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

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.