Background of study: Inhibition of DHFR is one of the potential targets for anticancer therapy. In this study, topochemical descriptor based calculations were carried out on substituted 4(3H)-quinazolinones to predict DHFR inhibition. A data set of 50 molecules of substituted 4(3H)-quinazolinones as potential DHFR inhibitors and its topological descriptors like Wiener__ampersandsignrsquo;s index, Balaban__ampersandsignrsquo;s index and Molecular topological index were investigated. Objective: The topological data was systematically analyzed and suitable descriptor based topochemical models were developed after the identification of the active ranges. Research methodology: Subsequently, the DHFR inhibition activity was predicted for all the data set of molecules using topochemical models. Then, the predicted activity was compared with the experimentally reported DHFR inhibitor activity. Results: The overall prediction of DHFR inhibition was found to be higher accuracy of 80% for the models based on Wiener__ampersandsignrsquo;s index as well as Molecular topological index and 74% for Balaban__ampersandsignrsquo;s index. Conclusions: The prediction of DHFR inhibition using topochemical descriptors can be useful to design newer analogues of quinazolines as potential DHFR inhibitors.