Neoruscogenin and its human metabolites as JAK2 ligands in hepatocellular carcinoma

Treatment options for hepatocellular carcinoma (HCC) remain unsatisfactory, and JAK2 continues to attract interest as a point of pharmacological intervention in this malignancy. The present work asked whether Neoruscogenin, a spirostanol sapogenin, and its predicted human metabolites can engage that node. Intersecting predicted compound targets with HCC-associated genes returned 97 shared proteins. Degree-based ranking placed IL2, SRC, STAT3, MTOR, PIK3CB, MAPK1, JAK2, MDM2, KIT and MAPK14 at the center of the resulting interaction network, and functional enrichment pointed toward cancer-related, MAPK, PI3K-Akt, apoptotic and immune-associated signaling. Four biotransformation products retaining the steroidal core were carried forward. Across ten prioritized receptors, the parent compound scored best against JAK2 (PDB 5AEP, -10.48 kcal/mol), among the metabolites, the 3-O-glucuronide BTM00003 came within 0.17 kcal/mol of it (-10.31 kcal/mol), and both exceeded the reference inhibitor QUP (-8.87 kcal/mol). All three complexes survived 200 ns of unrestrained dynamics, but the routes to stability differed: free energy landscape, principal component and cross-correlation analyses described a narrow, dynamically restrained basin for QUP, whereas Neoruscogenin and BTM00003 stayed bound while exploring wider conformational space. MMGBSA returned favorable binding free energies for all three, with QUP most negative. Predicted ADMET combined near-complete intestinal absorption for Neoruscogenin with poor aqueous solubility and a hepatotoxicity alert, and DFT descriptors described a hard, electronically stable scaffold. Neoruscogenin and its glucuronide therefore merit experimental follow-up in HCC as starting points rather than as optimized candidates.