Membrane interaction studies of isoniazid derivatives active against drug-resistant tuberculosis

• Isoniazid (INH) derivatives are promising agents against M. tuberculosis ( Mtb ); • Compounds tested did not disturb lipid rafts in mammalian cell membrane models; • They also interact with more fluid lipid bilayers like internal cellular membranes; • INH-C10 incorporates into rigid and compact gel membranes alike the Mtb cell wall; • Membrane interaction of the compounds corroborate their high therapeutic potential. Tuberculosis is one of the leading causes of mortality worldwide due to the growth of multi-drug resistant strains unsusceptible to currently available therapies. Four compounds, isoniazid (INH), and three derivatives, N'-decanoylisonicotinohydrazide (INH-C10), N'-(E)-(4-phenoxybenzylidene)isonicotinohydrazide (N34) and N’-(4-phenoxybenzyl)isonicotinohydrazide (N34red), were studied. Owing to their advantageous in vitro selectivity index against the primary mutation responsible for drug resistance in Mycobacterium tuberculosis (Mtb), as well as their suitable lipophilicity and interaction with human serum albumin, INH-C10 and N34 were deemed promising antitubercular compounds. N34red, despite differing from N34 only in the saturation of the N′ = C bond, presents a poor selectivity index. To delve deeper into the therapeutic potential of these compounds, their interaction with biomembrane models, mimicking biological barriers on the way to the target inside Mtb cells, was herein evaluated. All compounds, except N34red, weakened the packing of the acyl chains in the rigid lipid gel phase, especially INH-C10, which was the only compound disturbing liquid disordered membranes. Notably, all compounds except INH decreased membrane dipole potential, across all types of bilayers studied, but only N34red had a drastic effect. The insertion in gel phase bilayers suggests that the compounds may be able to penetrate the rigid cell wall of Mtb. Förster's resonance energy transfer (FRET) assays showed that, in ternary bilayers with liquid ordered/liquid disordered lateral heterogeneity mimicking human cell membranes, the compounds affected neither the size nor the organization of lipid domains. These results provide molecular insights into the low toxicity against human cell lines and improved activity against drug-resistant Mtb of INH-C10 and N34.