NSD2/H3K36me2 modulator-1 is an orally active modulator targeting NSD2/H3K36me2. It competitively binds to the SAM pocket of NSD2, effectively inhibiting NSD2 expression and H3K36me2 methylation. This compound can also reverse epithelial-mesenchymal transition (EMT), inhibit cell migration, and induce G0/G1 phase arrest and apoptosis (apoptosis). Additionally, NSD2/H3K36me2 modulator-1 reduces mitochondrial membrane potential (MMP) and promotes reactive oxygen species (ROS) generation. It is used in research for developing epigenetic anticancer strategies targeting NSD2 in hepatocellular carcinoma (HCC).

NSD2/H3K36me2 modulator-1 (Compound 5b) exhibits potent inhibitory activity with IC50 values below 5 μM across five cancer cell lines (A549, HepG-2, PC-3, HCT-116, and MCF-7, with respective IC50 values of 4.67, 2.12, 2.64, 4.56, and 4.99 μM), confirming its remarkable broad-spectrum anti-tumor efficacy. It shows higher tumor selectivity in HepG-2 cells compared to HaCaT cells (IC50 = 19.2 μM, selectivity index (SI) = 9.06). NSD2/H3K36me2 modulator-1 (2 μM, 24 hours, 37-62 ℃) effectively binds to NSD2 within HepG-2 cells. At concentrations of 2 and 4 μM for 24 hours, it inhibits NSD2 and selectively downregulates NSD2-mediated H3K36me2 in HepG-2 cells. It also reverses EMT at the molecular level, possibly through NSD2 inhibition followed by downregulation of TWIST1, altering the expression of key EMT regulatory factors. Additionally, NSD2/H3K36me2 modulator-1 (2, 4 μM, 18, 36 hours) significantly inhibits migration and proliferation of HepG-2 cells. Its multiple regulatory effects on HepG-2 cells (2, 4 μM, 24 hours) include effectively inducing apoptosis, triggering concentration-dependent G0/G1 phase arrest, dose-dependently reducing mitochondrial membrane potential (MMP), and efficiently promoting ROS production.

Compound NSD2/H3K36me2 modulator-1 (Compound 5b), administered orally at doses of 20 and 40 mg/kg daily over 18 days, demonstrated significant antitumor activity in a HepG-2 cell xenograft model. It effectively inhibited tumor growth without notable differences in body weight, organ index, or morphological characteristics, indicating good safety.