Benzimidazolyl-Phenylpropenones Synthesized and Evaluated for Selective Anticancer Activity in Prostate Colon and Breast Tumor Models

Benzimidazolyl-Phenylpropenones Synthesized and Evaluated for Selective Anticancer Activity in Prostate Colon and Breast Tumor Models | 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 Research Article Benzimidazolyl-Phenylpropenones Synthesized and Evaluated for Selective Anticancer Activity in Prostate Colon and Breast Tumor Models Aboudramane Koné, Adingra Francesco Kouassi, Souleymane Coulibaly, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7490108/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 14 You are reading this latest preprint version Abstract In this study, benzimidazolyl–phenylpropenone derivatives ( 3a–h ) were synthesized and fully characterized using 1 H, 13 C NMR spectroscopy and High-Resolution Mass Spectrometry (HRMS). Their in vitro anticancer activity was evaluated against human cancer cell lines, including prostate (PC3), colon (CaCo2 and HCT-116), and breast (MDA-MB-231 and MCF-7) tumors, alongside normal human skin fibroblasts. All compounds demonstrated promising cytotoxic activity against the cancer cell lines, with IC 50 values ranging from 1.78 to 8.83 µM. Toxicity toward normal fibroblasts was moderate, with IC 50 values between 3.23 and 8.02 µM. Compared with reference compounds Roscovitine and Paclitaxel (Taxol®), several derivatives showed higher activity than Roscovitine, though generally less potent than Paclitaxel. These findings position benzimidazolyl–phenylpropenones as promising scaffolds for the development of novel anticancer agents. Benzimidazolyl–phenylpropenone Anticancer activity Cytotoxicity Selective tumor targeting In vitro evaluation Figures Figure 1 Introduction Benzimidazole is a privileged heterocyclic scaffold found in many therapeutic agents, particularly anthelmintics [ 1 , 2 ] and widely recognized for its broad spectrum of biological activities, including antifungal [ 3 – 5 ], antibacterial [ 6 – 8 ], antiviral [ 9 – 11 ], anti-inflammatory [ 12 ], antioxidant [ 13 , 14 ], and anticancer effects [ 15 – 18 ]. Despite ongoing efforts to combat cancer, the World Health Organization reported approximately 20 million new cases and 9.7 million deaths worldwide in 2022 [ 19 ]. In Côte d’Ivoire, both cancer incidence and mortality remain high. Data from the Global Cancer Observatory (Globocan 2022) indicate 21,352 new cancer cases and 14,143 deaths in the country [ 20 ]. Prostate cancer is the most prevalent (18.9%), followed by breast (18.1%), liver (6%), and colon (5%) cancers. These statistics underscore that cancer remains a major public health challenge globally, and particularly in Côte d’Ivoire. Our previous work demonstrated the anticancer potential of benzimidazole–chalcone derivatives against breast and colon cancers, as well as their effects on normal skin fibroblasts [ 21 ] (Fig. 1 ). Chalcones are well known for their strong antimicrobial and anticancer activities. Building on the concept of combining biologically active moieties, the present study focuses on synthesizing benzimidazole–chalcone derivatives substituted at the 5-position of the benzimidazole ring. Their in vitro anticancer activity was evaluated against prostate, colon, and breast cancer cell lines, as well as normal skin fibroblasts (Fig. 1 ). Results and discussion The synthesis of benzimidazolyl–phenylpropenone derivatives ( 3a–h ) was achieved in three steps, following the strategy previously reported for benzimidazole–chalcone derivatives. In this work, seven compounds were successfully obtained, including derivatives 3a–c , which were previously evaluated against colon and breast cancer cell lines, as well as normal skin fibroblasts [ 21 ]. The synthetic route is outlined in Scheme 1 . Briefly, ortho -phenylenediamine derivatives were first reacted with glycolic acid in the presence of 4 N hydrochloric acid. The resulting intermediates ( 1a–d ) were then oxidized with manganese dioxide in ethanol. Finally, benzimidazole-2-carbaldehydes were condensed with substituted acetophenones via the Claisen–Schmidt reaction, affording the target benzimidazolyl–phenylpropenones ( 3a–h ) (Table 1 ). The synthesized benzimidazolyl–phenylpropenone derivatives ( 3a–h ) were fully characterized by 1 H and 13 C NMR spectroscopy and HRMS ( see Supporting Information ). In the 1 H NMR spectra, the ethylenic protons appeared as doublets between 7.50 and 8.50 ppm, with coupling constants ranging from 15.2 to 15.8 Hz, confirming the trans configuration of the synthesized compounds. In the 13 C NMR spectra, the carbonyl carbon (C=O) resonated between 187.9 and 195.5 ppm, while the ethylenic carbons appeared between 128 and 150 ppm. The cytotoxic activity of compounds 3a–h was evaluated against prostate, colon, and breast cancer cell lines, as well as normal human skin fibroblasts ( Table 2 ). According to the criteria proposed by Mahmoudi et al. [22] and Wilcox et al. [23], compounds are classified as highly active when IC 50 10 μM. The amplitude of activity was defined as the difference between the upper portion of the dose–response curve (no effect) and its lower portion (maximum effect). Table 2 : Cytotoxicity of synthesized compounds against a panel of cancer cell lines and normal skin fibroblasts. Prostate Colon Breast PC3 CaCo2 HCT-116 MDA-MB 231 MCF-7 Fibroblaste IC 50 amp IC 50 amp IC 50 amp IC 50 amp IC 50 amp IC 50 amp 3a 4.17 86 5.25 97 1.48 98 3.92 76 5.02 98 4.60 49 3b 3.01 88 2.64 92 2.73 97 2.58 83 3.22 91 2.00 38 3c 1.83 82 2.16 95 2.39 96 1.35 70 2.57 94 0.61 36 3d 2.67 85 4.06 97 4.99 97 2.80 76 2.50 96 7.54 47 3e 4.03 77 4.16 89 7.48 96 7.44 80 2.91 81 4.71 39 3f 3.13 87 3.32 97 3.92 98 2.26 75 3.60 98 3.23 42 3g 1.78 81 4.09 97 4.76 98 4.32 77 3.57 92 8.02 41 3h 3.89 74 5.52 93 3.66 98 6.56 78 5.45 85 5.95 39 Rosco 11 52 17 81 8 92 18 65 11 83 3 17 Taxol ® 0.001 43 0.03 59 0.004 82 0.04 47 0.01 66 >0.25 10 Compound 3a exhibited significant activity against PC3, HCT-116, and MDA-MB-231 cancer cell lines, with IC 50 values ranging from 1.48 to 4.17 μM, and moderate activity against CaCo2 and MCF-7, with IC 50 values of 5.25 μM and 7.77 μM, respectively. Its strongest effect was observed on HCT-116 colon cancer cells (IC 50 = 1.48 μM). Introduction of hydroxyl substituents at the ortho ( 3b ) and meta ( 3c ) positions of the phenyl ring markedly enhanced antitumor activity. Both compounds 3b and 3c were active across all tested cancer cell lines, with IC 50 values ranging from 1.35 to 3.01 μM. Among them, compound 3c displayed the highest potency, particularly against PC3 prostate (IC 50 = 1.83 μM) and MDA-MB-231 breast cancer cells (IC 50 = 1.35 μM). These results suggest that electron-donating hydroxyl groups on the phenyl ring favor enhanced antitumor activity. Substitution at the 5-position of the benzimidazole ring further influenced activity. Compounds bearing chloro, nitro, or benzoyl groups at this position were active against prostate cancer cells (IC 50 = 1.78–3.89 μM). Interestingly, the nitro derivative ( 3g ) was more effective against PC3 cells (IC 50 = 1.78 μM) compared with chloro ( 3d–f ) or benzoyl ( 3h ) analogs, suggesting that electron-withdrawing substituents such as nitro are favorable for activity in prostate cancer. However, in colon and breast cancer cell lines, chloro-substituted derivatives ( 3d–f ) did not lead to significant improvements, showing IC 50 values in the range of 2.26–4.99 μM (active) and 7.44–7.48 μM (moderately active). Similarly, nitro ( 3g ) and benzoyl ( 3h ) substitution at C-5 of the benzimidazole ring, combined with hydroxyl substitution on the phenyl ring, did not enhance activity in colon or breast cancer models. Regarding toxicity in normal human fibroblasts, all compounds showed measurable effects with IC 50 values between 3.23 and 8.02 μM. However, their amplitudes of activity were below 50%, indicating that only a limited fraction of dividing fibroblasts was affected. By contrast, the high amplitudes observed in cancer cell lines (74–98%) suggest that compounds 3a–h strongly target proliferating tumor cells, leaving only a small residual population unaffected. This selectivity supports their potential as antimitotic agents with reduced toxicity toward noncancerous cells. Comparison with reference drugs highlights the relative potency of these derivatives. Roscovitine was largely inactive across the panel, with IC 50 values >10 μM in four of the five cancer cell lines and only moderate activity in HCT-116 cells (IC 50 = 8 μM). In contrast, Paclitaxel (Taxol®) displayed exceptional potency, with IC 50 values between 0.001 and 0.04 μM across all cancer cell lines tested. In summary, benzimidazolyl–phenylpropenone derivatives demonstrated broad and selective anticancer activity, with hydroxyl-substituted analogs ( 3b , 3c ) and the nitro derivative ( 3g ) emerging as the most potent. By contrast, chloro-substituted derivatives, even when combined with hydroxyl groups, did not improve activity, a finding consistent with QSAR predictions and previous literature, 24-26 where the high lipophilicity of chloro substituents often compromises bioactivity rather than enhancing it. Overall, the present series was generally more active than Roscovitine while less potent than Paclitaxel, but their favorable selectivity toward cancer cells over normal fibroblasts underscores their promise as lead scaffolds for further optimization in anticancer drug development. The synthesized benzimidazolyl–phenylpropenone derivatives ( 3a–h ) were fully characterized by 1 H and 13 C NMR spectroscopy and HRMS ( see Supporting Information ). In the 1 H NMR spectra, the ethylenic protons appeared as doublets between 7.50 and 8.50 ppm, with coupling constants ranging from 15.2 to 15.8 Hz, confirming the trans configuration of the synthesized compounds. In the 13 C NMR spectra, the carbonyl carbon (C = O) resonated between 187.9 and 195.5 ppm, while the ethylenic carbons appeared between 128 and 150 ppm. The cytotoxic activity of compounds 3a–h was evaluated against prostate, colon, and breast cancer cell lines, as well as normal human skin fibroblasts (Table 2 ). According to the criteria proposed by Mahmoudi et al. [ 22 ] and Wilcox et al. [ 23 ], compounds are classified as highly active when IC 50 10 µM. The amplitude of activity was defined as the difference between the upper portion of the dose–response curve (no effect) and its lower portion (maximum effect). Table 2 Cytotoxicity of synthesized compounds against a panel of cancer cell lines and normal skin fibroblasts. Prostate Colon Breast PC3 CaCo2 HCT-116 MDA-MB 231 MCF-7 Fibroblaste IC 50 amp IC 50 amp IC 50 amp IC 50 amp IC 50 amp IC 50 amp 3a 4.17 86 5.25 97 1.48 98 3.92 76 5.02 98 4.60 49 3b 3.01 88 2.64 92 2.73 97 2.58 83 3.22 91 2.00 38 3c 1.83 82 2.16 95 2.39 96 1.35 70 2.57 94 0.61 36 3d 2.67 85 4.06 97 4.99 97 2.80 76 2.50 96 7.54 47 3e 4.03 77 4.16 89 7.48 96 7.44 80 2.91 81 4.71 39 3f 3.13 87 3.32 97 3.92 98 2.26 75 3.60 98 3.23 42 3g 1.78 81 4.09 97 4.76 98 4.32 77 3.57 92 8.02 41 3h 3.89 74 5.52 93 3.66 98 6.56 78 5.45 85 5.95 39 Rosco 11 52 17 81 8 92 18 65 11 83 3 17 Taxol® 0.001 43 0.03 59 0.004 82 0.04 47 0.01 66 > 0.25 10 Compound 3a exhibited significant activity against PC3, HCT-116, and MDA-MB-231 cancer cell lines, with IC 50 values ranging from 1.48 to 4.17 µM, and moderate activity against CaCo2 and MCF-7, with IC 50 values of 5.25 µM and 7.77 µM, respectively. Its strongest effect was observed on HCT-116 colon cancer cells (IC 50 = 1.48 µM). Introduction of hydroxyl substituents at the ortho ( 3b ) and meta ( 3c ) positions of the phenyl ring markedly enhanced antitumor activity. Both compounds 3b and 3c were active across all tested cancer cell lines, with IC 50 values ranging from 1.35 to 3.01 µM. Among them, compound 3c displayed the highest potency, particularly against PC3 prostate (IC 50 = 1.83 µM) and MDA-MB-231 breast cancer cells (IC 50 = 1.35 µM). These results suggest that electron-donating hydroxyl groups on the phenyl ring favor enhanced antitumor activity. Substitution at the 5-position of the benzimidazole ring further influenced activity. Compounds bearing chloro, nitro, or benzoyl groups at this position were active against prostate cancer cells (IC 50 = 1.78–3.89 µM). Interestingly, the nitro derivative ( 3g ) was more effective against PC3 cells (IC 50 = 1.78 µM) compared with chloro ( 3d–f ) or benzoyl ( 3h ) analogs, suggesting that electron-withdrawing substituents such as nitro are favorable for activity in prostate cancer. However, in colon and breast cancer cell lines, chloro-substituted derivatives ( 3d–f ) did not lead to significant improvements, showing IC 50 values in the range of 2.26–4.99 µM (active) and 7.44–7.48 µM (moderately active). Similarly, nitro ( 3g ) and benzoyl ( 3h ) substitution at C-5 of the benzimidazole ring, combined with hydroxyl substitution on the phenyl ring, did not enhance activity in colon or breast cancer models. Regarding toxicity in normal human fibroblasts, all compounds showed measurable effects with IC 50 values between 3.23 and 8.02 µM. However, their amplitudes of activity were below 50%, indicating that only a limited fraction of dividing fibroblasts was affected. By contrast, the high amplitudes observed in cancer cell lines (74–98%) suggest that compounds 3a–h strongly target proliferating tumor cells, leaving only a small residual population unaffected. This selectivity supports their potential as antimitotic agents with reduced toxicity toward noncancerous cells. Comparison with reference drugs highlights the relative potency of these derivatives. Roscovitine was largely inactive across the panel, with IC 50 values > 10 µM in four of the five cancer cell lines and only moderate activity in HCT-116 cells (IC 50 = 8 µM). In contrast, Paclitaxel (Taxol®) displayed exceptional potency, with IC 50 values between 0.001 and 0.04 µM across all cancer cell lines tested. In summary, benzimidazolyl–phenylpropenone derivatives demonstrated broad and selective anticancer activity, with hydroxyl-substituted analogs ( 3b , 3c ) and the nitro derivative ( 3g ) emerging as the most potent. By contrast, chloro-substituted derivatives, even when combined with hydroxyl groups, did not improve activity, a finding consistent with QSAR predictions and previous literature, 24–26 where the high lipophilicity of chloro substituents often compromises bioactivity rather than enhancing it. Overall, the present series was generally more active than Roscovitine while less potent than Paclitaxel, but their favorable selectivity toward cancer cells over normal fibroblasts underscores their promise as lead scaffolds for further optimization in anticancer drug development. Conclusion The benzimidazole derivatives designed and synthesized in this study demonstrated moderate to strong cytotoxic activity across prostate, colon, and breast cancer cell lines, while maintaining reduced effects on normal fibroblasts. Hydroxyl-substituted analogs ( 3b , 3c ) and the nitro derivative ( 3g ) emerged as the most potent, highlighting the importance of electronic effects in modulating anticancer activity. By contrast, substitution at the 5-position of the benzimidazole ring with chloro, nitro, or benzoyl groups generally did not enhance activity compared with their non-substituted counterparts. This observation is not surprising, as QSAR modeling and literature evidence suggest that the high lipophilicity of chloro substituents, even in combination with hydroxyl groups, often compromises bioactivity rather than improving it. Despite this, all 5-substituted derivatives displayed higher activity than the reference compound Roscovitine, though they remained less potent than Paclitaxel. These findings are encouraging, as they identify benzimidazolyl–phenylpropenones as promising scaffolds for further optimization. Future work will focus on refining the structure–activity relationships of benzimidazolyl–phenylpropenones through computational approaches, including molecular docking, and molecular dynamics simulations to better understand their interactions with potential biological targets. In addition, further structural modifications aimed at optimizing substituents on both the benzimidazole core and the phenyl ring may enhance potency and selectivity. Evaluation in 3D tumor spheroid models and subsequent in vivo studies will be crucial to validate their therapeutic potential. Ultimately, these efforts could accelerate the development of cost-effective anticancer agents particularly suited for addressing the growing cancer burden in low- and middle-income countries. Experimental section General For all compounds, the 1 H and 13 C NMR spectra were recorded at room temperature on a Bruker Avance AC300 (300 MHz for 1 H and 75 MHz for 13 C) or on a Bruker Avance AC300 (400 MHz for 1 H and 100 MHz for 13 C) in deuterated dimethylsulfoxide (DMSO- d 6 ). Tetramethylsilane (TMS) was used as a reference and chemical shifts are expressed in parts per million (ppm) while coupling constants ( J ) are expressed in Hertz (Hz). NMR spectra are described using the following symbols: s (singlet), d (doublet), t (triplet), q (quadruplet), m (massive), dd (doublet of doublet). Mass spectra were recorded using a ThermoScientific DSQII quadrupole instrument by electron impact (EI, 70 eV) or chemical ionization (CI, 500 eV). High-resolution mass spectroscopy (HRMS) was performed on a ThermoScientific LTQ-Orbitrap mass spectrometer in positive electrospray ionization mode. The progress of the reaction and the purity of the compounds were monitored by TLC on aluminum plates coated with silica gel (Kieselgel 60 F 254 , MERCK). After elution in the appropriate solvent or solvent mixture, the plates were revealed by UV fluorescence (λ = 254 nm) or by potassium permanganate (KMnO 4 ) solution followed by heating. The melting points of solid compounds were determined using a Köfler bench and are uncorrected. Solvents and reagents were purchased from Acros Organics (France) or Sigma-Aldrich (France). Synthesis Compounds 1a (1 H -benzimidazol-2-yl)methanol), 2a (1 H -benzimidazole-2-carbaldehyde), 3a (( E )-3-(1 H -Benzimidazol-2-yl)-1-phenylprop-2-en-1-one), 3b (( E )-3-(1 H -Benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one) and (( E )-3-(1 H -Benzimidazol-2-yl)-1-(2-hydroxy phenyl)prop-2-en-1-one) have already been reported [21]. Synthesis of (1 H -benzimidazol-2-yl)methanol derivatives 1b-d To a solution of o -phenylenediamine derivatives (1 eq, 5 g) in HCl 4N (50 mL), was added glycolic acid (3 eq). The reaction mixture was refluxed for 6 h then cooled to room temperature and quenched with ammonia solution 25% at 0 °C. The product was collected pure by filtration and dried at room temperature to afford benzimidazolyl-methanol derivatives. (5-Chloro-1 H -benzimidazol-2-yl)methanol 1b Brown solid, yield = 97%, 1 H NMR: (300 MHz, DMSO- d6 , δ ppm) 12.46 (br s, 1H, NH), 7.56-7.47 (m, 2H, H4 and H7) 7.15 (dd, J6-7 = 8.8, J6-4 = 2.1 Hz, 1H, H6), 5.76 (br s, 1H, OH), 4.69 (s, 2H, H1). 13 C NMR: (75 MHz, DMSO- d6 , δ ppm) 156.9 (CAr), 133.0 (CAr), 121.6 (CAr), 119.6 (CAr), 117.8 (CAr), 112.5 (CAr), 110.9 (CAr), 57.6 (CH2). MS : [EI, 70 eV, m/z (rel. Int)] : 184 (29), 183 (24), 182 (100), 181 (40), 165 (29). (5-Nitro-1 H -benzimidazol-2-yl)methanol ( 1c ) Brown solid, yield = 97%, 1 H NMR : (300 MHz, DMSO- d 6 , δ ppm) 12.71 (br s, 1H, NH), 8.68-8.15 (m, 1H, H 4 ), 8.08 (dd, J 6-7 = 8.9 Hz, J 6-4 = 2.3 Hz, 1H, H 6 ), 7.67 (d, J 7-6 = 8.8 Hz, 1H, H 7 ), 5.93 (s, 1H, OH), 4.77 (s, 2H, H 1 ). 13 C NMR: (75 MHz, DMSO- d 6 , δ ppm) 159.8 (C Ar ), 142.3 (C Ar ), 118.5 (C Ar ), 117.6 (C Ar ), 114.5 (C Ar ), 111.5 (C Ar ), 107.9 (C Ar ), 57.7 (CH 2 ). MS: [EI, 70 eV, m/z (rel. Int)]: 194 (16), 193 (100), 192 (28), 147 (18). (5-Benzoyl-1 H -benzimidazol-2-yl)methanol ( 1d ) Beige solide, yield = 97%, 1 H NMR: (300 MHz, DMSO- d 6 , δ ppm) 12.72 (br s, 1H, NH), 7.88 (t, J = 1.2 Hz, 1H, H 4 ), 7.76-7.70 (m, 2H, H 6 and H 7 ), 7.70-7.52 (m, 5H, H Ar ), 5.83 (br s, 1H, OH), 4.74 (s, 2H, H 1 ). 13 C NMR: (75 MHz, DMSO- d 6 , δ ppm) 195.7 (C=O), 138.2 (2C Ar ), 132.0 (C Ar ), 130.3 (2C Ar ), 129.3 (2C Ar ), 128.3 (4C Ar ), 123.6 (2C Ar ), 57.7 (CH 2 ). MS: [EI, 70 eV, m/z (rel. Int)] : 253 (20), 252 (100), 251 (14), 175 (76). Synthesis of 1 H -benzimidazole-2-carbaldehyde (2b-d) To a solution of benzimidazol-2-yl methanol and derivatives (1 eq, 1 g) in 10 mL of EtOH, was added activated manganese dioxide (8.5 eq). The reaction mixture was stirred at room temperature overnight and concentrated. 20 mL of boiling DMF was added, filtered over a pad of celite and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (elution: DCM/MeOH 98: 2) to afford benzimidazole-2-carbaldehydes derivatives 5-Chloro-1 H -benzimidazole-2-carbaldehyde (2b) Yellow solid, yield = 52%, 1 H NMR : (300 MHz, DMSO- d6 , δ ppm) 13.82 (br s, 1H, NH), 9.95 (s, 1H, CHO), 7.79-7.35 (m, 3 H, H4, H6 and H7) 13 C NMR : (75 MHz, DMSO- d6 , δ ppm) 183.4 (CH=O), 141.7 (CAr), 140.6 (CAr), 136.9 (CAr), 129.5 (CAr), 123.8 (CAr), 116.3 (CAr), 115.5 (CAr) MS : [EI, 70 eV, m/z (rel. Int)] : 180 (12), 154 (28), 152 (100), 151 (11). 5-Nitro-1 H -benzimidazole-2-carbaldehyde ( 2c ) Yellow solid, yield = 55%, 1 H NMR: (300 MHz, DMSO- d 6 , δ ppm) 14.04 (br s, 1H, NH), 9.97 (s, 1H, CHO), 8.50-7.73 (m, 3H, H 4 , H 6 and H 7 ). 13 C NMR: (75 MHz, DMSO- d 6 , δ ppm) 183.1 (CH=O), 145.1 (C Ar ), 144.4 (C Ar ), 141.5 (C Ar ), 139.8 (C Ar ), 118.3 (C Ar ), 116.6 (C Ar ), 113.0 (C Ar ). MS: [EI, 70 eV, m/z (rel. Int)]: 192 (16), 191 (100), 190 (28), 145 (18). 5-Benzoyl-1 H -benzimidazole-2-carbaldehyde ( 2d ) Yellow solid, yield = 57%, 1 H NMR : (400 MHz, DMSO- d 6 , δ ppm) 13.83 (s, 1H, NH), 10.01 (s, 1H, CHO), 7.80-7.75 (m, 3H, H Ar ), 7.72-7.68 (m, 2H, H Ar ), 7.61-7.57 (m, 3H, H Ar ). 13 C NMR: (75 MHz, DMSO- d 6 , δ ppm) 194.0 (C=O), 183.6 (CH=O), 142.6 (C Ar ), 141.5 (C Ar ), 138.6 (C Ar ), 138.4 (C Ar ), 132.4 (C Ar ), 131.2 (C Ar ), 130.3 (2C Ar ), 128.4 (2C Ar ), 124.1 (C Ar ), 119.0 (C Ar ), 114.9 (C Ar ). MS: [EI, 70 eV, m/z (rel. Int)] : 251 (19), 250 (100), 249 (14), 173 (82), 145 (17). Synthesis of ( E )-3-(1 H -benzimidazol-2-yl)-1-phenylprop-2-en-1-one derivatives (3d-h) To a solution of NaOH (7.5 eq) in ethanol (10 mL), were added benzimidazole-2- carbaldehyde derivatives 2b-d (1 eq, 500 mg) and appropriate acetophenone (1 eq). The mixture was stirred at room temperature overnight then quenched with acetic acid 20% at 0 °C. The crude was collected by filtration, dried and purified by flash-chromatography on silica gel (DCM/MeOH 95 :5) to afford 3-(1 H -benzimidazol-2-yl)-1-arylprop-2-en-1-one derivatives. ( E )-3-(5-Chloro-1 H -benzimidazol-2-yl)-1-phenylprop-2-en-1-one ( 3d ) Yellow solid, yield = 71%, M.p = 158-160 °C, 1 H NMR : (400 MHz, DMSO- d 6 , δ ppm) 12.04 (br s, 1H, NH), 8.50 (d, J 3-2 = 15.8 Hz, 1H, H 3 ), 8.16 (d, J = 7.3 Hz, 2H, H Ar ), 7.80-7.72 (m, 3H, H 2 and 2H Ar ), 7.65-7.59 (m, 3H, H Ar ), 7.40 (dd, J = 1.6 Hz, J = 8.7 Hz, 1H, H Ar ). 13 C NMR: (100 MHz, DMSO- d 6 , δ ppm) 188.2 (C=O), 149.0 (C Ar ), 137.6 (C Ar ), 136.7 (C Ar ), 135.5 (C Ar ), 133.9 (C Ar ), 129.5 (2C Ar ), 129.0 (2C Ar ), 128.8 (C Ar ), 128.7 (C Ar ), 128.6 (C Ar ), 124.7 (C Ar ), 116.5 (C Ar ), 114.8 (C Ar ). HRMS: (ES+) Calculated for C 16 H 12 N 2 OCl [M + H] + : 283.0638, found : 283.0634. IR: (ATR, υ cm -1 ) 3453.2 (N-H), 2919.1 (=C-H), 1673.1 (C=O), 1441.9 (C=C), 710.9 (C-Cl). ( E )-3-(5-Chloro-1 H -benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one ( 3e ) Yellow solid, yield = 69%, M.p = 116-118 °C, 1 H NMR : (400 MHz, DMSO- d 6 , δ ppm) 11.94 (s, 1H, NH), 8.26 (d, J 3-2 = 15.2 Hz, 1H, H 3 ), 8.00 (dd, J = 1.2 Hz, J = 8.1 Hz, 1H, H Ar ), 7.72-7.52 (m, 5H, 4H Ar and H 2 ), 7.29 (d, J = 8.3 Hz, 1H, H Ar ), 7.06-702 (m, 2H, H Ar ). 13 C NMR : (100 MHz, DMSO- d 6 , δ ppm) 192.2 (C=O), 170.2 (C Ar ), 161.0 (C Ar ), 138.3 (C Ar ), 136.3 (C Ar ), 131.2 (C Ar ), 130.4 (2C Ar ), 127.8 (C Ar ), 123.9 (C Ar ), 123.6 (C Ar ), 121.4 (C Ar ), 119.4 (2C Ar ), 117.7 (C Ar ), 113.5 (C Ar ). MS : [EI, 70 eV, m/z (rel. Int)] : 299 (23), 298 (100) [CI, NH 3, m/z] : 299 [M+H] + . HRMS: (ES+) Calculated for C 16 H 12 N 2 O 2 35 Cl [M + H] + : 299.0587, found : 299.0579. IR: (ATR, υ cm -1 ) 3425.4 (O-H), 2922.7 (=C-H), 1647.4 (C=O), 1583.0 (C=N), 748.7 (C-Cl). ( E )-3-(5-Chloro-1 H -benzimidazol-2-yl)-1-(3-hydroxyphenyl)prop-2-en-1-one ( 3f ) Yellow solid, yield = 63%, M.p = 210-212 °C, 1 H NMR : (400 MHz, DMSO- d 6 , δ ppm) 12.01 (br s, 1H, NH), 8.52 (d, J 3-2 = 15.7 Hz, 1H, H 3 ), 7.84 (s, 1H, H Ar ), 7.77 (d, J = 8.6 Hz, 1H, H Ar ), 7.68-7.55 (m, 3H, H 2 and 2H Ar ), 7.53-7.38 (m, 3H, H Ar ), 7.15 (dd, J = 2.1 Hz, J = 7.9 Hz, 1H, H Ar ). 13 C NMR : (100 MHz, DMSO- d 6 , δ ppm) 187.9 (C=O), 158.0 (C Ar ), 138.0 (C Ar ), 130.7 (C Ar ), 130.1 (C Ar ), 129.3 (2C Ar ), 127.4 (C Ar ), 125.3 (C Ar ), 121.2 (2C Ar ), 119.8 (2C Ar ), 116.4 (C Ar ), 114.6 (2C Ar ). HRMS: (ES+) Calculated for C 16 H 12 N 2 O 2 35 Cl [M + H] + : 299.0587, found : 299.0583. IR: (ATR, υ cm -1 ) 3321.4 (O-H), 2920.6 (=C-H), 1653.6 (C=O), 1585.0 (C=N), 778.4 (C-Cl). ( E )-3-(5-Nitro-1 H -benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one ( 3g ) Yellow solid, yield = 77%, M.p = 225-226 °C, 1 H NMR : (400 MHz, DMSO- d 6 , δ ppm) 11.90 (br s, 1H, NH), 8.47 (d, J 3-2 = 15.2 Hz, 1H, H 3 ), 8.21 (dd, J = 1.2 Hz, J = 8.1 Hz, 1H, H Ar ), 7.95-7.77 (m, 5H, 4H Ar and H 2 ), 7.65 (d, J = 8.3 Hz, 1H, H Ar ), 7.52-7.40 (m, 2H, H Ar ). 13 C NMR : (100 MHz, DMSO- d 6 , δ ppm) 195.5 (C=O), 184.7 (C Ar ), 158.9 (C Ar ), 150.3 (C Ar ), 144.1 (C Ar ), 137.7 (C Ar ), 137.6 (C Ar ), 132.4 (C Ar ), 132.3 (C Ar ), 129.5 (2C Ar ), 128.5 (C Ar ), 125.0 (2C Ar ), 119.5 (C Ar ), 115.7 (C Ar ). MS: [CI, NH 3, m/z] : 309 [M] + , 310 [M + H] + . HRMS: (ES+) Calculated for C 16 H 12 N 3 O 4 [M + H] + : 310.0828, found : 310.0824. IR: (ATR, υ cm -1 ) 3102.1 (O-H), 1670.2 (C=O), 1587.5 (C=N), 1340.1 (-NO 2 ). ( E )-3-(5-Benzoyl-1 H -benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one ( 3h ) Yellow solid, yield = 65%, M.p = 195-196 °C, 1 H NMR : (400 MHz, DMSO- d 6 , δ ppm) 11.91 (s, 1H, NH), 8.52 (d, J 3-2 = 15.8 Hz, 1H, H 3 ), 8.08-8.05 (m, 1H, H Ar ), 8.03 (s, 1H, H Ar ), 7.90-7.57 (m, 9H, H Ar ), 7.69 (d, J 2-3 = 15.8 Hz, 1H, H 2 ), 7.08-7.03 (m, 2H, H Ar ). 13 C NMR: (100 MHz, DMSO- d 6 , δ ppm) 195.3 (C=O), 192.0 (C=O), 161.0 (C Ar ), 150.4 (C Ar ), 137.6 (C Ar ), 136.5 (C Ar ), 132.6 (C Ar ), 132.4 (C Ar ), 130.6 (2C Ar ), 130.3 (C Ar ), 129.6 (2C Ar ), 128.5 (C Ar ), 125.6 (C Ar ), 121.5 (C Ar ), 119.5 (C Ar ), 118.3 (2C Ar ), 117.8 (C Ar ), 115.1 (2C Ar ), 114.3 (C Ar ). MS : [EI, 70 eV, m/z (rel. Int)] : 369 (35), 368 (100), 367(16) [CI, NH 3, m/z] : 368 [M] + , 369 [M + H] + . HRMS: (ES+) Calculated for C 23 H 17 N 2 O 3 [M + H] + : 369.1239, found : 369.1241. IR: (ATR, υ cm -1 ) 3287.4 (O-H), 3171.2 (N-H), 1682.6 (C=O), 1564.7 (C=N), 1478.1 (C=C). Biological material Normal human skin fibroblasts were purchased from Gibco (Thermo Fisher Scientific). The cancer cell lines Caco-2, MDA-MB-231, HCT-116, PC3, and MCF-7 were obtained from the European Collection of Authenticated Cell Cultures (ECACC, Porton, UK). Cells were maintained under the recommended conditions: Caco-2, MDA-MB-231, and MCF-7 in Dulbecco’s Modified Eagle Medium (DMEM); HCT-116 in McCoy’s medium; and PC3 in RPMI medium. All culture media were supplemented with 10% fetal bovine serum (FBS), 1% penicillin–streptomycin, and 2 mM glutamine. Cultures were incubated at 37 °C in a humidified atmosphere containing 5% CO 2 . All cell lines are maintained in liquid nitrogen as both master and working banks. Routine mycoplasma testing was performed to confirm the absence of contamination throughout the study. Declarations Acknowledgements The authors would like to thank laboratory Chemistry and Interdisciplinarity: Synthesis, Analysis, Modelling (CEISAM), University of Nantes for chemical reagents and spectroscopic analyses; the ‘‘ImPACcell’’ platform and Dr Rémy Le Guével of the University of Rennes 1 for cancer tests. Author contributions A.K conceived and designed the chemistry experiments, performed them, analyzed the data, prepared figures and/or tables, reviewed drafts of the article, and approved the final draft. A.F.K analyzed the data, prepared figures and/or tables, and approved the final draft. S.C analyzed the data, and wrote the final draft of the article. D. S analyzed the data, supervised, and approved the final draft. S. C conceived the work and designed the biological experiments and approved the final draft. Clinical trial number Not registered even if research protocols were performed in accordance with French legal guidelines (French Ministry of Health). Consent to participate Not applicable Consent to publish Not applicable Conflicts of interest The authors declare no conflicts of interest. Ethical approval The authors declare no competing interests. Data availability statements The data that support the findings of this study are available in the supplementary information of this article. The human-derived cell lines used in our study (colon and breast cancer cell lines, as well as normal human skin fibroblasts) were obtained from commercial suppliers (Gibco, CLS, ATCC, or ECACC) and were fully de-identified. No primary human tissues were collected specifically for this study. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. References Généviève Z, Elisabeth O, Boureima T, Basile T, Adama K, Balé B, Hamidou H. T, and Adrien M.G. M (2023) Anti-parasitic activities of four synthetic chemicals anthelmintics on Haemonchus contortus. World J. Adv. Res. Rev. , 20 (2), 683–689. Chung, N.T., Dung, V.C., and Duc, D.X. (2023) Recent achievements in the synthesis of benzimidazole derivatives. RSC Adv. , 13 (46), 32734–32771. Chandrika, N.T., Shrestha, S.K., Ngo, H.X., and Garneau-Tsodikova, S. (2016) Synthesis and investigation of novel benzimidazole derivatives as antifungal agents. Bioorganic Med. Chem. , 24 (16), 3680–3686. Kaplancikli, Z.A., Turan-Zitouni, G., Revial, G., and Guven, K. (2004) Synthesis and study of antibacterial and antifungal activities of Novel 2-[[(benzoxazole/benzimidazole-2-yl)sulfanyl] acetylamino]thiazoles. Arch. Pharm. Res. , 27 (11), 1081–1085. Bai, S., Zhang, M., Tang, S., Li, M., Wu, R., Wan, S., Chen, L., Wei, X., and Li, F. (2024) Research Progress on Benzimidazole Fungicides: A Review. Molecules , 29 (6), 1–14. Marinescu, M. (2021) Synthesis of antimicrobial benzimidazole–pyrazole compounds and their biological activities. Antibiotics , 10 (8), 1–29. Mahmood, K., Hashmi, W., Ismail, H., Mirza, B., Twamley, B., Akhter, Z., Rozas, I., and Baker, R.J. (2019) Synthesis, DNA binding and antibacterial activity of metal(II) complexes of a benzimidazole Schiff base. Polyhedron , 157 (Ii), 326–334. AlDifar, H.A., Baaiu, B.S., Darwish, K.M.A., Ali, M.F., Dakhil, O.O., Abd-Alsalam, M., and Al Difar, H. (2023) Synthesis of Benzimidazole and Phthaloylamino Acid derivatives and Antibacterial Activity. J. Med. Chem. Sci. , 6 (9), 1975–1984. Tomei, L., Altamura, S., Bartholomew, L., Biroccio, A., Ceccacci, A., Pacini, L., Narjes, F., Gennari, N., Bisbocci, M., Incitti, I., Orsatti, L., Harper, S., Stansfield, I., Rowley, M., De Francesco, R., and Migliaccio, G. (2003) Mechanismof Action and Antiviral Activity of Benzimidazole-Based AllostericInhibitors of the Hepatitis C Virus RNA-Dependent RNAPolymerase. J. Virol. , 77 (24), 13225–13231. Shaker, Y.M., Omar, M.A., Mahmoud, K., Elhallouty, S.M., El-Senousy, W.M., Ali, M.M., Mahmoud, A.E., Abdel-Halim, A.H., Soliman, S.M., and El Diwani, H.I. (2015) Synthesis, in vitro and in vivo antitumor and antiviral activity of novel 1-substituted benzimidazole derivatives. J. Enzyme Inhib. Med. Chem. , 30 (5), 826–845. Ahmad, N., Azad, M.I., Khan, A.R., and Azad, I. (2021) Benzimidazole As a Promising Antiviral Heterocyclic Scaffold: a Review. J. Sci. Arts , 21 (1), 273–284. Veerasamy, R., Roy, A., Karunakaran, R., and Rajak, H. (2021) Structure–activity relationship analysis of benzimidazoles as emerging anti-inflammatory agents: An overview. Pharmaceuticals , 14 (7). Mavrova, A.T., Yancheva, D., Anastassova, N., Anichina, K., Zvezdanovic, J., Djordjevic, A., Markovic, D., and Smelcerovic, A. (2015) Synthesis, electronic properties, antioxidant and antibacterial activity of some new benzimidazoles. Bioorganic Med. Chem. , 23 (19), 6317–6326. Aroua, L.M., Almuhaylan, H.R., Alminderej, F.M., Messaoudi, S., Chigurupati, S., Al-mahmoud, S., and Hamdoon, M.H. (2021) A facile approach synthesis of benzoylaryl benzimidazole as potential α-amylase and α-glucosidase inhibitor with antioxidant activity. Bioorg. Chem. , 114 (March), 105073. El-Gohary, N.S., and Shaaban, M.I. (2017) Synthesis and biological evaluation of a new series of benzimidazole derivatives as antimicrobial, antiquorum-sensing and antitumor agents. Eur. J. Med. Chem. , 131 , 255–262. Onnis, V., Demurtas, M., Deplano, A., Balboni, G., Baldisserotto, A., Manfredini, S., Pacifico, S., Liekens, S., and Balzarini, J. (2016) Design, synthesis and evaluation of antiproliferative activity of new benzimidazolehydrazones. Molecules , 21 (5), 4–12. Wang, Z., Deng, X., Xiong, S., Xiong, R., Liu, J., Zou, L., Lei, X., Cao, X., Xie, Z., Chen, Y., Liu, Y., Zheng, X., and Tang, G. (2018) Design, synthesis and biological evaluation of chrysin benzimidazole derivatives as potential anticancer agents. Nat. Prod. Res. , 32 (24), 2900–2909. Sharma, P., Reddy, T.S., Kumar, N.P., Senwar, K.R., Bhargava, S.K., and Shankaraiah, N. (2017) Conventional and microwave-assisted synthesis of new 1H-benzimidazole-thiazolidinedione derivatives: A potential anticancer scaffold. Eur. J. Med. Chem. , 138 , 234–245. Centre international de recherche sur le cancer (CIRC) (2024) Cancer : une charge toujours plus lourde dans le monde et des besoins en services croissants. l’Organisation Mond. la Santé , 1–5. Organization, W.H. (2022) Global Cancer Observatory. Int. Agency Res. Cancer. , 149 , 1–2. Koné, A., Souleymane, C., Kalo, M., Tchambaga Etienne, C., Collet, S., and Sissouma, D. (2025) Synthesis of novel benzimidazole-based retrochalcones and their anticancer activity against breast and colon cancer. Synth. Commun. , 55 (2), 175–182. Mahmoudi, N., de Julián-Ortiz, J.V., Ciceron, L., Gálvez, J., Mazier, D., Danis, M., Derouin, F., and García-Domenech, R. (2006) Identification of new antimalarial drugs by linear discriminant analysis and topological virtual screening. J. Antimicrob. Chemother. , 57 (3), 489–497. Willcox, M.L., and Bodeker, G. (2004) Traditional herbal medicines for malaria. Br. Med. J. , 329 (7475), 1156–1159. Dietrich. H., (1994) Toxizität chlororganischer Verbindungen: Einfluß der Einführung von Chlor in organische Moleküle. Angew. Chem . 106 , 1997. Dietrich. H., (1994) Toxicity of Chlorinated Organic Compounds: Effects of the Introduction of Chlorine in Organic Molecules. Angew. Chem. Int. Ed. Engl., 33 : 1920-1935. Šegan, S., Krunić, M. J., Andrić, D. B., Šukalović, V. B., Penjišević, J. Z., and Jevtić, I. I. ( 2024) Evaluation of lipophilicity and drug-likeness of donepezil-like compounds using reversed-phase thin-layer chromatography. Biomedical Chromatography ,38(7), e5867. Table Table 1 is available in the Supplementary Files section. Scheme Scheme 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. 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1","display":"","copyAsset":false,"role":"figure","size":82896,"visible":true,"origin":"","legend":"\u003cp\u003eDesign of benzimidazolyl-propenone structures\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7490108/v1/b77c48512beb5d5973bc2330.png"},{"id":95230581,"identity":"9076e321-32b1-4f1c-b191-87abbb9d5ea6","added_by":"auto","created_at":"2025-11-05 16:38:02","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1874110,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7490108/v1/d2e43143-c34e-45a9-a82a-bdfd48b7fc1f.pdf"},{"id":95226052,"identity":"3471bc6f-c50e-4108-9833-f2381dd904fd","added_by":"auto","created_at":"2025-11-05 16:26:07","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":4604964,"visible":true,"origin":"","legend":"","description":"","filename":"graphicalabstrat.tif","url":"https://assets-eu.researchsquare.com/files/rs-7490108/v1/703062d3cdd568bbe7891972.tif"},{"id":95225366,"identity":"60c6088e-684e-44f8-aa8e-3ddfd42e69d1","added_by":"auto","created_at":"2025-11-05 16:24:55","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":2353302,"visible":true,"origin":"","legend":"","description":"","filename":"SIKone.docx","url":"https://assets-eu.researchsquare.com/files/rs-7490108/v1/d8217fc11fb81cc73652aa79.docx"},{"id":95125894,"identity":"eded51f6-b557-4add-ab6e-44b33f95cbc9","added_by":"auto","created_at":"2025-11-04 14:59:17","extension":"png","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":53309,"visible":true,"origin":"","legend":"","description":"","filename":"scheme1.png","url":"https://assets-eu.researchsquare.com/files/rs-7490108/v1/34e8c481bdce4baba83e6060.png"},{"id":95125893,"identity":"c9e6ac78-9b65-46d7-b38b-9fa7fbece770","added_by":"auto","created_at":"2025-11-04 14:59:17","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":29712,"visible":true,"origin":"","legend":"","description":"","filename":"Table1.docx","url":"https://assets-eu.researchsquare.com/files/rs-7490108/v1/38291def202cddbba0bc018f.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Benzimidazolyl-Phenylpropenones Synthesized and Evaluated for Selective Anticancer Activity in Prostate Colon and Breast Tumor Models","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBenzimidazole is a privileged heterocyclic scaffold found in many therapeutic agents, particularly anthelmintics [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e] and widely recognized for its broad spectrum of biological activities, including antifungal [\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], antibacterial [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], antiviral [\u003cspan additionalcitationids=\"CR10\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], anti-inflammatory [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], antioxidant [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], and anticancer effects [\u003cspan additionalcitationids=\"CR16 CR17\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Despite ongoing efforts to combat cancer, the World Health Organization reported approximately 20\u0026nbsp;million new cases and 9.7\u0026nbsp;million deaths worldwide in 2022 [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. In C\u0026ocirc;te d\u0026rsquo;Ivoire, both cancer incidence and mortality remain high. Data from the Global Cancer Observatory (Globocan 2022) indicate 21,352 new cancer cases and 14,143 deaths in the country [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Prostate cancer is the most prevalent (18.9%), followed by breast (18.1%), liver (6%), and colon (5%) cancers. These statistics underscore that cancer remains a major public health challenge globally, and particularly in C\u0026ocirc;te d\u0026rsquo;Ivoire. Our previous work demonstrated the anticancer potential of benzimidazole\u0026ndash;chalcone derivatives against breast and colon cancers, as well as their effects on normal skin fibroblasts [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Chalcones are well known for their strong antimicrobial and anticancer activities. Building on the concept of combining biologically active moieties, the present study focuses on synthesizing benzimidazole\u0026ndash;chalcone derivatives substituted at the 5-position of the benzimidazole ring. Their \u003cem\u003ein vitro\u003c/em\u003e anticancer activity was evaluated against prostate, colon, and breast cancer cell lines, as well as normal skin fibroblasts (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e"},{"header":"Results and discussion","content":"\u003cp\u003eThe synthesis of benzimidazolyl\u0026ndash;phenylpropenone derivatives (\u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e) was achieved in three steps, following the strategy previously reported for benzimidazole\u0026ndash;chalcone derivatives. In this work, seven compounds were successfully obtained, including derivatives \u003cstrong\u003e3a\u0026ndash;c\u003c/strong\u003e, which were previously evaluated against colon and breast cancer cell lines, as well as normal skin fibroblasts [\u003cspan class=\"CitationRef\"\u003e21\u003c/span\u003e]. The synthetic route is outlined in \u003cstrong\u003eScheme 1\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003eBriefly, \u003cem\u003eortho\u003c/em\u003e-phenylenediamine derivatives were first reacted with glycolic acid in the presence of 4 N hydrochloric acid. The resulting intermediates (\u003cstrong\u003e1a\u0026ndash;d\u003c/strong\u003e) were then oxidized with manganese dioxide in ethanol. Finally, benzimidazole-2-carbaldehydes were condensed with substituted acetophenones \u003cem\u003evia\u003c/em\u003e the Claisen\u0026ndash;Schmidt reaction, affording the target benzimidazolyl\u0026ndash;phenylpropenones (\u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e) (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe synthesized benzimidazolyl\u0026ndash;phenylpropenone derivatives (\u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e) were fully characterized by \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC NMR spectroscopy and HRMS (\u003cem\u003esee Supporting Information\u003c/em\u003e). In the \u003csup\u003e1\u003c/sup\u003eH NMR spectra, the ethylenic protons appeared as doublets between 7.50 and 8.50 ppm, with coupling constants ranging from 15.2 to 15.8 Hz, confirming the \u003cem\u003etrans\u003c/em\u003e configuration of the synthesized compounds. In the \u003csup\u003e13\u003c/sup\u003eC NMR spectra, the carbonyl carbon (C=O) resonated between 187.9 and 195.5 ppm, while the ethylenic carbons appeared between 128 and 150 ppm.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe cytotoxic activity of compounds \u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e was evaluated against prostate, colon, and breast cancer cell lines, as well as normal human skin fibroblasts (\u003cstrong\u003eTable 2\u003c/strong\u003e). According to the criteria proposed by Mahmoudi et \u003cem\u003eal.\u003c/em\u003e [22] and Wilcox et \u003cem\u003eal.\u0026nbsp;\u003c/em\u003e[23], compounds are classified as highly active when IC\u003csub\u003e50\u003c/sub\u003e \u0026lt; 0.06 \u0026mu;M, active when 0.06 \u0026mu;M \u0026le; IC\u003csub\u003e50\u003c/sub\u003e \u0026le; 5 \u0026mu;M, moderately active when 5 \u0026mu;M \u0026le; IC\u003csub\u003e50\u003c/sub\u003e \u0026le; 10 \u0026mu;M, and inactive when IC\u003csub\u003e50\u003c/sub\u003e \u0026gt; 10 \u0026mu;M. The amplitude of activity was defined as the difference between the upper portion of the dose\u0026ndash;response curve (no effect) and its lower portion (maximum effect).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e:\u0026nbsp;Cytotoxicity of synthesized compounds against a panel of cancer cell lines and normal skin fibroblasts.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"633\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" style=\"width: 60px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 94px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eProstate\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" style=\"width: 188px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eColon\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" style=\"width: 188px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBreast\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 94px;\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 94px;\"\u003e\n \u003cp\u003eCaCo2\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 94px;\"\u003e\n \u003cp\u003eHCT-116\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 94px;\"\u003e\n \u003cp\u003eMDA-MB 231\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 94px;\"\u003e\n \u003cp\u003eMCF-7\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 104px;\"\u003e\n \u003cp\u003eFibroblaste\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e5.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e1.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e5.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e38\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e1.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e1.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e7.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e7.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e7.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3f\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e2.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e1.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e4.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e8.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e5.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e6.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e5.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e5.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRosco\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTaxol\u003c/strong\u003e\u003cstrong\u003e\u0026reg;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026gt;0.25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCompound \u003cstrong\u003e3a\u003c/strong\u003e exhibited significant activity against PC3, HCT-116, and MDA-MB-231 cancer cell lines, with IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues ranging from 1.48 to 4.17 \u0026mu;M, and moderate activity against CaCo2 and MCF-7, with IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues of 5.25 \u0026mu;M and 7.77 \u0026mu;M, respectively. Its strongest effect was observed on HCT-116 colon cancer cells (IC\u003csub\u003e50\u003c/sub\u003e= 1.48 \u0026mu;M). Introduction of hydroxyl substituents at the \u003cem\u003eortho\u003c/em\u003e (\u003cstrong\u003e3b\u003c/strong\u003e) and \u003cem\u003emeta\u003c/em\u003e (\u003cstrong\u003e3c\u003c/strong\u003e) positions of the phenyl ring markedly enhanced antitumor activity. Both compounds \u003cstrong\u003e3b\u003c/strong\u003e and \u003cstrong\u003e3c\u003c/strong\u003e were active across all tested cancer cell lines, with IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues ranging from 1.35 to 3.01 \u0026mu;M. Among them, compound \u003cstrong\u003e3c\u003c/strong\u003e displayed the highest potency, particularly against PC3 prostate (IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003e= 1.83 \u0026mu;M) and MDA-MB-231 breast cancer cells (IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003e= 1.35 \u0026mu;M). These results suggest that electron-donating hydroxyl groups on the phenyl ring favor enhanced antitumor activity. Substitution at the 5-position of the benzimidazole ring further influenced activity. Compounds bearing chloro, nitro, or benzoyl groups at this position were active against prostate cancer cells (IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003e= 1.78\u0026ndash;3.89 \u0026mu;M). Interestingly, the nitro derivative (\u003cstrong\u003e3g\u003c/strong\u003e) was more effective against PC3 cells (IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003e= 1.78 \u0026mu;M) compared with chloro (\u003cstrong\u003e3d\u0026ndash;f\u003c/strong\u003e) or benzoyl (\u003cstrong\u003e3h\u003c/strong\u003e) analogs, suggesting that electron-withdrawing substituents such as nitro are favorable for activity in prostate cancer. However, in colon and breast cancer cell lines, chloro-substituted derivatives (\u003cstrong\u003e3d\u0026ndash;f\u003c/strong\u003e) did not lead to significant improvements, showing IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues in the range of 2.26\u0026ndash;4.99 \u0026mu;M (active) and 7.44\u0026ndash;7.48 \u0026mu;M (moderately active). Similarly, nitro (\u003cstrong\u003e3g\u003c/strong\u003e) and benzoyl (\u003cstrong\u003e3h\u003c/strong\u003e) substitution at C-5 of the benzimidazole ring, combined with hydroxyl substitution on the phenyl ring, did not enhance activity in colon or breast cancer models. Regarding toxicity in normal human fibroblasts, all compounds showed measurable effects with IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues between 3.23 and 8.02 \u0026mu;M. However, their amplitudes of activity were below 50%, indicating that only a limited fraction of dividing fibroblasts was affected. By contrast, the high amplitudes observed in cancer cell lines (74\u0026ndash;98%) suggest that compounds \u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e strongly target proliferating tumor cells, leaving only a small residual population unaffected. This selectivity supports their potential as antimitotic agents with reduced toxicity toward noncancerous cells.\u003c/p\u003e\n\u003cp\u003eComparison with reference drugs highlights the relative potency of these derivatives. Roscovitine was largely inactive across the panel, with IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues \u0026gt;10 \u0026mu;M in four of the five cancer cell lines and only moderate activity in HCT-116 cells (IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003e= 8 \u0026mu;M). In contrast, Paclitaxel (Taxol\u0026reg;) displayed exceptional potency, with IC\u003csub\u003e50\u0026nbsp;\u003c/sub\u003evalues between 0.001 and 0.04 \u0026mu;M across all cancer cell lines tested. In summary, benzimidazolyl\u0026ndash;phenylpropenone derivatives demonstrated broad and selective anticancer activity, with hydroxyl-substituted analogs (\u003cstrong\u003e3b\u003c/strong\u003e, \u003cstrong\u003e3c\u003c/strong\u003e) and the nitro derivative (\u003cstrong\u003e3g\u003c/strong\u003e) emerging as the most potent. By contrast, chloro-substituted derivatives, even when combined with hydroxyl groups, did not improve activity, a finding consistent with QSAR predictions and previous literature,\u003csup\u003e24-26\u003c/sup\u003e where the high lipophilicity of chloro substituents often compromises bioactivity rather than enhancing it. Overall, the present series was generally more active than Roscovitine while less potent than Paclitaxel, but their favorable selectivity toward cancer cells over normal fibroblasts underscores their promise as lead scaffolds for further optimization in anticancer drug development.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe synthesized benzimidazolyl\u0026ndash;phenylpropenone derivatives (\u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e) were fully characterized by \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC NMR spectroscopy and HRMS (\u003cem\u003esee Supporting Information\u003c/em\u003e). In the \u003csup\u003e1\u003c/sup\u003eH NMR spectra, the ethylenic protons appeared as doublets between 7.50 and 8.50 ppm, with coupling constants ranging from 15.2 to 15.8 Hz, confirming the \u003cem\u003etrans\u003c/em\u003e configuration of the synthesized compounds. In the \u003csup\u003e13\u003c/sup\u003eC NMR spectra, the carbonyl carbon (C\u0026thinsp;=\u0026thinsp;O) resonated between 187.9 and 195.5 ppm, while the ethylenic carbons appeared between 128 and 150 ppm.\u003c/p\u003e\n\u003cp\u003eThe cytotoxic activity of compounds \u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e was evaluated against prostate, colon, and breast cancer cell lines, as well as normal human skin fibroblasts (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). According to the criteria proposed by Mahmoudi et \u003cem\u003eal.\u003c/em\u003e [\u003cspan class=\"CitationRef\"\u003e22\u003c/span\u003e] and Wilcox et \u003cem\u003eal.\u003c/em\u003e [\u003cspan class=\"CitationRef\"\u003e23\u003c/span\u003e], compounds are classified as highly active when IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.06 \u0026micro;M, active when 0.06 \u0026micro;M\u0026thinsp;\u0026le;\u0026thinsp;IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;\u0026le;\u0026thinsp;5 \u0026micro;M, moderately active when 5 \u0026micro;M\u0026thinsp;\u0026le;\u0026thinsp;IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;\u0026le;\u0026thinsp;10 \u0026micro;M, and inactive when IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;\u0026gt;\u0026thinsp;10 \u0026micro;M. The amplitude of activity was defined as the difference between the upper portion of the dose\u0026ndash;response curve (no effect) and its lower portion (maximum effect).\u003c/p\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCytotoxicity of synthesized compounds against a panel of cancer cell lines and normal skin fibroblasts.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eProstate\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eColon\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eBreast\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eCaCo2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eHCT-116\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eMDA-MB 231\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eMCF-7\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eFibroblaste\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eamp\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3a\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3b\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e38\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3f\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3g\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eRosco\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTaxol\u0026reg;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026gt;\u0026thinsp;0.25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cp\u003eCompound \u003cstrong\u003e3a\u003c/strong\u003e exhibited significant activity against PC3, HCT-116, and MDA-MB-231 cancer cell lines, with IC\u003csub\u003e50\u003c/sub\u003e values ranging from 1.48 to 4.17 \u0026micro;M, and moderate activity against CaCo2 and MCF-7, with IC\u003csub\u003e50\u003c/sub\u003e values of 5.25 \u0026micro;M and 7.77 \u0026micro;M, respectively. Its strongest effect was observed on HCT-116 colon cancer cells (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.48 \u0026micro;M). Introduction of hydroxyl substituents at the \u003cem\u003eortho\u003c/em\u003e (\u003cstrong\u003e3b\u003c/strong\u003e) and \u003cem\u003emeta\u003c/em\u003e (\u003cstrong\u003e3c\u003c/strong\u003e) positions of the phenyl ring markedly enhanced antitumor activity. Both compounds \u003cstrong\u003e3b\u003c/strong\u003e and \u003cstrong\u003e3c\u003c/strong\u003e were active across all tested cancer cell lines, with IC\u003csub\u003e50\u003c/sub\u003e values ranging from 1.35 to 3.01 \u0026micro;M. Among them, compound \u003cstrong\u003e3c\u003c/strong\u003e displayed the highest potency, particularly against PC3 prostate (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.83 \u0026micro;M) and MDA-MB-231 breast cancer cells (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.35 \u0026micro;M). These results suggest that electron-donating hydroxyl groups on the phenyl ring favor enhanced antitumor activity. Substitution at the 5-position of the benzimidazole ring further influenced activity. Compounds bearing chloro, nitro, or benzoyl groups at this position were active against prostate cancer cells (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.78\u0026ndash;3.89 \u0026micro;M). Interestingly, the nitro derivative (\u003cstrong\u003e3g\u003c/strong\u003e) was more effective against PC3 cells (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.78 \u0026micro;M) compared with chloro (\u003cstrong\u003e3d\u0026ndash;f\u003c/strong\u003e) or benzoyl (\u003cstrong\u003e3h\u003c/strong\u003e) analogs, suggesting that electron-withdrawing substituents such as nitro are favorable for activity in prostate cancer. However, in colon and breast cancer cell lines, chloro-substituted derivatives (\u003cstrong\u003e3d\u0026ndash;f\u003c/strong\u003e) did not lead to significant improvements, showing IC\u003csub\u003e50\u003c/sub\u003e values in the range of 2.26\u0026ndash;4.99 \u0026micro;M (active) and 7.44\u0026ndash;7.48 \u0026micro;M (moderately active). Similarly, nitro (\u003cstrong\u003e3g\u003c/strong\u003e) and benzoyl (\u003cstrong\u003e3h\u003c/strong\u003e) substitution at C-5 of the benzimidazole ring, combined with hydroxyl substitution on the phenyl ring, did not enhance activity in colon or breast cancer models. Regarding toxicity in normal human fibroblasts, all compounds showed measurable effects with IC\u003csub\u003e50\u003c/sub\u003e values between 3.23 and 8.02 \u0026micro;M. However, their amplitudes of activity were below 50%, indicating that only a limited fraction of dividing fibroblasts was affected. By contrast, the high amplitudes observed in cancer cell lines (74\u0026ndash;98%) suggest that compounds \u003cstrong\u003e3a\u0026ndash;h\u003c/strong\u003e strongly target proliferating tumor cells, leaving only a small residual population unaffected. This selectivity supports their potential as antimitotic agents with reduced toxicity toward noncancerous cells.\u003c/p\u003e\n\u003cp\u003eComparison with reference drugs highlights the relative potency of these derivatives. Roscovitine was largely inactive across the panel, with IC\u003csub\u003e50\u003c/sub\u003e values\u0026thinsp;\u0026gt;\u0026thinsp;10 \u0026micro;M in four of the five cancer cell lines and only moderate activity in HCT-116 cells (IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;8 \u0026micro;M). In contrast, Paclitaxel (Taxol\u0026reg;) displayed exceptional potency, with IC\u003csub\u003e50\u003c/sub\u003e values between 0.001 and 0.04 \u0026micro;M across all cancer cell lines tested. In summary, benzimidazolyl\u0026ndash;phenylpropenone derivatives demonstrated broad and selective anticancer activity, with hydroxyl-substituted analogs (\u003cstrong\u003e3b\u003c/strong\u003e, \u003cstrong\u003e3c\u003c/strong\u003e) and the nitro derivative (\u003cstrong\u003e3g\u003c/strong\u003e) emerging as the most potent. By contrast, chloro-substituted derivatives, even when combined with hydroxyl groups, did not improve activity, a finding consistent with QSAR predictions and previous literature,\u003csup\u003e24\u0026ndash;26\u003c/sup\u003e where the high lipophilicity of chloro substituents often compromises bioactivity rather than enhancing it. Overall, the present series was generally more active than Roscovitine while less potent than Paclitaxel, but their favorable selectivity toward cancer cells over normal fibroblasts underscores their promise as lead scaffolds for further optimization in anticancer drug development.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe benzimidazole derivatives designed and synthesized in this study demonstrated moderate to strong cytotoxic activity across prostate, colon, and breast cancer cell lines, while maintaining reduced effects on normal fibroblasts. Hydroxyl-substituted analogs (\u003cb\u003e3b\u003c/b\u003e, \u003cb\u003e3c\u003c/b\u003e) and the nitro derivative (\u003cb\u003e3g\u003c/b\u003e) emerged as the most potent, highlighting the importance of electronic effects in modulating anticancer activity. By contrast, substitution at the 5-position of the benzimidazole ring with chloro, nitro, or benzoyl groups generally did not enhance activity compared with their non-substituted counterparts. This observation is not surprising, as QSAR modeling and literature evidence suggest that the high lipophilicity of chloro substituents, even in combination with hydroxyl groups, often compromises bioactivity rather than improving it. Despite this, all 5-substituted derivatives displayed higher activity than the reference compound Roscovitine, though they remained less potent than Paclitaxel. These findings are encouraging, as they identify benzimidazolyl\u0026ndash;phenylpropenones as promising scaffolds for further optimization. Future work will focus on refining the structure\u0026ndash;activity relationships of benzimidazolyl\u0026ndash;phenylpropenones through computational approaches, including molecular docking, and molecular dynamics simulations to better understand their interactions with potential biological targets. In addition, further structural modifications aimed at optimizing substituents on both the benzimidazole core and the phenyl ring may enhance potency and selectivity. Evaluation in 3D tumor spheroid models and subsequent \u003cem\u003ein vivo\u003c/em\u003e studies will be crucial to validate their therapeutic potential. Ultimately, these efforts could accelerate the development of cost-effective anticancer agents particularly suited for addressing the growing cancer burden in low- and middle-income countries.\u003c/p\u003e"},{"header":"Experimental section","content":"\u003cp\u003e\u003cstrong\u003eGeneral\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor all compounds, the \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC NMR spectra were recorded at room temperature on a Bruker Avance AC300 (300 MHz for \u003csup\u003e1\u003c/sup\u003eH and 75 MHz for \u003csup\u003e13\u003c/sup\u003eC) or on a Bruker Avance AC300 (400 MHz for \u003csup\u003e1\u003c/sup\u003eH and 100 MHz for \u003csup\u003e13\u003c/sup\u003eC) in deuterated dimethylsulfoxide (DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e). Tetramethylsilane (TMS) was used as a reference and chemical shifts are expressed in parts per million (ppm) while coupling constants (\u003cem\u003eJ\u003c/em\u003e) are expressed in Hertz (Hz). NMR spectra are described using the following symbols: s (singlet), d (doublet), t (triplet), q (quadruplet), m (massive), dd (doublet of doublet). Mass spectra were recorded using a ThermoScientific DSQII quadrupole instrument by electron impact (EI, 70 eV) or chemical ionization (CI, 500 eV). High-resolution mass spectroscopy (HRMS) was performed on a ThermoScientific LTQ-Orbitrap mass spectrometer in positive electrospray ionization mode. The progress of the reaction and the purity of the compounds were monitored by TLC on aluminum plates coated with silica gel (Kieselgel 60 F\u003csub\u003e254\u003c/sub\u003e, MERCK). After elution in the appropriate solvent or solvent mixture, the plates were revealed by UV fluorescence (\u0026lambda; = 254 nm) or by potassium permanganate (KMnO\u003csub\u003e4\u003c/sub\u003e) solution followed by heating. The melting points of solid compounds were determined using a K\u0026ouml;fler bench and are uncorrected. Solvents and reagents were purchased from Acros Organics (France) or Sigma-Aldrich (France).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCompounds \u003cstrong\u003e1a\u0026nbsp;\u003c/strong\u003e(1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)methanol), \u003cstrong\u003e2a\u0026nbsp;\u003c/strong\u003e(1\u003cem\u003eH\u003c/em\u003e-benzimidazole-2-carbaldehyde), \u003cstrong\u003e3a\u0026nbsp;\u003c/strong\u003e((\u003cem\u003eE\u003c/em\u003e)-3-(1\u003cem\u003eH\u003c/em\u003e-Benzimidazol-2-yl)-1-phenylprop-2-en-1-one), \u003cstrong\u003e3b\u003c/strong\u003e ((\u003cem\u003eE\u003c/em\u003e)-3-(1\u003cem\u003eH\u003c/em\u003e-Benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one) and ((\u003cem\u003eE\u003c/em\u003e)-3-(1\u003cem\u003eH\u003c/em\u003e-Benzimidazol-2-yl)-1-(2-hydroxy phenyl)prop-2-en-1-one) \u0026nbsp;have already been reported [21].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of (1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)methanol derivatives 1b-d\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo a solution of \u003cem\u003eo\u003c/em\u003e-phenylenediamine derivatives (1 eq, 5 g) in HCl 4N (50 mL), was added glycolic acid (3 eq). The reaction mixture was refluxed for 6 h then cooled to room temperature and quenched with ammonia solution 25% at 0 \u0026deg;C. The product was collected pure by filtration and dried at room temperature to afford benzimidazolyl-methanol derivatives.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e(5-Chloro-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)methanol 1b\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBrown solid, yield = 97%, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003e\u003c/strong\u003e\u003cstrong\u003eH NMR: (300 MHz, DMSO-\u003cem\u003ed6\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e12.46 (br s, 1H, NH), 7.56-7.47 (m, 2H, H4 and H7) 7.15 (dd, \u003cem\u003eJ6-7\u0026nbsp;\u003c/em\u003e= 8.8, \u003cem\u003eJ6-4\u0026nbsp;\u003c/em\u003e= 2.1 Hz, 1H, H6), 5.76 (br s, 1H, OH), 4.69 (s, 2H, H1). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (75 MHz, DMSO-\u003cem\u003ed6\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e156.9 (CAr), 133.0 (CAr), 121.6 (CAr), 119.6 (CAr), 117.8 (CAr), 112.5 (CAr), 110.9 (CAr), 57.6 (CH2). \u003cstrong\u003eMS : [EI, 70 eV, m/z (rel. Int)] :\u0026nbsp;\u003c/strong\u003e184 (29), 183 (24), 182 (100), 181 (40), 165 (29).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(5-Nitro-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)methanol\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e1c\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eBrown solid, yield = 97%, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR\u0026nbsp;: (300 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e12.71 (br s, 1H, NH), 8.68-8.15 (m, 1H, H\u003csub\u003e4\u003c/sub\u003e), 8.08 (dd, \u003cem\u003eJ\u003csub\u003e6-7\u003c/sub\u003e\u0026nbsp;\u003c/em\u003e= 8.9 Hz, \u003cem\u003eJ\u003csub\u003e6-4\u003c/sub\u003e\u003c/em\u003e = 2.3 Hz, 1H, H\u003csub\u003e6\u003c/sub\u003e), 7.67 (d, \u003cem\u003eJ\u003csub\u003e7-6\u003c/sub\u003e\u0026nbsp;\u003c/em\u003e= 8.8 Hz, 1H, H\u003csub\u003e7\u003c/sub\u003e), 5.93 (s, 1H, OH), 4.77 (s, 2H, H\u003csub\u003e1\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (75 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e159.8 (C\u003csub\u003eAr\u003c/sub\u003e), 142.3 (C\u003csub\u003eAr\u003c/sub\u003e), 118.5 (C\u003csub\u003eAr\u003c/sub\u003e), 117.6 (C\u003csub\u003eAr\u003c/sub\u003e), 114.5 (C\u003csub\u003eAr\u003c/sub\u003e), 111.5 (C\u003csub\u003eAr\u003c/sub\u003e), 107.9 (C\u003csub\u003eAr\u003c/sub\u003e), 57.7 (CH\u003csub\u003e2\u003c/sub\u003e). \u003cstrong\u003eMS: [EI, 70 eV, m/z (rel. Int)]:\u0026nbsp;\u003c/strong\u003e194 (16), 193 (100), 192 (28), 147 (18).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(5-Benzoyl-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)methanol\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e1d\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eBeige solide, yield = 97%, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR: (300 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e12.72 (br s, 1H, NH), 7.88 (t, \u003cem\u003eJ\u003c/em\u003e = 1.2 Hz, 1H, H\u003csub\u003e4\u003c/sub\u003e), 7.76-7.70 (m, 2H, H\u003csub\u003e6\u003c/sub\u003e and H\u003csub\u003e7\u003c/sub\u003e), 7.70-7.52 (m, 5H, H\u003csub\u003eAr\u003c/sub\u003e), 5.83 (br s, 1H, OH), 4.74 (s, 2H, H\u003csub\u003e1\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (75 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e195.7 (C=O), 138.2 (2C\u003csub\u003eAr\u003c/sub\u003e), 132.0 (C\u003csub\u003eAr\u003c/sub\u003e), 130.3 (2C\u003csub\u003eAr\u003c/sub\u003e), 129.3 (2C\u003csub\u003eAr\u003c/sub\u003e), 128.3 (4C\u003csub\u003eAr\u003c/sub\u003e), 123.6 (2C\u003csub\u003eAr\u003c/sub\u003e), 57.7 (CH\u003csub\u003e2\u003c/sub\u003e). \u003cstrong\u003eMS:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e[EI, 70 eV, m/z (rel. Int)] :\u0026nbsp;\u003c/strong\u003e253 (20), 252 (100), 251 (14), 175 (76).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of 1\u003cem\u003eH\u003c/em\u003e-benzimidazole-2-carbaldehyde (2b-d)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo a solution of benzimidazol-2-yl methanol and\u0026nbsp;derivatives\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e(1 eq, 1 g) in 10 mL of EtOH, was added activated manganese dioxide (8.5 eq). The reaction mixture was stirred at room temperature overnight and concentrated. 20 mL of boiling DMF was added, filtered over a pad of celite and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (elution: DCM/MeOH 98: 2) to afford benzimidazole-2-carbaldehydes derivatives\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5-Chloro-1\u003cem\u003eH\u003c/em\u003e-benzimidazole-2-carbaldehyde (2b)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 52%, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003e\u003c/strong\u003e\u003cstrong\u003eH NMR : (300 MHz, DMSO-\u003cem\u003ed6\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e13.82 (br s, 1H, NH), 9.95 (s, 1H, CHO), 7.79-7.35 (m, 3 H, H4, H6 and H7) \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR : (75 MHz, DMSO-\u003cem\u003ed6\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e183.4 (CH=O), 141.7 (CAr), 140.6 (CAr), 136.9 (CAr), 129.5 (CAr), 123.8 (CAr), 116.3 (CAr), 115.5 (CAr) \u003cstrong\u003eMS : [EI, 70 eV, m/z (rel. Int)] :\u0026nbsp;\u003c/strong\u003e180 (12), 154 (28), 152 (100), 151 (11).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5-Nitro-1\u003cem\u003eH\u003c/em\u003e-benzimidazole-2-carbaldehyde\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e2c\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 55%, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR: (300 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e14.04 (br s, 1H, NH), 9.97 (s, 1H, CHO), 8.50-7.73 (m, 3H, H\u003csub\u003e4\u003c/sub\u003e, H\u003csub\u003e6\u003c/sub\u003e and H\u003csub\u003e7\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (75 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e183.1 (CH=O), 145.1 (C\u003csub\u003eAr\u003c/sub\u003e), 144.4 (C\u003csub\u003eAr\u003c/sub\u003e), 141.5 (C\u003csub\u003eAr\u003c/sub\u003e), 139.8 (C\u003csub\u003eAr\u003c/sub\u003e), 118.3 (C\u003csub\u003eAr\u003c/sub\u003e), 116.6 (C\u003csub\u003eAr\u003c/sub\u003e), 113.0 (C\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003eMS: [EI, 70 eV, m/z (rel. Int)]:\u0026nbsp;\u003c/strong\u003e192 (16), 191 (100), 190 (28), 145 (18).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5-Benzoyl-1\u003cem\u003eH\u003c/em\u003e-benzimidazole-2-carbaldehyde\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e2d\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 57%, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR\u0026nbsp;: (400 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e13.83 (s, 1H, NH), 10.01 (s, 1H, CHO), 7.80-7.75 (m, 3H, H\u003csub\u003eAr\u003c/sub\u003e), 7.72-7.68 (m, 2H, H\u003csub\u003eAr\u003c/sub\u003e), 7.61-7.57 (m, 3H, H\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (75 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e194.0 (C=O), 183.6 (CH=O), 142.6 (C\u003csub\u003eAr\u003c/sub\u003e), 141.5 (C\u003csub\u003eAr\u003c/sub\u003e), 138.6 (C\u003csub\u003eAr\u003c/sub\u003e), 138.4 (C\u003csub\u003eAr\u003c/sub\u003e), 132.4 (C\u003csub\u003eAr\u003c/sub\u003e), 131.2 (C\u003csub\u003eAr\u003c/sub\u003e), 130.3 (2C\u003csub\u003eAr\u003c/sub\u003e), 128.4 (2C\u003csub\u003eAr\u003c/sub\u003e), 124.1 (C\u003csub\u003eAr\u003c/sub\u003e), 119.0 (C\u003csub\u003eAr\u003c/sub\u003e), 114.9 (C\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003eMS: [EI, 70 eV, m/z (rel. Int)] :\u0026nbsp;\u003c/strong\u003e251 (19), 250 (100), 249 (14), 173 (82), 145 (17).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of (\u003cem\u003eE\u003c/em\u003e)-3-(1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-phenylprop-2-en-1-one derivatives (3d-h)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo a solution of NaOH (7.5 eq) in ethanol (10 mL), were added benzimidazole-2- carbaldehyde derivatives \u003cstrong\u003e2b-d\u0026nbsp;\u003c/strong\u003e(1 eq, 500 mg) and appropriate acetophenone (1 eq). The mixture was stirred at room temperature overnight then quenched with acetic acid 20% at 0 \u0026deg;C. The crude was collected by filtration, dried and purified by flash-chromatography on silica gel (DCM/MeOH 95 :5) to afford 3-(1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-arylprop-2-en-1-one derivatives.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(\u003cem\u003eE\u003c/em\u003e)-3-(5-Chloro-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-phenylprop-2-en-1-one\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e3d\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 71%, M.p = 158-160 \u0026deg;C, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR : (400 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e12.04 (br s, 1H, NH), 8.50 (d, \u003cem\u003eJ\u003csub\u003e3-2\u003c/sub\u003e\u003c/em\u003e = 15.8 Hz, 1H, H\u003csub\u003e3\u003c/sub\u003e), 8.16 (d, \u003cem\u003eJ\u003c/em\u003e = 7.3 Hz, 2H, H\u003csub\u003eAr\u003c/sub\u003e), 7.80-7.72 (m, 3H, H\u003csub\u003e2\u003c/sub\u003e and 2H\u003csub\u003eAr\u003c/sub\u003e), 7.65-7.59 (m, 3H, H\u003csub\u003eAr\u003c/sub\u003e), 7.40 (dd, \u003cem\u003eJ\u003c/em\u003e = 1.6 Hz, \u003cem\u003eJ\u003c/em\u003e = 8.7 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (100 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e188.2 (C=O), 149.0 (C\u003csub\u003eAr\u003c/sub\u003e), 137.6 (C\u003csub\u003eAr\u003c/sub\u003e), 136.7 (C\u003csub\u003eAr\u003c/sub\u003e), 135.5 (C\u003csub\u003eAr\u003c/sub\u003e), 133.9 (C\u003csub\u003eAr\u003c/sub\u003e), 129.5 (2C\u003csub\u003eAr\u003c/sub\u003e), 129.0 (2C\u003csub\u003eAr\u003c/sub\u003e), 128.8 (C\u003csub\u003eAr\u003c/sub\u003e), 128.7 (C\u003csub\u003eAr\u003c/sub\u003e), 128.6 (C\u003csub\u003eAr\u003c/sub\u003e), 124.7 (C\u003csub\u003eAr\u003c/sub\u003e), 116.5 (C\u003csub\u003eAr\u003c/sub\u003e), 114.8 (C\u003csub\u003eAr\u003c/sub\u003e).\u0026nbsp;\u003cstrong\u003eHRMS: (ES+)\u0026nbsp;\u003c/strong\u003eCalculated for C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eOCl [M + H]\u003csup\u003e+\u003c/sup\u003e : 283.0638, found : 283.0634. \u003cstrong\u003eIR: (ATR,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026upsilon;\u003c/strong\u003e\u003cstrong\u003e\u003cspan id=\"_Toc527564623\"\u003e cm\u003csup\u003e-1\u003c/sup\u003e)\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e3453.2 (N-H), 2919.1 (=C-H), 1673.1 (C=O), 1441.9 (C=C), 710.9 (C-Cl).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(\u003cem\u003eE\u003c/em\u003e)-3-(5-Chloro-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e3e\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 69%, M.p = 116-118 \u0026deg;C, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR : (400 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e11.94 (s, 1H, NH), 8.26 (d, \u003cem\u003eJ\u003csub\u003e3-2\u003c/sub\u003e\u003c/em\u003e = 15.2 Hz, 1H, H\u003csub\u003e3\u003c/sub\u003e), 8.00 (dd, \u003cem\u003eJ\u003c/em\u003e = 1.2 Hz, \u003cem\u003eJ\u003c/em\u003e = 8.1 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.72-7.52 (m, 5H, 4H\u003csub\u003eAr\u003c/sub\u003e and H\u003csub\u003e2\u003c/sub\u003e), 7.29 (d, \u003cem\u003eJ\u003c/em\u003e = 8.3 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.06-702 (m, 2H, H\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR : (100 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e192.2 (C=O), 170.2 (C\u003csub\u003eAr\u003c/sub\u003e), 161.0 (C\u003csub\u003eAr\u003c/sub\u003e), 138.3 (C\u003csub\u003eAr\u003c/sub\u003e), 136.3 (C\u003csub\u003eAr\u003c/sub\u003e), 131.2 (C\u003csub\u003eAr\u003c/sub\u003e), 130.4 (2C\u003csub\u003eAr\u003c/sub\u003e), 127.8 (C\u003csub\u003eAr\u003c/sub\u003e), 123.9 (C\u003csub\u003eAr\u003c/sub\u003e), 123.6 (C\u003csub\u003eAr\u003c/sub\u003e), 121.4 (C\u003csub\u003eAr\u003c/sub\u003e), 119.4 (2C\u003csub\u003eAr\u003c/sub\u003e), 117.7 (C\u003csub\u003eAr\u003c/sub\u003e), 113.5 (C\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003eMS : [EI, 70 eV, m/z (rel. Int)] :\u003c/strong\u003e 299 (23), 298 (100) \u003cstrong\u003e[CI, NH\u003csub\u003e3,\u0026nbsp;\u003c/sub\u003em/z] :\u0026nbsp;\u003c/strong\u003e299 [M+H]\u003csup\u003e+\u003c/sup\u003e.\u0026nbsp;\u003cstrong\u003eHRMS: (ES+)\u0026nbsp;\u003c/strong\u003eCalculated for C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e35\u003c/sup\u003eCl [M + H]\u003csup\u003e+\u003c/sup\u003e : 299.0587, found : 299.0579. \u003cstrong\u003eIR: (ATR,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026upsilon;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;cm\u003csup\u003e-1\u003c/sup\u003e)\u0026nbsp;\u003c/strong\u003e3425.4 (O-H), 2922.7 (=C-H), 1647.4 (C=O), 1583.0 (C=N), 748.7 (C-Cl).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(\u003cem\u003eE\u003c/em\u003e)-3-(5-Chloro-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-(3-hydroxyphenyl)prop-2-en-1-one\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e3f\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 63%, M.p = 210-212 \u0026deg;C, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR : (400 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e12.01 (br s, 1H, NH), 8.52 (d, \u003cem\u003eJ\u003csub\u003e3-2\u003c/sub\u003e\u003c/em\u003e = 15.7 Hz, 1H, H\u003csub\u003e3\u003c/sub\u003e), 7.84 (s, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.77 (d, \u003cem\u003eJ\u003c/em\u003e = 8.6 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.68-7.55 (m, 3H, H\u003csub\u003e2\u003c/sub\u003e and 2H\u003csub\u003eAr\u003c/sub\u003e), 7.53-7.38 (m, 3H, H\u003csub\u003eAr\u003c/sub\u003e), 7.15 (dd, \u003cem\u003eJ\u003c/em\u003e = 2.1 Hz, \u003cem\u003eJ\u003c/em\u003e = 7.9 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR : (100 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e187.9 (C=O), 158.0 (C\u003csub\u003eAr\u003c/sub\u003e), 138.0 (C\u003csub\u003eAr\u003c/sub\u003e), 130.7 (C\u003csub\u003eAr\u003c/sub\u003e), 130.1 (C\u003csub\u003eAr\u003c/sub\u003e), 129.3 (2C\u003csub\u003eAr\u003c/sub\u003e), 127.4 (C\u003csub\u003eAr\u003c/sub\u003e), 125.3 (C\u003csub\u003eAr\u003c/sub\u003e), 121.2 (2C\u003csub\u003eAr\u003c/sub\u003e), 119.8 (2C\u003csub\u003eAr\u003c/sub\u003e), 116.4 (C\u003csub\u003eAr\u003c/sub\u003e), 114.6 (2C\u003csub\u003eAr\u003c/sub\u003e).\u0026nbsp;\u003cstrong\u003eHRMS: (ES+)\u0026nbsp;\u003c/strong\u003eCalculated for C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e35\u003c/sup\u003eCl [M + H]\u003csup\u003e+\u003c/sup\u003e : 299.0587, found : 299.0583. \u003cstrong\u003eIR: (ATR,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026upsilon;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;cm\u003csup\u003e-1\u003c/sup\u003e)\u0026nbsp;\u003c/strong\u003e3321.4 (O-H), 2920.6 (=C-H), 1653.6 (C=O), 1585.0 (C=N), 778.4 (C-Cl).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(\u003cem\u003eE\u003c/em\u003e)-3-(5-Nitro-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e3g\u003c/strong\u003e)\u003c/p\u003e\n\u003cp id=\"_Toc527564624\"\u003eYellow solid, yield = 77%, M.p = 225-226 \u0026deg;C, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR : (400 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e11.90 (br s, 1H, NH), 8.47 (d, \u003cem\u003eJ\u003csub\u003e3-2\u003c/sub\u003e\u003c/em\u003e = 15.2 Hz, 1H, H\u003csub\u003e3\u003c/sub\u003e), 8.21 (dd, \u003cem\u003eJ\u003c/em\u003e = 1.2 Hz, \u003cem\u003eJ\u003c/em\u003e = 8.1 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.95-7.77 (m, 5H, 4H\u003csub\u003eAr\u003c/sub\u003e and H\u003csub\u003e2\u003c/sub\u003e), 7.65 (d, \u003cem\u003eJ\u003c/em\u003e = 8.3 Hz, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.52-7.40 (m, 2H, H\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR : (100 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e195.5 (C=O), 184.7 (C\u003csub\u003eAr\u003c/sub\u003e), 158.9 (C\u003csub\u003eAr\u003c/sub\u003e), 150.3 (C\u003csub\u003eAr\u003c/sub\u003e), 144.1 (C\u003csub\u003eAr\u003c/sub\u003e), 137.7 (C\u003csub\u003eAr\u003c/sub\u003e), 137.6 (C\u003csub\u003eAr\u003c/sub\u003e), 132.4 (C\u003csub\u003eAr\u003c/sub\u003e), 132.3 (C\u003csub\u003eAr\u003c/sub\u003e), 129.5 (2C\u003csub\u003eAr\u003c/sub\u003e), 128.5 (C\u003csub\u003eAr\u003c/sub\u003e), 125.0 (2C\u003csub\u003eAr\u003c/sub\u003e), 119.5 (C\u003csub\u003eAr\u003c/sub\u003e), 115.7 (C\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003eMS:\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e[CI, NH\u003csub\u003e3,\u0026nbsp;\u003c/sub\u003em/z] :\u0026nbsp;\u003c/strong\u003e309 [M]\u003csup\u003e+\u003c/sup\u003e, 310 [M + H]\u003csup\u003e+\u003c/sup\u003e. \u003cstrong\u003eHRMS: (ES+)\u0026nbsp;\u003c/strong\u003eCalculated for C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e [M + H]\u003csup\u003e+\u003c/sup\u003e : 310.0828, found : 310.0824. \u003cstrong\u003eIR: (ATR,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026upsilon;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;cm\u003csup\u003e-1\u003c/sup\u003e)\u0026nbsp;\u003c/strong\u003e3102.1 (O-H), 1670.2 (C=O), 1587.5 (C=N), 1340.1 (-NO\u003csub\u003e2\u003c/sub\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(\u003cem\u003eE\u003c/em\u003e)-3-(5-Benzoyl-1\u003cem\u003eH\u003c/em\u003e-benzimidazol-2-yl)-1-(2-hydroxyphenyl)prop-2-en-1-one\u0026nbsp;\u003c/strong\u003e(\u003cstrong\u003e3h\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eYellow solid, yield = 65%, M.p = 195-196 \u0026deg;C, \u003cstrong\u003e\u003csup\u003e1\u003c/sup\u003eH NMR : (400 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e11.91 (s, 1H, NH), 8.52 (d, \u003cem\u003eJ\u003csub\u003e3-2\u0026nbsp;\u003c/sub\u003e\u003c/em\u003e= 15.8 Hz, 1H, H\u003csub\u003e3\u003c/sub\u003e), 8.08-8.05 (m, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 8.03 (s, 1H, H\u003csub\u003eAr\u003c/sub\u003e), 7.90-7.57 (m, 9H, H\u003csub\u003eAr\u003c/sub\u003e), 7.69 (d, \u003cem\u003eJ\u003csub\u003e2-3\u003c/sub\u003e\u003c/em\u003e\u003csub\u003e\u0026nbsp;\u003c/sub\u003e= 15.8 Hz, 1H, H\u003csub\u003e2\u003c/sub\u003e), 7.08-7.03 (m, 2H, H\u003csub\u003eAr\u003c/sub\u003e). \u003cstrong\u003e\u003csup\u003e13\u003c/sup\u003eC NMR: (100 MHz, DMSO-\u003cem\u003ed\u003csub\u003e6\u003c/sub\u003e\u003c/em\u003e,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026delta;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;ppm)\u0026nbsp;\u003c/strong\u003e195.3 (C=O), 192.0 (C=O), 161.0 (C\u003csub\u003eAr\u003c/sub\u003e), 150.4 (C\u003csub\u003eAr\u003c/sub\u003e), 137.6 (C\u003csub\u003eAr\u003c/sub\u003e), 136.5 (C\u003csub\u003eAr\u003c/sub\u003e), 132.6 (C\u003csub\u003eAr\u003c/sub\u003e), 132.4 (C\u003csub\u003eAr\u003c/sub\u003e), 130.6 (2C\u003csub\u003eAr\u003c/sub\u003e), 130.3 (C\u003csub\u003eAr\u003c/sub\u003e), 129.6 (2C\u003csub\u003eAr\u003c/sub\u003e), 128.5 (C\u003csub\u003eAr\u003c/sub\u003e), 125.6 (C\u003csub\u003eAr\u003c/sub\u003e), 121.5 (C\u003csub\u003eAr\u003c/sub\u003e), 119.5 (C\u003csub\u003eAr\u003c/sub\u003e), 118.3 (2C\u003csub\u003eAr\u003c/sub\u003e), 117.8 (C\u003csub\u003eAr\u003c/sub\u003e), 115.1 (2C\u003csub\u003eAr\u003c/sub\u003e), 114.3 (C\u003csub\u003eAr\u003c/sub\u003e).\u0026nbsp;\u003cstrong\u003eMS : [EI, 70 eV, m/z (rel. Int)] :\u0026nbsp;\u003c/strong\u003e369 (35), 368 (100), 367(16) \u003cstrong\u003e[CI, NH\u003csub\u003e3,\u0026nbsp;\u003c/sub\u003em/z] :\u0026nbsp;\u003c/strong\u003e368 [M]\u003csup\u003e+\u003c/sup\u003e, 369 [M + H]\u003csup\u003e+\u003c/sup\u003e.\u0026nbsp;\u003cstrong\u003eHRMS: (ES+)\u0026nbsp;\u003c/strong\u003eCalculated for C\u003csub\u003e23\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e [M + H]\u003csup\u003e+\u003c/sup\u003e : 369.1239, found : 369.1241. \u003cstrong\u003eIR: (ATR,\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026upsilon;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;cm\u003csup\u003e-1\u003c/sup\u003e)\u0026nbsp;\u003c/strong\u003e3287.4 (O-H), 3171.2 (N-H), 1682.6 (C=O), 1564.7 (C=N), 1478.1 (C=C).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBiological material\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNormal human skin fibroblasts were purchased from Gibco (Thermo Fisher Scientific). The cancer cell lines Caco-2, MDA-MB-231, HCT-116, PC3, and MCF-7 were obtained from the European Collection of Authenticated Cell Cultures (ECACC, Porton, UK). Cells were maintained under the recommended conditions: Caco-2, MDA-MB-231, and MCF-7 in Dulbecco\u0026rsquo;s Modified Eagle Medium (DMEM); HCT-116 in McCoy\u0026rsquo;s medium; and PC3 in RPMI medium. All culture media were supplemented with 10% fetal bovine serum (FBS), 1% penicillin\u0026ndash;streptomycin, and 2 mM glutamine. Cultures were incubated at 37 \u0026deg;C in a humidified atmosphere containing 5% CO\u003csub\u003e2\u003c/sub\u003e. All cell lines are maintained in liquid nitrogen as both master and working banks. Routine mycoplasma testing was performed to confirm the absence of contamination throughout the study.\u003c/p\u003e"},{"header":"Declarations","content":"\u003col\u003e\n \u003cli\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eThe authors would like to thank laboratory Chemistry and Interdisciplinarity: Synthesis, Analysis, Modelling (CEISAM), University of Nantes for chemical reagents and spectroscopic analyses; the \u0026lsquo;\u0026lsquo;ImPACcell\u0026rsquo;\u0026rsquo; platform and Dr R\u0026eacute;my Le Gu\u0026eacute;vel of the University of Rennes 1 for cancer tests.\u003c/p\u003e\n\u003col start=\"2\"\u003e\n \u003cli\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eA.K conceived and designed the chemistry experiments, performed them, analyzed the data, prepared figures and/or tables, reviewed drafts of the article, and approved the final draft. A.F.K analyzed the data, prepared figures and/or tables, and approved the final draft. S.C analyzed the data, and wrote the final draft of the article. D. S analyzed the data, supervised, and approved the final draft. S. C conceived the work and designed the biological experiments and approved the final draft.\u003c/p\u003e\n\u003col start=\"3\"\u003e\n \u003cli\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eNot registered even if research protocols were performed in accordance with French legal guidelines (French Ministry of Health).\u003c/p\u003e\n\u003col start=\"4\"\u003e\n \u003cli\u003e\u003cstrong\u003eConsent to participate\u0026nbsp;\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003col start=\"5\"\u003e\n \u003cli\u003e\u003cstrong\u003eConsent to publish\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003col start=\"6\"\u003e\n \u003cli\u003e\u003cstrong\u003eConflicts of interest\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eThe authors declare no conflicts of interest.\u0026nbsp;\u003c/p\u003e\n\u003col start=\"7\"\u003e\n \u003cli\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003col start=\"8\"\u003e\n \u003cli\u003e\u003cstrong\u003eData availability statements\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eThe data that support the findings of this study are available in the supplementary information of this article.\u0026nbsp;The human-derived cell lines used in our study (colon and breast cancer cell lines, as well as normal human skin fibroblasts) were obtained from commercial suppliers (Gibco, CLS, ATCC, or ECACC) and were fully de-identified. No primary human tissues were collected specifically for this study.\u003c/p\u003e\n\u003col start=\"9\"\u003e\n \u003cli\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eG\u0026eacute;n\u0026eacute;vi\u0026egrave;ve Z, Elisabeth O, Boureima T, Basile T, Adama K, Bal\u0026eacute; B, Hamidou H. T, and Adrien M.G. M (2023) Anti-parasitic activities of four synthetic chemicals anthelmintics on Haemonchus contortus. \u003cem\u003eWorld J. Adv. Res. Rev.\u003c/em\u003e, \u003cstrong\u003e20\u003c/strong\u003e (2), 683\u0026ndash;689.\u003c/li\u003e\n\u003cli\u003eChung, N.T., Dung, V.C., and Duc, D.X. (2023) Recent achievements in the synthesis of benzimidazole derivatives. \u003cem\u003eRSC Adv.\u003c/em\u003e, \u003cstrong\u003e13\u003c/strong\u003e (46), 32734\u0026ndash;32771.\u003c/li\u003e\n\u003cli\u003eChandrika, N.T., Shrestha, S.K., Ngo, H.X., and Garneau-Tsodikova, S. (2016) Synthesis and investigation of novel benzimidazole derivatives as antifungal agents. \u003cem\u003eBioorganic Med. Chem.\u003c/em\u003e, \u003cstrong\u003e24\u003c/strong\u003e (16), 3680\u0026ndash;3686.\u003c/li\u003e\n\u003cli\u003eKaplancikli, Z.A., Turan-Zitouni, G., Revial, G., and Guven, K. (2004) Synthesis and study of antibacterial and antifungal activities of Novel 2-[[(benzoxazole/benzimidazole-2-yl)sulfanyl] acetylamino]thiazoles. \u003cem\u003eArch. Pharm. Res.\u003c/em\u003e, \u003cstrong\u003e27\u003c/strong\u003e (11), 1081\u0026ndash;1085.\u003c/li\u003e\n\u003cli\u003eBai, S., Zhang, M., Tang, S., Li, M., Wu, R., Wan, S., Chen, L., Wei, X., and Li, F. (2024) Research Progress on Benzimidazole Fungicides: A Review. \u003cem\u003eMolecules\u003c/em\u003e, \u003cstrong\u003e29\u003c/strong\u003e (6), 1\u0026ndash;14.\u003c/li\u003e\n\u003cli\u003eMarinescu, M. (2021) Synthesis of antimicrobial benzimidazole\u0026ndash;pyrazole compounds and their biological activities. \u003cem\u003eAntibiotics\u003c/em\u003e, \u003cstrong\u003e10\u003c/strong\u003e (8), 1\u0026ndash;29.\u003c/li\u003e\n\u003cli\u003eMahmood, K., Hashmi, W., Ismail, H., Mirza, B., Twamley, B., Akhter, Z., Rozas, I., and Baker, R.J. (2019) Synthesis, DNA binding and antibacterial activity of metal(II) complexes of a benzimidazole Schiff base. \u003cem\u003ePolyhedron\u003c/em\u003e, \u003cstrong\u003e157\u003c/strong\u003e (Ii), 326\u0026ndash;334.\u003c/li\u003e\n\u003cli\u003eAlDifar, H.A., Baaiu, B.S., Darwish, K.M.A., Ali, M.F., Dakhil, O.O., Abd-Alsalam, M., and Al Difar, H. (2023) Synthesis of Benzimidazole and Phthaloylamino Acid derivatives and Antibacterial Activity. \u003cem\u003eJ. Med. Chem. Sci.\u003c/em\u003e, \u003cstrong\u003e6\u003c/strong\u003e (9), 1975\u0026ndash;1984.\u003c/li\u003e\n\u003cli\u003eTomei, L., Altamura, S., Bartholomew, L., Biroccio, A., Ceccacci, A., Pacini, L., Narjes, F., Gennari, N., Bisbocci, M., Incitti, I., Orsatti, L., Harper, S., Stansfield, I., Rowley, M., De Francesco, R., and Migliaccio, G. (2003) Mechanismof Action and Antiviral Activity of Benzimidazole-Based AllostericInhibitors of the Hepatitis C Virus RNA-Dependent RNAPolymerase. \u003cem\u003eJ. Virol.\u003c/em\u003e, \u003cstrong\u003e77\u003c/strong\u003e (24), 13225\u0026ndash;13231.\u003c/li\u003e\n\u003cli\u003eShaker, Y.M., Omar, M.A., Mahmoud, K., Elhallouty, S.M., El-Senousy, W.M., Ali, M.M., Mahmoud, A.E., Abdel-Halim, A.H., Soliman, S.M., and El Diwani, H.I. (2015) Synthesis, in vitro and in vivo antitumor and antiviral activity of novel 1-substituted benzimidazole derivatives. \u003cem\u003eJ. Enzyme Inhib. Med. Chem.\u003c/em\u003e, \u003cstrong\u003e30\u003c/strong\u003e (5), 826\u0026ndash;845.\u003c/li\u003e\n\u003cli\u003eAhmad, N., Azad, M.I., Khan, A.R., and Azad, I. (2021) Benzimidazole As a Promising Antiviral Heterocyclic Scaffold: a Review. \u003cem\u003eJ. Sci. Arts\u003c/em\u003e, \u003cstrong\u003e21\u003c/strong\u003e (1), 273\u0026ndash;284.\u003c/li\u003e\n\u003cli\u003eVeerasamy, R., Roy, A., Karunakaran, R., and Rajak, H. (2021) Structure\u0026ndash;activity relationship analysis of benzimidazoles as emerging anti-inflammatory agents: An overview. \u003cem\u003ePharmaceuticals\u003c/em\u003e, \u003cstrong\u003e14\u003c/strong\u003e (7).\u003c/li\u003e\n\u003cli\u003eMavrova, A.T., Yancheva, D., Anastassova, N., Anichina, K., Zvezdanovic, J., Djordjevic, A., Markovic, D., and Smelcerovic, A. (2015) Synthesis, electronic properties, antioxidant and antibacterial activity of some new benzimidazoles. \u003cem\u003eBioorganic Med. Chem.\u003c/em\u003e, \u003cstrong\u003e23\u003c/strong\u003e (19), 6317\u0026ndash;6326.\u003c/li\u003e\n\u003cli\u003eAroua, L.M., Almuhaylan, H.R., Alminderej, F.M., Messaoudi, S., Chigurupati, S., Al-mahmoud, S., and Hamdoon, M.H. (2021) A facile approach synthesis of benzoylaryl benzimidazole as potential \u0026alpha;-amylase and \u0026alpha;-glucosidase inhibitor with antioxidant activity. \u003cem\u003eBioorg. Chem.\u003c/em\u003e, \u003cstrong\u003e114\u003c/strong\u003e (March), 105073.\u003c/li\u003e\n\u003cli\u003eEl-Gohary, N.S., and Shaaban, M.I. (2017) Synthesis and biological evaluation of a new series of benzimidazole derivatives as antimicrobial, antiquorum-sensing and antitumor agents. \u003cem\u003eEur. J. Med. Chem.\u003c/em\u003e, \u003cstrong\u003e131\u003c/strong\u003e, 255\u0026ndash;262.\u003c/li\u003e\n\u003cli\u003eOnnis, V., Demurtas, M., Deplano, A., Balboni, G., Baldisserotto, A., Manfredini, S., Pacifico, S., Liekens, S., and Balzarini, J. (2016) Design, synthesis and evaluation of antiproliferative activity of new benzimidazolehydrazones. \u003cem\u003eMolecules\u003c/em\u003e, \u003cstrong\u003e21\u003c/strong\u003e (5), 4\u0026ndash;12.\u003c/li\u003e\n\u003cli\u003eWang, Z., Deng, X., Xiong, S., Xiong, R., Liu, J., Zou, L., Lei, X., Cao, X., Xie, Z., Chen, Y., Liu, Y., Zheng, X., and Tang, G. (2018) Design, synthesis and biological evaluation of chrysin benzimidazole derivatives as potential anticancer agents. \u003cem\u003eNat. Prod. Res.\u003c/em\u003e, \u003cstrong\u003e32\u003c/strong\u003e (24), 2900\u0026ndash;2909.\u003c/li\u003e\n\u003cli\u003eSharma, P., Reddy, T.S., Kumar, N.P., Senwar, K.R., Bhargava, S.K., and Shankaraiah, N. (2017) Conventional and microwave-assisted synthesis of new 1H-benzimidazole-thiazolidinedione derivatives: A potential anticancer scaffold. \u003cem\u003eEur. J. Med. Chem.\u003c/em\u003e, \u003cstrong\u003e138\u003c/strong\u003e, 234\u0026ndash;245.\u003c/li\u003e\n\u003cli\u003eCentre international de recherche sur le cancer (CIRC) (2024) Cancer : une charge toujours plus lourde dans le monde et des besoins en services croissants. \u003cem\u003el\u0026rsquo;Organisation Mond. la Sant\u0026eacute;\u003c/em\u003e, 1\u0026ndash;5.\u003c/li\u003e\n\u003cli\u003eOrganization, W.H. (2022) Global Cancer Observatory. \u003cem\u003eInt. Agency Res. Cancer.\u003c/em\u003e, \u003cstrong\u003e149\u003c/strong\u003e, 1\u0026ndash;2.\u003c/li\u003e\n\u003cli\u003eKon\u0026eacute;, A., Souleymane, C., Kalo, M., Tchambaga Etienne, C., Collet, S., and Sissouma, D. (2025) Synthesis of novel benzimidazole-based retrochalcones and their anticancer activity against breast and colon cancer. \u003cem\u003eSynth. Commun.\u003c/em\u003e, \u003cstrong\u003e55\u003c/strong\u003e (2), 175\u0026ndash;182.\u003c/li\u003e\n\u003cli\u003eMahmoudi, N., de Juli\u0026aacute;n-Ortiz, J.V., Ciceron, L., G\u0026aacute;lvez, J., Mazier, D., Danis, M., Derouin, F., and Garc\u0026iacute;a-Domenech, R. (2006) Identification of new antimalarial drugs by linear discriminant analysis and topological virtual screening. \u003cem\u003eJ. Antimicrob. Chemother.\u003c/em\u003e, \u003cstrong\u003e57\u003c/strong\u003e (3), 489\u0026ndash;497.\u003c/li\u003e\n\u003cli\u003eWillcox, M.L., and Bodeker, G. (2004) Traditional herbal medicines for malaria. \u003cem\u003eBr. Med. J.\u003c/em\u003e, \u003cstrong\u003e329\u003c/strong\u003e (7475), 1156\u0026ndash;1159.\u003c/li\u003e\n\u003cli\u003eDietrich. H., (1994) Toxizit\u0026auml;t chlororganischer Verbindungen: Einflu\u0026szlig; der Einf\u0026uuml;hrung von Chlor in organische Molek\u0026uuml;le. \u003cem\u003eAngew. Chem\u003c/em\u003e. \u003cstrong\u003e106\u003c/strong\u003e, 1997. \u003c/li\u003e\n\u003cli\u003eDietrich. H., (1994) Toxicity of Chlorinated Organic Compounds: Effects of the Introduction of Chlorine in Organic Molecules. \u003cem\u003eAngew. Chem. Int. Ed. Engl.,\u003c/em\u003e \u003cstrong\u003e33\u003c/strong\u003e: 1920-1935. \u003c/li\u003e\n\u003cli\u003e\u0026Scaron;egan, S., Krunić, M. J., Andrić, D. B., \u0026Scaron;ukalović, V. B., Penji\u0026scaron;ević, J. Z., and Jevtić, I. I.\u003cstrong\u003e (\u003c/strong\u003e2024) Evaluation of lipophilicity and drug-likeness of donepezil-like compounds using reversed-phase thin-layer chromatography. \u003cem\u003eBiomedical Chromatography\u003c/em\u003e,38(7), e5867. \u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003eTable 1 is available in the Supplementary Files section.\u003c/p\u003e"},{"header":"Scheme ","content":"\u003cp\u003eScheme 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-chemistry","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Chemistry](https://link.springer.com/journal/44371)","snPcode":"44371","submissionUrl":"https://submission.nature.com/new-submission/44371/3","title":"Discover Chemistry","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Benzimidazolyl–phenylpropenone, Anticancer activity, Cytotoxicity, Selective tumor targeting, In vitro evaluation","lastPublishedDoi":"10.21203/rs.3.rs-7490108/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7490108/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn this study, benzimidazolyl\u0026ndash;phenylpropenone derivatives (\u003cb\u003e3a\u0026ndash;h\u003c/b\u003e) were synthesized and fully characterized using \u003csup\u003e1\u003c/sup\u003eH, \u003csup\u003e13\u003c/sup\u003eC NMR spectroscopy and High-Resolution Mass Spectrometry (HRMS). Their \u003cem\u003ein vitro\u003c/em\u003e anticancer activity was evaluated against human cancer cell lines, including prostate (PC3), colon (CaCo2 and HCT-116), and breast (MDA-MB-231 and MCF-7) tumors, alongside normal human skin fibroblasts. All compounds demonstrated promising cytotoxic activity against the cancer cell lines, with IC\u003csub\u003e50\u003c/sub\u003e values ranging from 1.78 to 8.83 \u0026micro;M. Toxicity toward normal fibroblasts was moderate, with IC\u003csub\u003e50\u003c/sub\u003e values between 3.23 and 8.02 \u0026micro;M. Compared with reference compounds Roscovitine and Paclitaxel (Taxol\u0026reg;), several derivatives showed higher activity than Roscovitine, though generally less potent than Paclitaxel.\u003c/p\u003e\u003cp\u003eThese findings position benzimidazolyl\u0026ndash;phenylpropenones as promising scaffolds for the development of novel anticancer agents.\u003c/p\u003e","manuscriptTitle":"Benzimidazolyl-Phenylpropenones Synthesized and Evaluated for Selective Anticancer Activity in Prostate Colon and Breast Tumor Models","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-04 14:59:12","doi":"10.21203/rs.3.rs-7490108/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-05T09:07:57+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-31T15:46:32+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"40215438070278198738683581424347826455","date":"2025-10-28T06:42:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"114918982699209653200972426370798865034","date":"2025-10-27T17:29:25+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-26T13:30:46+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-25T05:12:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"339784093242124074771823462198271987094","date":"2025-10-23T10:52:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"179621490668360876729658949002881848871","date":"2025-10-23T06:34:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"45800777481773860928230832633916613975","date":"2025-10-23T04:59:08+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-23T04:52:59+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-10-13T06:02:37+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-07T10:09:20+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-05T11:17:34+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Chemistry","date":"2025-10-05T10:40:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-chemistry","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Chemistry](https://link.springer.com/journal/44371)","snPcode":"44371","submissionUrl":"https://submission.nature.com/new-submission/44371/3","title":"Discover Chemistry","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e776739e-f8df-4d9f-9d43-d57bd04ec897","owner":[],"postedDate":"November 4th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-09T14:55:25+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-04 14:59:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7490108","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7490108","identity":"rs-7490108","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}