Discovery of Potent 1,2,4-Triazole-Derived HIV Capsid Modulators with Dual-Stage Antiviral Activity via Targeting the Unexplored GTNQ Sub-Pocket Using a Multi-Site Binding Strategy

HIV capsid (CA) is a promising target for novel anti-HIV drugs. Using a structure-guided multisite binding strategy, we designed and synthesized 42 novel 1,2,4-triazole derivatives targeting the unexplored GTNQ subpocket within the CA hexamer. Compound 5-33d exhibited potent antiviral activity (EC50 = 0.14 μM), 12-fold more active than the lead compound PF74 (EC50 = 1.7 μM). Surface plasmon resonance (SPR) assay indicated that 5-33d exhibited potent binding affinities to the CA hexamer (KD = 59.0 nM), rather than the CA monomer (KD = 175.7 μM). The cocrystal structure demonstrated that the N-ethyl cyclopropanesulfonylamide moiety of 5-33d forms an extensive hydrogen-bond and water-mediated interaction network with key residues surrounding the GTNQ subpocket. Besides, 5-33d exerts dual-stage inhibition by disrupting CA-host binding and modulating capsid assembly. These findings lay a structural and mechanistic foundation for the development of anti-HIV candidates targeting the GTNQ subpocket.