Integrated Stability and Metabolomic Investigation of New Rifampicin and Isoniazid co-loaded Liposome against Tuberculosis

This study aimed to develop and evaluate rifampicin (RIF) and isoniazid (INH) co-loaded liposome for sustained drug delivery to enhance therapeutic efficacy against tuberculosis (TB) and overcome challenges associated with prolonged treatment and drug resistance. A novel biocompatible liposomal system that enables sustained co-delivery of RIF and INH, thereby enhancing therapeutic efficacy and reducing drug resistance in TB treatment. However, the formulation offers a stable and scalable delivery platform with sustained release, improved antimicrobial efficacy, and significant potential for advancing tuberculosis therapy. Liposome were prepared using soybean lecithin and cholesterol (L-CH) via rotary evaporator-assisted thin film hydration, optimized by Box–Behnken design, and characterized for size, PDI, entrapment efficiency, and physicochemical properties (FT-IR, DSC, HR-TEM). In vitro release, accelerated stability, antimicrobial efficacy against M. smegmatis and M. tuberculosis H ₃₇ Rv, and LC-MS/MS-based metabolomic profiling were systematically evaluated. The optimised liposome exhibited a mean size of 129.5 ± 2.20 nm, PDI of 0.369 ± 0.06, and entrapment efficiencies of 63.84 ± 1.62% (RIF) and 56.92 ± 1.69% (INH). Following Higuchi diffusion-controlled kinetics, sustained release was observed, with INH (~ 92%) and RIF (~ 85%) over 45 hours. Accelerated stability studies showed minimal drug degradation. Antimicrobial assays revealed a twofold reduction in minimum inhibitory concentration compared to free drugs. Metabolomic analysis demonstrated modulation of glutathione, citric acid, and tyrosine pathways, suggesting improved redox balance and antimicrobial effect. The co-loaded RIF–INH liposomal system offers a promising, clinically translatable approach for sustained drug release and improved tuberculosis therapy.