Investigation of 4-(4-Chlorophenyl)-2,4-dihydro-5-(4-pyridyl)-3H-1,2,4-triazole-3-thione in morphine induced dependence model augmented by simulation and molecular docking

Opioid dependence is a serious worldwide health issue. The available treatments are frequently constrained by side effects and inadequate neuroprotection. New therapeutic drugs that can both address opioid-induced oxidative neurotoxicity and lessen withdrawal symptoms are therefore urgently needed. This study aimed to assess the neurotherapeutic potential of 4-(4-chlorophenyl)-2,4-dihydro-5-(4-pyridyl)-3H-1,2,4-triazole-3-thione (CPTT) in the treatment of morphine withdrawal and related oxidative damage. ADMET analysis was used to evaluate pharmacokinetic and toxicity profiles. The molecular docking and 100-ns molecular dynamics simulations were used to investigate the interaction with the NR2B subunit of the NMDA receptor, a key target in opioid dependency. Male albino mice were made morphine dependent by increasing intraperitoneal dosages over four days, followed by naloxone-induced withdrawal. The impact of CPTT on withdrawal behavior and oxidative stress markers was examined.In silico predictions showed excellent gastrointestinal absorption (89%), moderate solubility and permeability. No interaction was observed with P-glycoprotein, indicating good oral bioavailability. The compound was non-mutagenic and hepatotoxic, though hERG II inhibition flagged a potential cardiotoxicity risk. Metabolism analysis revealed involvement of CYP3A4, with inhibition of CYP1A2, CYP2C19, and CYP3A4, highlighting possible drug-drug interactions. Molecular docking demonstrated strong binding affinity for the NMDA NR2B subunit, and simulation data confirmed the structural stability of the ligand-receptor complex. In vivo , CPTT significantly alleviated naloxone-precipitated withdrawal symptoms. Biochemically, it restored depleted glutathione (GSH) levels, normalized glutathione-S-transferase (GST) and catalase (CAT) activities, and suppressed lipid peroxidation (LPO). It also reduced hippocampal nitrite levels, suggesting inhibition of nitric oxide (NO) production, a key mediator of morphine-induced neuroinflammation and oxidative stress. These findings indicate that CPTT has multi-targeted neuroprotective effects, which are most likely mediated by antioxidant characteristics and glutamatergic transmission regulation. The drug's favorable pharmacokinetic profile, robust NMDA receptor interaction, and in vivo efficacy warrant further preclinical and clinical research as a potential treatment for opioid withdrawal and neurotoxicity.