Fragment-based drug design was used to identify Mycobacterium tuberculosis (Mtb) dihydrofolate reductase (DHFR) inhibitors. Screening of ligands against the Mtb DHFR enzyme resulted in the identification of multiple fragment hits with IC₅₀ values in the range of 38–90 mM versus Mtb DHFR and minimum inhibitory concentration (MIC) values in the range of 31.5–125µg/mL. These fragment scaffolds would be useful for anti-tubercular drug design.