BRD4/CK2-IN-1 is the first highly effective and oral active dual-target inhibitor of BRD4/CK2 (bromodomain-containing protein 4/casein kinase 2), with IC50s of 180 nM and 230 nM for BRD4 and CK2, respectively. BRD4/CK2-IN-1 has strong anticancer activity without obvious toxicities. BRD4/CK2-IN-1 induces apoptosis and autophagy-associated cell death in triple-negative breast cancer (TNBC)[1]
BRD4/CK2-IN-1 (compound 44e) (0-25 μM; 24 hours) has anti-proliferation effect with IC50s of 2.66 and 3.52 μM in MDA-MB-231 and MDA-MB-468 cells, respectively[1].
BRD4/CK2-IN-1 (0-10 μM; 24 hours) dose-dependently induces apoptosis of MDA-MB-231 and MDA-MB-468 cells[1].
BRD4/CK2-IN-1 (0-10 μM; 24 hours) dose-dependently downregulates Bcl-2 but upregulates Bax and cleaved caspase-3[1].
BRD4/CK2-IN-1 (0-10 μM; 24 hours) significantly downregulates the autophagy substrate p62 and up-regulated beclin-1 and LC3II in MDA-MB-231 and MDA-MB[1].
Cell Viability Assay[1]
| Cell Line: | MDA-MB-231, MDA-MB-468 cells |
| Concentration: | 0, 0.19, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, 25 μM |
| Incubation Time: | 24 hours |
| Result: | Showed anti-proliferative rates in MDA-MB-231 and MDA-MB-468 cells (IC50 = 2.66 and 3.52 μM, respectively). |
Apoptosis Analysis[1]
| Cell Line: | MDA-MB-231, MDA-MB-468 cells |
| Concentration: | 0, 2.5, 5, 10 μM |
| Incubation Time: | 24 hours |
| Result: | Dose-dependently induced apoptosis of MDA-MB-231 and MDA-MB-468 cells. |
Western Blot Analysis[1]
| Cell Line: | MDA-MB-231, MDA-MB-468 cells |
| Concentration: | 0, 2.5, 5, 10 μM |
| Incubation Time: | 24 hours |
| Result: | Dose-dependently downregulated Bcl-2 but upregulated Bax and cleaved caspase-3. |
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| Result: |
BRD4/CK2-IN-1 (25 and 50 mg/kg; intragastric administration; daily for 19 days) inhibits tumor growth in TNBC xenograft models[1].
BRD4/CK2-IN-1 (25 and 50mg/kg; intragastric administration; daily for 19 days) shows weak toxicity measured by body weight loss in the MDA-MB-231 and MDA-MB-468 xenograft models[1].
Preliminary Assessment of Pharmacokinetics (PK) profile of BRD4/CK2-IN-1[1].
| Parameter | iv (1 mg/kg) | po (10 mg/kg) |
| T1/2 (h) | 4.21±0.57 | 5.14±0.71 |
| Cmax (ng/mL) | 237±11 | 206±6 |
| AUC0-t (ng.h/mL) | 579±49 | 2079±130 |
| AUC0-∞ (ng.h/mL) | 588±36 | 2090±146 |
| VZ (L/kg) | 21.1±2.6 | |
| CL ((mL/min)/kg) | 57.4±1.3 | |
| F (%) | 32.5 |
| Animal Model: | Female nude mice (BALB/c, 6-8 weeks, 20-22 g) bearing MDA-MB-231 cells[1] |
| Dosage: | 25 and 50 mg/kg |
| Administration: | Intragastric administration; daily for 19 days |
| Result: | Had the most pronounced tumor growth inhibition (TGI) (63.8%) in the MDA-MB-231 xenograft tumor model at 50 mg/kg. |