Label-free affinity screening, design and synthesis of inhibitors targeting the Mycobacterium tuberculosis L-alanine dehydrogenase
Fig 1
(a) DLAC screening tests reveal N6-methyl adenosine (D30) as a putative hit component against AlaDH. D28, C2-methyl adenosine; D29, cyclohexyl 2-[[5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazol-2-yl]sulfanyl]acetate; D30, N6-methyl adenosine; D31, 4-[[5-(4-methylphenyl)-1,3,4-oxadiazol-2-yl]sulfanyl]but-2-ynyl furan-2-carboxylate; D32, N-[4-(4-butanoylpiperazin-1-yl)-3-chlorophenyl]-5-nitrofuran-2-carboxamide; D33, 5-(1,3-benzodioxol-5-yl)-N-(5-chloro-2-hydroxyphenyl)-7-(trifluoromethyl)-1,5,6,7-tetrahydro pyrazolo[1,5-a]pyrimidine-2-carboxamide; D34, 5-(4-bromophenyl)-N-[(1,5-dimethylpyrazol-4-yl)methyl]-7-(trifluoromethyl)-1,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide; D35, 4-methyl-N-(5-pyridin-4-yl-1,3,4-oxadiazol-2-yl)benzamide; D36, benzyl (2S)-4-methyl-2-[[4-[(3-methylphenyl) carbamoyl]-1H-imidazole-5-carbonyl]amino]pentanoate. The gels are cropped from the full-length gels which are presented in S2a and S2c Fig in S1 File. ATP is used as a positive control based on our previous data [25], where NADH and ATP (but not NAD) bound strongly with this assay (b) Inhibition effect of N6-methyl adenosine is confirmed using enzyme kinetic studies.