CCG-222740 (CCG222740) is a novel and potent Rho/myocardin-related transcription factor (MRTF)/SRF inhibitor. CCG-222740 is also a potent inhibitor of the expression of the alpha-smooth muscle actin protein.

Physicochemical Properties

Molecular Formula	C23H19CLF2N2O3
Molecular Weight	444.858371973038
Exact Mass	444.105
Elemental Analysis	C, 62.10; H, 4.31; Cl, 7.97; F, 8.54; N, 6.30; O, 10.79
CAS #	1922098-69-8
Related CAS #	1922098-69-8
PubChem CID	121317937
Appearance	Light yellow to yellow solid powder
LogP	4.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	31
Complexity	657
Defined Atom Stereocenter Count	0
InChi Key	PMTPYUTZAJWGPE-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H19ClF2N2O3/c24-18-6-8-19(9-7-18)27-21(29)17-12-23(25,26)14-28(13-17)22(30)16-4-1-3-15(11-16)20-5-2-10-31-20/h1-11,17H,12-14H2,(H,27,29)
Chemical Name	N-(4-chlorophenyl)-5,5-difluoro-1-[3-(furan-2-yl)benzoyl]piperidine-3-carboxamide
Synonyms	CCG222740; CCG 222740; CCG-222740
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	Rho/MRTF pathway
ln Vitro	CCG-222740 causes a decrease in cyclin D1 and an increase in the protein p27. Using the MTT assay, CCG-222740 treatment reduced CAFs' cell viability with an IC50 of about 10 μM. One of the primary sources of matrix proteins, including multiple isoforms of collagen, in pancreatic cancer is CAFs. In CAFsIn cancer cells, the Rho/MRTF pathway inhibitor CCG-222740 reduces the amounts of collagens I, 2a, and IV as well as α-SMA. Treatment with CCG-222740 causes MRTF to be excluded from the nucleus.[1]
ln Vivo	CCG-222740 dramatically lowers the levels of α-SMA in the pancreas of KC mice stimulated with caerulein.[1]
Cell Assay	The culture medium used for murine cancer-associated fibroblasts (CAFs) contains 10% fetal bovine serum (FBS). After treating CAFs with various concentrations of CCG-222740 (740) for 72 hours, the levels of p27 and Cyclin D1 are assessed using western blotting.
Animal Protocol	Nine-week old KC mice 100 mg/kg Oral gavage
References	[1]. The Rho/MRTF pathway inhibitor CCG-222740 reduces stellate cell activation and modulates immune cell populations in KrasG12D; Pdx1-Cre (KC) mice. Sci Rep. 2019 May 8;9(1):7072. [2]. Local delivery of novel MRTF/SRF inhibitors prevents scar tissue formation in a preclinical model of fibrosis. Sci Rep. 2017 Mar 31;7(1):518.

Solubility Data

Solubility (In Vitro)

DMSO: ~125 mg/mL (~281 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.25 mg/mL (5.06 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 22.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.25 mg/mL (5.06 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 22.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.25 mg/mL (5.06 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 22.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2479 mL	11.2395 mL	22.4790 mL
	5 mM	0.4496 mL	2.2479 mL	4.4958 mL
	10 mM	0.2248 mL	1.1239 mL	2.2479 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.