Computational, In Vitro, and In Vivo Evaluation of Benzimidazole–Pyrazole Hybrid Scaffolds as Multi‐Target Anti‐Ulcer Agents

Abstract

Peptic ulcer disease (PUD) continues to be an important gastrointestinal condition attributed to Helicobacter pylori infection, usage of nonsteroidal anti‐inflammatory drugs, oxidative stress, and inflammatory mediators. Although there are available treatments for this condition, side effects, relapse, and safety issues call for development of new multi‐target anti‐ulcer compounds. In this research, a group of benzimidazole‐pyrazole analogs (5a–5j) was designed, synthesized, and studied with the aid of computational and experimental methods. Molecular docking into H+/K+‐ATPase (PDB ID: 5YLU) and COX‐2 (PDB ID: 3LN1) enzymes predicted high affinity of these compounds with docking scores of −9.4 and −9.3 kcal/mol for compounds 5i and 5e, respectively, higher than those obtained for the reference drugs omeprazole and celecoxib. The inhibition of the gastric H+/K+‐ATPase enzyme was also observed in vitro, with compounds 5i and 5e having better inhibitory effects. Studies on the ethanol and indomethacin‐induced gastric ulceration models demonstrated good inhibitory effects of these compounds. Biochemical analysis showed that these analogs significantly decreased the levels of TNF‐α, COX‐2, myeloperoxidase, and malondialdehyde and increased prostaglandin E2. Acute oral toxicity studies indicated no observable toxicity at 100 mg/kg. Collectively, these findings identify compounds 5i and 5e as promising multifunctional gastroprotective candidates warranting further pharmacological investigation.