Identification\nof Potential Inhibitors of Mycobacterium tuberculosis Amidases: An Integrated\nIn Silico and Experimental Study

Virtual screening is a crucial tool in early stage drug\ndiscovery\nfor identifying potential hit candidates. Here, we present an integrated\napproach that combines theoretical and experimental techniques to\nidentify, for the first time, inhibitors of amidases (Ami1–Ami4)\nfrom Mycobacterium tuberculosis. Through\ncomputational methods, we proposed a set of potential inhibitors,\nwhich were subsequently evaluated experimentally using differential\nscanning fluorimetry. This led to the identification of two promising\nhits: a carbohydrazide core (hit <b>1</b>) and a tetrazole core\n(hit <b>5</b>). We further developed a small collection of compounds\nderived from hit <b>1</b>, which demonstrated improved affinity\nfor Ami1. Additionally, we determined the crystallographic structure\nof the Ami1–hit <b>5</b> complex at a resolution of 1.45\nÅ, providing molecular-level insights into the interaction of\nthis compound within the catalytic site. The findings of this study\ncontribute to the advancement of drug discovery against tuberculosis\nand propose new targets for therapeutic development.