WSB1 Degrader 1 is a potent orally bioactive WSB1 (WD repeat and SOCS box-containing 1) degrader. WSB1 Degrader 1 has anti-cancer metastasis effects.

Physicochemical Properties

Molecular Formula	C21H22N2O2
Molecular Weight	334.4116
Exact Mass	334.168
CAS #	2306039-66-5
PubChem CID	149706874
Appearance	Off-white to light yellow solid powder
LogP	4.5
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	25
Complexity	419
Defined Atom Stereocenter Count	0
SMILES	O(C([H])([H])[H])C1C([H])=C([H])C(=C([H])C=1C([H])([H])[H])C1C([H])=C([H])C(N([H])[H])=NC=1C1C([H])=C([H])C(=C(C([H])([H])[H])C=1[H])OC([H])([H])[H]
InChi Key	AUHCLHXXUVPUBD-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H22N2O2/c1-13-11-15(5-8-18(13)24-3)17-7-10-20(22)23-21(17)16-6-9-19(25-4)14(2)12-16/h5-12H,1-4H3,(H2,22,23)
Chemical Name	5,6-bis(4-methoxy-3-methylphenyl)pyridin-2-amine
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

ln Vitro	The treatment of H1299-WSB1 cells with WSB1 Degrader 1 (compound 4; 0.25-2500 nM; 2-24 hours) causes WSB1 degradation in a dose- and time-dependent manner[1]. In KHOS and H460 cell lines, WSB1 Degrader 1 (compound 4) demonstrates strong antimigration efficacy with IC50 values of 39.1 μM and 24.47 μM, respectively[1]. In KHOS cells, WSB1 Degrader 1 (compound 4) dramatically reduces the migration of cancer cells in both normoxia and hypoxia. When KHOS cells are exposed to hypoxia, WSB1 Degrader 1 (5 μM) treatment increases the levels of RhoGDI2 protein[1]. Applying WSB1 Degrader 1 to H1299-WSB1 cells significantly reduces their ability to heal wounds. The A2780-WSB1/KO cells are not able to have their wound-healing ability blocked by WSB1 Degrader 1; only the wild-type A2780 (A2780-WT) cells are[1].
ln Vivo	Treatment with WSB1 Degrader 1 (compound 4; 100 mg/kg; po; daily; for 28 days) can successfully prevent cancer cells from metastasizing to the lungs[1]. WSB1 Degrader 1 (compound 4) was administered to rats orally at a dose of 100 mg/kg or intraperitoneally at a dose of 160 mg/kg. Both methods of administration were shown to have rapid absorption (Tmax) but rapid clearance (T1/2) for the oral dose. Additionally, appropriate blood exposure was demonstrated by the Cmax and AUC0-t values of WSB1 Degrader 1 in the oral or intraperitoneal dose groups[1].
Cell Assay	Western Blot Analysis[1] Cell Types: H1299-WSB1 cells Tested Concentrations: 0.25 nM, 2.5 nM, 25 nM, 250 nM, 2500 nM Incubation Duration: 2 hrs (hours), 4 hrs (hours), 6 hrs (hours), 8 hrs (hours), 12 hrs (hours), 24 hrs (hours) Experimental Results: Induced WSB1 degradation in time-dependent and dose-dependent manners.
Animal Protocol	Animal/Disease Models: Balb/c (nu/nu) mice bearing highly metastatic 4T1 breast cancer cells[1]. Doses: 100 mg/kg/day Route of Administration: po; daily ; for 28 days Experimental Results: Effectively inhibited the pulmonary metastasis of cancer cells.
References	[1]. Discovery of 5,6-Bis(4-methoxy-3-methylphenyl)pyridin-2-amine as a WSB1 Degrader to Inhibit Cancer Cell Metastasis. J Med Chem. 2021 Jun 1.

Solubility Data

Solubility (In Vitro)

DMSO: 25 mg/mL (74.76 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (6.22 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 20.8 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.9903 mL	14.9517 mL	29.9034 mL
	5 mM	0.5981 mL	2.9903 mL	5.9807 mL
	10 mM	0.2990 mL	1.4952 mL	2.9903 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.