DHFR-IN-22 (Compound 8) is a DHFR (Dihydrofolate Reductase) inhibitor that falls under the category of 2,4-diaminopyrimidine compounds. It demonstrates significant inhibitory activity against isolated DHFR enzyme and major nontuberculous mycobacteria (NTM) species, specifically Mycobacterium avium and Mycobacterium abscessus. The compound has an IC50 of 1.1 nM and a MIC of 1.5 μg/mL for M. abscessus, and an IC50 of 6.3 nM and a MIC of 0.1 μg/mL for M. avium, while its IC50 for human DHFR is 2100 nM. DHFR-IN-22 may be promising for developing new strategies to combat NTM infections.