CWP232228 sodium is a novel and selective Wnt/β-Catenin inhibitor that antagonizes binding of β-catenin to T-cell factor (TCF) in the nucleus. It preferentially inhibits the growth of breast cancer stem-like cells. CWP232228 targets liver cancer stem cells through Wnt/β- a novel therapeutic approach for liver cancer treatment.

Physicochemical Properties

Molecular Formula	C33H34N7NA2O7P
Molecular Weight	717.619069576263
Exact Mass	717.205
CAS #	1144044-02-9
Related CAS #	1144044-02-9 (sodium);1144045-59-9 (free acid);
PubChem CID	71509158
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	9
Heavy Atom Count	50
Complexity	1210
Defined Atom Stereocenter Count	2
SMILES	P(=O)([O-])([O-])OC1C=CC(=CC=1)C[C@H]1C(N(CC2=CC=CC3=CN(C)N=C23)C[C@@H]2N(C(NCC3C=CC=CC=3)=O)N(CC=C)CC(N21)=O)=O.[Na+].[Na+]
InChi Key	KKMKZLQVDAGSOA-GDUXWEAWSA-L
InChi Code	InChI=1S/C33H36N7O7P.2Na/c1-3-16-38-22-30(41)39-28(17-23-12-14-27(15-13-23)47-48(44,45)46)32(42)37(20-26-11-7-10-25-19-36(2)35-31(25)26)21-29(39)40(38)33(43)34-18-24-8-5-4-6-9-24/h3-15,19,28-29H,1,16-18,20-22H2,2H3,(H,34,43)(H2,44,45,46)/q2*+1/p-2/t28-,29-/m0../s1
Chemical Name	Sodium 4-(((6S,9aS)-2-Allyl-1-(benzylcarbamoyl)-8-((2-methyl-2H-indazol-7-yl)methyl)-4,7-dioxooctahydro-2H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenyl phosphate
Synonyms	CWP232228 CWP-232228 CWP 232228 CWP232228 sodium,
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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

ln Vitro	CWP232228 (0.01-100 μM; 48 hours) suppresses cell growth with IC50 values of 2 and 0.8 μM in mouse (4T1) and human (MDA-MB-435) breast cancer cell lines, respectively [1]. CWP232228 (0.01-10 μM; 48 hours) suppresses cell growth in Hep3B, Huh7, and HepG2 cells with IC50 of 2.566, 2.630, and 2.596 μM, respectively [2].
ln Vivo	Tumor volume was significantly reduced by CWP232228 (100 mg/kg, i.p.; daily; 21 days for mice with 4T1 cell tumors; 60 days for mice with MDA-MB-435 cell tumors) [1].
Cell Assay	Cell proliferation assay[1] Cell Types: mouse (4T1) and human (MDA-MB-435) Breast cancer cell lines Tested Concentrations: 0.01, 0.1, 1, 10, 100 μM Incubation Duration: 48 hrs (hours) Experimental Results: IC50 is 2 respectively and 0.8 μM for 4T1 and MDA-MB-435 cell lines respectively. Cell proliferation assay [2] Cell Types: Hepatoma cell lines HepG2, Huh7 and Hep3B Tested Concentrations: 0.01, 0.1, 0.5, 1, 5, 10 μM Incubation Duration: 48 hrs (hours) Experimental Results: IC50 are 2.566, 2.630 and 2.596 μM respectively for Hep3B , Huh7 and HepG2 cells.
Animal Protocol	Animal/Disease Models: 7weeks old female Balb/c and NOD/SCID (severe combined immunodeficient) mouse harboring 4T1 or MDA-MB-435 cell tumors [1] Doses: 100 mg/kg Route of Administration: intraperitoneal (ip) injection; daily; for 4T1 cell-bearing tumors It was 21 days for mice with tumors and 60 days for mice with MDA-MB-435 cell tumors. Experimental Results: Treatment resulted in a significant reduction in tumor volume.
References	[1]. Wnt/β-Catenin Small-Molecule Inhibitor CWP232228 Preferentially Inhibits the Growth of Breast Cancer Stem-like Cells. Cancer Res. 2015 Apr 15;75(8):1691-702. [2]. Oncotarget. 2016 Apr 12;7(15):20395-409. CWP232228 targets liver cancer stem cells through Wnt/β-catenin signaling: a novel therapeutic approach for liver cancer treatment.

Solubility Data

Solubility (In Vitro)	H2O : ~62.5 mg/mL (~87.09 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 50 mg/mL (69.67 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.3935 mL	6.9675 mL	13.9350 mL
	5 mM	0.2787 mL	1.3935 mL	2.7870 mL
	10 mM	0.1393 mL	0.6967 mL	1.3935 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.