Comprehensive drug-like assessment of pyridine carbothioamide analogs: from molecular modeling to <i>in-vivo</i> evaluation

To evaluate the anti-inflammatory potential of novel class of chemical compounds designed by the linkage of carbothioamide moiety with pyridine. In silico analysis was conducted using molecular docking followed by an in vitro cytotoxicity assay and evaluation of anti-inflammatory activity. Subsequently, in vivo performance was determined using the Complete Freund’s Adjuvant-induced inflammatory model, employing macroscopic, histopathological, and protein expression analyses. Molecular interaction studies revealed that compound R2 displayed the most favorable binding mode with human nitric oxide synthase, cyclooxygenase-1, and cycloxygenase-2. All compounds exhibit dose-dependent cytotoxicity. Notably, compound R4 was safer at higher concentration, whereas compound R2 was comparatively toxic. The in vitro anti-inflammatory activity demonstrated half maximal inhibitory concentration (IC₅₀) values ranging from 10.25 ± 0.0 to 23.15 ± 4.24 µM, with compound R6 exhibiting the lowest IC₅₀ value and compound R3 showing the highest. The in vivo results corroborated the anti-inflammatory effects, with a significant reduction in paw size (<i>p</i> &lt; 0.001). Among the tested compounds, compound R4 exhibited the most potent anti-inflammatory activity, whereas R2 exhibited the least potency. The study highlights the promise of discovering new anti-inflammatory drugs containing pyridine moiety with proven potency, efficacy, and reduced side effects.

本研究旨在评估通过碳硫酰胺（carbothioamide）基团与吡啶（pyridine）连接设计的新型化学化合物的抗炎潜能。本研究首先通过分子对接（molecular docking）开展计算机模拟分析，继而进行体外（in vitro）细胞毒性实验与抗炎活性评估。随后，采用完全弗氏佐剂（Complete Freund’s Adjuvant）诱导的炎症模型，结合宏观观察、组织病理学与蛋白质表达分析，评估其体内（in vivo）抗炎效果。分子相互作用研究表明，化合物R2与人一氧化氮合酶（nitric oxide synthase）、环氧合酶-1（cyclooxygenase-1）及环氧合酶-2（cyclooxygenase-2）均呈现出最优异的结合模式。所有受试化合物均表现出剂量依赖性细胞毒性。值得注意的是，化合物R4在高浓度下安全性更佳，而化合物R2则表现出相对更强的细胞毒性。体外抗炎活性测试结果显示，半数抑制浓度（half maximal inhibitory concentration，IC₅₀）值介于10.25±0.0至23.15±4.24 µM之间，其中化合物R6的IC₅₀值最低，化合物R3的IC₅₀值最高。体内实验结果证实了其抗炎效果，受试动物的足肿胀程度显著降低（*p* < 0.001）。在所有受试化合物中，化合物R4的抗炎活性最强，而化合物R2的抗炎活性最弱。本研究凸显了开发含吡啶基团的新型抗炎药物的潜力，这类药物已被证实具有良好的活性、疗效且副作用更低。