Design, Synthesis, and Biological Evaluation of Novel Tetrandrine Derivatives Targeting AKT1 for Hepatocellular Carcinoma Therapy: Integration of Network Pharmacology, Molecular Dynamics Simulation, and Experimental Validation

To discover novel and effective potential Anticancer agents, 24 novel 14N-substituted tetrandrine (Tet) derivatives were synthesized and investigated for their effects on antiproliferative activities against human Cancer cell lines HepG2 and A549 by the MTT method. Compound 17 exhibited the strongest cytotoxic effect against HepG2 cells with an IC₅₀ value of 2.09 μM and a satisfactory SI value of 11.5, which was 5.3- and 6.4-fold higher than the activity of parental tetrandrine and adriamycin, respectively. Further in vitro experiments (flow cytometry assay, transwell assay, and scratching assay) were designed to validate the significant therapeutic effect of representative compound 17 on HCC. Network pharmacology was used to examine the mechanism of 17 in HCC treatment and the Akt signaling pathway was identified as the key pathway. Molecular docking and molecular dynamics simulations showed good affinity between the active components and core targets of RGW, with stable binding within 200 nanoseconds. Cellular thermal shift assay (CETSA) and ADP-Gl kinase enzyme inhibition assay experiments further demonstrated that 17 could specifically bind to Akt and inhibit its phosphorylation, thereby activating the proapoptotic protein Bax, inhibiting the antiapoptosis protein Bcl-2, and activating Caspase 3, so as to induce Apoptosis. Moreover, it indicates a potent in vivo killing effect against liver cancers, orthotopically transplanted HCC in an AKT1-dependent manner, with a safety profile. Taken together, compound 17 shows therapeutic potential as a safe Anticancer agent through Apoptosis induction, worthy of further development.