Integrated stability and metabolomic investigation of new rifampicin and isoniazid co-loaded liposome against tuberculosis

Objectives 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. Significance The novel biocompatible liposomal system enables sustained co-delivery of RIF and INH, providing a scalable and stable platform with enhanced antimicrobial efficacy and strong potential to advance tuberculosis therapy. Methods 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. Results The optimized 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). The release study indicated sustained diffusion-controlled kinetics consistent with the Higuchi model, achieving cumulative releases of approximately 92% for INH and 85% for RIF over a 45-hour period. The accelerated stability studies confirmed negligible drug degradation, while antimicrobial assays demonstrated a twofold reduction in MIC relative to free drugs, and metabolomic profiling indicated modulation of glutathione, citric acid, and tyrosine pathways associated with enhanced redox balance and antimicrobial activity. Conclusions The co-loaded RIF-INH liposomal system offers a promising, clinically translatable approach for sustained drug release and improved tuberculosis therapy.