Abstract
Virtual screening is a crucial tool in early stage drug discovery for identifying potential hit candidates. Here, we present an integrated approach that combines theoretical and experimental techniques to identify, for the first time, inhibitors of amidases (Ami1-Ami4) from Mycobacterium tuberculosis. Through computational methods, we proposed a set of potential inhibitors, which were subsequently evaluated experimentally using differential scanning fluorimetry. This led to the identification of two promising hits: a carbohydrazide core (hit 1) and a tetrazole core (hit 5). We further developed a small collection of compounds derived from hit 1, which demonstrated improved affinity for Ami1. Additionally, we determined the crystallographic structure of the Ami1-hit 5 complex at a resolution of 1.45 Å, providing molecular-level insights into the interaction of this compound within the catalytic site. The findings of this study contribute to the advancement of drug discovery against tuberculosis and propose new targets for therapeutic development.