GKI-1 is a Greatwall (GWL) kinase inhibitor (antagonist) with IC50s of 4.9 and 2.5 µM for hGWLFL and hGWL-KinDom, respectively. GKI-1 can effectively inhibit ROCK1 with IC50 of 11 µM, but only weakly inhibits PKA.

Physicochemical Properties

Molecular Formula	C15H12CLN3
Molecular Weight	269.73
Exact Mass	269.071
CAS #	2444764-03-6
PubChem CID	137333439
Appearance	White to off-white solid powder
Density	1.320±0.06 g/cm3(Predicted)
Boiling Point	504.5±45.0 °C(Predicted)
LogP	4
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	3
Heavy Atom Count	19
Complexity	279
Defined Atom Stereocenter Count	0
InChi Key	GPXJDHFWTOJHQQ-UHFFFAOYSA-N
InChi Code	InChI=1S/C15H12ClN3/c16-13-4-6-14(7-5-13)19-15-3-1-2-11(8-15)12-9-17-18-10-12/h1-10,19H,(H,17,18)
Chemical Name	N-(4-chlorophenyl)-3-(1H-pyrazol-4-yl)aniline
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 (e.g. under nitrogen), 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

Targets	IC50: 4.9 µM (hGWLFL), 4.9 µM (hGWL-KinDom)[1]
ln Vitro	GKI-1 exhibits cellular effectiveness and inhibits human GWL in its entirety[1]. GKI-1 treatment of HeLa cells at 25 and 50 µM for 0-8.5 hours lowers the levels of ENSA/ARPP19 phosphorylation, which in turn lowers mitotic events, mitotic arrest/cell death, and cytokinesis failure[1].
References	[1]. A first generation inhibitor of human Greatwall kinase, enabled by structural and functional characterisation of a minimal kinase domain construct. Oncotarget. 2016 Nov 1;7(44):71182-71197.

Solubility Data

Solubility (In Vitro)

DMSO: 200 mg/mL (741.48 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 5 mg/mL (18.54 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 50.0 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: ≥ 5 mg/mL (18.54 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 50.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: ≥ 2.17 mg/mL (8.05 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 21.7 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.7074 mL	18.5371 mL	37.0741 mL
	5 mM	0.7415 mL	3.7074 mL	7.4148 mL
	10 mM	0.3707 mL	1.8537 mL	3.7074 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.