JNJ-7706621

Biological Activity

JNJ-7706621 is a novel cell cycle inhibitor that showed potent inhibition of several cyclin-dependent kinases (CDK) and Aurora kinases and selectively blocked proliferation of tumor cells of various origins but was about 10-fold less effective at inhibiting normal human cell growth in vitro. In human cancer cells, treatment with JNJ-7706621 inhibited cell growth independent of p53, retinoblastoma, or P-glycoprotein status; activated apoptosis; and reduced colony formation. At low concentrations, JNJ-7706621 slowed the growth of cells and at higher concentrations induced cytotoxicity. Inhibition of CDK1 kinase activity, altered CDK1 phosphorylation status, and interference with downstream substrates such as retinoblastoma were also shown in human tumor cells following compound treatment. Flow cytometric analysis of DNA content showed that JNJ-7706621 delayed progression through G1 and arrested the cell cycle at the G2-M phase. Additional cellular effects due to inhibition of Aurora kinases included endoreduplication and inhibition of histone H3 phosphorylation. JNJ-7706621 is a unique inhibitor regulating cell cycle progression at multiple points, suggesting that it could be useful for cell cycle analysis and therapy of various cancers, including Ewing's sarcoma.

Customer Product Validations & Biological Datas

	Source	Oncotarget (2015). Figure 4. JNJ-7706621
	Method	MTT assay
	Cell Lines	Ovarian cancer cells
	Concentrations	0.5 or 2μM
	Incubation Time	4 days
	Results	This reveals a less than 4-fold difference in the sensitivity to JNJ-7706621 between cell lines with and without CCNE1 overexpression

Protocol (for reference only)

Cell Experiment
Cell lines	HeLa cells
Preparation method	Colony formation assay. HeLa cells were plated at various densities in 90-mm diameter Petri dishes and exposed to JNJ-7706621 for 48 hours. Cells were then washed in PBS and incubated an additional 7 days in drug-free medium. Cells were fixed in 95% ethanol, stained with 0.5% crystal violet, and colonies containing >50 cells manually counted.
Concentrations	0~3μM
Incubation time	48 h

Animal Experiment
Animal models	A375 melanoma human tumor xenograft model in female nu/nu mice
Formulation	normal saline
Dosages	100 and 125 mg/kg daily
Administration	i.p.

Chemical Information

Molecular Weight	394.36
Formula	C15H12F2N6O3S
CAS Number	443797-96-4
Solubility (25°C)	DMSO 69 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Emanuel S, et al. Cancer Res. The in vitro and in vivo effects of JNJ-7706621: a dual inhibitor of cyclin-dependent kinases and aurora kinases.