Simulation Data related to the publication: Design, synthesis and biological activity of novel substituted 3-benzoic acid derivatives as MtDHFR inhibitors The files include raw trajectory files of the Desmond MD simulations of different MtDHFR inhibitors and substrates within the active site (trajectory format is out.cms and the full trj files, Schrödinger, LLC, New York, NY, 2019, more details on the materials and methods section of the respective publication). Article Abstract: The enzyme dihydrofolate reductase from M. tuberculosis (MtDHFR) have high untapped potential to be a target for new drugs against tuberculosis, due to its importance and uniqueness for this pathogen. Preliminary studies have obtained fragment-like molecules with low affinity to MtDHFR which can potentially become lead compounds. Taking this into account, the fragment MB872 was used as a prototype for analogue development by bioisosterism/retro-bioisosterism, which resulted in 20 new substituted 3-benzoic acid derivatives. Compounds were active against MtDHFR, with IC50 ranging from 7 to 40 μM, where compound 4e not only had the best inhibitory activity (IC50 = 7 μM), but also was 71-fold more active than the original fragment MB872. The 4e inhibition kinects revealed a known uncompetitive mechanism, which was supported by molecular modeling suggestion that the compounds can access an unique back-pocket. Thus, based on these results, substituted 3-benzoic acid derivatives have strong potential to be developed as novel MtDHFR inhibitors and also anti-TB agents.