Library screening of chemical compounds prepared in the laboratory for TNF-α inhibitory activity

Abstract

Introduction: TNFα is an inflammatory cytokine that plays a significant role in the progress of inflammatory and autoimmune diseases. Therefore, TNFα inhibitors can be effective in the treatment of these diseases. In this regard, synthetic small molecules can be considered as TNFα inhibitors.Objective: The goal of the present study is to identify new compounds from in house compound library that inhibit TNFα activity.Method: MTT assay was used for screening of library compounds against TNFα. The compounds that showed inhibitory effects on TNFα at a concentration of 50 µM were serially diluted and investigated in the case of dose-response effects in MTT assay. Further evaluation was carried out by in silico methodologies. For that, the identified compounds were docked into TNFα binding site using gold program and the resulted complexes were introduced to molecular dynamics simulation studies.Results:Among the available compounds, seven compounds at a concentration of 50 µM were able to inhibit the cytotoxic effects of TNFα on L929 cells, and among them, three compounds were able to dose-dependently inhibit TNFα with IC50 values of 43.70 (compound 55), 36.69 (compound 61) and 9.50 (compound 81). The results obtained in the molecular dynamics studies confirmed the laboratory studies in which compound 81 with a binding energy of -47.08 showed greater binding affinity to TNFα compared to compounds 61 and 55, and comparable to the binding energy of the control compound UCB-5307.Conclusion:Compound 81, which showed appropriate inhibitory effects on TNFα in laboratory studies and molecular dynamics studies comparable to the control compound, can be proposed as a TNFα inhibitor candidate for alleviating inflammatory and autoimmune diseases.