CCG-1423

Biological Activity

CCG-1423 is a small-molecule inhibitor of RhoA transcriptional signaling. CCG-1423 potently (<1 mumol/L) inhibits lysophosphatidic acid-induced DNA synthesis in PC-3 prostate cancer cells, and whereas it inhibits the growth of RhoC-overexpressing melanoma lines (A375M2 and SK-Mel-147) at nanomolar concentrations, it is less active on related lines (A375 and SK-Mel-28) that express lower levels of Rho.

In H9c2 cells, CCG-1423 inhibits MRTF nuclear localization, and completely blocks STARS proximal reporter activity. In vivo, pharmacological SRF inhibition by CCG-1423 reduced nuclear MKL1 and improved glucose uptake and tolerance in insulin-resistant mice.

Protocol

Cell Experiment
Cell lines	Overexpress RhoC melanoma lines (A375M2 and SK-Mel-147), low RhoC-expressing lines (A375 and SK-Mel-28), transformed (SW962, PC-3 and SKOV-3) and nontransformed (WI-38) cell lines
Preparation method	Cells in normal culture medium are plated (2,000 per well) in a 96-well plate coated with laminin. After attachment, the medium is replaced with serum-free medium (0% FBS) with 30 μmol/L LPA with or without 300 nM CCG-1423. Fresh LPA with or without CCG-1423 is added at day 5 to ensure that LPA and compound are present throughout the experiment. On day 8, WST-1 reagent is added to the wells for 1 h and absorbance at 450 nm is read using a Victor plate reader.
Concentrations	300 nM
Incubation time	72 hours

Animal Experiment
Animal models
Formulation
Dosages
Administration

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

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.

Chemical Information

References

Novel Rho/MRTF/SRF inhibitors block matrix-stiffness and TGF-β-induced fibrogenesis in human colonic myofibroblasts.
Johnson LA, et al. Inflamm Bowel Dis. 2014 Jan;20(1):154-65. PMID: 24280883.

STARS is essential to maintain cardiac development and function in vivo via a SRF pathway.
Chong NW, et al. PLoS One. 2012;7(7):e40966. PMID: 22815879.

Combination of small molecules enhances differentiation of mouse embryonic stem cells into intermediate mesoderm through BMP7-positive cells.
Mae S, et al. Biochem Biophys Res Commun. 2010 Mar 19;393(4):877-82. PMID: 20171952.

CCG-1423: a small-molecule inhibitor of RhoA transcriptional signaling.
Evelyn CR, et al. Mol Cancer Ther. 2007 Aug;6(8):2249-60. PMID: 17699722.