A series of six novel benzimidazole-pyrazole hybrid molecules was synthesized and characterized using elemental analysis (CHN) and spectroscopic methods (1HNMR, FT-IR). All the synthesized compounds were evaluated for their in vivo anti ulcerogenic activity using Albino rats (weighing 180–220 g). The interactions between the compounds and active site residues of H+/K+ ATPase were investigated by molecular docking studies using autodock vina 4.0. SCH28080 was used to validate the docking results. Also the drug likeliness of these compounds was predicted using Molinspiration server in light of Lipinski’s rule of five. All the six synthesized compounds exhibited higher anti-ulcer activity as compared to omeprazole. These novel hybrid compounds showed comparable anti-ulcer potential of 72–83% at dose level of 500 µg/kg, whereas omeprazole showed 83% anti-ulcer activity at dose level of 30 mg/kg. The results clearly indicate that these novel benzimidazole-pyrazole hybrids can present a new class of potential anti ulcer agents and can serve as new anti-ulcer drugs after further investigation. Graphical abstract An overveiw of synthesis, in silico and in vivo antiulcer screening of benzimidazole pyrazole hybrids

2-Mercaptobenzothiazole and its derivatives are widely known for their diverse biological activities, particularly antimicrobial and anticancer potential. In the present study, a series of new hybrid compounds consisting of 2-mercaptobenzothiazole and different aryl amines 2(a-j) were synthesized and characterized by Fourier transform infrared (FTIR), ¹H NMR, and ¹³C NMR spectral data. The synthesized compounds were screened for in vitro antibacterial activities through agar well diffusion assay. Among the series, 2b, 2c, and 2i exhibited significant antibacterial activity comparable to the standard drug levofloxacin. Based on their antibacterial potential, these compounds were further tested for their antibiofilm activity. All of the three compounds showed promising antibiofilm potential, even better than the standard drug cefadroxil at 100 μg/100 μL concentration. Molecular docking studies were performed to explore the antibacterial mechanism of these compounds. Strikingly, the molecule 2i shared the same hydrophobic pockets as those of levofloxacin in case of bacterial kinases and DNA gyrases. In addition, 2i exhibited satisfactory antibiofilm activity in comparison to the standard. Our study therefore suggested that the synthetic compound 2i possesses remarkable antibacterial activity and may serve as a lead molecule for the discovery of potent antibacterial agents.

Alkaline phosphatase (ALP) is a zinc-dependent metalloenzyme that catalyzes the hydrolysis of phosphate monoesters and is essential to several important physiological processes, such as bone mineralization and signal transduction. The clinical correlation of dysregulated ALP activity with a wide range of disease types, including bone disorders, hepatobiliary diseases, and certain malignancies, highlights the therapeutic importance of developing and identifying potent calf intestinal alkaline phosphatase (CIALP) inhibitors. Eleven new chalcone–benzamide derivatives (3a–3k), containing structurally different substituents, were synthesized and extensively investigated by spectroscopic methods such as IR, 1H NMR, 13C NMR, and low-resolution EI-MS analysis. All synthesized compounds were screened for DPPH radical-scavenging activity, with ascorbic acid as the standard (IC50 = 18.68 μM), and CIALP inhibitory activity, with KH2PO4 as the standard (IC50 = 2.80 ± 0.065 μM). In the DPPH assay, compound 3k showed the highest antioxidant activity (IC50 = 21.21 μM), followed by 3h (29.94 μM), 3j (39.92 μM), and 3c (53.22 μM), closest to the reference standard. In the enzyme inhibition assay, compounds 3d (IC50 = 3.93 ± 0.314 μM), 3h (IC50 = 4.49 ± 1.312 μM), and 3c (IC50 = 5.69 ± 1.124 μM) demonstrated the most potent enzyme inhibitory activities. Docking of the synthesized compounds was performed against the crystal structure of placental alkaline phosphatase (PLAP, PDB ID: 1EW2), used as a structural homologue in the absence of a deposited CIALP crystal structure, which showed that all synthesized compounds had good binding energies ranging from −7.9 to −8.3 kcal/mol; this narrow range is within the resolution limits of AutoDock scoring and did not correlate with experimental enzyme-inhibitory potency across the series. Prioritized lead candidates were found to have good oral bioavailability (expected to be above 90% absorption), acceptable physicochemical properties (compliant with Lipinski’s rule of five), and manageable toxicity risk. Together, the integrated in vitro biological and in silico computational data suggested that compounds 3h, 3c, and 3j are promising dual-function leads with both antioxidant and enzyme-inhibitory activity, while compound 3d stands out as a potent, structurally distinct enzyme inhibitor─making the benzamide scaffold a promising structural framework for the rational optimization of multitarget therapeutics.

Background/Objectives: Indoles and benzimidazoles are important nitrogen-containing heterocyclic scaffolds and are a cornerstone of synthetic drugs. In this study, benzamides of 2-aminophenylindoles and 2-aminophenylbenzimidazoles were synthesized and evaluated for their potential. Further, they were also subjected to computational studies for their ADMET profiling and binding with target proteins. Methods: The synthesized compounds were characterized by ATR-FTIR, 1H NMR, 13C NMR and electrospray ionization mass spectroscopy (ESI-MS). They were screened for their in vitro antibacterial activity by the Microplate Alamar Blue Assay (MABA) and for their anticancer potential by the MTT assay against the HeLa, PC3 and 3T3 cell lines. Further, the compounds were assessed for their ADMET profiles by the Deep-PK platform, and binding with selected kinases was assessed by AutoDock Vina v1.2.7, followed by MD simulations in GROMACS. Density functional theory (DFT) calculations were also performed to investigate the electronic properties of the synthesized compounds. Results: All synthesized compounds were inactive in antibacterial assays and mildly active against cancer cells. N-[4-(1H-benzimidazol-2-yl-phenyl]benzamide (4-APB-B) exhibited good activity against the HeLa cell line (IC50: 5.48 ± 0.01 µM) while showing very low cytotoxicity against the 3T3 cell line (selectivity index: 10.7), against which doxorubicin was highly active, indicating selectivity towards specific cancer cells. Docking studies indicated favorable binding affinities towards the selected kinase targets. Further, ligands with the best binding affinities in docking studies were subjected to MD simulations of 100 ns, and DFT studies were also performed to assess the electronic properties of the synthesized compounds. Conclusions: Our study provides a pathway for the synthesis of 2-phenylbenzimidazoles and 2-phenylindoles and demonstrates that the synthetic compound 4-APB-B possesses remarkable selective cytotoxic activity and can serve as a lead molecule for further development into a successful anticancer agent.