Discovery of Diaryl Pyrimidine-Guanidine Derivatives as Jumonji C Domain-Containing Lysine Histone Demethylase (KDM) Inhibitors with In Vivo Anti-Cervical Cancer Activity.

JMJD family histone demethylases are highly expressed in cervical cancer and promote tumor progression, highlighting their potential as therapeutic targets for this malignancy. Using a scaffold-hopping-based virtual screening, we discovered a series of diaryl pyrimidine-guanidine derivatives as KDM6B inhibitors. The optimized compound 40 exhibited potent KDM6B inhibitory activity comparable to that of GSK-J4, but differs mechanistically by being non-competitive with 2-OG. Although 40 showed pan-KDM inhibition in enzymatic level, it selectively elevated H3K27me3 levels in C33A cells without affecting H3K4me3 or H3K9me3. In C33A cells, 40 suppressed proliferation, induced cell-cycle arrest and apoptosis. In a xenograft mouse model, 40 inhibited tumor growth, downregulated Ki-67, and upregulated cleaved caspase-3, p-H2A.X, and H3K27me3. Collectively, 40 represents a promising chemical probe for investigating JMJD KDMs and H3K27 modifications in cervical cancer and establishes a foundation for developing targeted agents.