Synthesis, Characterization, and Multi-ModeI Evaluation of Anti-Inflammatory Activity of Four Porphyrin Derivatives: In Vivo, In Vitro, and In Silico Approaches

Abstract

This study is part of ongoing efforts to develop effective alternative treatments against inflammation. It focuses on evaluating the anti-inflammatory activity of four porphyrin derivatives synthesized in the laboratory and assessed through three complementary approaches: in vitro, in vivo, and in silico. Chronic inflammation poses a major medical challenge today, being linked to several serious diseases such as cancer, type 2 diabetes, multiple sclerosis, and neurodegenerative disorders, which highlights the need for new, safer, and more effective compounds. This study evaluated the anti-inflammatory potential of four derivatives: NiTPPH₂, TbiPPH₂, TPPH₂(o-methyl), and ZnTPPH₂, compared to diclofenac as a reference drug, using a glyphosate-induced inflammation model in rats, alongside cellular and computational assessments to determine efficacy, toxicity, and pharmacokinetic behavior. The results of both in vitro and in vivo evaluations demonstrated a marked effectiveness of porphyrin derivatives as anti-inflammatory agents. In the bovine serum albumin (BSA) denaturation assay, these derivatives exhibited strong protein stabilization capacity, with inhibition rates ranging from 79.86% to 90.22% and IC₅₀ values between 68.91 and 92.71 µg/mL. These findings indicate significant anti-inflammatory activity at the cellular level. In the rat inflammation model, treatment with porphyrin derivatives led to notable biological improvements, including a reduction in hepatic malondialdehyde (MDA) levels from 8.06 to 4.0 nmol/mg protein following administration of TPPH₂(o-methyl), and an increase in brain reduced glutathione (GSH) levels from 5.8 to 10.53 nmol/mg protein with NiTPPH₂. The treatment also stabilized liver enzyme levels (ALT and AST) close to control values and normalized the weights of affected organs, particularly the liver and kidneys, compared to the glyphosate-only group. These results reflect the promising potential of porphyrin derivatives in modulating inflammation associated with oxidative stress. Histological analysis supported these findings, showing clear structural improvement in liver, kidney, and brain tissues, with reduced inflammation. In silico studies confirmed strong pharmacological potential, especially for TPPH₂(o-methyl) and NiTPPH₂, with COX-1 binding energies reaching (–10.3 kcal/mol), and favorable ADMET profiles in terms of absorption, distribution, and safety.