BRD4/NAMPT-IN-1 (Compound A2) exhibits strong inhibitory effects on NAMPT and BRD4, with IC50 values of 35 nM (NAMPT) and 58 nM (BRD4). This compound significantly suppresses the growth and migration of liver cancer cells while promoting apoptosis. Additionally, BRD4/NAMPT-IN-1 demonstrates potent anticancer activity in HCCLM3 xenograft mouse models without noticeable toxicity.

BRD4 BD2:41 nM, BRD4 BD1:12 nM, BRD4 BD1/2:58 nM

BRD4/NAMPT-IN-1 exhibits IC 50 values of 12 nM for BRD4(BD1) and 41 nM for BRD4(BD2) against other members of the BET family. It inhibits cancer cell proliferation with IC 50 values of 2.37 μM in Hep3B, 6.49 μM in Huh7, 5.44 μM in HCCLM3, and 9.51 μM in LX-2 cells. Treating Hep3B cells with BRD4/NAMPT-IN-1 (1-10 μM; 72 h) suppresses the expression of BRD4-upregulated oncogenes, reduces NAPRT and NAMPT levels, significantly increases cell arrest in the G0/G1 phase, induces apoptosis in a dose-dependent manner, and inhibits cell migration ability. Additionally, BRD4/NAMPT-IN-1 (1-10 μM; 72 h) reduces NAD+ concentration in Hep3B and HCCLM3 cells in a dose-dependent fashion.

Administered intraperitoneally (i.p.) at 40 mg/kg/day and 80 mg/kg/day for 27 consecutive days, BRD4/NAMPT-IN-1 shows a dose-dependent tumor suppressive effect in HCCLM3 xenograft nude mice.