IN SILICO ANALYSIS OF MUTATIONAL EFFECTS ON HUMAN DHFR IN RELATION TO THE RESPONSE TO METHOTREXATE AND EXPLORATION OF NEW POTENTIAL NON-TOXIC INHIBITORS

Abstract

Dihydrofolate reductase (DHFR) is a vital enzyme involved in folate metabolism, DNA synthesis, and cell growth. Dysregulation of DHFR has been linked to several malignancies, making it an attractive target for targeted cancer therapy. Methotrexate, a commonly used anticancer drug, is known to inhibit DHFR activity. However, resistance to methotrexate treatment is a significant challenge in cancer therapy. In this study, we performed an in silico analysis to investigate the effects of mutations on human DHFR in relation to the response to methotrexate and explored potential non-toxic inhibitors. DHFR mutations were collected from databases, and their impact on enzymatic activity was assessed using computational methods. Additionally, we examined the binding affinity of methotrexate and novel non-toxic inhibitors to mutant DHFR proteins through molecular docking simulations. Our results demonstrated that specific mutations influenced the enzymatic activity of DHFR and disrupted critical interactions, potentially contributing to the development of cancer. Furthermore, we observed that methotrexate exhibited favorable binding affinity towards wild-type DHFR and certain types of mutations, providing insights into methotrexate resistance. Interestingly, one of the newly identified non-toxic inhibitors, CID21453, displayed superior binding affinity compared to other ligands. However, further experimental validation is necessary to determine the efficacy and therapeutic potential of CID21453 as a targeted cancer therapy. Overall, our in silico analysis contributes to a better understanding of the mutational effects on DHFR and provides potential avenues for developing novel inhibitors with improved therapeutic outcomes.