - CEE 6075 Stochastic Simulation methods in Engineering and Bayesian Computation,” Environ. Eng. , vol. 1, no. 3, pp. 1–3, 2009, doi: 10.1158/1078-0432.CCR-11-1000.Hsp90 . F. H. Schopf, M. M. Biebl, and J. Buchner, “The HSP90 chaperone machinery,” Nat. Rev. Mol. Cell Biol., vol. 18, no. 6, pp. 345–360, 2017, doi: 10.1038/nrm.2017.20 . T. E. M. M. Costa, N. M. Raghavendra, and C. Penido, “Natural heat shock protein 90 inhibitors in cancer and inflammation,” Eur. J. Med. Chem., vol. 189, p. 112063, 2020, doi: 10.1016/j.ejmech.2020.112063 . M. Serhan et al. , “Total iron measurement in human serum with a smartphone,” AIChE Annu. Meet. Conf. Proc. , vol. 2019-Novem, pp. 0–1, 2019, doi: 10.1039/x0xx00000x . M. G. Marcu, A. Chadli, I. Bouhouche, M. Catelli, and L. M. Neckers, “The heat shock protein 90 antagonist novobiocin interacts with a previously unrecognized ATP-binding domain in the carboxyl terminus of the chaperone,” J. Biol. Chem., vol. 275, no. 47, pp. 37181–37186, 2000, doi: 10.1074/jbc.M003701200 . J. A. Burlison, L. Neckers, A. B. Smith, A. Maxwell, and B. S. J. Blagg, “Novobiocin: Redesigning a DNA gyrase inhibitor for selective inhibition of Hsp90,” J. Am. Chem. Soc., vol. 128, no. 48, pp. 15529–15536, 2006, doi: 10.1021/ja065793p . K. H. Yim et al. , “Gambogic acid identifies an isoform-specific druggable pocket in the middle domain of Hsp90β,” Proc. Natl. Acad. Sci. U. S. A. , vol. 113, no. 33, pp. E4801–E4809, 2016, doi: 10.1073/pnas.1606655113 . J. Dvorák and V. Dvorák, “Orthopedic examination of the foot: refresher course,” Sportverletz. Sportschaden, vol. 9, no. 3, pp. 253–270, 1995, doi: 10.1055/s-2007-993444 . G. Garg, A. Khandelwal, and B. S. J. Blagg, Anticancer Inhibitors of Hsp90 Function: Beyond the Usual Suspects , 1st ed., vol. 129. Elsevier Inc., 2016. doi: 10.1016/bs.acr.2015.12.001 . J. A. H. Kowah et al. , “European Journal of Medicinal Chemistry Reports Matrine family derivatives: Synthesis, reactions procedures, mechanism, and application in medicinal, agricultural, and materials chemistry,” vol. 7, no. December 2022, 2023, doi: 10.1016/j.ejmcr.2022.100098 . Q. Sun, W. Ma, Y. Gao, W. Zheng, B. Zhang, and Y. Peng, “Meta-analysis: Therapeutic effect of transcatheter arterial chemoembolization combined with Compound Kushen Injection in hepatocellular carcinoma,” African J. Tradit. Complement. Altern. Med., vol. 9, no. 2, 2012, doi: 10.4314/ajtcam.v9i2.1 . D. Quan et al. , “Structure-Based Design of Novel Alkynyl Thio-Benzoxazepinone Receptor-Interacting Protein Kinase-1 Inhibitors: Extending the Chemical Space from the Allosteric to ATP Binding Pockets,” J. Med. Chem. , vol. 1, no. Figure 1, 2022, doi: 10.1021/acs.jmedchem.2c02067 . L. Wu et al. , “Synthesis and biological evaluation of matrine derivatives containing benzo-α-pyrone structure as potent anti-lung cancer agents,” Sci. Rep. , vol. 6, no. 30 mL, 2016, doi: 10.1038/srep35918 . M. Hossain, “A Review on Heterocyclic: Synthesis and Their Application in Medicinal Chemistry of Imidazole Moiety,” Sci. J. Chem., vol. 6, no. 5, p. 83, 2018, doi: 10.11648/j.sjc.20180605.12 . C. Li, H. Zhu, H. Zhang, Y. Yang, and F. Wang, “Synthesis of 2H-Chromenones from Salicylaldehydes and Arylacetonitriles,” Molecules, vol. 22, no. 7, Jul. 2017, doi: 10.3390/molecules22071197 . A. Kolbus, A. Danel, D. Grabka, M. Kucharek, and K. Szary, “Spectral Properties of Highly Emissive Derivative of Coumarin with N,N-Diethylamino, Nitrile and Tiophenecarbonyl Moieties in Water-Methanol Mixture,” J. Fluoresc., vol. 29, no. 6, pp. 1393–1399, 2019, doi: 10.1007/s10895-019-02446-5 . H. Valizadeh and H. Gholipour, “Imidazolium-based phosphinite ionic liquid (IL-OPPh2) as reusable catalyst and solvent for the knoevenagel condensation reaction,” Synth. Commun., vol. 40, no. 10, pp. 1477–1485, 2010, doi: 10.1080/00397910903097310 . M. M. Heravi, S. Khaghaninejad, and M. Mostofi, “Pechmann reaction in the synthesis of coumarin derivatives,” in Advances in Heterocyclic Chemistry , vol. 112, Academic Press Inc., 2014, pp. 1–50. doi: 10.1016/B978-0-12-800171-4.00001-9 . M. Maheswara, V. Siddaiah, G. L. V. Damu, Y. K. Rao, and C. V. Rao, “A solvent-free synthesis of coumarins via Pechmann condensation using heterogeneous catalyst,” J. Mol. Catal. A Chem., vol. 255, no. 1–2, pp. 49–52, 2006, doi: 10.1016/j.molcata.2006.03.051 . M. Lončarić, D. G. Sokač, S. Jokić, and M. Molnar, “Recent advances in the synthesis of coumarin derivatives from different starting materials,” Biomolecules , vol. 10, no. 1. MDPI AG, Jan. 01, 2020. doi: 10.3390/biom10010151 . F. Chao, D. E. Wang, R. Liu, Q. Tu, J. J. Liu, and J. Wang, “Synthesis, characterization and activity evaluation of matrinic acid derivatives as potential antiproliferative agents,” Molecules, vol. 18, no. 5, pp. 5420–5433, 2013, doi: 10.3390/molecules18055420 . D. D. Li, L. L. Dai, N. Zhang, and Z. W. Tao, “Synthesis, structure-activity relationship and biological evaluation of novel nitrogen mustard sophoridinic acid derivatives as potential anticancer agents,” Bioorganic Med. Chem. Lett. , vol. 25, no. 19, pp. 4092–4096, Oct. 2015, doi: 10.1016/j.bmcl.2015.08.035 . S. Tang et al. , “Structure–activity relationship and hypoglycemic activity of tricyclic matrines with advantage of treating diabetic nephropathy,” Eur. J. Med. Chem., vol. 201, p. 112315, 2020, doi: 10.1016/j.ejmech.2020.112315 . Y. Xu et al. , “Design, synthesis, and biological evaluation of matrine derivatives possessing piperazine moiety as antitumor agents,” Med. Chem. Res., vol. 28, no. 10, pp. 1618–1627, 2019, doi: 10.1007/s00044-019-02398-2 . Z. Shi et al. , “Novel NO-releasing scopoletin derivatives induce cell death via mitochondrial apoptosis pathway and cell cycle arrest,” Eur. J. Med. Chem., vol. 200, 2020, doi: 10.1016/j.ejmech.2020.112386 . D. K. Das, S. Sarkar, M. Khan, M. Belal, and A. T. Khan, “A mild and efficient method for large scale synthesis of 3-aminocoumarins and its further application for the preparation of 4-bromo-3-aminocoumarins,” Tetrahedron Lett., vol. 55, no. 35, pp. 4869–4874, 2014, doi: 10.1016/j.tetlet.2014.07.035 . G. Brahmachari, “Room Temperature One-Pot Green Synthesis of Coumarin-3-carboxylic Acids in Water: A Practical Method for the Large-Scale Synthesis,” ACS Sustain. Chem. Eng. , vol. 3, no. 9, pp. 2350–2358, Sep. 2015, doi: 10.1021/acssuschemeng.5b00826 . S. Fiorito, S. Genovese, V. A. Taddeo, and F. Epifano, “Microwave-assisted synthesis of coumarin-3-carboxylic acids under ytterbium triflate catalysis,” Tetrahedron Lett., vol. 56, no. 19, pp. 2434–2436, 2015, doi: 10.1016/j.tetlet.2015.03.079 . N. Suzuki et al. , “Design, synthesis, and biological activity of boronic acid-based histone deacetylase inhibitors,” J. Med. Chem., vol. 52, no. 9, pp. 2909–2922, 2009, doi: 10.1021/jm900125m . S. Tang et al. , “Synthesis and biological evaluation of 12-benzyl matrinic amide derivatives as a novel family of anti-HCV agents,” Chinese Chem. Lett., vol. 27, no. 7, pp. 1052–1057, 2016, doi: 10.1016/j.cclet.2016.03.006 . R. Mathew, A. K. Kruthiventi, J. V. Prasad, S. P. Kumar, G. Srinu, and D. Chatterji, “Inhibition of mycobacterial growth by plumbagin derivatives,” Chem. Biol. Drug Des., vol. 76, no. 1, pp. 34–42, 2010, doi: 10.1111/j.1747-0285.2010.00987.x . M. J. Matos et al. , “Study of coumarin-resveratrol hybrids as potent antioxidant compounds,” Molecules , vol. 20, no. 2, pp. 3290–3308, Feb. 2015, doi: 10.3390/molecules20023290 . S. Tang et al. , “SAR evolution and discovery of benzenesulfonyl matrinanes as a novel class of potential coxsakievirus inhibitors,” Future Med. Chem. , vol. 8, no. 5, pp. 495–508, Apr. 2016, doi: 10.4155/fmc-2015-0019 . A. Thakur, R. Singla, and V. Jaitak, “Coumarins as anticancer agents: A review on synthetic strategies, mechanism of action and SAR studies,” Eur. J. Med. Chem., vol. 101, pp. 476–495, 2015, doi: 10.1016/j.ejmech.2015.07.010 . Y. T. Huang and B. S. J. Blagg, “A library of noviosylated coumarin analogues,” J. Org. Chem., vol. 72, no. 10, pp. 3609–3613, 2007, doi: 10.1021/jo062083t . S. Tang et al. , “Structure–activity relationship and hypoglycemic activity of tricyclic matrines with advantage of treating diabetic nephropathy,” Eur. J. Med. Chem., vol. 201, Sep. 2020, doi: 10.1016/j.ejmech.2020.112315 . S. Paul, P. Roy, P. Saha Sardar, and A. Majhi, “Design, Synthesis, and Biophysical Studies of Novel 1,2,3-Triazole-Based Quinoline and Coumarin Compounds,” ACS Omega , vol. 4, no. 4, pp. 7213–7230, Apr. 2019, doi: 10.1021/acsomega.9b00414 . M. Li, X. She, Y. Ou, J. Liu, Z. Yuan, and Q. shi Zhao, “Design, synthesis and biological evaluation of a new class of Hsp90 inhibitors vibsanin C derivatives,” Eur. J. Med. Chem., vol. 244, pp. 1–96, 2022, doi: 10.1016/j.ejmech.2022.114844 . X. Wang, S. Zhang, K. Han, L. Wang, and X. Liu, “Induction of Apoptosis by Matrine Derivative ZS17 in Human Hepatocellular Carcinoma BEL-7402 and HepG2 Cells through ROS-JNK-P53 Signalling Pathway Activation,” 2022. J. Zou et al. , “A novel oral camptothecin analog, gimatecan, exhibits superior antitumor efficacy than irinotecan toward esophageal squamous cell carcinoma in vitro and in vivo,” Cell Death Dis., vol. 9, no. 6, 2018, doi: 10.1038/s41419-018-0700-0 . Y. Sun et al. , “Discovery of CZS-241: A Potent, Selective, and Orally Available Polo-Like Kinase 4 Inhibitor for the Treatment of Chronic Myeloid Leukemia,” J. Med. Chem., 2022, doi: 10.1021/acs.jmedchem.2c02124 . “RCSB PDB: Homepage.” https://www.rcsb.org / (accessed Aug. 30, 2023). Tables Tables 1-3 are available in the Supplementary Files section. Schemes Schemes1-3 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files SupplementaryInformation.pdf Graphicalabstract.tif Schemes.docx Tables.docx 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 Our Team In Review Editorial Policies Advisory Board 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-4632508","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":335314842,"identity":"21d91034-8ea8-4b21-a19d-eba39054df11","order_by":0,"name":"Jamal A.H Kowah","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Jamal","middleName":"A.H","lastName":"Kowah","suffix":""},{"id":335314843,"identity":"8abfb3b7-5019-4819-8781-030bd4925201","order_by":1,"name":"Chenxi Guan","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Chenxi","middleName":"","lastName":"Guan","suffix":""},{"id":335314844,"identity":"113855bd-7317-4355-9ff3-d256b803c349","order_by":2,"name":"Meiyan Jiang","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Meiyan","middleName":"","lastName":"Jiang","suffix":""},{"id":335314845,"identity":"11aaf5c6-77b8-40cb-996b-ed6d9f28c231","order_by":3,"name":"Ruobing Gao","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Ruobing","middleName":"","lastName":"Gao","suffix":""},{"id":335314846,"identity":"4c186427-6a5a-491a-b5a9-ba2624ddda10","order_by":4,"name":"Yufang Li","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Yufang","middleName":"","lastName":"Li","suffix":""},{"id":335314847,"identity":"26c45811-a147-40bf-be24-ee490c42016c","order_by":5,"name":"Keyan Han","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Keyan","middleName":"","lastName":"Han","suffix":""},{"id":335314848,"identity":"21062269-7f2e-403e-b1bb-5521c8ff4719","order_by":6,"name":"Gan Qiu","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Gan","middleName":"","lastName":"Qiu","suffix":""},{"id":335314849,"identity":"85f3f821-c8e9-4d81-9c42-13cc568c118a","order_by":7,"name":"Suzhen Yan","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Suzhen","middleName":"","lastName":"Yan","suffix":""},{"id":335314850,"identity":"f06358ef-e4ee-4ad2-8df3-bf0dbd08e02f","order_by":8,"name":"Lisheng Wang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAx0lEQVRIiWNgGAWjYBACPghlA6F4iNHCBqHSSNdymBQt/GcMPxf8Om+34UYC44O3bQzy5gS1SOQYS8/su508c0YCs+HcNgbDnQ0EtfBukObtuZ3ML5HAJs3bxpBgcICgw85u/s3bcy6ZTSKB/TdxWhhyt0nz/DhgB7KFmTgtEvnfrHkbkhMkex42S845J2G4gZAWfv5jybd5/tjZGxxPPvjhTZmNPEFbwICxjSGxgYGxAciUIEY9CPxhsCdW6SgYBaNgFIxAAAC5Fzn9zoVZhgAAAABJRU5ErkJggg==","orcid":"","institution":"Guangxi University","correspondingAuthor":true,"prefix":"","firstName":"Lisheng","middleName":"","lastName":"Wang","suffix":""},{"id":335314851,"identity":"94fbf225-d68b-41ac-9472-7f38158511a6","order_by":9,"name":"Xu Liu","email":"","orcid":"","institution":"Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Xu","middleName":"","lastName":"Liu","suffix":""}],"badges":[],"createdAt":"2024-06-24 23:23:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4632508/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4632508/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":61856703,"identity":"79f5445d-d087-452d-802d-63c293a6c0a5","added_by":"auto","created_at":"2024-08-06 09:57:48","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":60285,"visible":true,"origin":"","legend":"\u003cp\u003eImportant Compounds Containing Coumarin Derivatives of Representative Hsp90 Inhibitors\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/a4da3683d12194b025585d7e.jpg"},{"id":61856692,"identity":"51253c5f-ad73-4b6a-8e12-3ce4f5245a4b","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":65703,"visible":true,"origin":"","legend":"\u003cp\u003eDesign of Matrine Contains Coumarins Derivatives as Hsp90 (NTD\u0026amp;CTD) Isoform selective Inhibitors\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/332029aa664aefb32c7eeb57.jpg"},{"id":61858500,"identity":"1095544c-fe22-4653-9963-f4da069e8072","added_by":"auto","created_at":"2024-08-06 10:21:47","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":51876,"visible":true,"origin":"","legend":"\u003cp\u003eSummary of the SARs of the Matrine Contain Coumarin Derivatives\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/5ce6335caae1f6fcbffb1942.jpg"},{"id":61856689,"identity":"f78d1a44-a5d3-4bcd-8f64-8765823bf4de","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":39907,"visible":true,"origin":"","legend":"\u003cp\u003eThe Inhibitory Effect of \u003cstrong\u003e5a\u003c/strong\u003eon the Proliferation of Human Lung Cancer A549 Cells\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/f2686201999aabe47f4e39ea.jpg"},{"id":61856705,"identity":"77bfa2db-d80a-42d2-8e9c-19d08ef1eb40","added_by":"auto","created_at":"2024-08-06 09:57:49","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":30977,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e5a\u003c/strong\u003e Drug Toxicity to Human Normal Lung Epithelial Cells BEAS-2B Cells\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/2e67c8ebd7809048e76f41e3.jpg"},{"id":61856701,"identity":"11455e75-f008-4d37-bc31-991d3adc1fd2","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":60392,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e5a \u003c/strong\u003eInhibited Colony Formation of Human Lung Cancer Cell (A549. (A) Representative images of A549 cell colonies after treatment with \u003cstrong\u003e5a\u003c/strong\u003eat indicated doses 48 hours; (B) Numbers of A549 cell colonies after treatment with \u003cstrong\u003e5a\u003c/strong\u003e at different doses.\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/58202264f23ee532898502e9.jpg"},{"id":61856702,"identity":"3643fdca-a302-4893-b939-a391e4a5a81a","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":74941,"visible":true,"origin":"","legend":"\u003cp\u003eAntimigration Activity of Compound 5a on A549 Cell Lines. The wound-healing assay was used to evaluate the migration of A549 cells, and images were captured at 0, 24 and 48 h after treatments with \u003cstrong\u003e5a\u003c/strong\u003e.\u003c/p\u003e","description":"","filename":"7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/4ebcb0e8243dbf479d3bfbbf.jpg"},{"id":61856691,"identity":"c8da756e-b532-48e3-9d06-8bf6bd7080dd","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":63023,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Compound\u003cstrong\u003e 5a \u003c/strong\u003eon Cell Apoptosis in A549 Cells. (\u003cstrong\u003eA\u003c/strong\u003e) A549 cell lines resulted from apoptosis by DAPI staining and apoptosis morphological analysis (\u003cstrong\u003eB\u003c/strong\u003e)The cells were treated with compound \u003cstrong\u003e5a \u003c/strong\u003eat (0, 2, 4 and 8 μM) for 24 h and the percentages of cells were detected after Annexin-V/PI staining by flow cytometry analysis. The diverse cell stages were given as live (Q4), early apoptotic (Q3), late apoptotic (Q2) and necrotic cells (Q1).\u003c/p\u003e","description":"","filename":"8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/76103218465f403d93a9b387.jpg"},{"id":61858124,"identity":"6eaab841-add7-43da-ba79-06f2303ac94b","added_by":"auto","created_at":"2024-08-06 10:13:47","extension":"jpg","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":84558,"visible":true,"origin":"","legend":"\u003cp\u003eIn vivo toxicity of vehicle, 5-Fluorouracil, Matrine, and compound \u003cstrong\u003e5a\u003c/strong\u003e. (\u003cstrong\u003eA\u003c/strong\u003e) Vehicle, 5-Fluorouracil, Matrine, and compound \u003cstrong\u003e5a\u003c/strong\u003e were administered just on the first day and the curves represent the body weight of mice (n = 6 per group) over 28 days. (\u003cstrong\u003eB\u003c/strong\u003e) HE staining of the heart, spleen, kidney, lung, liver and tumor of the mice. No abnormality of these organs was observed.\u003c/p\u003e","description":"","filename":"10.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/4d12d350f96b89512cc48d26.jpg"},{"id":61859053,"identity":"ea181b6b-d7bd-409b-9c2a-3013c791e15c","added_by":"auto","created_at":"2024-08-06 10:29:47","extension":"jpg","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":76497,"visible":true,"origin":"","legend":"\u003cp\u003eIn vivo antitumor activity of compound\u003cstrong\u003e 5a\u003c/strong\u003e in A549 cell line xenograft mouse model. Tumor volume and changes in body weight were measured for each group. (\u003cstrong\u003eA\u003c/strong\u003e) Images of A549 tumors from the mice at 28 days after initiation of treatment (n = 6). (\u003cstrong\u003eB\u003c/strong\u003e) Weight of the excised tumors of each group (\u003csup\u003e*\u003c/sup\u003eP＜0.0, \u003csup\u003e**\u003c/sup\u003eP＜0.01). (\u003cstrong\u003eC\u003c/strong\u003e) Tumor growth and body weight changes of the mice during the dosage period; \u003cstrong\u003e(D) \u003c/strong\u003eRelationship curves of tumor volumes at various times over 28 days after administration data points represent mean tumor volume X±SD.\u0026nbsp;\u003c/p\u003e","description":"","filename":"11.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/e4d1392e7b7cbdf62a1b63b8.jpg"},{"id":61856704,"identity":"9eb3a661-9f98-4752-9df3-7a80c75144ac","added_by":"auto","created_at":"2024-08-06 09:57:49","extension":"jpg","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":103785,"visible":true,"origin":"","legend":"\u003cp\u003e(\u003cstrong\u003eA\u003c/strong\u003e) Docking mode of Hsp90 ATP-binding site of NTD (PDB ID: 3T0Z) with relevant side chains in stick representation. Hydrogen bonds are depicted as green dashed lines. (\u003cstrong\u003eB\u003c/strong\u003e) 2D representation of the binding pocket interactions of Hsp90 ATP-binding site of NTD (PDB ID: 3T0Z). (\u003cstrong\u003eC\u003c/strong\u003e) Docking mode of Hsp90 ATP-binding active sites of CTD (PDB ID: 2CG9) with relevant side chains in stick representation. (\u003cstrong\u003eD\u003c/strong\u003e) 2D representation of the binding pocket interactions of Hsp90 ATP-binding site of NTD (PDB ID: 3T0Z). The docking mode and 2D ligand interaction diagram were generated in Discovery Studio 19.\u003c/p\u003e","description":"","filename":"12.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/47c85ee132ef57b86f00d33e.jpg"},{"id":76174061,"identity":"5118b233-b9a4-4c8f-8508-d33d33924191","added_by":"auto","created_at":"2025-02-13 06:05:05","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2672842,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/668aada6-bfd3-4f8b-9a72-32f23bbf0ff8.pdf"},{"id":61856700,"identity":"ce6514a9-896a-4800-a5a9-ec2c64f65cce","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":7560263,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/5b842b8b58967db10936e61b.pdf"},{"id":61858122,"identity":"a37187b1-ea3e-43b3-9fa4-9a4589f66fd9","added_by":"auto","created_at":"2024-08-06 10:13:47","extension":"tif","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":319898,"visible":true,"origin":"","legend":"","description":"","filename":"Graphicalabstract.tif","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/fca75ae037de2647b7aaf360.tif"},{"id":61857219,"identity":"4a5c3d7d-2459-4fe5-b6db-7a6ea7e01e26","added_by":"auto","created_at":"2024-08-06 10:05:47","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":549268,"visible":true,"origin":"","legend":"","description":"","filename":"Schemes.docx","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/826ff9213de9dd6fef142339.docx"},{"id":61856690,"identity":"2cc63a82-2f99-4cf0-9e89-da087f4d6b4f","added_by":"auto","created_at":"2024-08-06 09:57:47","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":357036,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-4632508/v1/63b4aa3a18eae17fe0b81d22.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Rational Design and Synthesis of Matrine Containing Coumarin Derivatives as Hsp90 (NTD\u0026CTD) Isoform selective Inhibitors for the Treatment of Lung Carcinoma","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCancer represents a significant global health risk, despite the remarkable strides made in the field of medical science for cancer treatment. Consequently, substantial health concerns persist among individuals affected by this devastating disease[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The incidence and mortality of cancer have continued to rise over time [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Chemotherapy remains an essential component in the clinical treatment of solid tumors. However, various challenges, including drug resistance and unfavorable side effects, limit the widespread application of chemotherapy. It is vitally necessary to develop new anticancer medicines that are more effective and less toxic to healthy cells. Type II programmed cell death or autophagy is new and vital for preserving cellular homeostasis via lysosome-dependent degradation of cytoplasmic proteins and organelles. Recent research has demonstrated that activating autophagic cell death has a significant effect on cancer treatment [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e] and autophagic cell death has become more and more widely used in the field of tumor treatment [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eLong identified as an anticancer target, Hsp90 is an ATP-dependent chaperone necessary for various protein assembly and folding processes. Recently, scientists discovered that Hsp90 plays a significant role in inflammation, neurodegenerative disease and viral infection. Developing drugs that inhibit this chaperone might help treat these previously untreatable conditions. Numerous client proteins implicated in cancer pathogenesis have been identified several of which possess carcinogenic properties. Notable examples include the tumor protein 53 (p53), the human epidermal growth factor receptor 2 (HER2/neu), signal transduction proteins Braf and Akt, and cell cycle regulators CDK4/6. These proteins play pivotal roles in critical cellular processes associated with cancer development, progression, and therapeutic response. [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. A promising target for anticancer medicines has been identified as Hsp90 due to its involvement in crucial cancer activities [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Hsp90 is essential for the development of tumors and the expression of such an enzyme is significantly correlated with tumor malignancy and a poor prognosis. Hsp90 inhibitors affect more than 30 oncoproteins directly impacting several oncogenic processes [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. As a result of Hsp90's prominence, it has become an attractive drug target for treating multiple diseases [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Several studies have discovered that it can be used as a potential strategy for cancer treatment [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. According to various interaction sites, Hsp90 inhibitors are generally categorized as N-terminal, C-terminal and middle-domain inhibitors. The main clinical downside of Hsp90 N-terminal domain (NTD) inhibitors is the heat shock response (HSR). This pro-survival mechanism is activated by N-terminal Hsp90 inhibitors leading to increased Hsp70 transcription [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Due to NTD inhibitors' disadvantages, CTD and MD inhibitors were developed. The issues posed by NTD inhibitors motivated the development of C-terminal domain (CTD) and middle domain (MD) inhibitors. The discovery of the Hsp90 CTD ATP-binding site has led to the discovery of specific CTD inhibitors as potent anticancer therapeutic options including Novobiocin, a natural drug that contains coumarin [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] and Gambogic acid was also a selective inhibitor that binds to Hsp90 middle domain Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Twenty Hsp90 inhibitors have enrolled in clinical studies since this time [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Therefore, the imperative to create new Hsp90 inhibitors is still very important [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTraditional Chinese medicine is a rich natural product library containing several natural medicines with pharmacological effects. Matrine, a primary constituent found in leguminous plants like Sophora flavescens and Sophora alopecuroides, has demonstrated clinical potential. Studies have indicated that the combination of matrine with other anticancer drugs notably enhances patients' immune capacity and treatment efficacy [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Currently, the scientific literature on the synthesis of matrine derivatives is significant, indicating that matrine has various pharmacological properties and clinical potential. Moreover, matrine has several advantages over traditional Chinese medicine injections in terms of quality control as a one-component drug [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Synthesis of anticancer drugs such as matrine-containing coumarins derivatives is very attractive[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Previously, our group focused on matrine derivatives bearing benzo-α-pyrone structures scaffold, a privileged structure with a broad pharmacological activity that exerted potent anticancer activity by inhibiting lung cancer cell proliferation in vitro and in vivo with no apparent side effects [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. This study builds upon the findings of our previous investigations, as we continue our research efforts in designing and synthesizing novel matrine-containing coumarin derivatives with a specific focus on targeting Hsp90 inhibitors for the treatment of Lung Carcinoma. The primary objective of our current work is to enhance the antiproliferative efficacy of these compounds against cancer cells. To achieve this, we have implemented strategic structural modifications to the matrine scaffold, aiming to optimize their pharmacological properties and bolster the potency of these Hsp90 inhibitors in combating cancer, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eThe importance of coumarin has led to many efforts to develop new synthetic procedures. There is a wide range of applications of coumarins in biology, medicine, and polymer science [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Modern and recent medicine uses several coumarins as anticancer activities[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Organic and medicinal chemists have synthesized bioactive natural products containing coumarin heterocyclic nucleus. A variety of methods can be used to synthesize coumarins including the Perkin reaction [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], Knoevenagel condensation[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], Pechmann condensation [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], Wittig reaction, Baylis Hillman reaction, Claisen rearrangement, Vilsmeier-Haack and Suzuki cross‐coupling reaction[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. The medicinal properties of coumarins have been studied by several authors[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn our quest to develop Hsp90 inhibitory autophagy activators with heightened anticancer potential, we undertook a series of modifications on matrine-based coumarin compounds. These alterations encompassed substituting the 12-N atom with various benzoyl halides, acyl halides, or sulfonyl halides, alongside exploring variations in the carboxyl group at the C-11 position. To evaluate the cytotoxicity of the synthesized compounds, we conducted the 3-(4,5-dimethylthiazol-2-diphenyltetrazolium) bromide (MTT) assay using three distinct cancer cell lines: human cervical carcinoma (HeLa), human hepatocellular carcinoma (HepG-2), and human lung adenocarcinoma (A549) cells. Additionally, we carried out structure-activity relationship (SAR) analysis, providing valuable insights into the correlation between compound structure and anticancer activity, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. To deepen our comprehension of the most effective compound's anticancer mechanism, we explored its potential as an autophagy-inducing agent. We further evaluated the compound's impact on colony formation, cellular migration, and apoptosis, as well as investigated in vivo antitumor efficacy and toxicity. Moreover, docking studies were conducted to assess the design of the target compounds (a, b, and c). Importantly, to the best of our knowledge [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e"},{"header":"2. Matrine Coumarin Derivatives Design of an Hsp90(NTD\u0026CTD) Isoform Selective Inhibitor.","content":"\u003cp\u003eEmploying matrine as the foundational scaffold, our investigation was centered on targeting the N-terminal domain (NTD) and C-terminal domain (CTD) of the Hsp90 protein. Informed by insights derived from the two-dimensional crystal structures of the Hsp90 protein NTD (PDB code: 3T0Z) and CTD (PDB code: 2CG9), recognized for their upregulated expression in tumor cells, a specific array of matrine inhibitors was meticulously crafted to interact with both the NTD and CTD domains. This deliberate strategy resulted in the development of a group of matrine compounds intricately designed to engage with and modulate both the NTD and CTD regions of the Hsp90 protein.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"3. Results and discussion","content":"\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Chemistry\u003c/h2\u003e \u003cp\u003eAs a part of this investigation, we synthesized different series of new compounds containing other various moieties in one structure. To obtain primary groups important for Hsp90 inhibitory activity target compounds, we first designed three matrine-based coumarin derivatives as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The matrinic acid derivatives were efficiently synthesized according to the protocol outlined in Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The detailed chemical structures of compounds are organized in Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, and \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The experimental section describes a novel target compound synthesis using commercially available matrine as the initial material. The primary synthetic pathway involved the hydrolysis of matrine \u003cb\u003e1\u003c/b\u003e with aqueous KOH to obtain intermediate \u003cb\u003e2\u003c/b\u003e, which was then subjected to esterification through the addition of SOCl\u003csub\u003e2\u003c/sub\u003e and an alcohol reaction, resulting in intermediate \u003cb\u003e3\u003c/b\u003e. Subsequent substitution reactions with acyl, sulfonyl, or benzoyl halides and anhydrous potassium carbonate yielded intermediate \u003cb\u003e4\u003c/b\u003e, further hydrolyzed to produce intermediate \u003cb\u003e5\u003c/b\u003e with a 90% yield [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eReduction reactions were instrumental in transforming functional groups, with selective reduction of the carbonyl group yielding allyl alcohol compounds \u003cb\u003e6\u003c/b\u003e, \u003cb\u003e9\u003c/b\u003e, and \u003cb\u003e10\u003c/b\u003e using reducing LiAlH\u003csub\u003e4\u003c/sub\u003e agent. The drop-wise addition of LiAlH\u003csub\u003e4\u003c/sub\u003e to the esters significantly enhanced product yield, as validated by literature reports [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Additionally, matrinic acid \u003cb\u003e2\u003c/b\u003e was processed to yield protected 12 N-Fmoc Matrinic acid \u003cb\u003e7\u003c/b\u003e and 12 N-Boc Matrinic acid \u003cb\u003e8\u003c/b\u003e with a higher yield of 85% [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] as depicted in Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eIn another synthesis pathway, 3-aminocoumarins \u003cb\u003e14\u003c/b\u003e were synthesized through Dakin's procedure from N-acetylglycine 11 and various salicylaldehyde derivatives 12. Hydrolysis of 3-acetamidocoumarins 13 was conducted using H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e, with 70% H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e exclusively yielding 3-aminocoumarin within \u003cb\u003e15\u003c/b\u003e minutes [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Coumarin-3-carboxylic acids \u003cb\u003e16\u003c/b\u003e were efficiently synthesized through Knoevenagel condensation between salicylaldehyde derivatives \u003cb\u003e12\u003c/b\u003e and Meldrum's acid \u003cb\u003e15\u003c/b\u003e utilizing sodium azide or potassium carbonate as catalysts [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] as depicted in Scheme \u003cspan refid=\"Sch2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eThe synthesis of target compounds (\u003cb\u003ea\u003c/b\u003e) involved the reaction of matrinic acid (\u003cb\u003e5, 7, 8\u003c/b\u003e) with 3-aminocoumarins 14 in the presence of EDC, HOBt, and NMM, resulting in targets (\u003cb\u003e1a-21a\u003c/b\u003e) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Furthermore, DCC esterification emerged as a pivotal methodology in the esterification reactions of Coumarin-3-carboxylic acids \u003cb\u003e16\u003c/b\u003e and matrine alcohols (\u003cb\u003e6, 9, 10)\u003c/b\u003e. The addition of a small quantity of 4-methylaminopyridine (DMAP) as a catalyst in the reaction system accelerated the reaction, enhancing the conversion rate and yielding compound (\u003cb\u003e1b-8b\u003c/b\u003e) effectively [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Additionally, the synthesis of compound (\u003cb\u003ec\u003c/b\u003e) from matrinic acid (\u003cb\u003e5, 7, 8\u003c/b\u003e) and 2-hydroxybenzaldehyde derivatives using anhydrous potassium acetate and acetic anhydride yielded target compounds (\u003cb\u003e1c-16c\u003c/b\u003e) with high yields of 60\u0026ndash;70% as depicted in Scheme \u003cspan refid=\"Sch3\" class=\"InternalRef\"\u003e3\u003c/span\u003e[\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Biological evaluation and SAR study\u003c/h2\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e3.2.1. In vitro biological activity\u003c/h2\u003e \u003cp\u003eIn the context of in vitro cell growth activities, all synthesized matrine-based coumarin derivatives (\u003cb\u003ea, b\u003c/b\u003e, and \u003cb\u003ec\u003c/b\u003e) were evaluated against three human cancer cell lines, including HeLa (human cervical cancer cell line), A549 (Human lung cancer cells) and HepG-2 (Human Hepatoma Cells) serving as subjects for the MTT assay, with matrine employed as the positive control. The IC\u003csub\u003e50\u003c/sub\u003e values of target matrine derivatives (\u003cb\u003ea\u003c/b\u003e, \u003cb\u003eb\u003c/b\u003e and \u003cb\u003ec\u003c/b\u003e) were illustrated in Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. According to Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the parent matrine displayed unsatisfactory growth activities across these three cancer cell lines, all target compounds exhibited moderate to potent growth activities against the tested cell lines. In an effort to identify more potent inhibitors of Hsp90 a series of matrine-based coumarin derivatives were synthesized and initial structure-activity relationships (SARs) were established. The SARs observed between matrine-based coumarin compounds and their Hsp90 inhibitory activity is summarized in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The SAR investigation involved modifications inspired by previous studies [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e] particularly the opening of the D-ring from matrine. Chemical modifications of the nitrogen atoms at the 12-N position were substituted with benzoyl halides, acyl halides, or sulfonyl halides and the focus shifted to variations of the carboxyl group at the C-11 attachment aiming to enhance the activity of matrine [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Significantly, among these derivatives, compounds \u003cb\u003e5a\u003c/b\u003e and \u003cb\u003e4c\u003c/b\u003e, modified by 4-tert-butylbenzenesulfonyl chloride at the 12-N atom and carboxyl group by 3-aminocoumarins for compound \u003cb\u003e5a\u003c/b\u003e, and 2-hydroxy-4-methylbenzaldehyde for compound \u003cb\u003e4c\u003c/b\u003e, exhibited the most potent cytotoxicity with IC\u003csub\u003e50\u003c/sub\u003e values of A549, HepG-2, and HeLa (7.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.097, 7.727\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10, 8.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.065 \u0026micro;M) and (10.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88, 12.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36, 14.62\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3), respectively. Additionally, compound \u003cb\u003e1b\u003c/b\u003e, modified by 4-bromobenzoyl chloride at the 12-N atom and carboxyl group by coumarin-3-carboxylic acids, also demonstrated significant cytotoxicity with IC\u003csub\u003e50\u003c/sub\u003e values of A549, HepG-2, and HeLa (8.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58, 10.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07, 11.22\u0026thinsp;\u0026plusmn;\u0026thinsp;1.25) respectively. Among the compounds tested derivative \u003cb\u003e5a\u003c/b\u003e exhibited the highest potency. Considering these observations, the antiproliferative efficacy of the three newly designed matrine-based coumarin derivatives on the aforementioned cell lines prompted further investigation of their cytotoxicity profile against additional cancer types. Furthermore, the most effective compound, \u003cb\u003e5a\u003c/b\u003e was evaluated for its cytotoxic effect on normal lung epithelial cells (BEAS-2B). As presented in Table\u0026nbsp;5S, 5a exhibited a substantially higher IC\u003csub\u003e50\u003c/sub\u003e value against normal lung epithelial cells (BEAS-2B) indicating its exceptional selectivity between A549 (human lung cancer cells) and normal lung epithelial cells (BEAS-2B). Given its potent efficacy and increased selectivity for cancer cells compound \u003cb\u003e5a\u003c/b\u003e was selected for further investigation of its mechanism of growth inhibition in cancer cells.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section4\"\u003e \u003ch2\u003e3.2.1.1. Cell viability of compound \u003cb\u003e5a\u003c/b\u003e against A549 cell lines\u003c/h2\u003e \u003cp\u003eIn the MTT assay, the A549 cells were studied to determine the potential anticancer mechanism of compound \u003cb\u003e5a\u003c/b\u003e. To evaluate its effectiveness, different concentrations of drug \u003cb\u003e5a\u003c/b\u003e (0, 1, 2, 4, 8, 16, and 32 \u0026micro;M) were administered to the A549 cells for durations of 24, 48, and 72 h. The results are depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e and Table\u0026nbsp;4S. demonstrate that the proliferation of human lung cancer cells A549 was considerably diminished in a dose and time-dependent manner by \u003cb\u003e5a\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section4\"\u003e \u003ch2\u003e3.2.1.2. Antiproliferative activity of compound \u003cb\u003e5a\u003c/b\u003e on the normal cell line BEAS-2B and A549\u003c/h2\u003e \u003cp\u003eThe objective of using drugs to cure malignant tumors is to destroy the cancer cells without causing harm or toxicity to the healthy cells or overall health. Consequently, the effect of \u003cb\u003e5a\u003c/b\u003e on the survival of normal lung epithelial cells (BEAS-2B) and lung cancer cells (A549) was assessed by the MTT assay at various concentrations (0, 1, 2, 4, 8, 16 and 32 \u0026micro;M) after a 48 h treatment. As illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e and Table\u0026nbsp;5S. the findings establish that \u003cb\u003e5a\u003c/b\u003e is comparatively less harmful to normal cells in contrast to lung cancer cells.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section4\"\u003e \u003ch2\u003e3.2.1.3. Effect of compound \u003cb\u003e5a\u003c/b\u003e on cancer cell colony formation of A549 cell lines\u003c/h2\u003e \u003cp\u003eCancer cell colony formation was widely used to estimate neoplastic transformation. After being exposed to \u003cb\u003e5a\u003c/b\u003e at various concentrations for 48 h human lung cancer cell A549 exhibited evidence of cell cloning. According to Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e, the number of cell clones in A549 cells was significantly reduced after treatment with \u003cb\u003e5a\u003c/b\u003e at various concentrations (0, 2, 4, 8, 16, and 32 \u0026micro;M) compared to the negative control and the inhibitory effect grew more assertive with the increase in concentration.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section4\"\u003e \u003ch2\u003e3.2.1.4. Antimigration activity of compound \u003cb\u003e5a\u003c/b\u003e on A549 cell lines\u003c/h2\u003e \u003cp\u003eSeveral matrine inhibitors demonstrated anti-angiogenesis activities against tumor endothelium and angiogenesis is crucial for the progression development and metastasis of human cancer. Therefore, utilizing human lung cancer A549 cells as a model, we evaluated \u003cb\u003e5a\u003c/b\u003e at various concentrations (0, 2, 4 and 8 \u0026micro;M) for 24 and 48 h to see how it affected endothelial cells. We also observed cell migration and calculated the scratch area which is a standard method to confirm anti-angiogenesis effects. We first evaluated \u003cb\u003e5a\u003c/b\u003e ability to inhibit A549 migration which is a crucial stage in the development of new blood vessels as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e and Table\u0026nbsp;6S. The study's findings illustrated that within 48 hours, untreated cells repopulated the initially scraped area. Conversely, compound 5a distinctly and dose-dependently inhibited the migration of A549 cells, as indicated by the results [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section4\"\u003e \u003ch2\u003e3.2.1.5. Cell apoptosis assay of \u003cb\u003e5a\u003c/b\u003e on A549.\u003c/h2\u003e \u003cp\u003eWe first evaluated its anticancer activities to determine whether derivative \u003cb\u003e5a\u003c/b\u003e inhibitory effects on the A549 cell lines resulted from apoptosis by DAPI staining and apoptosis morphological analysis. Derivative \u003cb\u003e5a\u003c/b\u003e caused apoptosis after 48 h of incubation with it at various concentrations (0, 1, 2, 4, 6 and 8 \u0026micro;M) in DAPI staining, as demonstrated by the micronuclei of proliferating A549 cells. \u003cb\u003e5a\u003c/b\u003e caused a significant amount of dense blue staining and the development of apoptotic bodies as the nuclear membrane was ruptured, indicating that \u003cb\u003e5a\u003c/b\u003e had an anticancer activity on A549 cells as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eA.\u003c/p\u003e \u003cp\u003eThe specific cytotoxic effects of A549 cells were also evaluated utilizing flow cytometry for the apoptotic analysis method and double stained with annexin V-FITC/PI. As illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eB and Table\u0026nbsp;7S. we evaluated the apoptosis induced by \u003cb\u003e5a\u003c/b\u003e in A549 and compared it with the standard. As a result, different concentrations of \u003cb\u003e5a\u003c/b\u003e (0, 2, 4 and 8 \u0026micro;M) were used to analyze human lung cancer A549 cells for apoptosis using Annexin V-FITC. The fluorescence of Annexin V labeled with FITC can be detected by flow cytometry when it binds to necrotic cells; PI (propidium iodide) is a nucleic acid dye and normal cells and early apoptosis can be detected. PI cannot penetrate the cell membrane in normal cells and early apoptosis. However, it can penetrate the membrane of necrotic and apoptotic cells to form a red color. Annexin V-FITC and PI can be used to detect early apoptosis, late apoptosis, and necrosis. As a result, flow cytometry can be used to quantitatively determine the apoptosis rate of A549 cells. Based on the flow cytometry results shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eB, \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea can induce early and late apoptosis in A549 cells. With the increase of dose, the proportion of A549 cell apoptosis induced by \u003cb\u003e5a\u003c/b\u003e low medium and high dose groups was 6.89% \u0026plusmn;1.12%, 8.645\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12%, and 58.79\u0026thinsp;\u0026plusmn;\u0026thinsp;7.35%, which were significantly different from the control group 4.244\u0026thinsp;\u0026plusmn;\u0026thinsp;0.80% (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (* p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, ** p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, *** p\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section4\"\u003e \u003ch2\u003e3.2.1.6. The effect of compound \u003cb\u003e5a\u003c/b\u003e on the apoptosis-related client proteins Hsp90, Bcl-2 and Bax.\u003c/h2\u003e \u003cp\u003eHuman lung cancer cells A549 were treated with \u003cb\u003e5a\u003c/b\u003e (0, 2, 4 and 8 \u0026micro;M) for 48 h and apoptosis-related proteins were determined to understand the effects of various concentrations of drugs on related proteins. The client protein Hsp90, Bcl-2 and Bax family members play a crucial role in the process of cell apoptosis, which can be divided into two main categories. The changes in the expression levels of Hsp90, Bcl-2 and Bax in the A549 cells were detected by Western blot. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e and Table\u0026nbsp;8S. the experiment confirmed significant changes in the expression levels. The expression of Bcl-2 and Hsp90 proteins that inhibit apoptosis decreased and the presentation of Bax a protein that promotes apoptosis increased.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e \u003ch2\u003e3.2.2. In Vivo antiproliferative activity\u003c/h2\u003e \u003cdiv id=\"Sec14\" class=\"Section4\"\u003e \u003ch2\u003e3.2.2.1. Acute Toxicity Study of Compound \u003cb\u003e5a\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eAccording to the Sub-Acute toxicity assay results, compound \u003cb\u003e5a\u003c/b\u003e did not cause death in mice and the body weight of the treatment group gradually increased while remaining identical to that of the vehicle, Matrine and 5-Fluorouracil groups as positive control Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003eA and Table\u0026nbsp;9S. In addition, HE staining revealed that mice receiving 40 mg/kg of the drug did not exhibit overt toxicity in their hearts, spleens, kidneys, lungs, livers and tumors Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003eB. Overall, molecule \u003cb\u003e5a\u003c/b\u003e had reasonable pharmacokinetic parameters and was safe for oral administration in vivo.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section4\"\u003e \u003ch2\u003e3.2.2.2. Compound \u003cb\u003e5a\u003c/b\u003e inhibited A549 tumor growth in vivo\u003c/h2\u003e \u003cp\u003eTo evaluate compound \u003cb\u003e5a\u003c/b\u003e antitumor activity in vivo we established the A549 cell inoculated xenograft mouse model in which female Balb/c nude mice were divided into four groups: vehicle, 5-Fluorouracil, Matrine and compound \u003cb\u003e5a\u003c/b\u003e (40 mg/kg intraperitoneal QD). In each group, mice were given an oral treatment after three days. Following 28 days of continuous treatment as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e11\u003c/span\u003eA, B compound \u003cb\u003e5a\u003c/b\u003e exhibited more potent antitumor activity than 5-Fluorouracil and Matrine as the dosage increased its antitumor activity improved. The tumor growth inhibition (TGI) value of \u003cb\u003e5a\u003c/b\u003e, Matrine and 5-Fluorouracil was 72.4%, 64.3% and 46.8% respectively Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e11\u003c/span\u003eC. In contrast the body weight of the \u003cb\u003e5a\u003c/b\u003e-treated group did not differ from that of the vehicle, 5-Fluorouracil and Matrine groups further demonstrating the good safety profile of compound \u003cb\u003e5a\u003c/b\u003e in vivo. The relationship curves of tumor volumes at various times of treatments over 28 days after administration as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e11\u003c/span\u003eD.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"4. Molecular Docking","content":"\u003cp\u003eIn order to comprehensively assess the potency of Matrine Contains Coumarins derivatives and to provide guidance for further Structure-Activity Relationship (SAR) studies, [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] conducted molecular docking of the target compounds, elucidating their action mechanisms on the N-terminal domain (NTD) and C-terminal domain (CTD) action targets of the Hsp90 protein.[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Molecular docking of various designed compounds with corresponding action targets is established to establish the basis of various inhibitor molecules [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. Demonstrates compound docking into the Hsp90 ATP-binding site of NTD (PDB ID: 3T0Z) the poses of compounds \u003cb\u003e5a\u003c/b\u003e into the active site of protein were generated based on the scores. As seen in Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e12\u003c/span\u003eA; B the resulting docking model with minimum relative binding energy is -10.5 kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicates that compound \u003cb\u003e5a\u003c/b\u003e has interaction with NTD action targets of Hsp90. The interaction between compound \u003cb\u003e5a\u003c/b\u003e and protein was formed through hydrogen bonds from GLY137; GLY135; ASN51 and LYS112 in protein with the carbonyl group of the amide bond in compound \u003cb\u003e5a\u003c/b\u003e as well as CH-π interaction and also the interaction between compound \u003cb\u003e5a\u003c/b\u003e and protein was formed through N-H donor bond from GLY135. The binding model of compound \u003cb\u003e5a\u003c/b\u003e into the Hsp90 ATP-binding site of NTD revealed several molecular interactions thought to be responsible for the observed affinity: pi\u0026ndash;anion interaction between the benzene ring of coumarin and ASP 54 and other interactions, including Alkyl MET 98; LEU 107; ALA 55 and Van der Waals. As seen in Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e12\u003c/span\u003eC; D compound \u003cb\u003e5a\u003c/b\u003e docking into the Hsp90 ATP-binding active sites of CTD (PDB ID: 2CG9) formed interactions with the key amino acid residues, the resulting docking model with minimum relative binding energy is -10.2 kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicates that compound \u003cb\u003e5a\u003c/b\u003e has interaction with CTD action targets of Hsp90. The interaction between compound \u003cb\u003e5a\u003c/b\u003e and protein was formed through hydrogen bonds from LEU454; GLU453; ASN446 and SER419 in protein with carbonyl group of the amide bond in compound \u003cb\u003e5a\u003c/b\u003e as well as CH-π interaction, and interaction also was formed through N-H donor bond from GLU415. The binding model of compound \u003cb\u003e5a\u003c/b\u003e into Hsp90 ATP-binding active sites revealed several molecular interactions thought to be responsible for the observed affinity: including carbon-hydrogen bond -OCH\u003csub\u003e3\u003c/sub\u003e from ASP503 and Van der Waals. This result suggests that 4-tert-Butylbenzenesulfonyl introduced to the nitrogen atom at the 12-position and 3-amino-5-methoxy-coumarin through the carboxyl group might be beneficial for anticancer activity.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eIn summary, Matrine Contains Coumarins derivatives were designed, synthesized and evaluated for their cytotoxic activity against three cancer cell lines (A549, HepG-2 and HeLa cells) as Hsp90 inhibitors for the treatment of lung carcinoma. SAR studies indicated that compound \u003cb\u003e5a\u003c/b\u003e with 4-tert-butylbenzenesulfonyl chloride at the 12-N atom and carboxyl group by 3-amino-5-methyl-2H-chromen-2-one, had the best anti-cancer activity. Based on the bioactivity results derivative \u003cb\u003e5a\u003c/b\u003e was the most potent compound exhibiting excellent antiproliferative activity against A549, HepG-2 and Hela cells with IC\u003csub\u003e50\u003c/sub\u003e values of 7.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.097, 7.727\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 and 8.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.065 \u0026micro;M respectively. Additionally, derivative \u003cb\u003e5a\u003c/b\u003e demonstrated the most potent activity against A549 cells and was non-toxic to normal lung epithelial cells BEAS-2B. Further mechanism investigations confirmed that \u003cb\u003e5a\u003c/b\u003e inhibited A549 cell proliferation, and suppressed colony formation and migration. In vivo, experiments showed that \u003cb\u003e5a\u003c/b\u003e possessed different mechanisms against A549 cancer cells and exhibited more potent antitumor activity than Matrine and 5-Fluorouracil with a tumor growth inhibition (TGI) value of 72.4% compared to 64.3% and 46.8% respectively. Based on the in vitro and in vivo experiments, compound \u003cb\u003e5a\u003c/b\u003e showed promise as an antitumor agent and further studies including pharmacokinetic profiles and prophase tests are encouraged. Molecular docking analysis revealed that the interactions between \u003cb\u003e5a\u003c/b\u003e functionalities and residues were crucial for binding compounds to proteins. Enhancing these interactions could positively impact the inhibitory activity of the Hsp90 ATP-binding at both CTD and NTD sites. This work provides useful information for further structural modifications of these compounds and the synthesis of new, potent antitumor agents.\u003c/p\u003e"},{"header":"6. Experimental section","content":"\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e6.1. General Chemistry\u003c/h2\u003e \u003cp\u003eReagents and solvents were purchased from commercial sources and used without further purification. Reactions were monitored by thin-layer chromatography (TLC) using precoated silica gel plates (silica gel GF/UV 254), and spots were visualized under UV light (254 nm). The products were purified by flash column chromatography equipped with commercial silica gel (300\u0026ndash;400 mesh). All the compounds were characterized with \u003csup\u003e1\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR and MS. \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC nuclear magnetic resonance (NMR) data were recorded on Bruker Avance 500 spectrometers operating at \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC frequencies of 500 and 126 MHz respectively, in the indicated solvents (DMSO-d6 or CDCl\u003csub\u003e3\u003c/sub\u003e, TMS as internal standard). Chemical shifts (δ) are in ppm relative to the residual solvent signal (DMSO-d6 with 2.48 and 39.52 ppm and CDCl\u003csub\u003e3\u003c/sub\u003e with 7.26 and 77.16 ppm for \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC, respectively). As an internal standard, chemical shifts were given in ppm (d) relative to SiMe4. Coupling constants (J) were in hertz (Hz) and signals were designated as follows: s, singlet; d, doublet; t, triplet; m, multiple; br, broad singlet, etc. Mass spectra were obtained from a ThermoFisher LCQ Fleet (ESI). Melting points were determined in open capillary tubes on the X-4 melting point apparatus without correction. Synthesis of intermediate 1\u0026ndash;14 is shown in the supplementary information file.\u003c/p\u003e \u003cdiv id=\"Sec20\" class=\"Section3\"\u003e \u003ch2\u003e6.1.1. Synthesis of compound (1a-20a)\u003c/h2\u003e \u003cp\u003eMatrinic acid derivatives \u003cb\u003e(5,7,8)\u003c/b\u003e (2.10 mmol), HOBt (2.10 mmol) and EDCI (2.10 mmol) were dissolved in DCM (10 mL). After 10 min of stirring NMM (3.15 mmol) and 3-aminocoumarins \u003cb\u003e12\u003c/b\u003e (3.15 mmol) were added and stirred for 12 h. and monitored the reaction by TLC. Their action mixture was poured into water and extracted with EtOAc. The organic layer was washed with saturated bicarbonate solution followed by brine solution. It was dried under anhydrous Na\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e and the solvent was evaporated. The gained residue was purified by flash column chromatography on silica gel with CH\u003csub\u003e2\u003c/sub\u003eCl\u003csub\u003e2\u003c/sub\u003e /CH\u003csub\u003e3\u003c/sub\u003eOH as the eluent to afford the title compounds \u003cb\u003e1a-21a\u003c/b\u003e[\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003etert-butyl1-(4-((6-chloro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e \u003cb\u003e(1a)\u003c/b\u003eYield 64%; Mp 185\u0026ndash;188\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.29 (s, 1H, NH), 8.02 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1.4 Hz, 1H, CH), 7.99 (s, 1H, ArH), 7.39 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5, 1.4 Hz,1H, ArH), 7.30 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5 Hz, 1H, ArH), 4.29 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.4, 7.7 Hz, 1H, CH), 3.21 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.4, 7.7 Hz, 1H), 2.80 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.8, 4.0 Hz, 1H), 2.51 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.3, 5.5 Hz, 2H), 2.41 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.4, 5.4 Hz, 2H), 2.30\u0026ndash;2.23 (m, 1H), 2.19\u0026ndash;2.09 (m, 1H), 2.04 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;5.0 Hz, 3H), 1.92\u0026ndash;1.84 (m, 2H), 1.83\u0026ndash;1.78 (m, 1H), 1.62 (td, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.8, 4.1 Hz, 3H), 1.57\u0026ndash;1.49 (m, 2H), 1.47\u0026ndash;1.43 (m, 2H), 1.42 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.40\u0026ndash;1.28 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 157.65, 155.50, 149.07, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 81.20, 67.56, 54.76, 53.21, 46.17, 40.85, 37.70, 36.88, 30.89, 28.31, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eClN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 543.2500, found 544.2949. Anal: C, 64.02; H, 7.04; Cl, 6.52; N, 7.72; O, 14.70.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(6-nitro-2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e\u003cb\u003e(2a)\u003c/b\u003eYield 60%; Light yellow oil. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.33 (s, 1H, NH), 8.58 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.9 Hz,1H, ArH), 8.21 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.9, 3.0 Hz, 1H, ArH), 8.05 (s, 1H, ArH), 7.71\u0026ndash;7.50 (m, 5H, Ar-5H), 3.47 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.7, 13.7 Hz, 1H), 2.91\u0026ndash;2.81 (m, 2H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.9 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.8 Hz, 2H), 2.04 (td, J\u0026thinsp;=\u0026thinsp;16.2, 1.8 Hz, 2H), 1.96\u0026ndash;1.82 (m, 2H), 1.82\u0026ndash;1.69 (m, 3H),1.63 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;21.8 Hz, 1H), 1.57\u0026ndash;1.49 (m, 2H), 1.50\u0026ndash;1.43 (m, 4H), 1.44\u0026ndash;1.35 (m, 2H), 1.32 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.32\u0026ndash;1.26 (m, 1H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 155.50, 154.56, 154.40, 144.74, 133.92, 128.79, 128.32, 126.05, 125.20, 124.51, 121.90, 117.69, 113.24, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 650.2774, found 650.2900. Anal: C, 62.75; H, 6.51; N, 8.61; O, 17.21; S, 4.93.\u003c/p\u003e \u003cp\u003e \u003cem\u003etert-butyl1-(4-((6-nitro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e\u003cb\u003e(3a)\u003c/b\u003eYield 49%; brown oil. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.27 (s, 1H, NH), 8.59 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, ArH), 8.24 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15.0, 2.9 Hz, 1H, ArH), 8.04 (s, 1H, CH), 7.62 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 4.37 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 14.7 Hz,1H, CH), 3.04 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 14.6 Hz, 1H, CH), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.9 Hz, 3H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.8 Hz, 2H), 2.07 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15.6, 2.7 Hz, 3H), 1.95 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;16.2, 5.8 Hz, 1H), 1.86\u0026ndash;1.76 (m, 2H), 1.77\u0026ndash;1.68 (m, 2H), 1.67\u0026ndash;156 (m, 3H), 1.55\u0026ndash;1.49 (m, 2H), 1.48\u0026ndash;1.43 (m, 2H), 1.42 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.39\u0026ndash;1.27 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 157.65, 155.50, 154.56, 144.74, 126.05, 125.20, 124.51, 121.90, 117.69, 113.24, 81.20, 67.56, 54.76, 53.21, 46.17, 40.85, 37.70, 36.88, 30.89, 28.31, 26.28, 23.44, 23.25, 22.61 (s). HRMS (ESI): Calcd. C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 554.2740, found 555.2471. Anal: C, 62.80; H, 6.91; N, 10.10; O, 20.19.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-cinnamoyldecahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(5-methoxy-2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e\u003cb\u003e(4a)\u003c/b\u003eYield 57%; Mp 189\u0026ndash;192\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, DMSO) δ 9.22 (s, 1H, NH), 7.97 (s, 1H, CH), 7.92 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;4.7, 2.1 Hz, 1H, ArH), 7.90 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3.1, 1.5 Hz, 1H, ArH), 7.59 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;13.7 Hz, 2H), 7.50 (ddd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;17.6, 6.3, 3.2 Hz, 1H), 7.40 (s, 1H, ArH), 7.36 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;20.0 Hz, 1H, ArH), 7.32 (s, 1H, CH), 7.23 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;13.6, 4.4 Hz, 1H, ArH), 7.03 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;30.2 Hz, 1H, CH), 3.83 (s, 3H, Ar-OCH\u003csub\u003e3\u003c/sub\u003e), 3.61 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 14.6 Hz, 1H), 3.05 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 14.5 Hz, 1H), 2.63 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;22.2, 8.2 Hz, 1H), 2.57\u0026ndash;2.48 (m, 2H), 2.47\u0026ndash;2.37 (m, 2H), 2.34\u0026ndash;2.22 (m, 1H), 2.21\u0026ndash;2.11 (m, 1H), 2.10\u0026ndash;2.06 (m, 1H), 2.04 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;16.3, 12.4 Hz, 1H), 1.90\u0026ndash;1.71 (m, 3H), 1.66\u0026ndash;1.55 (m, 3H), 1.55\u0026ndash;1.47 (m, 2H), 1.46\u0026ndash;1.40 (m, 2H), 1.38\u0026ndash;1.25 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, DMSO) δ 171.53, 163.97, 155.06, 145.89, 139.85, 138.48, 136.15, 128.70 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;16.4 Hz), 128.05, 124.68, 124.14, 123.52, 119.86, 119.20, 115.18, 114.05, 67.56, 58.77, 56.83, 53.21, 46.87, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e39\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 569.2890, found 569.2653. Anal: C, 71.68; H, 6.90; N, 7.38; O, 14.04.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(5-methoxy-2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e\u003cb\u003e(5a\u003c/b\u003e)Yield 59%; Mp 195\u0026ndash;196\u0026deg;C. 1H NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.21 (s, 1H, NH), 7.93 (s, 1H, CH), 7.65\u0026ndash;7.58 (m, 4H, ArH), 7.42\u0026ndash;7.31 (m, 2H, ArH), 7.04 (dd, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, Ar-1H), 3.83 (s, 3H, Ar-OCH\u003csub\u003e3\u003c/sub\u003e), 3.82 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 13.7 Hz, 1H, CH), 3.37 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 13.7 Hz, 1H), 3.17 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;22.0, 8.0 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;23.2, 12.4 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;14.0, 10.8 Hz, 2H), 2.05 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15.7, 5.4 Hz, 2H),1.85\u0026ndash;1.70 (m, 4H), 1.60 (d, J\u0026thinsp;=\u0026thinsp;21.8 Hz, 1H), 1.54\u0026ndash;1.48 (m, 3H), 1.47\u0026ndash;1.42 (m, 3H), 1.41\u0026ndash;1.35 (m, 2H), 1.33 (s, 9H), 1.28 (dd, J\u0026thinsp;=\u0026thinsp;11.7, 4.1 Hz, 1H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 161.08, 155.50, 154.40, 150.91, 133.92, 128.79, 128.32, 127.48, 124.19, 116.58, 114.16, 111.15, 102.14, 67.56, 59.57, 56.08, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e35\u003c/sub\u003eH\u003csub\u003e45\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 635.3029, found 635.2773. Anal: C, 66.12; H, 7.13; N, 6.61; O, 15.10; S, 5.04.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(6a)\u003c/b\u003e Yield 55%; Mp 175\u0026ndash;178\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.25 (s, 1H, NH), 8.12 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5, 5.2 Hz, 2H, Ar-2H), 7.95 (s, 1H, CH), 7.68 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5, 1.4 Hz,1H, ArH), 7.55 (td, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5, 1.5 Hz, 1H, ArH), 7.44 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.6, 6.9 Hz, 2H, Ar-2H), 7.29 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.0, 5.6 Hz, 2H, Ar-2H), 3.36 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.4, 7.3 Hz, 1H), 2.99 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.4, 7.3 Hz, 1H), 2.68 (q, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;11.0, 4.1 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;12.3, 5.4 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;12.4, 5.4 Hz, 2H), 2.34\u0026ndash;2.23 (m, 1H), 2.21\u0026ndash;2.10 (m, 1H), 2.04 (dd, J\u0026thinsp;=\u0026thinsp;5.0 Hz, 3H), 1.87\u0026ndash;1.80 (m, 2H), 1.80\u0026ndash;1.72 (m, 1H), 1.72\u0026ndash;1.60 (m, 3H), 1.57\u0026ndash;1.49 (m, 2H), 1.48\u0026ndash;1.38 (m, 2H), 1.38\u0026ndash;1.27 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 173.57, 153.33, 149.07, 133.61, 131.63, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 115.98, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eFN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 531.2533, found 532.2344. Analysis: C, 70.04; H, 6.45; F, 3.57; N, 7.90; O, 12.04.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(7a)\u003c/b\u003e Yield 50%; Mp 190\u0026ndash;192\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, DMSO) δ 9.25 (s, 1H, NH), 7.96 (s, 1H, CH), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.3, 2.8 Hz, 1H, ArH), 7.61\u0026ndash;7.50 (m, 1H),1H, ArH), 7.38\u0026ndash;7.30 (m, 2H, Ar-2H), 3.01 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.9 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;12.3, 5.5 Hz, 2H), 2.46 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.3 Hz, 1H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;12.4, 5.4 Hz, 2H), 2.17\u0026ndash;2.07 (m, 2H), 2.04 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.0, 4.6 Hz, 2H), 1.86\u0026ndash;1.66 (m, 5H), 1.61\u0026ndash;1.48 (m, 2H), 1.51\u0026ndash;1.45 (m, 4H), 1.45\u0026ndash;1.29 (m, 4H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, DMSO) δ 171.53, 155.50, 149.07, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 68.36, 58.08, 53.21, 47.89, 41.90, 37.70, 36.11, 32.24, 28.36, 26.36, 23.44, 23.25, 21.56. HRMS (ESI): Calcd. C\u003csub\u003e24\u003c/sub\u003eH\u003csub\u003e31\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 409.2365, found 409.7183. Anal: C, 70.39; H, 7.63; N, 10.26; O, 11.72.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(4-cyanobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(8a)\u003c/b\u003e Yield 49%; Light yellow oil. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, DMSO) δ 9.25 (s, 1H, NH), 8.19 (dd, J\u0026thinsp;=\u0026thinsp;15.4, 2.9 Hz, 2H, Ar-2H), 8.09 (dd, J\u0026thinsp;=\u0026thinsp;15.4, 2.9 Hz, 2H, Ar-2H), 7.95 (s, 1H, Ar-CH), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.5, 2.7 Hz, 1H, Ar-H), 7.55 (dd, J\u0026thinsp;=\u0026thinsp;14.6, 12.2 Hz, 1H, Ar-H), 7.36 (dt, J\u0026thinsp;=\u0026thinsp;14.0, 3.1 Hz, 2H, Ar-H), 3.34 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H), 3.00 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.3 Hz, 1H), 2.69 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;22.0, 8.1 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.8 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.8 Hz, 2H), 2.34\u0026ndash;2.14 (m, 2H), 2.10\u0026ndash;1.97, 1.88\u0026ndash;1.72 (m, 3H), 1.72\u0026ndash;1.57 (m, 3H), 1.59\u0026ndash;1.44 (m, 3H), 1.44\u0026ndash;1.22 (m, 3H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, DMSO) δ 171.53, 170.91, 155.50, 149.07, 139.16, 132.92, 129.44, 128.88, 125.69, 125.46, 124.19, 123.65, 118.94, 117.56, 114.76, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61 HRMS (ESI): Calcd. C\u003csub\u003e32\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 538.2580, found 538.6747. Anal: C, 71.35; H, 6.36; N, 10.40; O, 11.88.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(6-methyl-2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(9a)\u003c/b\u003e Yield 56%; Mp 190\u0026ndash;193\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.28 (s, 1H, NH), 7.96 (s, 1H, CH), 7.66\u0026ndash;7.54 (m, 4H, Ar-4H), 7.52 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, Ar-H), 7.28 (d, J\u0026thinsp;=\u0026thinsp;14.9 Hz, 1H, Ar-H), 7.19 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 2.8 Hz, 1H, Ar-H), 3.31 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 13.7 Hz, 1H, CH), 2.91 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 13.7 Hz, 1H), 2.60\u0026ndash;2.51 (m, 1H), 2.51\u0026ndash;2.41 (m, 2H), 2.41 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.38 (dd, J\u0026thinsp;=\u0026thinsp;17.7, 7.0 Hz, 1H), 2.19\u0026ndash;2.13 (m, 3H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;10.6 Hz, 3H), 1.87\u0026ndash;1.78 (m, 1H), 1.75\u0026ndash;1.66 (m, 3H), 1.63\u0026ndash; 158 (m, 3H), 1.57\u0026ndash;1.47 (m, 2H), 1.47\u0026ndash;1.41 (m, 1H), 1.40\u0026ndash;1.35 (m, 1H), 1.32 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.29 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;13.1, 8.9 Hz, 1H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 155.50, 154.40, 148.30, 134.77, 133.92, 128.79, 128.32, 127.63, 125.71, 124.51, 121.57, 117.43, 113.24, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61, 21.23. HRMS (ESI): Calcd. C\u003csub\u003e35\u003c/sub\u003eH\u003csub\u003e45\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 619.3080, found 620.3302. Anal: C, 67.82; H, 7.32; N, 6.78; O, 12.91; S, 5.17.\u003c/p\u003e \u003cp\u003e \u003cem\u003eN-(6-chloro-2-oxo-2H-chromen-3-yl)-4-(2-(3-chlorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butanamide\u003c/em\u003e \u003cb\u003e(10a)\u003c/b\u003e Yield 47%; Mp 188\u0026ndash;190\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.25 (s, 1H, NH), 8.02 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, ArH), 8.01 (s, 1H, CH), 7.87 (t, J\u0026thinsp;=\u0026thinsp;2.9 Hz,1H, ArH), 7.85 (dt, J\u0026thinsp;=\u0026thinsp;14.7, 3.2 Hz, 1H, ArH), 7.69 (dt, J\u0026thinsp;=\u0026thinsp;15.0, 3.1 Hz, 1H, ArH), 7.58 (t, J\u0026thinsp;=\u0026thinsp;14.8 Hz, 1H, ArH), 7.37 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 2.9 Hz, 1H, ArH), 7.30 (d, J\u0026thinsp;=\u0026thinsp;14.9 Hz, 1H, ArH), 3.42 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H, CH), 3.10\u0026ndash;2.86 (m, 2H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.6 Hz, 2H), 2.24\u0026ndash;2.10 (m, 1H), 2.04 (q, J\u0026thinsp;=\u0026thinsp;12.9 Hz, 3H), 2.00\u0026ndash;1.85 (m, 1H), 1.76 (t, J\u0026thinsp;=\u0026thinsp;7.1 Hz, 3H), 1.65 (ddd, J\u0026thinsp;=\u0026thinsp;8.3 Hz, 3H), 1.62\u0026ndash;1.54 (m, 2H), 1.45\u0026ndash;1.39 (m, 1H), 1.37\u0026ndash;1.26 (m, 3H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl3) δ 174.53, 169.06, 155.50, 148.29, 137.27, 134.68, 132.01, 130.13, 129.96, 129.61, 126.77, 126.37, 124.51, 123.35, 122.38, 118.80, 113.24, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e33\u003c/sub\u003eCl\u003csub\u003e2\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 581.1848, found 581.2474. Anal: C, 67.94; H, 6.25; Cl, 6.47; N, 7.67; O, 11.68.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(3-chlorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(11a)\u003c/b\u003e Yield 45%; Mp 178\u0026ndash;180\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.25 (s, 1H, NH), 7.95 (s, 1H, CH), 7.86 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;12.7, 3.0 Hz, 1H, ArH), 7.82 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3.2 Hz, 1H, ArH), 7.70 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.2, 3.1 Hz, 1H, ArH), 7.67 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.6, 3.4 Hz, 1H, ArH), 7.59 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.8 Hz, 1H, ArH), 7.54 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.3, 2.9 Hz, 1H, ArH), 7.34 (ddd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;18.0, 8.2, 2.0 Hz, 2H, Ar-2H), 3.38 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H, CH), 2.98 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.3 Hz, 1H), 2.68 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;22.0, 8.2 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.9 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.7 Hz, 2H), 2.35\u0026ndash;2.12 (m, 2H), 2.11\u0026ndash;1.98 (m, 3H), 1.88\u0026ndash;1.73 (m, 3H), 1.71\u0026ndash;1.61 (m, 3H), 1.60\u0026ndash;1.53 (m, 2H), 1.52\u0026ndash;1.41 (m, 2H), 1.42\u0026ndash;1.27 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 169.06, 155.50, 149.07, 137.27, 134.68, 130.13, 129.96 (s), 129.61, 128.88, 126.37, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eClN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 547.2238, found 548.2740. Anal: C, 67.94; H, 6.25; Cl, 6.47; N, 7.67; O, 11.68.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(3-bromobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(12a)\u003c/b\u003e Yield 39%; Mp 187\u0026ndash;188\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, DMSO) δ 9.25 (s, 1H, NH), 8.14 (t, J\u0026thinsp;=\u0026thinsp;3.0 Hz, 1H, ArH), 7.95 (s, 1H, CH), 7.89 (dt, J\u0026thinsp;=\u0026thinsp;14.8, 2.9 Hz, 1H, ArH), 7.79 (dt, J\u0026thinsp;=\u0026thinsp;14.8, 3.0 Hz, 1H, ArH), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.3, 2.8 Hz, 1H, ArH), 7.57 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;13.4, 2.4 Hz, 1H, ArH), 7.50 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.9 Hz, 1H, ArH) 7.37\u0026ndash;7.30 (m, 2H, Ar-2H), 3.35 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.5 Hz, 1H, CH), 2.96 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H), 2.66 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;22.0, 8.1 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.9 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.7 Hz, 2H), 2.28\u0026ndash;2.11 (m, 2H), 2.08 (dd, J\u0026thinsp;=\u0026thinsp;7.1, 2.7 Hz, 1H), 2.03 (dd, J\u0026thinsp;=\u0026thinsp;16.1, 12.2 Hz, 2H), 1.88\u0026ndash;1.73 (m, 3H), 1.71\u0026ndash;1.61 (m, 3H), 1.60\u0026ndash;1.53 (m, 2H), 1.52\u0026ndash;1.41 (m, 2H), 1.42\u0026ndash;1.27 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, DMSO) δ 171.53, 169.06, 155.50, 149.07, 136.35, 133.80, 131.78, 130.77, 128.88, 126.18, 125.69, 125.46, 124.19, 123.65, 123.31, 117.56, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eBrN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 591.1733, found 592.2801. Anal: C, 62.84; H, 5.78; Br, 13.49; N, 7.09; O, 10.80.\u003c/p\u003e \u003cp\u003e \u003cem\u003eN-(2-oxo-2H-chromen-3-yl)-4-(2-(phenylsulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butanamide\u003c/em\u003e \u003cb\u003e(13a)\u003c/b\u003e Yield 51%; Mp 199\u0026ndash;201\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, DMSO) δ 9.33 (s, 1H, NH), 7.92 (s, 1H, CH), 7.89 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;5.8, 2.5 Hz, 1H, ArH), 7.68 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.3, 2.8 Hz, 1H, ArH), 7.64 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;9.3, 3.2 Hz, 3H, Ar-3H), 7.59\u0026ndash;7.52 (m, 1H, ArH), 7.38\u0026ndash;7.30 (m, 2H, Ar-2H), 3.33 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 13.8 Hz, 1H, CH), 2.93 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.9, 13.7 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.49 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;20.3, 6.2 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 20.9, 10.8 Hz, 2H), 2.20\u0026ndash;2.09 (m, 1H), 2.05 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.6 Hz, 2H), 1.87\u0026ndash;1.77 (m, 4H), 1.65 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;21.6 Hz, 2H), 1.52\u0026ndash;1.47 (m, 2H), 1.46\u0026ndash;1.42 (m, 2H), 1.41\u0026ndash;1.35 (m, 2H), 1.36\u0026ndash;1.27 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, DMSO) δ 171.53, 155.50, 149.07, 138.88, 134.48, 129.76, 129.05, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e35\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u0026thinsp;+\u0026thinsp;m/z: 549.2297, found 549.2999. Anal: C, 65.55; H, 6.42; N, 7.64; O, 14.55; S, 5.83.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-benzoyldecahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(14a)\u003c/b\u003e Yield 61%; Mp 196\u0026ndash;199\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.25, (s, 1H, NH), 7.97 (d, J\u0026thinsp;=\u0026thinsp;3.3 Hz, 1H, ArH), 7.96 (s, 1H, Ar-1H), 7.93(dd, J\u0026thinsp;=\u0026thinsp;3.3, 1.7 Hz,1H, CH ), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.6, 3.1 Hz, 1H, ArH), 7.60 (ddd, J\u0026thinsp;=\u0026thinsp;8.2, 7.3, 4.0 Hz, 1H, ArH), 7.57\u0026ndash;7.55 (m, 1H, Ar-1H), 7.52 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.9, 2.7 Hz, 1H, Ar-1H), 7.38\u0026ndash;7.30 (m, 2H), 3.44 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.5 Hz, 1H, CH), 3.08\u0026ndash;2.94 (m, 2H), 2.51 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 2H), 2.41 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 10.6 Hz, 2H), 2.14 (ddd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;17.0, 11.7, 1.5 Hz, 1H), 2.05 (t, J\u0026thinsp;=\u0026thinsp;12.2 Hz, 2H), 2.00\u0026ndash;1.88 (m, 3H), 1.80\u0026ndash;1.71 (m, 3H), 1.69\u0026ndash;1.63 (m, 2H), 1.58 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;11.9, 8.7 Hz, 2H), 1.50 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;20.9, 10.2 Hz, 1H), 1.45\u0026ndash;1.40 (m, 1H), 1.39\u0026ndash;1.28 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 170.91, 155.50, 149.07, 135.71, 130.18, 128.88, 128.26, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e35\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 513.2628, found 513.1701. Anal: C, 72.49; H, 6.87; N, 8.18; O, 12.46.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(4-cyanobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(6-nitro-2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(15a)\u003c/b\u003e Yield 53%; Mp 198\u0026ndash;200\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.14 (s, 1H, NH), 8.58 (d, J\u0026thinsp;=\u0026thinsp;3.1 Hz, 1H, ArH), 8.25 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, ArH), 8.21 (dd, J\u0026thinsp;=\u0026thinsp;6.1, 2.7 Hz, 1H, ArH), 8.18\u0026ndash;8.17 (m, 1H), 8.09 (dd, J\u0026thinsp;=\u0026thinsp;15.5, 2.8 Hz, 2H, ArH), 8.01 (s, 1H, CH), 7.62 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 3.66 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 11.4 Hz, 1H), 3.50 (dt, J\u0026thinsp;=\u0026thinsp;16.8, 13.6 Hz, 1H), 2.51 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.0 Hz, 2H), 2.48 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.9 Hz, 1H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 20.9, 10.8 Hz, 2H), 2.17\u0026ndash;1.96 (m, 4H), 1.86 (s, 1H), 1.78\u0026ndash;1.67 (m, 1H), 1.68\u0026ndash;1.59 (m, 4H), 1.61\u0026ndash;1.51 (m, 2H), 1.50\u0026ndash;1.42 (m, 2H), 1.41\u0026ndash;1.33 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 170.91, 155.50, 154.56, 144.74, 139.16, 132.92, 129.44, 126.05, 125.20, 124.51, 121.90, 118.94, 117.69, 114.76, 113.24, 67.56, 58.99, 53.21, 47.57, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e32\u003c/sub\u003eH\u003csub\u003e33\u003c/sub\u003eN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 583.2431, found 584.2520. Anal: C, 65.85; H, 5.70; N, 12.00; O, 16.45.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)-N-(6-chloro-2-oxo-2H-chromen-3-yl)butanamide\u003c/em\u003e \u003cb\u003e(16a)\u003c/b\u003e Yield 49%; Mp 195\u0026ndash;198\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.28 (s, 1H, NH), 8.00 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz,, 1H, ArH), 7.98 (s, 1H, CH), 7.67\u0026ndash;7.57 (m, 4H, Ar-4H), 7.38 (dd, J\u0026thinsp;=\u0026thinsp;14.8, 2.9 Hz, 1H, ArH), 7.28 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 3.32 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 13.8 Hz, 1H, CH), 2.91 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 13.7 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.9 Hz, 2H), 2.49 (t, J\u0026thinsp;=\u0026thinsp;10.8 Hz, 1H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 10.7 Hz, 2H), 2.18\u0026ndash;2.09 (m, 1H), 2.05 (td, J\u0026thinsp;=\u0026thinsp;10.7, 0.6 Hz, 2H), 1.87\u0026ndash;1.76 (m, 2H), 1.76\u0026ndash;1.66 (m, 2H), 1.60 (d, J\u0026thinsp;=\u0026thinsp;2.1 Hz, 1H), 1.56\u0026ndash;1.50 (m, 2H), 1.56\u0026ndash;1.50 (m, 2H), 1.49\u0026ndash;1.45 (m, 3H), 1.41\u0026ndash;1.34 (m, 3H), 1.33 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.31\u0026ndash;1.23 (m, 1H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 155.50, 154.40, 148.29, 133.92, 132.01, 128.79, 128.32, 126.77, 124.51, 123.35, 122.38, 118.80, 113.24, 67.56, 59.57, 53.21, 48.99, 41.06, 37.70, 36.19, 34.58, 31.36, 29.06, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eClN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 639.2534, found 640.3938. Anal: C, 63.78; H, 6.61; Cl, 5.54; N, 6.56; O, 12.49; S, 5.01.\u003c/p\u003e \u003cp\u003e \u003cem\u003eN-(6-chloro-2-oxo-2H-chromen-3-yl)-4-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butanamide\u003c/em\u003e \u003cb\u003e(17a)\u003c/b\u003e Yield 44%; Mp 187\u0026ndash;189\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.28 (s, 1H, NH), 8.12 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 9.9 Hz, 2H, Ar-2H), 8.02 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, ArH), 8.00 (s, 1H, CH), 7.40 (q, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3.2 Hz, 1H, ArH), 7.36 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.9, 6.0 Hz, 2H, Ar-2H), 7.29 (d, J\u0026thinsp;=\u0026thinsp;14.9 Hz, 1H, ArH), 3.40 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.5 Hz, 1H, CH), 3.11\u0026ndash;2.94 (m, 2H), 2.51 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.7, 10.7 Hz, 2H), 2.41 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 10.6 Hz, 2H), 2.19\u0026ndash;2.10 (m, 1H), 2.05 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15.8, 9.1 Hz, 3H), 2.00\u0026ndash;1.86 (m, 1H), 1.81\u0026ndash;1.71 (m, 3H), 1.72\u0026ndash;1.62 (m, 3H), 1.62\u0026ndash;1.46 (m, 3H), 1.46\u0026ndash;1.39 (m, 2H), 1.39\u0026ndash;1.29 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 173.30, 154.16, 148.29, 133.61, 132.01, 131.63, 126.77, 124.51, 123.35, 122.38, 118.80, 115.98, 113.24, 67.56, 58.99 (s), 53.21 (s), 47.57 (s), 40.85 (s), 37.70 (s), 36.88 (s), 30.89 (s), 28.28 (s), 26.28 (s), 23.44 (s), 23.25 (s), 22.61 (s). HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e33\u003c/sub\u003eClFN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u0026thinsp;+\u0026thinsp;m/z: 565.2144, found 566.2206. Anal: C, 65.78; H, 5.88; Cl, 6.26; F, 3.36; N, 7.42; O, 11.31.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003e4-(decahydro-1H,4H-pyrido[3,2,1-ij][, ]naphthyridin-1-yl)-N-(6-methyl-2-oxo-2H-chromen-3-yl)butanamide\u003c/h3\u003e\n\u003cp\u003e \u003cb\u003e(18a)\u003c/b\u003e Yield 39%; Mp 200\u0026ndash;202\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.24 (s, 1H, NH), 7.97 (s, 1H, CH), 7.52 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, ArH), 7.31 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 7.22 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 2.9 Hz, 1H, ArH), 3.01 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.9 Hz, 1H, CH), 2.51 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.7, 10.7 Hz, 2H), 2.42 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e), 2.41 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 10.6 Hz, 2H), 2.11 (ddd, J\u0026thinsp;=\u0026thinsp;15.9, 13.8, 8.0 Hz, 2H), 2.03 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;16.4, 9.6 Hz, 2H), 1.82\u0026ndash;1.68 (m, 5H), 1.56\u0026ndash;1.49 (m, 2H), 1.46 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;5.5 Hz, 3H), 1.43\u0026ndash;1.40 (m, 2H), 1.39\u0026ndash;1.31 (m, 1H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 155.50, 148.30, 134.77, 127.63, 125.71, 124.51, 121.57, 117.43, 113.24, 68.36, 58.08, 53.21, 47.89, 41.90, 37.70, 36.11, 32.24, 28.36, 26.36, 23.44, 23.25, 21.56, 21.23. HRMS (ESI): Calcd. C\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e33\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 423.2522, found 423.3215. Anal: C, 70.89; H, 7.85; N, 9.92; O, 11.33.\u003c/p\u003e \u003cp\u003e \u003cem\u003etert-butyl 1-(4-oxo-4-((2-oxo-2H-chromen-3-yl)amino)butyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e \u003cb\u003e(19a)\u003c/b\u003eYield 47%; Light yellow oil. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.30 (s, 1H, NH), 7.96 (s, 1H, CH), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.6, 3.1 Hz, 1H, ArH), 7.59\u0026ndash;7.52 (m, 1H, ArH), 7.34 (td, J\u0026thinsp;=\u0026thinsp;14.9, 7.6 Hz, J\u0026thinsp;=\u0026thinsp;10.0 Hz, 2H, Ar-2H), 4.30 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 15.2 Hz, 1H), 3.21 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.8, 15.2 Hz, 1H), 2.83 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;21.6, 8.0 Hz, 1H), 2.57\u0026ndash;2.46 (m, 2H), 2.46\u0026ndash;2.35 (m, 2H), 2.24\u0026ndash;2.10 (m, 2H), 2.10\u0026ndash;1.98 (m, 3H), 1.93\u0026ndash;1.78 (m, 3H), 1.68\u0026ndash;1.59 (m, 3H), 1.60\u0026ndash;1.50 (m, 2H), 1.51\u0026ndash;1.43 (m, 2H), 1.42 (s, 9H), 1.40\u0026ndash;1.25 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 157.65, 155.50, 149.07, 128.88, 125.69, 125.46, 124.19, 123.65, 117.56, 114.16, 81.20, 67.56, 54.76, 53.21, 46.17, 40.85, 37.70, 36.88, 30.89, 28.31, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e39\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 509.2890, found 509.8866. Anal: C, 68.35; H, 7.71; N, 8.25; O, 15.70.\u003c/p\u003e \u003cp\u003e \u003cem\u003e(9H-fluoren-9-yl)methyl1-(4-((5-chloro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e\u003cb\u003e(20a)\u003c/b\u003eYield 52%; Mp 196\u0026ndash;199\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.29 (s, 1H, NH), 8.27 (s, 1H, CH), 7.90 (dd, J\u0026thinsp;=\u0026thinsp;14.7, 3.2 Hz, 2H, Ar-2H), 7.57 (t, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 7.41 (dd, J\u0026thinsp;=\u0026thinsp;14.6, 3.4 Hz, 2H, Ar-2H), 7.34 (td, J\u0026thinsp;=\u0026thinsp;14.9, 3.4 Hz, 2H, Ar-2H),7.26 (dd, J\u0026thinsp;=\u0026thinsp;7.1, 3.1 Hz, 1H, ArH), 7.24 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3.1 Hz, 2H, Ar-2H), 7.17 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 3.0 Hz, 1H, ArH), 5.31 (d, J\u0026thinsp;=\u0026thinsp;11.8 Hz, 2H, CH2), 4.46 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.3 Hz, 1H, CH), 3.99 (t, J\u0026thinsp;=\u0026thinsp;11.8 Hz, 1H), 3.37 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.4 Hz, 1H), 2.82 (dt, J\u0026thinsp;=\u0026thinsp;21.6, 8.1 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.28\u0026ndash;2.13 (m, 2H), 2.07\u0026ndash;1.98 (m, 3H), 1.94\u0026ndash;1.79 (m, 3H), 1.71\u0026ndash;1.56 (m, 3H), 1.55\u0026ndash;1.53 (m, 2H), 1.55\u0026ndash;1.39 (m, 3H), 1.38\u0026ndash;1.24 (m, 1H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53 (s), 157.88 (s), 155.50 (s), 149.92 (s), 143.41 (s), 139.81 (s), 128.10 (s), 127.80 (s), 127.52 (s), 126.50\u0026ndash;126.07 (m), 125.07 (s), 120.89 (s), 117.47 (s), 116.92 (s), 116.65 (s), 67.56 (s), 67.18 (s), 54.76 (s), 53.21 (s), 48.95 (s), 46.17 (s), 40.85 (s), 37.70 (s), 36.88 (s), 30.89 (s), 28.28 (s), 26.28 (s), 23.44 (s), 23.25 (s), 22.61 (s). HRMS (ESI): Calcd. C\u003csub\u003e39\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eClN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 665.2656, found 666.4602. Anal: C, 70.31; H, 6.05; Cl, 5.32; N, 6.31; O, 12.01.\u003c/p\u003e \u003cp\u003e \u003cem\u003e(9H-fluoren-9-yl)methyl1-(4-((5-fluoro-2-oxo-2H-chromen-3-yl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e \u003cb\u003e(21a)\u003c/b\u003eYield 56%; Mp 195\u0026ndash;198\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 9.25 (s, 1H, NH), 8.06 (s, 1H, CH), 7.90 (dd, J\u0026thinsp;=\u0026thinsp;14.7, 3.2 Hz, 2H, ArH), 7.61 (td, J\u0026thinsp;=\u0026thinsp;15.0, 10.0 Hz, 1H, ArH), 7.41 (dt, J\u0026thinsp;=\u0026thinsp;12.1, 6.0 Hz, 2H, ArH), 7.33 (dd, J\u0026thinsp;=\u0026thinsp;14.8, 3.4 Hz, 2H, ArH), 7.24 (td, J\u0026thinsp;=\u0026thinsp;14.8, 3.2 Hz, 2H, ArH), 7.13 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 3.0 Hz, 1H, ArH), 6.91 (ddd, J\u0026thinsp;=\u0026thinsp;15.9, 14.9, 2.9 Hz, 1H, ArH), 5.31 (d, J\u0026thinsp;=\u0026thinsp;11.8 Hz, 2H, CH\u003csub\u003e2\u003c/sub\u003e), 4.46 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.4 Hz, 1H, CH), 3.99 (t, J\u0026thinsp;=\u0026thinsp;11.8 Hz, 1H, CH), 3.37 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.3 Hz, 1H), 2.82 (dt, J\u0026thinsp;=\u0026thinsp;21.4, 8.0 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.24\u0026ndash;2.10 (m, 2H), 2.10\u0026ndash;2.00 (m, 3H), 1.94\u0026ndash;1.78 (m, 3H), 1.72\u0026ndash;1.57 (m, 3H), 1.57\u0026ndash;1.49 (m, 2H), 1.48\u0026ndash;1.42 (m, 2H), 1.40\u0026ndash;1.23 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 174.53, 157.88, 155.50, 153.38, 149.70, 143.41, 139.81, 127.52, 126.18, 125.07, 120.89, 116.42, 115.03, 112.00, 110.97, 67.56, 67.18, 54.76, 53.21, 48.95, 46.17, 40.85, 37.70, 36.88, 30.89, 28.28, 26.28, 23.44, 23.25, 22.61. HRMS (ESI): Calcd. C\u003csub\u003e39\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eFN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 649.2952, found 650.7940. Anal: C, 72.09; H, 6.21; F, 2.92; N, 6.47; O, 12.31.\u003c/p\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003e6.1.2. Synthesis of compound (1b-7b)\u003c/h2\u003e \u003cp\u003eThe solution of Matrinic alcohol derivatives \u003cb\u003e(6,9,10)\u003c/b\u003e (1.0 eq) and coumarin-3-carboxylic acids derivatives \u003cb\u003e14 (\u003c/b\u003e2.0 eq) in 10 mL of anhydrous dichloromethane (DCM) was cooled to 0\u0026deg;C. DCC (77 mg, 1.5 eq) and pyridine (0.168 mL 2.0 eq) were added to this, and a catalytic amount of DMAP (0.1 eq). The reaction mixture was stirred overnight at room temperature diluted with DCM and washed with water and brine solution. The dried organic layer on column chromatography yielded the desired product [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(4-bromobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e \u003cb\u003e(1b)\u003c/b\u003e Yield 58%; Light yellow oil. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.55 (s, 1H, CH), 7.96 (d, J\u0026thinsp;=\u0026thinsp;2.5 Hz, 2H, Ar-2H), 7.93 (d, J\u0026thinsp;=\u0026thinsp;2.3 Hz, 2H), 7.69 (d, J\u0026thinsp;=\u0026thinsp;2.5 Hz, 1H), 7.67 (dd, J\u0026thinsp;=\u0026thinsp;4.2, 2.4 Hz, 1H, ArH), 7.65 (dd, J\u0026thinsp;=\u0026thinsp;3.8, 3.2 Hz, ArH, 1H), 7.39\u0026ndash;7.28 (m, 3H, Ar-3H), 3.97 (t, J\u0026thinsp;=\u0026thinsp;9.9 Hz, 2H), 3.36 (dt, J\u0026thinsp;=\u0026thinsp;21.6, 9.2 Hz, 1H), 3.18 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.7 Hz, 1H), 2.65 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.7 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.7 Hz, 2H), 2.16 (t, J\u0026thinsp;=\u0026thinsp;58.8 Hz, 1H), 2.08\u0026ndash;1.99 (m, 1H), 1.84\u0026ndash;1.70 (m, 2H), 1.71\u0026ndash;1.67 (m, 3H), 1.64 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.3 Hz, 2H), 1.62\u0026ndash;1.54 (m, 2H), 1.43 (ddd, J\u0026thinsp;=\u0026thinsp;19.2 Hz, 2H), 1.35 (dt, J\u0026thinsp;=\u0026thinsp;23.5, 7.5 Hz, 2H), 1.30\u0026ndash;1.19 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 170.91, 163.31, 160.42, 152.43, 136.97, 132.91, 131.51, 131.20, 130.81, 128.55, 125.32, 124.45, 122.09, 120.12, 117.43, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e32\u003c/sub\u003eH\u003csub\u003e35\u003c/sub\u003eBrN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 606.1729, found 606.3915. Anal: C, 63.26; H, 5.81; Br, 13.15; N, 4.61; O, 13.17.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(2-naphthoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e\u003cb\u003e(2b)\u003c/b\u003eYield 57%; Light yellow oil \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.55 (s, 1H, CH), 8.40 (t, J\u0026thinsp;=\u0026thinsp;1.4 Hz, 1H, ArH), 8.15 (dd, J\u0026thinsp;=\u0026thinsp;7.5, 1.4 Hz, 1H, ArH), 7.90 (ddt, J\u0026thinsp;=\u0026thinsp;6.7, 4.9, 1.5 Hz, 2H, Ar-2H), 7.83 (dd, J\u0026thinsp;=\u0026thinsp;7.5, 1.6 Hz, 1H, ArH), 7.68 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.5, 1.4 Hz, 1H, ArH), 7.64 (dd, J\u0026thinsp;=\u0026thinsp;7.4, 1.5 Hz, 1H, ArH), 7.60 (td, J\u0026thinsp;=\u0026thinsp;7.4, 1.6 Hz, 1H, ArH), 7.56 (ddd, J\u0026thinsp;=\u0026thinsp;9.0, 8.4, 1.6 Hz, 1H, ArH), 7.36 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.4, 1.2 Hz, 1H, ArH), 7.34 (dd, J\u0026thinsp;=\u0026thinsp;10.0 Hz, 1H, ArH), 3.97 (t, J\u0026thinsp;=\u0026thinsp;7.5 Hz, 2H), 3.49 (dd, J\u0026thinsp;=\u0026thinsp;12.3, 7.4 Hz, 1H), 3.02 (dd, J\u0026thinsp;=\u0026thinsp;12.5, 7.3 Hz, 1H), 2.72 (dt, J\u0026thinsp;=\u0026thinsp;11.0, 4.1 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;12.3, 5.4 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;12.4, 5.4 Hz, 2H), 2.27\u0026ndash;2.16 (m, 1H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;11.2 Hz, 1H), 1.96 (ddt, J\u0026thinsp;=\u0026thinsp;12.0, 8.2, 5.9 Hz, 1H), 1.80\u0026ndash;1.71 (m, 2H), 1.66 (ddd, J\u0026thinsp;=\u0026thinsp;11.8, 7.2, 3.5 Hz, 2H), 1.63\u0026ndash;1.59 (m, 2H), 1.53 (dq, J\u0026thinsp;=\u0026thinsp;11.1, 5.6 Hz, 2H), 1.43 (tt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;10.2, 5.2 Hz, 2H), 1.39\u0026ndash;1.30 (m, 2H), 1.25 (p, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.8 Hz, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 169.44, 163.31, 160.42, 152.43, 134.48, 134.11, 132.91, 131.20, 129.46, 128.60, 127.71, 127.31, 126.64, 125.32, 122.09, 120.12, 118.49, 117.43, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e36\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 578.2781, found 579.1513. Anal: C, 74.72; H, 6.62; N, 4.84; O, 13.82.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(2-methylbenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl-5-fluoro-2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e\u003cb\u003e(3b)\u003c/b\u003eYield 49%; Mp 201\u0026ndash;203\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.55 (s, 1H, CH), 7.76 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 2.9 Hz, 1H, ArH), 7.61 (td, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15.0, 10.0 Hz, 1H, ArH), 7.34 (td, J\u0026thinsp;=\u0026thinsp;14.8, 2.9 Hz, 1H, ArH), 7.24 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 3.3 Hz, 1H, ArH), 7.11 (ddd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;18.0, 10.6, 3.1 Hz, Ar-2H), 6.94 (ddd, J\u0026thinsp;=\u0026thinsp;15.8, 15.2, 3.0 Hz, 1H, ArH), 3.97 (t, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 2H), 3.33 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.9 Hz, 1H), 2.87 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.8 Hz, 1H), 2.69 (dt, J\u0026thinsp;=\u0026thinsp;21.4, 8.2 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.22 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.16\u0026ndash;2.02 (m, 2H), 2.00\u0026ndash;1.84 (m, 2H), 1.81\u0026ndash;1.71 (m, 1H), 1.73\u0026ndash;1.66 (m, 4H), 1.66\u0026ndash;1.60 (m, 2H), 1.48\u0026ndash;1.35 (m, 2H), 1.33\u0026ndash;1.27 (m, 2H), 1.26\u0026ndash;1.18 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 171.46, 164.39, 160.30, 153.40, 138.49, 136.46, 130.96, 130.26, 130.06, 128.72, 125.57, 124.80, 115.39, 110.48, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25, 20.26. HRMS (ESI): Calcd. C\u003csub\u003e33\u003c/sub\u003eH\u003csub\u003e37\u003c/sub\u003eFN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 560.2687, found 560.2420. Anal: C, 70.69; H, 6.65; F, 3.39; N, 5.00; O, 14.27.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(phenylsulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl-5-chloro-2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e\u003cb\u003e(4b)\u003c/b\u003eYield 59%; Mp 199\u0026ndash;201\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.52 (s, 1H, CH), 7.85 (dd, J\u0026thinsp;=\u0026thinsp;6.6, 2.8 Hz, 2H, ArH), 7.63\u0026ndash;7.59 (m, 3H, Ar-3H), 7.55 (dd, J\u0026thinsp;=\u0026thinsp;15.5, 14.3 Hz, 1H, ArH), 7.21 (d, J\u0026thinsp;=\u0026thinsp;1.7 Hz, 2H, Ar-2H), 3.96 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.8 Hz, 2H), 3.76 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.6 Hz, 1H), 3.33 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.6 Hz, 1H), 2.96 (dt, J\u0026thinsp;=\u0026thinsp;21.8, 12.5 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 3H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 3H), 1.85\u0026ndash;1.72 (m, 2H), 1.65 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;22.3 Hz, 1H), 1.59\u0026ndash;1.51 (m, 4H), 1.50\u0026ndash;1.42 (m, 4H), 1.43\u0026ndash;1.35 (m, 2H), 1.34\u0026ndash;1.19 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 163.31, 160.42, 152.74, 138.88, 138.69, 134.48, 131.17, 129.74, 129.05, 126.27, 125.07, 117.40, 116.61, 67.56, 66.85, 59.57, 53.21, 48.99, 41.06, 36.19, 31.85, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e35\u003c/sub\u003eClN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 598.1904, found 599.1418. Anal: C, 62.15; H, 5.89; Cl, 5.92; N, 4.68; O, 16.02; S, 5.35.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(2-methylbenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl-2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e\u003cb\u003e(5b)\u003c/b\u003eYield 67%; Mp 200\u0026ndash;202\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.53 (s, 1H, CH), 7.74 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 3.0 Hz, 1H, ArH), 7.66 (dd, J\u0026thinsp;=\u0026thinsp;14.3, 2.8 Hz, 1H, ArH), 7.57\u0026ndash;7.50 (m, 1H, ArH), 7.38\u0026ndash;7.31 (m, 2H, Ar-2H), 7.29 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.8 Hz, 1H), 7.22 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 3.4 Hz, 1H, ArH), 7.09 (td, J\u0026thinsp;=\u0026thinsp;14.7, 3.3 Hz, 1H, ArH), 3.96 (t, J\u0026thinsp;=\u0026thinsp;14.9 Hz, 2H), 3.35 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.7 Hz,1H), 2.83 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.7 Hz, 1H), 2.62 (dt, J\u0026thinsp;=\u0026thinsp;21.8, 8.0 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.21 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.17\u0026ndash;2.01 (m, 2H), 2.01\u0026ndash;1.88 (m, 1H), 1.79\u0026ndash;1.70 (m, 1H), 1.69\u0026ndash;1.60 (m, 5H), 1.58\u0026ndash;1.49 (m, 2H), 1.49\u0026ndash;1.42 (m, 2H), 1.41\u0026ndash;1.29 (m, 2H), 1.30\u0026ndash;1.21 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 171.46, 163.31, 160.42, 152.43, 136.46, 132.91, 131.20, 130.96, 130.26, 128.72, 128.55, 125.57, 125.32, 122.09, 120.12, 117.43, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25, 20.26. HRMS (ESI): Calcd. C\u003csub\u003e33\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 542.2781, found 542.2996. Anal: C, 73.04; H, 7.06; N, 5.16; O, 14.74.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-(2-methylbenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl-6-methyl-2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e\u003cb\u003e(6b)\u003c/b\u003eYield 48%; Mp 200\u0026ndash;203\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.55 (s, 1H, CH), 7.76 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 2.9 Hz, 1H, ArH), 7.52 (d, J\u0026thinsp;=\u0026thinsp;2.7 Hz, 1H, ArH), 7.36 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 2.5 Hz, 2H, ArH), 7.31 (dd, J\u0026thinsp;=\u0026thinsp;9.0, 6.0 Hz, 1H, ArH),, 7.24 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 3.3 Hz, 1H, ArH), 7.11 (td, J\u0026thinsp;=\u0026thinsp;14.7, 3.3 Hz, 1H, ArH), 3.97 (t, J\u0026thinsp;=\u0026thinsp;14.9 Hz, 2H), 3.35 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.8 Hz, 1H), 2.84 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.8 Hz, 1H), 2.63 (dt, J\u0026thinsp;=\u0026thinsp;21.8, 8.0 Hz, 1H), 2.52\u0026ndash;2.44 (m, 3H), 2.42 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.39 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;17.8, 7.0 Hz, 1H), 2.22 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.13\u0026ndash;2.02 (m, 2H), 2.01\u0026ndash;1.89 (m, 1H), 1.79\u0026ndash;1.71 (m, 3H), 1.70\u0026ndash;1.63 (m, 4H), 1.48\u0026ndash;1.42 (m, 3H), 1.41\u0026ndash;1.37 (m, 2H), 1.29\u0026ndash;1.21 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 171.46, 163.31, 160.42, 151.23, 136.46, 135.61, 131.32, 130.89, 130.26, 128.72, 128.18, 125.57, 123.00, 120.49, 117.16, 67.56, 66.85, 58.99, 53.21, 47.57, 40.85, 36.88, 32.94, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25, 21.23, 20.26. HRMS (ESI): Calcd. C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 556.2937, found 557.4050. Anal: C, 73.36; H, 7.24; N, 5.03; O, 14.37.\u003c/p\u003e \u003cp\u003e \u003cem\u003e4-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)butyl-5-bromo-2-oxo-2H-chromene-3-carboxylate\u003c/em\u003e\u003cb\u003e(7b)\u003c/b\u003eYield 51%; Mp 195\u0026ndash;198\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.55 (s, 1H, CH), 7.67\u0026ndash;7.60 (m, 4H, Ar-4H), 7.52 (t, J\u0026thinsp;=\u0026thinsp;14.8 Hz, 1H, ArH), 7.37 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 3.3 Hz, 1H, ArH), 7.30 (dd, J\u0026thinsp;=\u0026thinsp;14.7, 3.2 Hz, 1H, ArH), 3.97 (t, J\u0026thinsp;=\u0026thinsp;14.9 Hz, 2H), 3.88 (dd, J\u0026thinsp;=\u0026thinsp;24.1, 14.0 Hz, 1H), 3.33 (dd, J\u0026thinsp;=\u0026thinsp;24.4, 14.3 Hz, 1H), 3.05 (dt, J\u0026thinsp;=\u0026thinsp;21.4, 13.6 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;50.0 Hz, 2H), 1.87\u0026ndash;1.79 (m, 2H), 1.69\u0026ndash;1.60 (m, 4H), 1.54\u0026ndash;1.49 (m, 3H), 1.47\u0026ndash;1.41 (m, 4H), 1.41\u0026ndash;1.34 (m, 2H), 1.33 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.32\u0026ndash;1.18 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 163.31, 160.42, 154.40, 151.68, 133.92, 133.24, 131.14, 128.86, 128.32, 123.66, 118.81, 118.30, 117.05, 67.56, 66.85, 59.57, 53.21, 48.99, 41.06, 36.19, 34.58, 31.85, 31.36, 29.77, 28.28, 26.28, 24.17, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e35\u003c/sub\u003eH\u003csub\u003e43\u003c/sub\u003eBrN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 698.2025, found 699.4138. Anal: C, 60.08; H, 6.19; Br, 11.42; N, 4.00; O, 13.72; S, 4.58.\u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003e6.1.3. Synthesis of compound (1c-11c)\u003c/h2\u003e \u003cp\u003eA solution containing anhydrous potassium acetate (CH\u003csub\u003e3\u003c/sub\u003eCO\u003csub\u003e2\u003c/sub\u003eK, 2.94 mmol) the conveniently substituted matrinic acid derivatives (\u003cb\u003e5,7,8\u003c/b\u003e) (1.67 mmol) and the corresponding 2-hydroxybenzaldehyde (1.67 mmol) in acetic anhydride (Ac\u003csub\u003e2\u003c/sub\u003eO, 10 mL) was refluxed (138\u0026deg;C) for 16 h. The reaction mixture was cooled, neutralized with 10% aqueous sodium bicarbonate (NaHCO\u003csub\u003e3\u003c/sub\u003e), and extracted (3 \u0026times; 30 mL) with ethyl acetate (EtOAc). The organic layers were combined and washed with distilled water dried with anhydrous sodium sulfate (Na\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e) and evaporated under reduced pressure. The product was purified by recrystallization in ethanol (EtOH) and dried to afford the desired compound [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-6-nitro-2H-chromen-2-one\u003c/em\u003e \u003cb\u003e(1c)\u003c/b\u003eYield 53%; Mp 173\u0026ndash;174\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.76 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H), 8.28 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 2.9 Hz, 1H, ArH), 8.22 (s, 1H, ArH), 8.12 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.9, 10.0 Hz, 2H, Ar-2H), 7.62 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 7.42\u0026ndash;7.32 (m, 2H, Ar-2H), 3.72 (td, J\u0026thinsp;=\u0026thinsp;13.4, 12.0 Hz, 1H), 3.49 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.3 Hz, 1H), 2.64 (td, J\u0026thinsp;=\u0026thinsp;11.6, 2.0 Hz, 2H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 15.4, 12.6 Hz, 3H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 20.9, 10.0 Hz, 2H), 2.22\u0026ndash;2.10 (m, 1H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;21.7 Hz, 1H), 1.91\u0026ndash;1.75 (m, 4H), 1.72 (dt, J\u0026thinsp;=\u0026thinsp;13.4, 7.2 Hz, 2H), 1.53 (dd, J\u0026thinsp;=\u0026thinsp;12.8, 11.4 Hz, 2H), 1.50\u0026ndash;1.38 (m, 2H), 1.39\u0026ndash;1.20 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 167.32, 160.95, 156.78, 145.89, 144.19, 133.61, 131.63, 127.46, 127.16, 126.86, 122.40, 117.90, 115.98, 67.56, 58.76, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eFN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 519.2169, found 519.3663. Anal: C, 67.04; H, 5.82; F, 3.66; N, 8.09; O, 15.40.\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-6-nitro-2H-chromen-2-one\u003c/em\u003e \u003cb\u003e(2c)\u003c/b\u003eYield 59%; Mp 175\u0026ndash;177\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.56 (s, 1H, CH), 8.28 (dd, J\u0026thinsp;=\u0026thinsp;14.9, 3.0 Hz, 1H, ArH), 7.68\u0026ndash;7.59 (m, 6H, Ar-6H), 3.62 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.7 Hz, 1H), 3.20 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.7 Hz, 1H), 3.09 (dt, J\u0026thinsp;=\u0026thinsp;21.4, 12.4 Hz, 1H), 2.63\u0026ndash;2.55 (m, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 15.4, 12.6 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 20.9, 10.0 Hz, 2H), 2.39\u0026ndash;2.32 (m, 1H), 2.26 (ddd, J\u0026thinsp;=\u0026thinsp;12.5, 1.9 Hz, 2H), 1.90\u0026ndash;1.72 (m, 2H), 1.63 (d, J\u0026thinsp;=\u0026thinsp;22.3 Hz, 1H), 1.59\u0026ndash;1.52 (m, 4H), 1.46\u0026ndash;1.41 (m, 1H), 1.39\u0026ndash;1.34 (m, 2H), 1.33 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e ), 1.31\u0026ndash;1.21 (m, 1H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.95, 156.78, 154.40, 145.89, 144.19, 133.92, 128.79, 128.32, 127.46, 127.16, 126.86, 122.40, 117.90, 67.56, 57.82, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e32\u003c/sub\u003eH\u003csub\u003e39\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 593.2560, found 593.1752. Anal: C, 64.73; H, 6.62; N, 7.08; O, 16.17; S, 5.40.\u003c/p\u003e \u003cp\u003e3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij][\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]naphthyridin-1-yl)ethyl)-8-methoxy-2H-chromen-2-one(\u003cb\u003e3c\u003c/b\u003e)Yield 45%; Mp 170\u0026ndash;173\u0026deg;C. 1H NMR (500 MHz, CDCl3) δ 7.65 (t, J\u0026thinsp;=\u0026thinsp;10.6 Hz, 4H), 7.42 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H), 7.36 (dd, J\u0026thinsp;=\u0026thinsp;15.9, 5.3 Hz, 1H), 7.25 (s, 1H), 7.23 (dd, J\u0026thinsp;=\u0026thinsp;12.8, 3.6 Hz, 1H), 3.83 (s, 3H, Ar-OCH3), 3.79 (d, J\u0026thinsp;=\u0026thinsp;14.5 Hz, 1H), 3.42 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.6 Hz, 1H), 3.15 (dt, J\u0026thinsp;=\u0026thinsp;21.6, 12.3 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 15.4, 12.6 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 20.9, 10.0 Hz, 2H), 2.11 (td, J\u0026thinsp;=\u0026thinsp;14.2, 0.6 Hz, 2H), 1.86\u0026ndash;1.71 (m, 2H), 1.68\u0026ndash;1.62 (m, 2H), 1.61 (t, J\u0026thinsp;=\u0026thinsp;22.5 Hz, 1H), 1.48 (dd, J\u0026thinsp;=\u0026thinsp;13.7, 8.7 Hz, 6H), 1.39\u0026ndash;1.34 (m, 2H), 1.33 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.48, 154.40, 145.53, 144.99, 143.50, 133.92, 128.79, 128.32, 127.13, 125.21, 122.84, 121.33, 115.54, 67.56, 57.82, 56.83, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e33\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 578.2814, found 578.3625. Anal: C, 68.48; H, 7.31; N, 4.84; O, 13.82; S, 5.54.\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-6-methyl-2H-chromen-2-one\u003c/em\u003e\u003cb\u003e(4c)\u003c/b\u003eYield 59%; Mp 180\u0026ndash;183\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 7.80 (s, 1H, Ar-H), 7.63 (t, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 4H, Ar-4H), 7.52 (d, J\u0026thinsp;=\u0026thinsp;2.5 Hz, 1H, Ar-H), 7.29 (t, J\u0026thinsp;=\u0026thinsp;16.6 Hz, 2H, Ar-2H), 3.36 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 13.9 Hz, 1H), 2.97 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 13.9 Hz, 1H), 2.69 (t, J\u0026thinsp;=\u0026thinsp;8.0 Hz, 1H), 2.69\u0026ndash;2.61 (m, 2H), 2.59\u0026ndash;2.51 (m, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 15.4, 12.6 Hz, 2H), 2.42 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.8, 20.9, 10.0 Hz, 2H), 2.14\u0026ndash;2.02 (m, 1H), 1.92\u0026ndash;1.74 (m, 2H), 1.70\u0026ndash;1.62 (m, 2H), 1.74\u0026ndash;1.63 (m, 4H), 1.48\u0026ndash;1.42 (m, 2H), 1.41\u0026ndash;1.34 (m, 3H), 1.33 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.95, 154.40, 150.53, 145.89, 135.05, 133.92, 129.82, 128.79, 128.32, 127.46, 127.13, 122.25, 116.94, 67.56, 57.82, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25, 21.23. HRMS (ESI): Calcd. C\u003csub\u003e33\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 562.2865, found 563.5492. Anal: C, 70.43; H, 7.52; N, 4.98; O, 11.37; S, 5.70.\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-((4-(tert-butyl)phenyl)sulfonyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-2H-chromen-2-one\u003c/em\u003e\u003cb\u003e(5c)\u003c/b\u003eYield 57%; Mp 176\u0026ndash;178\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 7.68 (s, 1H, ArH), 7.63 (t, J\u0026thinsp;=\u0026thinsp;4.4 Hz, 3H, Ar-3H), 7.61\u0026ndash;7.54 (m, 1H, ArH), 7.53 (tdt, J\u0026thinsp;=\u0026thinsp;24.0 Hz, 2H, Ar-2H), 7.35 (ttd, J\u0026thinsp;=\u0026thinsp;10.0 Hz, 2H, Ar-2H), 3.83 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 14.6 Hz, 1H), 3.69 (dd, J\u0026thinsp;=\u0026thinsp;24.9, 14.5 Hz, 1H), 3.23 (dt, J\u0026thinsp;=\u0026thinsp;21.6, 12.5 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 1H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;10.0, 7.5 Hz, 1H), 2.39 (dt, J\u0026thinsp;=\u0026thinsp;10.0, 7.5 Hz, 2H), 2.10 (td, J\u0026thinsp;=\u0026thinsp;10.7, 1.8 Hz, 2H), 1.87\u0026ndash;1.71 (m, 2H), 1.63 (t, J\u0026thinsp;=\u0026thinsp;22.3 Hz, 1H), 1.54 (ddd, J\u0026thinsp;=\u0026thinsp;10.8, 6.1, 3.1 Hz, 4H), 1.43 (ddd, J\u0026thinsp;=\u0026thinsp;11.7, 7.1, 3.2 Hz, 2H), 1.43\u0026ndash;1.36 (m, 2H), 1.33 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.30\u0026ndash;1.16 (m, 1H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.95, 154.40, 151.59, 146.28, 133.92, 131.84, 128.79, 128.32, 127.52, 126.97, 124.91, 120.68, 117.32, 67.56, 57.82, 53.21, 48.99, 41.06, 36.19, 34.58, 31.36, 29.51, 28.28, 26.67, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e32\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eS, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 548.2709, found 549.2999. Anal: C, 70.04; H, 7.35; N, 5.11; O, 11.66; S, 5.84.\u003c/p\u003e \u003cp\u003e \u003cem\u003etert-butyl-1-(2-(6-methyl-2-oxo-2H-chromen-3-yl)ethyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e\u003cb\u003e(6c)\u003c/b\u003eYield 55%; Light brown viscous oil. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 7.91 (s, 1H, ArH), 7.52 (d, J\u0026thinsp;=\u0026thinsp;2.5 Hz, 1H, ArH), 7.31 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 7.27 (dd, J\u0026thinsp;=\u0026thinsp;19.3 Hz, 1H, ArH), 4.32 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.1 Hz, 1H), 3.24 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 15.2 Hz, 1H), 2.87 (dt, J\u0026thinsp;=\u0026thinsp;32.5 Hz, 3H), 2.80 (td, J\u0026thinsp;=\u0026thinsp;15.6, 2.0 Hz, 2H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 1H), 2.42 (s, 3H, Ar-CH\u003csub\u003e3\u003c/sub\u003e), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;10.0, 7.5 Hz, 1H), 2.26\u0026ndash;2.11 (m, 2H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;21.8 Hz,1H), 1.92\u0026ndash;1.74 (m, 1H), 1.74\u0026ndash;1.63 (m, 3H), 1.61\u0026ndash;1.49 (m, 2H), 1.49\u0026ndash;1.43 (m, 2H), 1.42 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.39\u0026ndash;1.28 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.95, 157.65, 150.53, 145.89, 135.05, 129.82, 127.46, 127.13, 122.25, 116.94, 81.20, 67.56, 54.22, 53.21, 46.17, 40.85, 36.88, 29.51, 28.31, 27.54, 26.28, 23.44, 23.25, 21.23. HRMS (ESI): Calcd. C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 466.2832, found 466.2039. Anal: C, 72.07; H, 8.21; N, 6.00; O, 13.71.\u003c/p\u003e \u003cp\u003e \u003cem\u003e(9H-fluoren-9-yl)methyl 1-(2-(2-oxo-2H-chromen-3-yl)ethyl)octahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridine-2(3H)-carboxylate\u003c/em\u003e\u003cb\u003e(7c)\u003c/b\u003eYield 49%; Mp 166\u0026ndash;169\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 7.90 (dd, J\u0026thinsp;=\u0026thinsp;14.7, 3.2 Hz, 2H, Ar-2H), 7.77 (s, 1H, CH), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.6, 3.1 Hz, 1H, ArH), 7.56 (ddd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;18.5, 10.7, 3.2 Hz, 3H, Ar-3H), 7.37 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3.2 Hz, 1H, ArH), 7.33 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.8, 3.3 Hz, 1H, ArH), 7.24 (td, J\u0026thinsp;=\u0026thinsp;14.9, 3.3 Hz, 2H, Ar-2H), 5.33 (d, J\u0026thinsp;=\u0026thinsp;11.4 Hz, 2H, CH\u003csub\u003e2\u003c/sub\u003e), 4.15 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 16.2 Hz, 1H), 4.05 (t, J\u0026thinsp;=\u0026thinsp;11.4 Hz, 1H), 3.56 (q, J\u0026thinsp;=\u0026thinsp;12.3 Hz, 1H), 3.44 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 16.1 Hz, 1H), 2.71 (td, J\u0026thinsp;=\u0026thinsp;15.8, 1.9 Hz, 2H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.9 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.9 Hz, 2H), 2.28\u0026ndash;2.12 (m, 1H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;6.3 Hz, 2H), 2.13\u0026ndash;1.79 (m, 2H), 1.68 (td, J\u0026thinsp;=\u0026thinsp;15.7, 12.2 Hz, 2H), 1.58\u0026ndash;1.47 (m, 4H), 1.46\u0026ndash;1.33 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.95, 157.88, 151.59, 146.28, 143.41, 139.81, 131.84, 127.52, 126.97, 126.18, 125.07, 124.91, 120.89, 120.68, 117.32, 67.56, 67.18, 54.22, 53.21, 48.95, 46.17, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e37\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 574.2832, found 574.2695. Anal: C, 77.33; H, 6.66; N, 4.87; O, 11.14.\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-2H-chromen-2-one\u003c/em\u003e\u003cb\u003e(8c)\u003c/b\u003eYield 54%; Mp 181\u0026ndash;183\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.17\u0026ndash;8.08 (m, 2H, Ar-2H), 7.73 (s, 1H, CH), 7.68 (dd, J\u0026thinsp;=\u0026thinsp;14.7, 3.0 Hz, 1H, ArH), 7.55 (dtd, J\u0026thinsp;=\u0026thinsp;11.8 Hz, 1H, ArH), 7.42\u0026ndash;7.28 (m, 2H, Ar-2H), 3.39 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H), 3.02 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H), 2.75 (dt, J\u0026thinsp;=\u0026thinsp;22.0, 8.2 Hz, 1H), 2.68 (td, J\u0026thinsp;=\u0026thinsp;15.6, 1.9 Hz, 1H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.6 Hz, 2H), 2.25\u0026ndash;2.09 (m, 2H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;22.0 Hz, 1H), 1.81\u0026ndash;1.58 (m, 4H), 1.60\u0026ndash;1.48 (m, 2H), 1.48\u0026ndash;1.42 (m, 2H), 1.41\u0026ndash; 1.31(m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 170.91, 162.34, 151.59, 146.28, 133.61, 131.84, 131.63, 127.52, 126.97, 124.91, 120.68, 117.32, 115.98, 67.56, 58.76, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e31\u003c/sub\u003eFN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 474.2319, found 475.3319. Analysis: C, 73.40; H, 6.58; F, 4.00; N, 5.90; O, 10.11.\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-(4-fluorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-8-methoxy-2H-chromen-2-one\u003c/em\u003e\u003cb\u003e(9c)\u003c/b\u003eYield 49%; Mp 173\u0026ndash;176\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 8.12 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;14.9, 10.0 Hz, 2H, Ar-2H ), 7.74 (s, 1H, CH), 7.40 (dd, J\u0026thinsp;=\u0026thinsp;4.0, 1.8 Hz, 1H, ArH), 7.37 (dd, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;4.5, 1.7 Hz, 2H, Ar-2H), 7.34 (t, J\u0026thinsp;=\u0026thinsp;3.1 Hz, 1H, ArH ), 7.23 (dd, J\u0026thinsp;=\u0026thinsp;13.6, 4.4 Hz, 1H, ArH), 3.83 (s, 3H, Ar-OCH\u003csub\u003e3\u003c/sub\u003e), 3.39 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H), 3.01 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 14.4 Hz, 1H), 2.75 (dt, J\u0026thinsp;=\u0026thinsp;22.0, 8.2 Hz, 1H), 2.67 (ddd, J\u0026thinsp;=\u0026thinsp;15.6, 1.9 Hz, 2H), 2.51 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.8 Hz, 2H), 2.41 (dt, J\u0026thinsp;=\u0026thinsp;24.7, 10.7 Hz, 2H), 2.25\u0026ndash;2.09 (m, 2H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;22.0 Hz, 1H), 1.83\u0026ndash;1.72 (m, 2H), 1.69 (dt, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.7, 4.7 Hz, 2H), 1.64\u0026ndash;1.51 (m, 2H), 1.48\u0026ndash;1.38 (m, 2H), 1.37\u0026ndash;1.29 (m, 2H).\u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 172.31, 164.23, 163.33, 160.25, 145.53, 144.99, 143.50, 133.61, 131.63, 127.13, 125.21, 122.84, 121.33, 115.98, 115.54, 67.56, 58.76, 56.83, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e33\u003c/sub\u003eFN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 504.2424, found 504.2653. Anal: C, 71.41; H, 6.59; F, 3.77; N, 5.55; O, 12.68.\u003c/p\u003e \u003cp\u003e \u003cem\u003etert-butyl-1-(2-(6-methoxy-2-oxo-2H-chromen-3-yl)ethyl)decahydro-1H-benzo[de]isoquinoline-2(3H)-carboxylate(\u003c/em\u003e \u003cb\u003e10c\u003c/b\u003e \u003cem\u003e)\u003c/em\u003eYield 53%; Light yellow oil; \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 7.92 (s, 1H, CH), 7.85 (d, J\u0026thinsp;=\u0026thinsp;15.0 Hz, 1H, ArH), 7.23 (dd, J\u0026thinsp;=\u0026thinsp;15.0, 2.9 Hz, 1H, ArH), 6.98 (d, J\u0026thinsp;=\u0026thinsp;2.9 Hz, 1H, ArH), 4.32 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.2 Hz, 1H), 3.77 (s, 3H, Ar-OCH\u003csub\u003e3\u003c/sub\u003e), 3.24 (dd, J\u0026thinsp;=\u0026thinsp;24.8, 15.1 Hz, 1H), 2.90\u0026ndash;2.83 (m, 1H), 2.80 (td, J\u0026thinsp;=\u0026thinsp;15.4, 2.0 Hz, 2H), 2.51 (td, J\u0026thinsp;=\u0026thinsp;23.2, 12.5 Hz, 2H), 2.41 (td, J\u0026thinsp;=\u0026thinsp;23.2, 12.5 Hz, 2H), 2.22\u0026ndash;2.09 (m, 2H), 2.04 (t, J\u0026thinsp;=\u0026thinsp;21.8 Hz, 1H), 1.89\u0026ndash;1.72 (m, 1H), 1.72\u0026ndash;1.62 (m, 3H), 1.60\u0026ndash;1.50 (m, 2H), 1.49\u0026ndash;1.44 (m, 2H), 1.42 (s, 9H, C-(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e), 1.39\u0026ndash;1.27 (m, 2H). \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 160.95, 157.65, 156.09, 146.95, 145.89, 127.46, 122.22, 117.51, 116.26, 109.66, 81.20, 67.56, 56.08, 54.22, 53.21, 46.17, 40.85, 36.88, 29.51, 28.31, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 482.2781, found 483.2837. Anal: C, 69.68; H, 7.94; N, 5.80; O, 16.58.\u003c/p\u003e \u003cp\u003e \u003cem\u003e3-(2-(2-(3-chlorobenzoyl)decahydro-1H,4H-pyrido[3,2,1-ij]\u003c/em\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003cem\u003enaphthyridin-1-yl)ethyl)-2H-chromen-2-on\u003c/em\u003e\u003cb\u003e(11c)\u003c/b\u003eYield 59%; Mp 189\u0026ndash;192\u0026deg;C. \u003csup\u003e1\u003c/sup\u003eH NMR (500 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 7.82 (dt, J\u0026thinsp;=\u0026thinsp;12.5, 3.0 Hz, 1H, ArH), 7.78 (t, J\u0026thinsp;=\u0026thinsp;3.2 Hz, 1H, ArH), 7.70 (s, 1H, ArH), 7.67 (dd, J\u0026thinsp;=\u0026thinsp;7.2, 3.2 Hz, 1H, ArH), 7.64 (dd, J\u0026thinsp;=\u0026thinsp;7.2, 3.2 Hz, 1H, Ar-1H), 7.56 (dd, J\u0026thinsp;=\u0026thinsp;6.6, 3.4 Hz, 1H, CH), 7.50 (dd, J\u0026thinsp;=\u0026thinsp;10.0 Hz, 1H, Ar-1H), 7.34 (d, J\u0026thinsp;=\u0026thinsp;14.8 Hz, 1H), 7.29 (td, J\u0026thinsp;=\u0026thinsp;1.9 Hz, 1H), 3.40 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.4 Hz,, 1H), 2.99 (dd, J\u0026thinsp;=\u0026thinsp;24.7, 14.5 Hz, 1H), 2.73 (td, J\u0026thinsp;=\u0026thinsp;13.8, 8.2 Hz, 1H), 2.66 (ddd, J\u0026thinsp;=\u0026thinsp;15.5, 1.8 Hz, 2H), 2.51 (td, J\u0026thinsp;=\u0026thinsp;23.2, 12.5 Hz, 2H), 2.41 (td, J\u0026thinsp;=\u0026thinsp;23.2, 12.5 Hz, 2H), 2.25\u0026ndash;2.10 (m, 2H), 2.03 (t, J\u0026thinsp;=\u0026thinsp;22.0 Hz, 1H), 1.84\u0026ndash;1.67 (m, 4H), 1.53 (ddd, J\u0026thinsp;=\u0026thinsp;20.0, 11.2, 6.8 Hz, 2H), 1.48\u0026ndash;1.41 (m, 2H), 1.40\u0026ndash;1.27 (m, 2H).\u003c/p\u003e \u003cp\u003e \u003csup\u003e13\u003c/sup\u003eC NMR (126 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ 169.06, 160.95, 151.59, 146.28, 137.27, 134.68, 131.84, 130.13, 129.96, 129.61, 127.52, 126.97, 126.37, 124.91, 120.68, 117.32, 67.56, 58.76, 53.21, 47.57, 40.85, 36.88, 29.51, 28.28, 27.54, 26.28, 23.44, 23.25. HRMS (ESI): Calcd. C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e31\u003c/sub\u003eClN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003em/z: 490.2023, found 491.2933. Anal: C, 70.94; H, 6.36; Cl, 7.22; N, 5.71; O, 9.77.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003e6.2. Biological evaluation assay\u003c/h2\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003e6.2.1. Cell culture\u003c/h2\u003e \u003cp\u003eA549 (Human lung cancer cells), HepG-2 (Human Hepatoma Cells), and HeLa (human cervical cancer cell line) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI-1640 (Solarbio) medium containing 10% Fetal Bovine Serum (FBS, GIBCO), 100U/ml Penicillin, and 100 mg/ml Streptomycin at 37\u0026deg;C under a 5% CO\u003csub\u003e2\u003c/sub\u003e humidified atmosphere.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003e6.2.2. Antiproliferative activity\u003c/h2\u003e \u003cp\u003eA549, HeLa and HepG-2 cells were subjected to an MTT assay to evaluate the investigated compounds' antiproliferative activity. In a CO\u003csub\u003e2\u003c/sub\u003e incubator at 37\u0026deg;C cells were cultured in RPMI-1640 or DMEM complete medium with 10% fetal bovine serum. 96-well plates with a cell density of 2 * 10\u003csup\u003e3\u003c/sup\u003e cells per well were employed to plate exponentially growing cells which were then incubated for 24 h at 37\u0026deg;C to evaluate for attachment. The test compound was dissolved in DMSO. Different concentrations of compounds (100, 50, 25, 12.5, and 6.25\u0026micro;M, respectively) were used to treat the cells for 48 h. Each group consisted of three replicate wells, and subsequent incubation of the plate was conducted at a temperature of 37 ◦C for a duration of 48 hours. MTT solution was added to each well and incubated at 37 ◦C for 4 h. Absorbance of each well was recorded at a wavelength of 570 nm with the multi-function microplate reader. Finally, the data are expressed as means of three independent experiments\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD). the growth inhibition data represented as IC\u003csub\u003e50\u003c/sub\u003e values were calculated through the prism statistical package [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003e6.2.3. Cell cycle assays\u003c/h2\u003e \u003cp\u003eA549 and BEAS-2B cell lines were added to a 6-well plate at a final concentration of 2 * 10\u003csup\u003e5\u003c/sup\u003e cells per well and various concentrations of \u003cb\u003e5a\u003c/b\u003e (0, 1, 2, 4, 8, 16, and 32 \u0026micro;M) were subsequently applied. After 48 h the wells were detached utilizing a trypsin/EDTA solution fixed using 70% ice-cold ethanol for 12 h and subsequently exposed to a 50 \u0026micro;L RNase (50 g/ml) treatment for an hour at 37\u0026deg;C. After that, 250 \u0026micro;l 50 g/ml PI was added and the cells were incubated at 4\u0026deg;C for 4 h. Cell Quest software (BD) and Flow Jo 4.8 were used to evaluate the effects on the cell cycle using a FACS Calibur (BD). In this manner, the cell cycle phase of 2 * 10\u003csup\u003e4\u003c/sup\u003e cells was examined.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section3\"\u003e \u003ch2\u003e6.2.4. Colony formation assay\u003c/h2\u003e \u003cp\u003eA549 cells were seeded in 6-well plates with a density of 1 * 10\u003csup\u003e3\u003c/sup\u003e cells/well and incubated for 48 h before the medium was shifted to a fresh medium containing \u003cb\u003e5a\u003c/b\u003e at concentrations of (0, 2, 4, 8, 16 and 32 \u0026micro;M). The cells were treated for 7 days before being washed twice with PBS, fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and imaged under a microscope (Nikon). After being dissolved in alcohol the violet crystals were read by a microplate reader (BioTek, USA) at an absorbance of 595 nm. GraphPad Prism 7.0 was utilized to analyze the readers.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section3\"\u003e \u003ch2\u003e6.2.5. Cell migration assay\u003c/h2\u003e \u003cp\u003eA549 cells were cultured on a six-well plate allowing space to adhere to other cells. With a 200 \u0026micro;L pipette tip, a wound was created by drawing a line through the middle of each well. Following three PBS washes to remove floating cells the wells were grown in DMEM with 2% FBS as an addition. After adding various doses of \u003cb\u003e5a\u003c/b\u003e (0, 2, 4 and 8 \u0026micro;M) to the wells the plate was incubated for 48 h. Following the removal of the medium three distinct areas were chosen randomly for imaging using a microscope (Nikon, Japan) (\u0026times;100 magnification). The following formula was used to determine the scratch area healing rate: Scratch area healing rate is calculated as follows: [initial scratch area - scratch area at a specific time point initial scratch area 100%].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec30\" class=\"Section3\"\u003e \u003ch2\u003e6.2.6. 4,6-Diamidino-2-phenylindole (DAPI) staining analysis.\u003c/h2\u003e \u003cp\u003eCells were seeded in 12-well plates, and exposed to \u003cb\u003e5a\u003c/b\u003e at a preset concentration (0, 1, 2, 4, 6 and 8) for 48 h and then the nutrient supernatant was discarded. The uncovered cells were fixed in 4% paraformaldehyde for 15 min and washed with PBS twice. Finally, the cells were stained in DAPI dye liquor (Southern Biotech Company, USA) at the final concentration of 1 mg/L for 15 min and washed in PBS once again. The samples were observed and photographed by fluorescence microscope (FM) (Olympus BX51, Olympus, Japan).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec31\" class=\"Section3\"\u003e \u003ch2\u003e6.2.7. Annexin V-FITC/PI apoptosis assay\u003c/h2\u003e \u003cp\u003eFor apoptosis determination, 3 * 10\u003csup\u003e5\u003c/sup\u003e cells/well were seeded in a six-well plate and treated with various concentrations of \u003cb\u003e5a\u003c/b\u003e (0, 2, 4 and 8 \u0026micro;M). After 24 h, the medium was removed and the cells were washed once with PBS, followed by detachment using trypsin/EDTA solution. The cells were stained with annexin V‐FITC/PI solution, and viability was measured on a FACS Calibur (BD) using Cell Quest software (BD) and analyzed using Flow Jo (Flow Jo LLC) as described before.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec32\" class=\"Section3\"\u003e \u003ch2\u003e6.2.8. Western blot analysis\u003c/h2\u003e \u003cp\u003eTo examine the effect of \u003cb\u003e5a\u003c/b\u003e on the protein expression level in A549 cells the various concentrations (0, 2, 4 and 8 \u0026micro;M) of \u003cb\u003e5a\u003c/b\u003e-treated cells were harvested and lysed in sodium dodecyl sulfate (SDS) buffer for 30 min at 4\u0026deg;C. The supernatant obtained by centrifugation at 12000 g for 20 min was used to determine protein concentration by the bicinchoninic acid assay. Proteins (20\u0026ndash;50 \u0026micro;g) were separated by SDS polyacrylamide gel electrophoresis in 8%-12% gels and transferred onto polyvinylidene difluoride membranes (Millipore) at a low temperature. After blocking the membrane for 1 h at room temperature in 5% skim milk in Tris‐buffered saline (TBS) containing 0.1% of Tween‐20 (TBST), they were incubated with an appropriate primary antibody (1:1000) at 4\u0026deg;C for 12 h. The membranes were then washed with TBST and probed with secondary antibodies (1:1000) for 1 h at room temperature. After washing protein expression was measured using an enhanced chemiluminescence kit.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec33\" class=\"Section2\"\u003e \u003ch2\u003e6.3. Animal Experiments\u003c/h2\u003e \u003cdiv id=\"Sec34\" class=\"Section3\"\u003e \u003ch2\u003e6.3.1 Ethics Approval\u003c/h2\u003e \u003cp\u003e This study was conducted by the guidelines of the Institutional Animal Care and Use Committee (IACUC). The protocol was approved by Animal Experimental Ethical Committee of Guangxi University under the No: Gxu-2024-258.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec35\" class=\"Section3\"\u003e \u003ch2\u003e6.3.2. Acute-Toxicity Assessment\u003c/h2\u003e \u003cp\u003eThe Balb/c nude mice were divided randomly (six mice for each group) into one treatment group and the control group. Compound \u003cb\u003e5a\u003c/b\u003e (40 mg/kg, 10% DMSO/10% Tween-80/80% normal saline) and Vehicle (10% DMSO/10% Tween 80/80% normal saline) were administered one day. Subsequently, mice were observed for 28 days and the Mice were placed in a chamber gradually filled with CO\u003csub\u003e2\u003c/sub\u003e gas until unconsciousness and death occurred and the weight was recorded daily.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec36\" class=\"Section3\"\u003e \u003ch2\u003e6.3.3. A549 Xenograft Tumor Model\u003c/h2\u003e \u003cp\u003ePrior to implantation, A549 cell suspension was collected at a concentration of 1 * 10\u003csup\u003e7\u003c/sup\u003e cells/ml, in PBS and 0.1 ml of each cell was inoculated subcutaneously in the right armpit of the Balb/c nude mice. The diameter of transplanted tumors in mice was measured with a vernier caliper, and when the tumors grew to 100 mm\u003csup\u003e3\u003c/sup\u003e the animals were randomly divided into 6 groups. At the same time, the mice in each group started to be administered. The method of measuring the tumor diameter was used to observe the anti-tumor effect of the test samples dynamically. Immediately after the end of the experiment, the mice were sacrificed and the tumor mass was surgically removed and weighed. Body weight and tumor size were measured every 3 days for 28 days after treatment. The formula for calculating tumor volume (TV) is as follows: TV\u0026thinsp;=\u0026thinsp;1/2\u0026times;a\u0026times;b\u003csup\u003e2\u003c/sup\u003e where a and b represent length and width, respectively [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec37\" class=\"Section3\"\u003e \u003ch2\u003e6.3.4. Euthanasia study\u003c/h2\u003e \u003cp\u003eThe experimental mice were introduced into a controlled environment within a CO\u003csub\u003e2\u003c/sub\u003e anesthesia chamber. The flow of CO\u003csub\u003e2\u003c/sub\u003e agent was regulated to gradually induce unconsciousness in the mice. Upon achieving unconsciousness, the concentration of CO\u003csub\u003e2\u003c/sub\u003e was elevated to 100%. During this phase, observations were made to confirm the absence of finger pinching reflex and muscle tone, indicative of complete unconsciousness. Following verification, ventilation was sustained for an additional 2 minutes to ensure the cessation of vital signs, thereby confirming the animals' death.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec38\" class=\"Section3\"\u003e \u003ch2\u003e6.3.5. Histological examination\u003c/h2\u003e \u003cp\u003eThe hearts, spleens, kidneys, lungs, livers, and tumor tissues were isolated to observe the integrity and injury in different groups by HE staining. Briefly, after ischemia and reperfusion, mice hearts, spleens, kidneys, lungs, livers, and tumors were isolated and the tissues were fixed in 4% paraformaldehyde overnight at room temperature, and then dehydrated by passing through gradient concentrations of ethanol (80% for 2 h, 90% for 2 h, 95% overnight, 100% for 0.5, 0.5 and 1 h) at room temperature, followed by embedding in paraffin wax. The paraffin‑embedded samples were then sectioned at 4 \u0026micro;m for staining with Mayer's Hematoxylin (H8070, Solarbio Life Sciences, Beijing, China) for 10 min and then by 0.5% aqueous eosin (DH0050, Leigene Biotech, Beijing, China) for 3 min at room temperature. With this method, the nucleus and other acidic structures are stained blue, while the cytoplasm is stained red. Images were acquired using a light microscope at x400 magnification [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec39\" class=\"Section2\"\u003e \u003ch2\u003e6.4. Molecular docking studies\u003c/h2\u003e \u003cp\u003eThe CTD (PDB ID: 3T0Z) and NTD (PDB ID: 2CG9) atomic coordinates have been retrieved from the RCSB PDB (protein data bank) database [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e] charge assignment, solution measurements, and fragment volumes to the protein have been performed using Autodock Vina PyRx and Discovery Studio 2019 Client for docking analysis. The ligand's 2D structure was drawn and analyzed using ChemDraw, standardized, and converted into PDBQT format using the PyRx Virtual Screening Tool. To show the binding modes 2D, 3D and ligand interaction we carried out using the Discovery Studio 2019 Client.\u003c/p\u003e \u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eHsp90 Heat Shock Protein 90\u003c/p\u003e \u003cp\u003eNTD N-terminal domain\u003c/p\u003e \u003cp\u003eMD Middle Domain\u003c/p\u003e \u003cp\u003eCTD C-Terminal Domain\u003c/p\u003e \u003cp\u003eDMAP Dimethylaminopyridine;\u003c/p\u003e \u003cp\u003eEDCI 1-ethyl-3-(3-(dimethylamino)propyl) carbodiimide hydrochloride;\u003c/p\u003e \u003cp\u003eDCC N, N-dicyclohexylcarbodiimide;\u003c/p\u003e \u003cp\u003eDCM Dichloromethane\u003c/p\u003e \u003cp\u003eSTD Saturation Transfer Difference\u003c/p\u003e \u003cp\u003eHSR Heat Shock Response\u003c/p\u003e \u003cp\u003eNMR Nuclear Magnetic Resonance\u003c/p\u003e \u003cp\u003eDMSO Dimethyl Sulfoxide;\u003c/p\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e Half Maximal Inhibitory Concentration\u003c/p\u003e \u003cp\u003eH\u0026amp;E Hematoxylin and Eosin\u003c/p\u003e \u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eW.L. Supervision, Funding acquisition, Writing - review \u0026amp; editing, Project administration.K.J. Conceptualization, Methodology, Formal analysis, Writing - original draft, Visualization. G.CH. Conceptualization, Methodology, Formal analysis, Visualization.J.M, G.R, L.Y, H.K, Q.G and Y.S Investigation, Data curation, review \u0026amp; editing.L.X. Software, Validation, Resources, Writing - review \u0026amp; editing.Each author contributed significantly to the study's conception, design, data analysis, and interpretation. All authors have read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThis work was supported by the local funding project for scientific and technological development under the guidance of the central government (GuiKe ZY21195012) and Guangxi Innovation-Driven Development Project (GuiKe AA18242040).\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe manuscript and supplementary information file contain the data. The original raw data can be obtained by contacting the Corresponding Author.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003e\u0026ldquo;About WHO.\u0026rdquo; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.who.int/about/\u003c/span\u003e\u003cspan address=\"https://www.who.int/about/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (accessed Mar. 10, 2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWHO, \u0026ldquo;IARC Research \u0026ndash; IARC,\u0026rdquo; \u003cem\u003eIARC GLOBOCAN cancer incidence and mortality rates\u003c/em\u003e, 2018. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.iarc.who.int/research-home/\u003c/span\u003e\u003cspan address=\"https://www.iarc.who.int/research-home/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (accessed Dec. 08, 2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eY. Xu \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Novel matrinic acid derivatives bearing 2-anilinothiazole structure for non-small cell lung cancer treatment with improved Hsp90 targeting effect.,\u0026rdquo; \u003cem\u003eDrug Dev. Res.\u003c/em\u003e, no. June, pp. 1\u0026ndash;21, 2022, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/ddr.21974\u003c/span\u003e\u003cspan address=\"10.1002/ddr.21974\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. Li, X. She, Y. Ou, J. Liu, Z. Yuan, and Q. shi Zhao, \u0026ldquo;Design, synthesis and biological evaluation of a new class of Hsp90 inhibitors vibsanin C derivatives,\u0026rdquo; Eur. J. Med. Chem., vol. 244, no. July, p. 114844, 2022, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2022.114844\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2022.114844\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eC. Prodromou and L. Pearl, \u0026ldquo;Structure and Functional Relationships of Hsp90,\u0026rdquo; Curr. Cancer Drug Targets, vol. 3, no. 5, pp. 301\u0026ndash;323, 2005, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2174/1568009033481877\u003c/span\u003e\u003cspan address=\"10.2174/1568009033481877\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eA. R\u0026ouml;hl, J. Rohrberg, and J. Buchner, \u0026ldquo;The chaperone Hsp90: Changing partners for demanding clients,\u0026rdquo; Trends Biochem. Sci., vol. 38, no. 5, pp. 253\u0026ndash;262, 2013, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.tibs.2013.02.003\u003c/span\u003e\u003cspan address=\"10.1016/j.tibs.2013.02.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Shantanam and MUELLER, \u0026ldquo;HHS Public Access,\u0026rdquo; Physiol. Behav., vol. 176, no. 1, pp. 139\u0026ndash;148, 2018, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/nrc2887.Targeting\u003c/span\u003e\u003cspan address=\"10.1038/nrc2887.Targeting\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eI. Ny, \u0026ldquo;Spring 2009 - CEE 6075 Stochastic Simulation methods in Engineering and Bayesian Computation,\u0026rdquo; \u003cem\u003eEnviron. Eng.\u003c/em\u003e, vol. 1, no. 3, pp. 1\u0026ndash;3, 2009, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1158/1078-0432.CCR-11-1000.Hsp90\u003c/span\u003e\u003cspan address=\"10.1158/1078-0432.CCR-11-1000.Hsp90\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eF. H. Schopf, M. M. Biebl, and J. Buchner, \u0026ldquo;The HSP90 chaperone machinery,\u0026rdquo; Nat. Rev. Mol. Cell Biol., vol. 18, no. 6, pp. 345\u0026ndash;360, 2017, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/nrm.2017.20\u003c/span\u003e\u003cspan address=\"10.1038/nrm.2017.20\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eT. E. M. M. Costa, N. M. Raghavendra, and C. Penido, \u0026ldquo;Natural heat shock protein 90 inhibitors in cancer and inflammation,\u0026rdquo; Eur. J. Med. Chem., vol. 189, p. 112063, 2020, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2020.112063\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2020.112063\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. Serhan \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Total iron measurement in human serum with a smartphone,\u0026rdquo; \u003cem\u003eAIChE Annu. Meet. Conf. Proc.\u003c/em\u003e, vol. 2019-Novem, pp. 0\u0026ndash;1, 2019, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1039/x0xx00000x\u003c/span\u003e\u003cspan address=\"10.1039/x0xx00000x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. G. Marcu, A. Chadli, I. Bouhouche, M. Catelli, and L. M. Neckers, \u0026ldquo;The heat shock protein 90 antagonist novobiocin interacts with a previously unrecognized ATP-binding domain in the carboxyl terminus of the chaperone,\u0026rdquo; J. Biol. Chem., vol. 275, no. 47, pp. 37181\u0026ndash;37186, 2000, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1074/jbc.M003701200\u003c/span\u003e\u003cspan address=\"10.1074/jbc.M003701200\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJ. A. Burlison, L. Neckers, A. B. Smith, A. Maxwell, and B. S. J. Blagg, \u0026ldquo;Novobiocin: Redesigning a DNA gyrase inhibitor for selective inhibition of Hsp90,\u0026rdquo; J. Am. Chem. Soc., vol. 128, no. 48, pp. 15529\u0026ndash;15536, 2006, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/ja065793p\u003c/span\u003e\u003cspan address=\"10.1021/ja065793p\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eK. H. Yim \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Gambogic acid identifies an isoform-specific druggable pocket in the middle domain of Hsp90β,\u0026rdquo; \u003cem\u003eProc. Natl. Acad. Sci. U. S. A.\u003c/em\u003e, vol. 113, no. 33, pp. E4801\u0026ndash;E4809, 2016, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1073/pnas.1606655113\u003c/span\u003e\u003cspan address=\"10.1073/pnas.1606655113\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJ. Dvor\u0026aacute;k and V. Dvor\u0026aacute;k, \u0026ldquo;Orthopedic examination of the foot: refresher course,\u0026rdquo; Sportverletz. Sportschaden, vol. 9, no. 3, pp. 253\u0026ndash;270, 1995, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1055/s-2007-993444\u003c/span\u003e\u003cspan address=\"10.1055/s-2007-993444\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eG. Garg, A. Khandelwal, and B. S. J. Blagg, \u003cem\u003eAnticancer Inhibitors of Hsp90 Function: Beyond the Usual Suspects\u003c/em\u003e, 1st ed., vol. 129. Elsevier Inc., 2016. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/bs.acr.2015.12.001\u003c/span\u003e\u003cspan address=\"10.1016/bs.acr.2015.12.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJ. A. H. Kowah \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;European Journal of Medicinal Chemistry Reports Matrine family derivatives: Synthesis, reactions procedures, mechanism, and application in medicinal, agricultural, and materials chemistry,\u0026rdquo; vol. 7, no. December 2022, 2023, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmcr.2022.100098\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmcr.2022.100098\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQ. Sun, W. Ma, Y. Gao, W. Zheng, B. Zhang, and Y. Peng, \u0026ldquo;Meta-analysis: Therapeutic effect of transcatheter arterial chemoembolization combined with Compound Kushen Injection in hepatocellular carcinoma,\u0026rdquo; African J. Tradit. Complement. Altern. Med., vol. 9, no. 2, 2012, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4314/ajtcam.v9i2.1\u003c/span\u003e\u003cspan address=\"10.4314/ajtcam.v9i2.1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD. Quan \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Structure-Based Design of Novel Alkynyl Thio-Benzoxazepinone Receptor-Interacting Protein Kinase-1 Inhibitors: Extending the Chemical Space from the Allosteric to ATP Binding Pockets,\u0026rdquo; \u003cem\u003eJ. Med. Chem.\u003c/em\u003e, vol. 1, no. Figure 1, 2022, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/acs.jmedchem.2c02067\u003c/span\u003e\u003cspan address=\"10.1021/acs.jmedchem.2c02067\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eL. Wu \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Synthesis and biological evaluation of matrine derivatives containing benzo-α-pyrone structure as potent anti-lung cancer agents,\u0026rdquo; \u003cem\u003eSci. Rep.\u003c/em\u003e, vol. 6, no. 30 mL, 2016, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/srep35918\u003c/span\u003e\u003cspan address=\"10.1038/srep35918\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. Hossain, \u0026ldquo;A Review on Heterocyclic: Synthesis and Their Application in Medicinal Chemistry of Imidazole Moiety,\u0026rdquo; Sci. J. Chem., vol. 6, no. 5, p. 83, 2018, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.11648/j.sjc.20180605.12\u003c/span\u003e\u003cspan address=\"10.11648/j.sjc.20180605.12\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eC. Li, H. Zhu, H. Zhang, Y. Yang, and F. Wang, \u0026ldquo;Synthesis of 2H-Chromenones from Salicylaldehydes and Arylacetonitriles,\u0026rdquo; Molecules, vol. 22, no. 7, Jul. 2017, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules22071197\u003c/span\u003e\u003cspan address=\"10.3390/molecules22071197\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eA. Kolbus, A. Danel, D. Grabka, M. Kucharek, and K. Szary, \u0026ldquo;Spectral Properties of Highly Emissive Derivative of Coumarin with N,N-Diethylamino, Nitrile and Tiophenecarbonyl Moieties in Water-Methanol Mixture,\u0026rdquo; J. Fluoresc., vol. 29, no. 6, pp. 1393\u0026ndash;1399, 2019, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10895-019-02446-5\u003c/span\u003e\u003cspan address=\"10.1007/s10895-019-02446-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eH. Valizadeh and H. Gholipour, \u0026ldquo;Imidazolium-based phosphinite ionic liquid (IL-OPPh2) as reusable catalyst and solvent for the knoevenagel condensation reaction,\u0026rdquo; Synth. Commun., vol. 40, no. 10, pp. 1477\u0026ndash;1485, 2010, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/00397910903097310\u003c/span\u003e\u003cspan address=\"10.1080/00397910903097310\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. M. Heravi, S. Khaghaninejad, and M. Mostofi, \u0026ldquo;Pechmann reaction in the synthesis of coumarin derivatives,\u0026rdquo; in \u003cem\u003eAdvances in Heterocyclic Chemistry\u003c/em\u003e, vol. 112, Academic Press Inc., 2014, pp. 1\u0026ndash;50. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/B978-0-12-800171-4.00001-9\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-12-800171-4.00001-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. Maheswara, V. Siddaiah, G. L. V. Damu, Y. K. Rao, and C. V. Rao, \u0026ldquo;A solvent-free synthesis of coumarins via Pechmann condensation using heterogeneous catalyst,\u0026rdquo; J. Mol. Catal. A Chem., vol. 255, no. 1\u0026ndash;2, pp. 49\u0026ndash;52, 2006, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.molcata.2006.03.051\u003c/span\u003e\u003cspan address=\"10.1016/j.molcata.2006.03.051\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. Lončarić, D. G. Sokač, S. Jokić, and M. Molnar, \u0026ldquo;Recent advances in the synthesis of coumarin derivatives from different starting materials,\u0026rdquo; \u003cem\u003eBiomolecules\u003c/em\u003e, vol. 10, no. 1. MDPI AG, Jan. 01, 2020. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/biom10010151\u003c/span\u003e\u003cspan address=\"10.3390/biom10010151\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eF. Chao, D. E. Wang, R. Liu, Q. Tu, J. J. Liu, and J. Wang, \u0026ldquo;Synthesis, characterization and activity evaluation of matrinic acid derivatives as potential antiproliferative agents,\u0026rdquo; Molecules, vol. 18, no. 5, pp. 5420\u0026ndash;5433, 2013, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules18055420\u003c/span\u003e\u003cspan address=\"10.3390/molecules18055420\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD. D. Li, L. L. Dai, N. Zhang, and Z. W. Tao, \u0026ldquo;Synthesis, structure-activity relationship and biological evaluation of novel nitrogen mustard sophoridinic acid derivatives as potential anticancer agents,\u0026rdquo; \u003cem\u003eBioorganic Med. Chem. Lett.\u003c/em\u003e, vol. 25, no. 19, pp. 4092\u0026ndash;4096, Oct. 2015, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bmcl.2015.08.035\u003c/span\u003e\u003cspan address=\"10.1016/j.bmcl.2015.08.035\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Tang \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Structure\u0026ndash;activity relationship and hypoglycemic activity of tricyclic matrines with advantage of treating diabetic nephropathy,\u0026rdquo; Eur. J. Med. Chem., vol. 201, p. 112315, 2020, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2020.112315\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2020.112315\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eY. Xu \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Design, synthesis, and biological evaluation of matrine derivatives possessing piperazine moiety as antitumor agents,\u0026rdquo; Med. Chem. Res., vol. 28, no. 10, pp. 1618\u0026ndash;1627, 2019, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s00044-019-02398-2\u003c/span\u003e\u003cspan address=\"10.1007/s00044-019-02398-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZ. Shi \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Novel NO-releasing scopoletin derivatives induce cell death via mitochondrial apoptosis pathway and cell cycle arrest,\u0026rdquo; Eur. J. Med. Chem., vol. 200, 2020, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2020.112386\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2020.112386\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD. K. Das, S. Sarkar, M. Khan, M. Belal, and A. T. Khan, \u0026ldquo;A mild and efficient method for large scale synthesis of 3-aminocoumarins and its further application for the preparation of 4-bromo-3-aminocoumarins,\u0026rdquo; Tetrahedron Lett., vol. 55, no. 35, pp. 4869\u0026ndash;4874, 2014, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.tetlet.2014.07.035\u003c/span\u003e\u003cspan address=\"10.1016/j.tetlet.2014.07.035\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eG. Brahmachari, \u0026ldquo;Room Temperature One-Pot Green Synthesis of Coumarin-3-carboxylic Acids in Water: A Practical Method for the Large-Scale Synthesis,\u0026rdquo; \u003cem\u003eACS Sustain. Chem. Eng.\u003c/em\u003e, vol. 3, no. 9, pp. 2350\u0026ndash;2358, Sep. 2015, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/acssuschemeng.5b00826\u003c/span\u003e\u003cspan address=\"10.1021/acssuschemeng.5b00826\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Fiorito, S. Genovese, V. A. Taddeo, and F. Epifano, \u0026ldquo;Microwave-assisted synthesis of coumarin-3-carboxylic acids under ytterbium triflate catalysis,\u0026rdquo; Tetrahedron Lett., vol. 56, no. 19, pp. 2434\u0026ndash;2436, 2015, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.tetlet.2015.03.079\u003c/span\u003e\u003cspan address=\"10.1016/j.tetlet.2015.03.079\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eN. Suzuki \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Design, synthesis, and biological activity of boronic acid-based histone deacetylase inhibitors,\u0026rdquo; J. Med. Chem., vol. 52, no. 9, pp. 2909\u0026ndash;2922, 2009, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/jm900125m\u003c/span\u003e\u003cspan address=\"10.1021/jm900125m\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Tang \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Synthesis and biological evaluation of 12-benzyl matrinic amide derivatives as a novel family of anti-HCV agents,\u0026rdquo; Chinese Chem. Lett., vol. 27, no. 7, pp. 1052\u0026ndash;1057, 2016, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.cclet.2016.03.006\u003c/span\u003e\u003cspan address=\"10.1016/j.cclet.2016.03.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR. Mathew, A. K. Kruthiventi, J. V. Prasad, S. P. Kumar, G. Srinu, and D. Chatterji, \u0026ldquo;Inhibition of mycobacterial growth by plumbagin derivatives,\u0026rdquo; Chem. Biol. Drug Des., vol. 76, no. 1, pp. 34\u0026ndash;42, 2010, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1747-0285.2010.00987.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1747-0285.2010.00987.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. J. Matos \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Study of coumarin-resveratrol hybrids as potent antioxidant compounds,\u0026rdquo; \u003cem\u003eMolecules\u003c/em\u003e, vol. 20, no. 2, pp. 3290\u0026ndash;3308, Feb. 2015, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules20023290\u003c/span\u003e\u003cspan address=\"10.3390/molecules20023290\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Tang \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;SAR evolution and discovery of benzenesulfonyl matrinanes as a novel class of potential coxsakievirus inhibitors,\u0026rdquo; \u003cem\u003eFuture Med. Chem.\u003c/em\u003e, vol. 8, no. 5, pp. 495\u0026ndash;508, Apr. 2016, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4155/fmc-2015-0019\u003c/span\u003e\u003cspan address=\"10.4155/fmc-2015-0019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eA. Thakur, R. Singla, and V. Jaitak, \u0026ldquo;Coumarins as anticancer agents: A review on synthetic strategies, mechanism of action and SAR studies,\u0026rdquo; Eur. J. Med. Chem., vol. 101, pp. 476\u0026ndash;495, 2015, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2015.07.010\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2015.07.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eY. T. Huang and B. S. J. Blagg, \u0026ldquo;A library of noviosylated coumarin analogues,\u0026rdquo; J. Org. Chem., vol. 72, no. 10, pp. 3609\u0026ndash;3613, 2007, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/jo062083t\u003c/span\u003e\u003cspan address=\"10.1021/jo062083t\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Tang \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Structure\u0026ndash;activity relationship and hypoglycemic activity of tricyclic matrines with advantage of treating diabetic nephropathy,\u0026rdquo; Eur. J. Med. Chem., vol. 201, Sep. 2020, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2020.112315\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2020.112315\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS. Paul, P. Roy, P. Saha Sardar, and A. Majhi, \u0026ldquo;Design, Synthesis, and Biophysical Studies of Novel 1,2,3-Triazole-Based Quinoline and Coumarin Compounds,\u0026rdquo; \u003cem\u003eACS Omega\u003c/em\u003e, vol. 4, no. 4, pp. 7213\u0026ndash;7230, Apr. 2019, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/acsomega.9b00414\u003c/span\u003e\u003cspan address=\"10.1021/acsomega.9b00414\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM. Li, X. She, Y. Ou, J. Liu, Z. Yuan, and Q. shi Zhao, \u0026ldquo;Design, synthesis and biological evaluation of a new class of Hsp90 inhibitors vibsanin C derivatives,\u0026rdquo; Eur. J. Med. Chem., vol. 244, pp. 1\u0026ndash;96, 2022, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ejmech.2022.114844\u003c/span\u003e\u003cspan address=\"10.1016/j.ejmech.2022.114844\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eX. Wang, S. Zhang, K. Han, L. Wang, and X. Liu, \u0026ldquo;Induction of Apoptosis by Matrine Derivative ZS17 in Human Hepatocellular Carcinoma BEL-7402 and HepG2 Cells through ROS-JNK-P53 Signalling Pathway Activation,\u0026rdquo; 2022.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJ. Zou \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;A novel oral camptothecin analog, gimatecan, exhibits superior antitumor efficacy than irinotecan toward esophageal squamous cell carcinoma in vitro and in vivo,\u0026rdquo; Cell Death Dis., vol. 9, no. 6, 2018, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41419-018-0700-0\u003c/span\u003e\u003cspan address=\"10.1038/s41419-018-0700-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eY. Sun \u003cem\u003eet al.\u003c/em\u003e, \u0026ldquo;Discovery of CZS-241: A Potent, Selective, and Orally Available Polo-Like Kinase 4 Inhibitor for the Treatment of Chronic Myeloid Leukemia,\u0026rdquo; J. Med. Chem., 2022, doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/acs.jmedchem.2c02124\u003c/span\u003e\u003cspan address=\"10.1021/acs.jmedchem.2c02124\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u0026ldquo;RCSB PDB: Homepage.\u0026rdquo; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.rcsb.org\u003c/span\u003e\u003cspan address=\"https://www.rcsb.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e/ (accessed Aug. 30, 2023).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTables 1-3 are available in the Supplementary Files section.\u003c/p\u003e"},{"header":"Schemes","content":"\u003cp\u003eSchemes1-3 are available in the Supplementary Files section.\u003c/p\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":"Matrine-based Coumarin, Anticancer Agents, Hsp90 Inhibitors, Lung Carcinoma","lastPublishedDoi":"10.21203/rs.3.rs-4632508/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4632508/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMatrine serves as the molecular backbone, targeting the Hsp90 protein N-terminal domain (NTD) and C-terminal domain (CTD), both highly expressed in lung tumor cells. In this study, Matrine Contains Coumarins derivatives were designed and synthesized based on our previously reported compound \u003cb\u003eC\u003c/b\u003e. Employing primary structure-activity relationships and docking analysis, a series of derivatives were biologically assessed for their antiproliferative effects against three cancer cell lines: A549, HepG-2, and HeLa cells. Based on the bioactivity results, derivative \u003cb\u003e5a\u003c/b\u003e emerged as the most potent, significantly enhancing antiproliferation against A549, HepG-2, and HeLa cells, with IC\u003csub\u003e50\u003c/sub\u003e values of 7.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.097, 7.727\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10, and 8.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.065 \u0026micro;M, respectively. Subsequent mechanistic investigations confirmed \u003cb\u003e5a\u003c/b\u003e's ability to inhibit A549 cell proliferation and suppress colony formation and migration. In in vivo studies utilizing a xenograft mouse model inoculated with A549 cells in female Balb/c nude mice, compound \u003cb\u003e5a\u003c/b\u003e displayed superior antitumor activity compared to reference compounds 5-Fluorouracil and Matrine. Notably, the tumor growth inhibition (TGI) values for \u003cb\u003e5a\u003c/b\u003e, 5-Fluorouracil, and Matrine were 72.4%, 64.3%, and 46.8%, respectively.\u003c/p\u003e","manuscriptTitle":"Rational Design and Synthesis of Matrine Containing Coumarin Derivatives as Hsp90 (NTD\u0026amp;CTD) Isoform selective Inhibitors for the Treatment of Lung Carcinoma","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-06 09:57:41","doi":"10.21203/rs.3.rs-4632508/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":"24f0a4ea-ea44-4a52-be4c-f0136d5adb72","owner":[],"postedDate":"August 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":35515027,"name":"Physical sciences/Chemistry/Biochemistry"},{"id":35515028,"name":"Physical sciences/Chemistry/Biosynthesis"},{"id":35515029,"name":"Biological sciences/Biochemistry"},{"id":35515030,"name":"Biological sciences/Cell biology"},{"id":35515031,"name":"Biological sciences/Chemical biology"},{"id":35515032,"name":"Biological sciences/Drug discovery"},{"id":35515033,"name":"Biological sciences/Evolution"},{"id":35515034,"name":"Biological sciences/Molecular biology"},{"id":35515035,"name":"Physical sciences/Chemistry"}],"tags":[],"updatedAt":"2025-02-13T05:55:49+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-06 09:57:41","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4632508","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4632508","identity":"rs-4632508","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}