Efficient separation of chemical constituents from shilajit by an inner-recycling counter-current chromatography and their molecular docking analysis for osteoporosis.

Shilajit (Asphaltum punjabianum), a herbomineral ethnomedicinal substance from layers of rocks in the Himalayas and Hindukush ranges, is reported to treat urinary bladder cancer, it reduces inflammation, oxidative stress and bone loss. To date, there are no such authentic reports on the detailed separation or isolation of the chemical constituents from shilajit. In the current study, the high-speed counter-current chromatography technique is used for the separation of chemical constituents from shilajit for the first time. Thus, the current investigation depicts an efficient separation of six components namely boscartin J (1), 3α-O-acetyl-11-keto-β-boswellic acid (2), boscartin K (3), hemerocallal A (4), 3α-hydroxytirucalla-8,24-dien-21-oic acid (5), and centrolobol (6) using HSCCC employing a solvent system MTBE/n-But/ACN/H2O (2:2:1:5, v/v). Using the molecular docking, compounds 2 and 5 exhibited a significant affinity for ERα (PDB 3OCB) and Cathepsin K (PDB 6WOK). It is worth mentioning that the simulated results reflect a profound alignment with established and renowned bone-protective pharmacological principles. 3α-O-Acetyl-11-keto-β-boswellic acid and 3α-hydroxytirucalla-8,24-dien-21-oic acid exhibit promising dual activity, modulation of ERα and inhibition of Cat. K for the treatment of osteoporosis.