NRX-252114 is a potent enhancer of the interaction between β-catenin and the homologous E3 ligase SCFβ-TrCP, enhancing the binding of pSer33/S37A β-catenin and β-TrCP with EC50 of 6.5 nM and a Kd of 0.4 nM. NRX-252114 can promote the degradation of mutant β-catenin.

Physicochemical Properties

Molecular Formula	C22H12CL2F3N3O2S
Molecular Weight	510.31579208374
Exact Mass	508.997
CAS #	2763260-39-3
PubChem CID	162640509
Appearance	White to off-white solid powder
LogP	4.6
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	3
Heavy Atom Count	33
Complexity	935
Defined Atom Stereocenter Count	0
SMILES	C1C2=C(C(C#N)=CC=C2)CN1C(C1=C(SC2=C(Cl)C=CC=C2Cl)C=C(C(F)(F)F)NC1=O)=O
InChi Key	BMQXGPBUQGPLOV-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H12Cl2F3N3O2S/c23-14-5-2-6-15(24)19(14)33-16-7-17(22(25,26)27)29-20(31)18(16)21(32)30-9-12-4-1-3-11(8-28)13(12)10-30/h1-7H,9-10H2,(H,29,31)
Chemical Name	2-[4-(2,6-dichlorophenyl)sulfanyl-2-oxo-6-(trifluoromethyl)-1H-pyridine-3-carbonyl]-1,3-dihydroisoindole-4-carbonitrile
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	EC50: 6.5 nM (β-catenin:β-TrCP Interaction)[1] Kd: 0.4 nM (β-catenin:β-TrCP Interaction)[1]
ln Vitro	The binding of pSer33/S37A β-catenin peptide to β-TrCP is enhanced by about 1500 times by NRX-252114 (20 μM) [1]. Long ubiquitin chains are formed when pSer33/S37A β-catenin peptide is ubiquitinated more heavily by NRX-252114 (0-25 μM) [1]. The ubiquitination of unphosphorylated Ser33/S37A β-catenin peptides and S33E/S37A β-catenin is enhanced by NRX-252114 (0-25 μM) [1]. In transfected HEK293T cells, NRX-252114 (20 μM, 6 hours) generates a strong S33E/S37A β-catenin:β-TrCP association [1].
References	[1]. Prospective discovery of small molecule enhancers of an E3 ligase-substrate interaction. Nat Commun. 2019 Mar 29;10(1):1402.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (195.96 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.90 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 25.0 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: 2.5 mg/mL (4.90 mM) in 10% DMSO + 90% Corn Oil (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 25.0 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	1.9596 mL	9.7978 mL	19.5955 mL
	5 mM	0.3919 mL	1.9596 mL	3.9191 mL
	10 mM	0.1960 mL	0.9798 mL	1.9596 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.