GSK-25 is a potent, selective, orally bioactive ROCK1 inhibitor (IC50=7 nM). GSK-25 maintains good selectivity for 31 kinases as well as RSK1 and p70S6K (RSK1: IC50=398 nM, p70S6K: IC50=1 μM). GSK-25 can inhibit P450 (IC50 of CYP2C9, CYP2D6, and CYP3A4 are 2.5, 5.2, and 2.5 μM, respectively).

Physicochemical Properties

Molecular Formula	C24H16N6OF2CL2
Molecular Weight	513.32624
Exact Mass	512.073
CAS #	874119-56-9
PubChem CID	11548023
Appearance	Light yellow to yellow solid powder
Density	1.6±0.1 g/cm3
Boiling Point	751.7±60.0 °C at 760 mmHg
Flash Point	408.4±32.9 °C
Vapour Pressure	0.0±2.5 mmHg at 25°C
Index of Refraction	1.720
LogP	4.18
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	4
Heavy Atom Count	35
Complexity	875
Defined Atom Stereocenter Count	0
InChi Key	KZYAHEIPPAPADQ-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H16Cl2F2N6O/c1-11-21(24(35)32-19-6-13-10-30-34-18(13)9-17(19)28)22(15-3-2-14(25)8-16(15)27)33-23(31-11)12-4-5-29-20(26)7-12/h2-10,22H,1H3,(H,30,34)(H,31,33)(H,32,35)
Chemical Name	4-(4-chloro-2-fluorophenyl)-2-(2-chloropyridin-4-yl)-N-(6-fluoro-1H-indazol-5-yl)-6-methyl-1,4-dihydropyrimidine-5-carboxamide
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 Vivo	GSK-25 was examined in a model of spontaneous hypertension (arterial aorta IC50=256 nM). GSK-25 has a 25 mmHg burst at 30 mg/kg (atm). It also has a good half-life (1.8 hours for a human, 2.2 hours for a monkey), with good pyramidal bioavailability (49% in the Sprague-Dawley cohort and 19% in monkeys)[1].
References	[1]. Potent, selective and orally bioavailable dihydropyrimidine inhibitors of Rho kinase (ROCK1) as potential therapeutic agents for cardiovascular diseases. J Med Chem. 2008 Nov 13;51(21):6631-4.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~97.40 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.75 mg/mL (5.36 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 27.5 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.87 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9481 mL	9.7403 mL	19.4806 mL
	5 mM	0.3896 mL	1.9481 mL	3.8961 mL
	10 mM	0.1948 mL	0.9740 mL	1.9481 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.