Unleashing potential of methotrexate-amino acid methyl ester-loaded lipidic nanocapsules as magic bullets against resistant breast cancer

Current research aims to investigate the augmentation of anticancer efficacy in MTX-resistant (MDA-MB-231) breast cancer cells through the formulation of MTX-amino acid methyl ester-loaded lipidic nanocapsules (LNCs) (MCME (6b), MADE (6c) LNCs) derived from MCME (6b), MADE (6c) conjugates. The conjugates MAME, MCME, and MADE (6a–6c) were synthesized via DCC/HOBt/TEA-based amidation and characterized using FTIR, 1H NMR, and Mass spectroscopy. Further, (6a–6c) conjugates were screened for cytotoxicity, selectivity (MDA-MB-231, A549, and A431 cancer cells), cytocompatibility (WI-38), docking (hDHFR PDB ID:1KMV), ADMET, and MD simulation. Lipidic nanocapsules of active conjugates (MCME (6b), MADE (6c) were developed by adding virgin coconut oil, soya lecithin, poloxamer 407, and glycerol through a high-pressure homogenizer, which was further processed for lyophilization. The resulting MCME (6b) and MADE (6c) LNCs exhibited a spherical shape, a uniform distribution of nanoparticles with a small size, exceptional stability during storage, and a controlled release pattern for MTX. The corresponding 6b and 6c LNCs were analyzed for cytotoxicity (MDA-MB-231), cytocompatibility (WI-38), cell cycle arrest, cellular uptake, apoptosis (Annexin V-FITC/PI), and DNA cleavage. Remarkably, LNCs 6b and 6c exhibited enhanced cytotoxicity towards MTX-resistant MDA-MB-231 cells, with notable impacts on the cell cycle, cellular uptake, apoptosis, and moderate DNA cleavage ability. The histo-toxicological evaluation of Cirrhinus mrigala fish fingerlings demonstrated a safer ecotoxicological profile of LNC 6b compared to LNC 6c and MTX. Therefore, the present research revealed the potential of MCME (6b) LNCs as targets for MTX-resistant breast cancer. [Display omitted] •MTX-α, γ-bis (amide)amino acid methyl ester conjugates MAME (6a), MCME (6b), and MADE(6c) were synthesized.•Most bioactive MCME (6b) and MADE (6c) conjugates were developed into lipidic nanocapsules.•MTT assay, cellular uptake, cell cycle, and apoptosis study indicated the increase in cytotoxicity of lipidic nanocapsules (LNCs) MCME (6b), MADE (6c).•MCME (6b) and MADE (6c) lipidic nanocapsules displayed controlled release of MTX.•Lipidic nanocapsules exhibited storage stability for six months at 4 ± 1 °C and safer eco-toxicological profile.