GW406108X is a specific Kif15 (Kinesin-12) inhibitor (antagonist) with IC50 of 0.82 uM. GW406108X is a potent autophagy inhibitor and ATP competitive inhibitor of ULK1 with pIC50 of 6.37 (427 nM). GW406108X inhibits ULK1 and autophagy without affecting mTOR or AMPK activity.

Physicochemical Properties

Molecular Formula	C20H11CL2NO4
Molecular Weight	400.211643457413
Exact Mass	399.006
CAS #	1644443-92-4
PubChem CID	44312236
Appearance	Light yellow to yellow solid powder
LogP	4.5
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	27
Complexity	628
Defined Atom Stereocenter Count	0
SMILES	ClC1C(=C(C=C(C=1)/C=C1\C(NC2C=CC(C(C3=CC=CO3)=O)=CC=2\1)=O)Cl)O
InChi Key	XKTUKRBLWOHYIL-MLPAPPSSSA-N
InChi Code	InChI=1S/C20H11Cl2NO4/c21-14-7-10(8-15(22)19(14)25)6-13-12-9-11(3-4-16(12)23-20(13)26)18(24)17-2-1-5-27-17/h1-9,25H,(H,23,26)/b13-6-
Chemical Name	(3Z)-3-[(3,5-dichloro-4-hydroxyphenyl)methylidene]-5-(furan-2-carbonyl)-1H-indol-2-one
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

Monitoring ULK1 pS758 (mTOR site) or pS556 (AMPK site) levels demonstrates that GW406108X operates on ULK1 and autophagy without changing mTOR or AMPK activity. With pIC50 values of 6.34 (457 nM) and 6.38 (417 nM), respectively, GW406108X inhibits both AMPK and VPS34. The presence of 5 μM GW406108X greatly reduces the increase in ATG13 phosphorylation that is caused by hunger [1]. Kif15-N420 ATPase activity is 76% inhibited by GW406108X[2].

References

[1]. The identification and characterisation of autophagy inhibitors from the published kinase inhibitor sets. Biochem J. 2020;477(4):801-814. [2]. Dual inhibition of Kif15 by oxindole and quinazolinedione chemical probes. Bioorg Med Chem Lett. 2019;29(2):148-154.

Solubility Data

Solubility (In Vitro)

DMSO : 50 mg/mL (124.93 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.08 mg/mL (5.20 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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	2.4987 mL	12.4934 mL	24.9869 mL
	5 mM	0.4997 mL	2.4987 mL	4.9974 mL
	10 mM	0.2499 mL	1.2493 mL	2.4987 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.