Mometasone furoate is a synthetic corticosteroid used in the treatment of skin inflammatory conditions, hay fever and asthma. The industrial manufacturing routes to mometasone furoate are generally accompanied by the formation of numerous process impurities that need to be detected and quantified, as requested by regulatory authorities. The ready availability of such impurities in the required quantity and purity is therefore essential for toxicological studies, analytical method development and process validation. Herein, we report the multi-gram scale preparation of 21′-chloro--furan-2-carboxylate (mometasone furoate EP impurity C), one of the known impurities of mometasone furoate. This study also includes the systematic investigation of the final acylation step, as well as the characterization of the difuroate enol ether intermediate and its conversion to the target impurity C. Pharmac1) in the European Pharmacopoeia . Be. Be2 standitions ,24 by tr mixture , step f.2Cl2 at 25 °C. Under these conditions, a partial conversion of 14 was observed affording the target product 2 in 27% isolated yield (entry 1). The use of Et3N (2 equiv.) instead of pyridine in the presence of DMAP (0.1 equiv.) as the acyl transfer reagent resulted in the formation of the target product 2 (46% yield) along with unreacted 14 after 24 h at 25 °C (entry 2). However, when the reaction was scaled-up at 1 g, a longer reaction time (up to 48 h) and higher temperature (reflux) were needed (entry 3). Interestingly, the use of higher amounts of Et3N (4 equiv.), DMAP (0.25 equiv.) and 2-furoyl chloride (2 equiv.) resulted in the almost quantitative consumption of 14 and in the formation of a different reaction product (76% yield) with a TLC retention factor similar to that of the target product 2 (entry 4).The reaction was initially performed using pyridine (2 equiv.) as the base and 2-furoyl chloride (1.2 equiv.) in CHed yield , entry 1roduct 2 , entry 4m/z 579.1785 that corresponds to the formula C32H31ClO8 in positive ion mode. Nuclear magnetic resonance (NMR) analysis confirmed the presence of a second 2-furoyl group at the side chain. In particular, the comparison of the 1H- and 13C-NMR data with that of compound 2 clearly evidenced the loss of 21-methylene protons at around 4 ppm and the formation of a double bond associated with a single vinylic proton (singlet at 6.31 ppm and CH at 111.5 ppm) of the chloro vinyl ether moiety bearing the second furoyl group. Moreover, key spatial correlations in the NOESY experiment confirmed the Z-geometry at the double bond of the chloro vinyl ether group , it is commercially available from some specialized suppliers of API impurities [15 to the desired mometasone furoate EP impurity C (2) (15 to MF (1) by treatment with concentrated HCl and AcOH at 15–25 °C [15 may undergo an acid-catalysed migration of the angular 18-CH3 group to the electron deficient C-17 carbon atom via a carbonium ion by either a stepwise or concerted mechanism (High resolution mass spectrometry (HRMS) analysis showed a mass of roup see . Based opurities . We therty C (2) . Heggie 15–25 °C . Howeverechanism  [25].15 (entries 1–3). Also, the use of 3 N HCl in AcOH at 0 °C and p-toluenesulfonic acid (pTSA) in H2O/acetone gave traces of the desired target product 2 along with unreacted 14 (entries 4 and 5). Interestingly, the use of HClO4 in CH2Cl2 at 0 °C afforded the target product 2 in 46% isolated yield (entry 6). Lower temperatures (−10 °C) and diluted conditions gave a better outcome (72% yield) (entry 7).In our case, the use of 12 N HCl resulted in the formation of unidentified side product(s) along with the almost quantitative consumption of % yield) , entry 72 was obtained in 33% yield from 8-DM (6) over eight steps at mg scale (15 to the target mometasone furoate EP impurity C (2) (step h) and evaluated the feasibility of the synthetic strategy shown in 6). At this point, in order to make the synthetic route easily applicable in industrial settings, we focused on improving work-up and isolation procedures by avoiding, when possible, tedious chromatographic purifications and aqueous extractive work-ups  was assessed by quantitative normal phase (NP) and reverse phase (RP) C18-thin layer chromatography, quantitative 1H-NMR in the presence of dimethyl sulfone (DMS) as the internal standard and high-performance liquid chromatography (HPLC) analysis , conforming the presence of a chlorine atom. The comparison of 1H- and 13C-NMR spectra of 2 with those of the starting material 8-DM (6) confirmed the presence of the conjugated dienone at the A ring of the steroid backbone . The presence of two additional carbonyl groups at C11 and C20 position was proved by the signals at δ 207.5 and 196.1, respectively, in the carbon spectrum. The presence of a methylene group bearing the chlorine atom at the side chain was confirmed by the presence of a multiplet at 4.03–4.11 ppm, which corresponds to the signal at 44.6 ppm in the 13C-NMR spectrum. Finally, the signals at δ 6.57, 7.29 and 7.64 in the 1H-NMR spectrum and the carbon nuclei at δ 112.5, 120.2, 142.8, 147.5 and 158.0 ppm are related to the presence of the 2-furoyl group at the C17 position (carbon at 96.8 ppm). Scalar and spatial correlation in homonuclear 1H-1H COSY and NOESY experiments confirmed the aforementioned atom arrangement  or Bruker AC 400 MHz . Chemical shifts are reported in parts per million (ppm) and are relative to CDCl3 (7.26 ppm and 77.0 ppm). The abbreviations used are as follows: s, singlet; brs, broad singlet; d, doublet; dd, double of doublets; dt, doublet of triplets; t, triplet; q, quartet; qui, quintet; m, multiplet; and brm, broad multiplet. Coupling constants (J) are reported in Hertz (Hz). Thin-layer chromatography (TLC) was performed on aluminium backed silica plates or RP-18 glass plates . Spots were visualized by UV detector (λ: 254 nm) and/or by staining and warming with phosphomolybdic acid (5% w/v solution in EtOH). When required, flash chromatographic purifications were performed using Biotage Isolera . Uncorrected melting points were determined using a Stuart SMP3 apparatus . IR spectra were recorded using an IRSpirit–Shimadzu FTIR/ATR apparatus  in the spectral range of 400–5000 cm−1 and with a resolution of 4 cm−1 at 50 scans. Purity of the target product 2 was assessed by quantitative NP- and RP-18 TLC using 8-DM (6) as the reference standard, qNMR in the presence of dimethyl sulfone  as the internal standard  and by HPLC analysis. HPLC analyses (method Ph. Eur. 8.0 for mometasone furoate) were performed on a Shimadzu  LC-20A Prominence equipped with a CBM-20A communication bus module, two LC-20AD dual piston pumps, an SPD-M20A photodiode array detector, and a Rheodyne 7725i injector  with a 20-μL stainless steel loop. A GraceSmart RP18 column  was used as the analytical column. The analysis was performed at a 25 °C column temperature using analytical grade MeCN and ultrapure water (σ = 18.3 MΩ × cm) obtained from a Milli-Q Plus185 system from Millipore . The HPLC analysis was performed at 1.0 mL min−1 eluent flow rate for a running time of 100 min and at 254 nm of detection wavelength, after previous conditioning by passing through the column the selected mobile phase  for at least 30 min at the same eluent flow rate. Before use, the mobile phase was filtered through a 0.22 mm Millipore filter  and then degassed with 20 min sonication. The column temperature was controlled through a heather/chiller thermostat . Samples of the target product were prepared by dissolving 2 at 1 mg mL−1 in analytical grade MeCN and 20 μL were injected. High resolution mass spectrometry (HRMS) measurements were performed on a UHPLC–MS/MS system consisting of an Agilent 1290 Infinity II module combined with an Agilent 6560 mass spectrometer . The chromatographic separation was performed using a Zorbax Rrhd Eclipse Plus C18 column . HPLC eluent A was water  with 0.1% (v/v) of formic acid , while eluent B was MeCN . The gradient was 50% B for 1 min, then a linear increase took place up to 97% B over 6 min and this condition was maintained for 5 min. Finally, the system returned to 50% B in 1 min and was re-equilibrated for 2 min. The column temperature was kept at 25 °C and the flow rate was 0.3 mL min−1. The injection volume was 5 µL. Samples were dissolved in MeOH  and sonicated for 5 min. MassHunter software version B.05.01 (Agilent Technology) was used to control the LC–MS/MS system, for data prediction, acquisition, analysis and processing. For MS detection, the Dual AJS ESI source operated in positive ion mode. The gas temperature was set at 300 °C with a flow of 5 L min−1 while the sheath gas temperature was 350 °C with a flow of 11 L min−1. The nebulizer pressure was set at 35 psi, while the capillary and fragmentation voltages were 3500 V and 400 V, respectively.Unless otherwise noted, chemicals were obtained from commercial suppliers and used without further purification. Reactions requiring anhydrous conditions were conducted in dried glass apparatus under a positive pressure of N6)  in N,N-dimethylacetamide (120 mL), AcOK  and Ac2O  were sequentially added. The reaction mixture was stirred for 1 h at 25 °C. The reaction mixture was then poured into ice/H2O (600 mL) and kept under vigorous stirring for 30 min. The precipitate thus formed was collected by filtration under vacuum and washed with H2O (300 mL). The solid was dried in a vacuum oven  affording the desired 9β,11β-epoxy-17α-hydroxy-16α-methyl-3,20-dioxo-pregna-1,4-dien-21-acetate  as a white solid (m.p.: 192–194 °C) that was used for the next step without further purification. 1H-NMR : δ 0.88 , 0.93 , 1.44 , 2.16 , 3.22 , 4.62–5.05 , 6.15 , 6.19 , 6.60 . 13C-NMR : δ 14.5, 17.3, 20.4, 23.7, 29.1, 29.9, 30.4, 33.2, 34.2, 35.3, 44.1, 47.6, 48.1, 62.7, 66.0, 67.5, 90.5, 125.0, 127.9, 152.2, 164.9, 170.6, 186.0, 204.6.To a solution of dexamethasone 9,11-epoxide  in AcOH (135 mL), 48% aqueous solution of HBr (8 mL) was added dropwise over 15 min at 0 °C. The resulting mixture was stirred at 0 °C for 30 min and then allowed to warm to 25 °C and stirred for a further 2 h. The reaction mixture was then poured into ice/H2O (600 mL) and kept under vigorous stirring for 30 min. The precipitate thus formed was collected by filtration under vacuum and washed with H2O (300 mL). The solid was dried in a vacuum oven  affording the desired 9α-bromo-11β,17α-dihydroxy-16α-methyl-3,20-dioxo-pregna-1,4-dien-21-acetate  as a white solid that was used for the next step without further purification. 1H-NMR : δ 0.86 , 0.97 , 1.67 , 2.14 , 4.68–4.70 , 4.86 , 6.02 , 6.29 , 7.28 . 13C-NMR : δ 14.6, 17.0, 20.6, 24.9, 28.8, 30.6, 31.8, 35.3, 35.5, 35.8, 45.2, 48.7, 50.5, 68.9, 75.5, 85.9, 91.1, 124.5, 128.7, 154.0, 167.6, 171.7, 187.1, 205.1.To a solution of 9β,11β-epoxy-17α-hydroxy-16α-methyl-3,20-dioxopregna-1,4-dien-21-yl acetate  in DMF (100 mL), DMSO (12 mL), CrCl3∙6H2O , Zn dust  and thioglycolic acid  were sequentially added at 0 °C. The resulting suspension was stirred at 0 °C for 4 h. The reaction mixture was then poured into ice/H2O (600 mL) and kept under vigorous stirring for 30 min. The precipitate thus formed was collected by filtration under vacuum and washed with H2O (300 mL). The solid was dried in a vacuum oven  affording the desired 11β,17α-dihydroxy-16α-methyl-3,20-dioxopregna-1,4-dien-21-acetate  as a greenish solid that was used for the next step without further purification. 1H-NMR : δ 0.90 , 1.06 , 1.44 , 2.17 , 4.47 , 4.87 , 6.00 , 6.26 , 7.26 . 13C-NMR : δ 14.7, 17.0, 20.6, 21.0, 31.2, 32.0, 32.6, 34.0, 35.9, 40.0, 44.2, 48.9, 50.7, 55.5, 68.3, 70.1, 91.2, 122.3, 127.7, 156.8, 170.7, 171.0, 186.8, 205.2.To a solution of 9α-bromo-11β,17α-dihydroxy-16α-methyl-3,20-dioxopregna-1,4-dien-21-yl acetate  in CH2Cl2 (270 mL), pyridinium chlorochromate  was added portionwise over 30 min at 0 °C. At the end of the addition, the resulting suspension was stirred at 25 °C for further 4 h. The suspension was filtered on a pad of celite, the filtrate was washed with 5% (w/v) aqueous solution of Na2S2O3 (2 × 200 mL) and the aqueous phase was extracted with CH2Cl2 (3 × 200 mL). The combined organic extracts were washed with 3 N aqueous HCl (200 mL), H2O (200 mL) and brine (200 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude was purified by crystallization from n-hexane/EtOAc  affording, after drying , the desired 17α-hydroxy-16α-methyl-3,11,20-trioxo-pregna-1,4-dien-21-acetate  as a white solid (m.p.: 186–188 °C). 1H-NMR : δ 0.75 , 0.96 , 1.42 , 2.15 , 4.60–5.02 , 6.06 , 6.20 , 7.68 . 13C-NMR : δ 14.5, 15.7, 18.8, 20.4, 31.7, 32.2, 33.6, 36.2, 36.8, 42.4, 49.0, 49.8, 52.1, 60.5, 67.6, 90.3, 124.6, 127.5, 155.4, 166.7, 170.5, 186.4, 204.7, 208.5.To a solution of 11β,17α-dihydroxy-16α-methyl-3,20-dioxopregna-1,4-dien-21-acetate  in MeOH (500 mL), an aqueous saturated solution of NaHCO3 (125 mL) was added. The resulting mixture was stirred at 80 °C for 4 h. The crude mixture was concentrated under reduced pressure and the residue was extracted with EtOAc (3 × 200 mL). The combined organic extracts were washed with H2O (200 mL) and brine (200 mL), dried over anhydrous with Na2SO4 and concentrated under reduced pressure. The desired 17α,21-dihydroxy-16α-methyl-3,11,20-trioxopregna-1,4-diene  was obtained as a white solid and used for the next step without further purification. 1H-NMR : δ 0.73 , 0.97 , 1.41 , 4.19–4.60 , 6.06 , 6.20 , 7.65 . 13C-NMR : δ 14.7, 16.0, 18.7, 31.8, 32.2, 33.5, 36.2, 37.2, 42.4, 49.0, 49.9, 52.5, 60.4, 67.8, 89.6, 124.5, 127.5, 155.4, 166.7, 186.4, 208.6, 211.7.To a solution of 17α-hydroxy-16α-methyl-3,11,20-trioxo-pregna-1,4-dien-21-acetate  in CH2Cl2 (150 mL), 4-dimethylaminopyridine  was added at 25 °C. The solution was cooled to 0 °C and methanesulfonyl chloride  was added dropwise over 10 min. After 30 min, the mixture was allowed to warm to 25 °C and stirred for a further 30 min. The mixture was then refluxed and stirred for a further 3 h. The reaction mixture was allowed to cool to room temperature and washed with aqueous saturated solution of NaHCO3 (200 mL), 3N aqueous HCl (200 mL), H2O (200 mL) and brine (200 mL), dried over anhydrous with Na2SO4 and concentrated under reduced pressure. The crude 21-chloro-17α-hydroxy-16α-methyl-3,11,20-trioxo-pregna-1,4-diene  was obtained as a white solid and used for the next step without further purification. 1H-NMR : δ 0.74 , 0.97 , 1.42 , 4.16–4.55 , 6.07 , 6.20 , 7.67 . 13C-NMR : δ 14.6, 16.1, 18.8, 31.7, 32.2, 33.5, 36.2, 37.0, 42.4, 47.8, 48.9, 50.1, 52.2, 60.4, 90.8, 124.6, 127.6, 155.3, 166.6, 186.4, 202.7, 208.4.To a solution of 17α,21-dihydroxy-16α-methyl-3,11,20-trioxo-pregna-1,4-diene  in CH2Cl2 (170 mL), Et3N , dimethylaminopyridine  and 2-furoyl chloride  were sequentially added at 0 °C and the resulting solution was stirred at 0 °C for 1 h and for a further 2 h at 25 °C. The mixture was washed with 3 N aqueous HCl (200 mL), aqueous saturated solution of NaHCO3 (200 mL), H2O (200 mL) and brine (200 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude was filtered on a short pad of silica  affording the desired 2-chloro-1-+ calcd. for C32H31ClO8: 579.1780, found: 579.1785, Δppm: +0.78; p + 1 (abund. 31.64%), calcd. 580.1814, found 580.1817, Δppm: +0.57. r.t.: 3.524 min.To a solution of 21-chloro-17α-hydroxy-16α-methyl-3,11,20-trioxo-pregna-1,4-diene  in CH2Cl2 (500 mL), HClO4 (11 mL) was slowly added dropwise over 30 min at −10 °C. The mixture was stirred at −10 °C for a further 30 min. Aqueous saturated solution of NaHCO3 (200 mL) was then slowly added and the phases were separated. The aqueous phase was extracted with CH2Cl2 (2 × 200 mL), and the combined organic extracts were washed with H2O (200 mL) and brine (200 mL), dried over anhydrous with Na2SO4 and concentrated under reduced pressure. The crude was purified by automated flash chromatography on silica gel  and the collected fractions were dried under vacuo and crystallized from acetone/MeOH  affording, after drying , the desired 21′-chloro- furan-2-carboxylate  as a white solid. . The product was fully characterized by NOESY and COSY bidimensional NMR experiments, 1H- and 13C-NMR, IR and HRMS. 1H-NMR : δ 0.83 , 1.00 , 1.45 , 2.97 ), 3.40–3.45 , 4.03–4.11 , 6.11 , 6.23 , 6.57 , 7.29 , 7.64 , 7.68 . 13C-NMR : δ 16.0, 16.8, 18.8, 32.1, 32.8, 33.4, 36.0, 36.7, 42.2, 44.6, 49.1, 50.2, 51.6, 60.4, 96.8, 112.5, 120.2, 124.3, 127.8, 142.8, 147.5, 154.8, 158.0, 165.9, 186.2, 196.1, 207.5. HRMS m/z [M + H]+ calcd. for C27H29ClO6: 485.1725, found: 485.1728, Δppm: +0.58; p + 1 (abund. 27.29%), calcd. 486.1759, found 486.1763, Δppm: +0.65. r.t.: 2.977 min. Purity: HPLC-DAD : 96.18% ; calibrated 1H-NMR : 95.8–96.3%; quantitative NP- and RP-18 TLC: ≥96%.To a solution of 2-chloro-1-[17′α-((furan-2″-carbonyl)oxy)-16′α-methyl-3′,11′,20′-trioxo-pregna-1′,4′-dienyl)vinyl furan-2-carboxylate (1) is a potent steroidal anti-inflammatory drug (SAID) that, after more than 30 years since its introduction on the market, still represents one of the most prescribed SAIDs and an essential medicine. During its industrial manufacturing, several API-related, process-related or stability-related impurities have been identified and listed in the corresponding monograph of the European Pharmacopoeia. Among them, mometasone furoate EP impurity C (2) is commercially available as a reference standard  at very high costs, but no synthetic procedure for its preparation has been reported so far. Considering the need to synthesise the target impurity 2 at the gram scale and in high purity grade for regulatory activities, in this work we successfully developed a straightforward multi-gram scale synthesis of mometasone furoate EP impurity C (2) starting from cheap and readily available steroidal precursor 6. The structural elucidation of the target product 2 was discussed along with the synthetic efforts aimed at optimizing both the acylation step and the acid-promoted hydrolysis of difuroate enol ether intermediate 15 to the target impurity 2. The proposed route resulted in good overall yield, minimized time-consuming and expensive purification extra-steps and a suitable purity of the final product, and it is therefore well suited to be applicable in R&D laboratories of steroid pharmaceutical companies.Mometasone furoate (MF,


H)-ones 1a–e with the active methylene compounds pentane-2,4-dione (2a), 1,3-diphenylpropane-1,3-dione (2b), and K2CO3 was investigated in this study. This approach afforded 4-quinolin-2(1H)-ones 3a–j in high yields and purity. All newly synthesized products’ structures were identified. Compounds 3a–j were tested for antiproliferative activity against a panel of four cancer cell lines. In comparison to the reference erlotinib (GI50 = 33), compounds 3f–j were the most potent derivatives, with GI50 values ranging from 22 nM to 31 nM. The most effective antiproliferative derivatives, 3f–j, were subsequently investigated as possible multi-target inhibitors of EGFR, BRAFV600E, and EGFRT790M. Compound 3h was the most potent inhibitor of the studied molecular targets, with IC50 values of 57 nM, 68 nM, and 9.70 nM, respectively. The apoptotic assay results demonstrated that compounds 3g and 3h function as caspase-3, 8, and Bax activators as well as down-regulators of the antiapoptotic Bcl2, and hence can be classified as apoptotic inducers. Finally, compounds 3g and 3h displayed promising antioxidant activity at 10 µM, with DPPH radical scavenging of 70.6% and 73.5%, respectively, compared to Trolox (77.6%).The reaction of 4-azido-quinolin-2(1 Despite scientific and social advancements, cancer remains one of the most prevalent diseases of concern and a primary cause of human suffering. Cancer deaths globally are expected to climb by more than 13.1 million by 2030, according to estimates . As a reCancer is a term used to describe a group of diseases caused by abnormalities in cell proliferation and replication . Cancer Although these strategies can achieve poly-pharmacological effects, they have encountered unavoidable challenges, such as the difficulty and length of time required to determine the best therapeutic pairings and timing, the difficulty in managing the bio-distribution characteristics and pharmacokinetics of a particular treatment, potential interactions between several drugs that lead to potential side effects, and low patient tolerance ,24. AlteSeveral antineoplastic drugs with various structures have been developed as a consequence of research efforts over the last few decades ,33. Due Moreover, small compounds, such as quinazoline derivatives, can suppress EGFR by inhibiting tyrosine kinase at ATP-binding sites ,44. The V600E. Compound I . Compound I inhibited both EGFR and BRAFV600E with IC50 values of 1.30 µM and 3.80 µM, respectively. Another study [II . Compound II demonstrated the best topo II inhibitory activity at the examined concentrations (100 µM = 47.6% and 20 µM = 19.5%) when compared to the positive control, etoposide (100 µM = 83.7% and 20 µM = 66.0%). Moreover, compound II exhibited promising dual inhibitory action against CDK2 and EGFR with IC50 values of 1.60 µM and 0.40 µM, respectively.In a recent publication , we descmpound I  was the er study  describetudy [II  was provAdditionally, 1,2,3-triazoles have found substantial uses in pharmaceutical chemistry ,51,52,533a–j  cell line). All of the newly synthesized analogues were tested for antiproliferative activity against four cancer cell lines. The most potent derivatives were studied further as multi-target inhibitors against wild-type EGFR, mutant-type EGFR (EGFRT790M), and BRAFV600E. The most active derivative’s potential apoptotic and antioxidant capabilities were also examined.Compounds 1a–e with active methylene compounds such as pentane-2,4-dione (2a) and 1,3-diphenylpropane-1,3-dione (2b) quninolin-2-ones via the click reaction and investigated their biological characteristics ,64. Howeone (2b) .H)-one (1a) and pentane-2,4-dione (2a) as active methylene compounds and various bases at room temperature, worming, and under refluxing temperature. When we employed other bases to optimize our conditions, such as Et3N, piperidin, NaOH, and KOH in ethanol as a solvent, we obtained compound 3a with a negligible yield of less than 20% when either cold or hot , we obtained the product 3f in an excellent yield as 82%. So, the best result was obtained when the reaction was conducted by starting with 4-azidoquinolin-2(1H)-one 1a–e (1 mmol), the active methylene compounds 2a,b (1 mmol), and 1.2 mmol of K2CO3 in ethanol according to a previous publication [To optimize the reaction condition, we performed the reaction by starting with 4-azidoquinolin-2-6-methylquinolin-2(1H)-one -ones (1b) and one molecule of acetyl acetone (2a) with the elimination of one molecule of H2O. The 1H NMR spectra for compound 3b showed five singlet signals in a ratio (1:1:3:3:3) with chemical shifts at δH 12.26, 7.40, 6.87, 3.32, 2.45, and 2.26 ppm, which were assigned as NH, H-5, COCH3, CH3 (H-5a’), and CH3 (H-6a), respectively. The three methyl groups were further confirmed from their 13C NMR spectra, which gave signals at δC 27.67 (COCH3), 20.38 (CH3-6a), and 9.29 ppm (CH3-5a’).The structures for all obtained products are fully consistent with their spectral data, such as (1H)-one . Mass sp13C NMR spectra clearly showed the presence of two downfield signals at δC 193.14 and 160.76 ppm, which were assigned as (COCH3) and quniolinone-C-2, respectively, as shown in 1H NMR spectra or in 13C NMR spectra were identical to those in mass spectrometry and elemental analysis, and to the structure, which was deduced exactly.Also, the 3a–j, the elemental analysis and mass spectrometry clearly show that the gross formula for all compounds came from the reaction between one mole of 4-azidoquinolin-2(1H)-ones (1a–e) and one mole of the active methylene  with the elimination of one H2O molecule. Also, the 1H NMR clearly showed the presence of a downfield singlet signal at δH = 6.86–6.96 ppm, which was assigned as quinolinon-H-3 and confirmed with 13C NMR with a chemical shift at δC = 120.38–122.24 ppm for all obtained products. Additionally, the presence of two carbonyl groups was confirmed by the 13C NMR with a chemical shift at δC = 160.20–160.97, 186.08–186.18, and 193.09–193.18 ppm, for all quinolinon-C-2 (for compounds 3a–j), COCH3 (for compounds 3a–e), and COPh (compounds 3f–j), respectively.Furthermore, in all obtained products 13C NMR spectroscopy. Through the preceding values of the different analyses, the validity of the chemical composition of all the compounds that were obtained is clear, and it is clear that the behavior of the 4-azidoquinoli-2(1H)-ones 1a–e with the active methylene 2a,b, whether aromatic or aliphatic, exhibited the same behavior during the reaction as shown in Finally, through the chemical shift’s value of the external carbonyl group COR as shown above, which refers to the nature of the group attached to the carbonyl, as the methyl group is a donor group and the phenyl group is withdrawn, the first appears at a lower value in the 3a–j can be rationalized with the following suggested mechanism -ones 1a–e and forming the intermediate 4, and then intra-nucleophilic addition from the N-3 atom of the 1a–e on the carbonyl group and cyclization resulting in the adduct 5 which accepted a hydrogen proton and the loss of a H2O molecule to give the final products 3a–j.The formation of compounds echanism  via the 3a–j. Compounds 3a–j were cultured on MCF-10A cells for four days before being evaluated for vitality using the MTT assay [The human mammary gland epithelial (MCF-10A) cell line was used to investigate the viability of the 1,2,3-triazole-based derivatives TT assay ,67. Acco3a–j was evaluated against four human cancer cell lines using the MTT assay [50) and the average of IC50 (GI50).The antiproliferative activity of TT assay ,68 and e3a–j had promising antiproliferative activity, with GI50 against the four cancer cell lines tested ranging from 22 nM to 65 nM, compared to erlotinib’s 33 nM. The newly synthesized compounds 3a–j can be divided into two major Scaffolds. Scaffold A compounds are 4-acetly, 5-methyl-1,2,3-triazoles 3a–e (R4 = CH3), and scaffold B compounds are 4-benzoyl, 5-phenyl-1,2,3-triazoles 3f–j (R4 = Ph). Compounds 3a–e (R4 = CH3) showed GI50 values ranging from 39 nM to 65 nM and were found to be less potent than congeners 3f–j (R4 = Ph) of GI50 values ranging from 22 nM to 31 nM, indicating the relevance of the phenyl group of 1,2,3-triazole moiety for the antiproliferative action.In general, compounds 3f, 3g, 3h, and 3i (Scaffold B) were revealed to be the most potent derivatives, with GI50 values of 28, 26, 22, and 31 nM, respectively. These compounds outperformed the reference drug erlotinib (GI50 = 33 nM). Compounds 3f, 3g, and 3h were found to be more effective than erlotinib against all cancer cell lines tested.Compounds 3h  was the most potent derivative among all synthesized derivatives, with a GI50 value of 22 nM, 1.5-fold more potent than the reference erlotinib. The phenyl ring substitution on the 1,2,3-triazole moiety results in compound 3c , which had a GI50 value of 39 nM and was 1.8-fold less potent than compound 3h, indicating the relevance of the phenyl ring substitution on the 1,2,3-triazole moiety.Compound 3g  and 3f . Compounds 3g and 3f have GI50 values of 26 nM and 28 nM, respectively, and are both less potent than 3h , demonstrating the importance of the substitution pattern of position five of the quinoline moiety on the antiproliferative activity of the newly synthesized compounds and that the activity increases in the order OCH3 > CH3 > H. Additionally, compound 3i  had a GI50 value of 35 nM, which was less potent than compound 3f , suggesting the relevance of the quinoline moiety’s Free N-1 atom for antiproliferative action.Another significant point to consider is the relevance of the substitution pattern at position five on the quinoline moiety, as seen in compounds 3a–e (R4 = CH3) were less potent than compounds 3f–j (R4 = Ph), with GI50 values ranging from 39 nM to 65 nM, where all parameters that affect the activity of compounds 3f–j were also present in compounds 3a–e. For example, compound 3c  had a GI50 value of 39 nM against the four cancer cell lines tested, being more potent than compounds 3b  and 3a , indicating the relevance of the methoxy group at the fifth position of the quinoline ring to antiproliferative action. Moreover, compound 3d  had a GI50 value of 62 nM, being 1.4-fold less potent than 3a, indicating the relevance of the quinoline moiety’s Free N-1 atom for antiproliferative action.Finally, as stated above, Scaffold A compounds 3f, 3g, 3h, and 3j, the most effective antiproliferative derivatives, were evaluated for their inhibitory action on EGFR as a potential target for their antiproliferative action [50 values versus erlotinib as a reference drug. The investigated compounds 3f, 3g, 3h, and 3j demonstrated promising EGFR inhibitory action with IC50 values of 72 nM, 64 nM, 57 nM, and 79 nM, respectively. The results of the EGFR inhibitory assay corroborate the results of the antiproliferative assay, with the most active antiproliferative derivatives also being the most active EGFR inhibitors, showing that EGFR may be a viable target for antiproliferative activity. Compounds 3f, 3g, and 3h demonstrated superior EGFR inhibitory action to the reference erlotinib, where compound 3j (IC50 = 79 nM) was equipotent to erlotinib (IC50 = 80 nM). The most potent antiproliferative agent, compound 3h, was also the most potent EGFR inhibitor, with an IC50 value of 57 nM, being 1.4-fold more potent than erlotinib.Compounds e action . Table 33f, 3g, 3h, and 3j were tested for their ability to inhibit mutant BRAF [50 values. With IC50 values ranging from 68 nM to 93 nM, the compounds examined showed potential inhibitory activity against BRAFV600E. In all cases, the investigated derivatives were shown to be less effective as BRAFV600E inhibitors than erlotinib (IC50 = 60 ± 5 nM). The most potent antiproliferative agent and EGFR inhibitor, compound 3h, was also the most potent BRAFV600E inhibitor, with an IC50 value of 68 ± 5 nM, comparable to erlotinib in BRAFV600E inhibitory action. Compounds 3f and 3g were shown to be the second and third most active, with IC50 values of 86 ± 6 nM and 73 ± 6 nM, respectively. Finally, compound 3j was the least effective BRAFV600E inhibitor, with an IC50 of 97 ± 7 nM, being 1.6-fold less potent than the reference erlotinib (IC50 = 60 ± 5 nM). Compounds 3g and 3h were discovered to be potential antiproliferative agents with dual EGFR and BRAFV600E inhibitory action, requiring significant structural modifications in their backbone structures to optimize their biological activity.Compounds ant BRAF . Table 33g and 3h’s encouraging results as antiproliferative agents with potential EGFR and BRAFV600E inhibitory activities motivated us to test their efficacy on the mutant EGFR (EGFR T790M) receptor type [50 values in 3g and 3h inhibited EGFRT790M with IC50 values of 8.40 ± 0.7 nM and 9.70 ± 0.8 nM, which were equivalent to the reference Osimertinib’s IC50 value of 8 nM. These findings add to the evidence of the investigated compounds’ antiproliferative activity, which can serve as multi-target inhibitors.Compounds tor type . Results3f, 3g, and 3h were tested for their capacity to activate the apoptosis cascade and disclose their proapoptotic potential.Controlling or perhaps terminating the uncontrolled proliferation of cancer cells is one method of treating cancer. Using the cell’s natural dying process is a very successful strategy. Apoptosis evasion is a hallmark of cancer and is not particular to the cause or kind of cancer, and hence targeting apoptosis is useful for many types of cancer. Many anticancer medications target different phases in both the intrinsic and extrinsic pathways ,71,72. C3f, 3g, and 3h, were tested as caspase-3 activators against the human epithelial cancer cell line (A-594) [3f, 3g, and 3h had promising caspase-3 protein overexpression levels of 524 ± 5, 587 ± 5, and 715 ± 6 pg/mL, respectively. They elevated the protein caspase-3 in the A-594 cancer cell line by approximately 8, 9, and 11 times when compared to untreated control cells. In all cases, the investigated compounds 3g, 3f, and 3h were shown to be more active than the standard staurosporine, which had a caspase-3 level overexpression of 465 ± 4 pg/mL. These findings demonstrated the investigated compounds’ apoptotic potential, which may explain their antiproliferative activity.Caspases play a crucial function in the induction and achievement of apoptosis. Caspase-3 is an essential caspase that cleaves different proteins in cells, resulting in apoptosis ,74. The  (A-594) , and the3g and 3h were investigated further for their effect on caspase-8, Bax, and antiapoptotic Bacl-2 levels against the human epithelial cancer cell line (A-594) using staurosporine as a control, as shown in 3g and 3h noticeably increased caspase-8 and Bax levels.Compounds 3h (2.35 ng/mL), followed by 3g (2.17 ng/mL) and standard staurosporine (1.85 ng/mL). In comparison to the control untreated cells, 3g and 3h increased caspase-8 levels by 24-fold and 26-fold, respectively.Caspase-8 overexpression was highest in compound 3g and 3h induced Bax 36- and 37-fold greater  than staurosporine . Finally, compared to staurosporine, compounds 3g and 3g elicited equipotent down-regulation of anti-apoptotic Bcl-2 protein levels in the A-594 cell line. These findings suggest that 3g and 3h function as caspase-3, 8, and Bax activators and down-regulators of the antiapoptotic Bcl2, and hence can be classified as apoptotic inducers.Moreover, when compared to untreated A-594 cancer cells, compounds 3f, 3g, and 3h, using Trolox as a control . Compounds 3g and 3h show comparable radical scavenging activity to Trolox at 100 and 50 µM, respectively . Compounds ectively . CompounGeneral information: refer to Additional information -ones 1a–e were prepared according to the published literature [2a) and 1,3-diphenylpropane-1,3-dione (2b), and they were used as purchased (Merck).4-azidoquinolin-2quinolin-2-ones (3a–j) [s (3a–j) ,791a–e (1 mmol), active methylene ketones 2a, b (1.0 mmol), and K2CO3 (1.2 mmol). For 20 h, the reaction mixture was stirred at room temperature. Compounds 3a–j were obtained after the reaction was completed by pouring them into a 500 mL beaker containing 200 gm of crushed ice (in the case of the reaction with 2b) or by evaporating the solvent (in the case of the reaction with 2a). All of the obtained 3a–j products were recrystallized from ethanol.In 30 mL of ethanol, we combined 4-azido-2-quinolinones 1H NMR (DMSO-6d): δH 12.28 , 7.63 , 7.50 , 7.18 , 6.90 , 6.89 , 3.32 , 2.35 ppm ; 13C NMR (DMSO-6d): δC 193.18 (CO), 160.97 , 142.60 (C-4), 142.24 (C-5’), 139.39 (C-4’), 139.24 (C-7), 132.15 (C-5), 123.46 (C-8a), 121.26 (C-6), 120.68 (C-3), 115.98 (C-8), 115.57 (C-4a), 27.74 (CH3), 9.31 ppm (CH3); EI-MS: m/z 268 . Anal. Calcd for C14H12N4O2: C, 62.68; H, 4.51; N, 20.88. Found: C, 62.77; H, 4.39; N, 20.69.This compound was found to be a pale yellow solid; mp 243-45 °C; yield 90%; 1H NMR (DMSO-6d): δH 12.26 , 7.48 , 7.45 , 7.40 , 6.87 , 3.33 , 2.67 , 2.26 ppm ; 13C NMR (DMSO-6d): δC 193.14 (CO), 160.76 , 142.57 (C-4), 142.02 (C-5’), 139.16 (C-4’), 138.16 (C-7), 137.42 (C-5), 122.56 (C-8a), 121.14 (C-3), 115.91 (C-6), 115.49 (C-8), 27.67 (CH3), 20.38 (6-CH3), 9.29 ppm (CH3); EI-MS: m/z 282. Anal. Calcd for C15H14N4O2: C, 63.82; H, 5.00; N, 19.85. Found: C, 63.71; H, 5.12; N, 20.02.This compound was found to be a pale yellow solid; mp 296-98 °C; yield 90%; 1H NMR (DMSO-6d): δH 12.25 , 7.42 , 7.33 , 7.30 , 6.90 , 3.78 , 3.33 , 2.47 ppm ; 13C NMR (DMSO-6d): δC 193.11 (CO), 160.40 , 154.73 (C-6), 142.62 (C-4), 141.71 (C-5’), 139.16 (C-4’), 133.99 (C-5), 121.55 (C-8a), 121.17 (C-3), 117.48 (C-4a), 55.54 (OCH3), 27.66 (COCH3), 9.32 ppm (CH3); EI-MS: m/z 298 . Anal. Calcd for C15H14N4O3: C, 60.40; H, 4.73; N, 18.78. Found: C, 60.51; H, 4.66; N, 18.91.This compound was found to be a colorless solid; mp 263-65 °C; yield 87%; 1H NMR (DMSO-6d): δH 12.76 , 7.79 , 7.25 , 6.93 , 3.73 , 3.45 , 2.45 ppm ; 13C NMR (DMSO-6d): Cδ 193.12 (CO), 160.55 , 142.49 (C-4), 141.12 (C-5’), 140.12 (C-4’), 139.60 (C-7), 137.49 (C-5), 123.41 (C-8a), 122.12 (C-6), 120.12 (C-3), 116.41 (C-8), 115.12 (C-4a), 27.64 (CH3), 20.31 (N-CH3), 9.28 ppm (CH3); EI-MS: m/z 282. Anal. Calcd for C15H14N4O2: C, 63.82; H, 5.00; N, 19.85. Found: C, 63.70; H, 4.87; N, 19.96.This compound was found to be a colorless solid; mp 255-57 °C; yield 71%; 1H NMR (DMSO-6d): δH 7.74 , 7.29 , 7.25 , 6.93 , 3.72 , 3.34 , 2.45 ppm ; 13C NMR (DMSO-6d): δC 193.09 (CO), 160.20 , 142.56 (C-4), 141.12 (C-5’), 140.12 (C-4’), 139.33 (C-7), 132.49 (C-5), 123.95 (C-8a), 122.91 (C-6), 120.38 (C-3), 116.41 (C-8), 115.60 (C-4a), 29.65 (N-CH3), 27.64 (COCH3), 9.22 ppm (CH3-5a’); EI-MS: m/z 282. Anal. Calcd for C15H14N4O2: C, 63.82; H, 5.00; N, 19.85. Found: C, 63.69; H, 5.13; N, 20.01.This compound was found to be a pale yellow solid; mp 188-90 °C; yield 88%; 1H NMR (DMSO-6d): δH 12.22 , 8.20-8.17 , 7.71-7.66 , 7.59-7.55 , 7.45-7.42 , 7.36-7.32 , 7.16-7.13 , 6.96 ppm ; 13C NMR (DMSO-6d): δC 186.18 (CO), 160.70 (C-2), 143.04 (C-4), 142.65 (C-4’), 142.65, 142.58 (Ph-C), 138.96 (C-5’), 136.88 (C-7), 133.38 (C-6), 132.13 (C-5), 130.27, 130.07, 129.72, 128.41, 128.32, 125.19 (Ar-CH), 123.63 (C-8a), 122.24 (C-3), 115.82 (C-8), 115.64 ppm (C-4a); EI-MS: m/z 392 (M+). Anal. Calcd for C24H16N4O2: C, 73.46; H, 4.11; N, 14.28. Found: C, 73.55; H, 4.17; N, 14.14.This compound was found to be a pale yellow solid; mp 250-52 °C; yield 82%; 1H NMR (DMSO-6d): δH 12.21 , 8.21-8.18 , 7.71-7.67 , 7.60-7.55 , 7.44-7.30 , 7.35-7.28 , 6.94 , 6.86 , 2.26 ppm ; 13C NMR (DMSO-6d): δC 186.08 (CO), 160.52 (C-2), 142.70 (C-4), 142.58 (C-4’), 137.03, 136.89 (Ph-C), 133.43 (C-7), 133.32 (C-5), 132.13 (C-7), 130.24 (C-6), 130.01, 129.70, 128.39, 128.24, 125.33 (Ar-CH), 122.84 (C-8a), 121.99 (C-3), 115.73 (C-8), 115.48 (C-4a), 20.33 ppm (CH3); EI-MS: m/z 406 . Anal. Calcd for C25H18N4O2: C, 73.88; H, 4.46; N, 13.78. Found: C, 74.03; H, 4.61; N, 13.90.This compound was found to be a pale yellow solid; mp 278-80 °C; yield 85%; 1H NMR (DMSO-6d): δH 12.18 , 8.17-8.14 , 7.71-7.66 , 7.59-7.54 , 7.44-7.42 , 7.36-7.34 , 7.23 , 6.89 , 3.89 ppm ; 13C NMR (DMSO-6d): δC 186.44 (CO), 160.46 (C-2), 154.87 (C-6), 142.73 (C-4), 137.06, 133.72 (Ph-C), (C-4’), 137.03, 136.89 (Ph-C), 133.43 (C-7), 133.32 (C-5), 132.13 (C-7), 130.45 (C-6), 130.35, 129.90, 128.61, 128.56, 125.33 (Ar-CH), 122.84 (C-8a), 121.70 (C-3), 117.84 (C-8), 116.18 (C-4a), 55.80 ppm (OCH3). EI-MS: m/z 422 (M+). Anal. Calcd for C25H18N4O3: C, 71.08; H, 4.29; N, 13.26. Found: C, 71.19; H, 4.13; N, 13.41.This compound was found to be a colorless solid; mp 290-92 °C; yield 87%; 1H NMR (DMSO-6d): δH 11.38 , 8.20-7.01 , 6.96 , 3.35 ppm ; 13C NMR (DMSO-6d): δC 186.14 (CO), 161.05 , 143.48 (C-4), 142.61 (C-5’), 137.36, 136.87 (Ph-C), 133.35 (C-4’), 130.23 (C-8a), 130.04, 129.70, 128.38, 128.29, 125.18 (Ar-CH), 124.41 (C-5), 122.56 (C-8), 122.02 (C-6), 121.48 (C-3), 115.88 (C-4a), 17.41 ppm (CH3); EI-MS: m/z 406 (M+). Anal. Calcd for C25H18N4O2: C, 73.88; H, 4.46; N, 13.78. Found: C, 73.69; H, 4.55; N, 13.60.This compound was found to be a colorless solid; mp 276-78 °C; yield 77%; 1H NMR (DMSO-6d): δH 8.20-8.18 , 7.75-7.06 , 6.77 , 3.65 ppm ; 13C NMR (DMSO-6d): δC 186.13 (CO), 159.95 (C-2), 142.74 (C-4), 142.62 (C-4′), 141.91, 141.58 (Ph-C), 139.70 (C-5′), 138.48 (C-7), 133.38 (C-6), 132.51 (C-5), 130.25, 130.10, 129.70, 128.99, 128.87, 126.36 (Ar-CH), 124.21 (C-8a), 122.96 (C-3), 116.48 (C-8), 115.54 (C-4a), 29.55 N-CH3); EI-MS: m/z 406 (M+). Anal. Calcd for C25H18N4O2: C, 73.88; H, 4.46; N, 13.78. Found: C, 73.72; H, 4.61; N, 13.66.This compound was found to be a pale yellow solid; mp 227-29 °C; yield 80%; 3a–j [The human mammary gland epithelial (MCF-10A) cell line was applied to assess the viability of the new derivatives 3a–j ,67. Refe3a–j to four human cancer cell lines: colon cancer (HT-29) cell line, pancreatic cancer (Panc-1) cell line, lung cancer (A-549) cell line, and breast cancer (MCF-7) cell line, against erlotinib as the reference [The MTT assay was used to compare the antiproliferative activity of eference . Refer t3f, 3g, 3h, and 3j, the most effective antiproliferative derivatives, were evaluated for their inhibitory action on EGFR as a potential target for their antiproliferative action [Compounds e action . Refer t3f, 3g, 3h, and 3j were tested for their ability to inhibit mutant BRAF (BRAFV600E) according to reported procedures [Compounds ocedures . Refer t3g and 3h were tested for their efficacy on the mutant EGFR (EGFR T790M) receptor type using the previously reported method [Compounds d method . Refer t3f, 3g, and 3h, were tested as caspase-3 activators against human epithelial cancer cell line (A-594) [The most potent derivatives in all in vitro studies, compounds  (A-594) . See Add3g and 3h were investigated further for their effect on caspase-8, Bax, and antiapoptotic Bacl-2 levels against the human epithelial cancer cell line (A-594) using staurosporine as a control [Compounds  control . See Add3f, 3g, and 3h, using Trolox as a control [The scavenging of stable free radicals by 2,2-diphenyl-1-picrylhydrazyl (DPPH) was used to investigate the potential antioxidant properties of compounds  control . See AddH)-ones (3a–j) was developed and synthesized in high yields and purity. The structures of all newly synthesized compounds were determined using elemental analysis, mass spectrometry, and NMR spectroscopy. The newly synthesized 3a–j showed potential antiproliferative efficacy against a panel of four cancer cell lines with compounds 3f–j being the most effective antiproliferative agents. Compounds 3f, 3g, 3h, and 3j were explored as possible multi-target inhibitors of EGFR, BRAFV600E, and EGFRT790M. Our findings demonstrated that compounds 3g and 3h can act as multi-target inhibitors. Furthermore, the apoptotic activity of compounds 3f, 3g, and 3h demonstrated that 3g and 3h activate caspase-3, 8, and Bax, as well as down-regulate the antiapoptotic Bcl2, and therefore can be classed as apoptotic inducers. Finally, DPPH radical scavenging activity demonstrated that compounds 3g and 3h are potent antiproliferative agents with promising antioxidant activity. After structural modifications, these newly synthesized compounds could constitute a new class of antiproliferative agents with multi-target inhibitory action.Through a 1,3-dipolar cycloaddition approach, a novel series of 4-quinolin-2(1