Ravoxertinib hydrochloride (GDC-0994; RG-7842), the hydrochloride salt of Ravoxertinib, is an orally bioavailable inhibitor of extracellular signal-regulated kinase (ERK1/2) being developed as an anticancer drug. It inhibits ERK1/2 with IC50 values of 1.1 nM and 0.3 nM, respectively. Following oral administration, GDC-0994 blocks ERK phosphorylation as well as the activation of ERK-mediated signal transduction pathways. This stops ERK-dependent tumor cell survival and proliferation. The mitogen-activated protein kinase (MAPK)/ERK pathway is activated in a variety of tumor cell types and is essential for tumor cell survival, proliferation, and differentiation.

Physicochemical Properties

Molecular Formula	C21H19CL2FN6O2
Molecular Weight	477.3190
Exact Mass	476.093
CAS #	2070009-58-2
Related CAS #	Ravoxertinib;1453848-26-4
PubChem CID	92044395
Appearance	Light yellow to yellow solid powder
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	32
Complexity	709
Defined Atom Stereocenter Count	1
SMILES	ClC1C([H])=C([H])C(=C([H])C=1F)[C@@]([H])(C([H])([H])O[H])N1C([H])=C([H])C(C2C([H])=C([H])N=C(N([H])C3=C([H])C([H])=NN3C([H])([H])[H])N=2)=C([H])C1=O.Cl[H]
InChi Key	RMNVBUVHPAETTJ-GMUIIQOCSA-N
InChi Code	InChI=1S/C21H18ClFN6O2.ClH/c1-28-19(5-8-25-28)27-21-24-7-4-17(26-21)13-6-9-29(20(31)11-13)18(12-30)14-2-3-15(22)16(23)10-14;/h2-11,18,30H,12H2,1H3,(H,24,26,27);1H/t18-;/m1./s1
Chemical Name	1-[(1S)-1-(4-chloro-3-fluorophenyl)-2-hydroxyethyl]-4-[2-[(2-methylpyrazol-3-yl)amino]pyrimidin-4-yl]pyridin-2-one;hydrochloride
Synonyms	RG 7842 HCl; GDC0994; GDC-0994; GDC 0994; RG7842; RG-7842; RG 7842
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, 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	ERK2 (IC50 = 3.1 nM); ERK1 (IC50 = 6.1 nM); p-RSK (IC50 = 12 nM)
ln Vitro	Ravoxertinib also inhibits p90RSK with IC50 of 12 nM[1]. Ravoxertinib has a biochemical potency of 1.1 nM and 0.3 nM, respectively, and is highly selective for ERK1 and ERK2[2]. Ravoxertinib (GDC0994; 50 nM, 0.5 µM, and 5 µM; 48 hours) reduces the viability of lung adenocarcinoma cell lines (A549, HCC827, and HCC4006)[4].
ln Vivo	To achieve the desired target coverage for at least 8 hours in CD-1 mice, a 10 mg/kg oral dose of Ravoxertinib is sufficient[1]. In numerous in vivo cancer models, such as KRAS- and BRAF-mutant human xenograft tumors in mice, daily oral dosing of ravoxertinib produces significant single-agent activity[2].
Enzyme Assay	Ravoxertinib (GDC-0994) is an orally bioavailable ERK kinase inhibitor with an IC50 of 6.1 nM and 3.1 nM for ERK1 and ERK2, respectively. With an IC50 of 12 nM, ravoxertinib (GDC-0994) also inhibits p90RSK. Ravoxertinib (GDC-0994) has a biochemical potency of 1.1 nM and 0.3 nM for ERK1 and ERK2, respectively.
Cell Assay	GDC-0994 potently inhibits phospho-p90RSK in tumor cells.
Animal Protocol	Mice: Ravoxertinib PK/PD data in the HCT116 mouse xenograft model. In nude mice, HCT116 tumors grow to a tumor volume of 400–600 mm3. When compared to the vehicle control alone (40% PEG400/60% (10% HPβCD)), mice are given a single oral dose of 22 at 15, 30, or 100 mg/kg. Tumor and plasma samples are then collected at 2, 8, 16, and 24 hours after the dose. By using a quantitative Western blot, tumor levels of phosphorylated p90RSK (pRSK) and relative total p90RSK (tRSK) are determined. At 2 hours after the dose, these levels are normalized to the vehicle control (set to 100%). By using LC-MS, concentrations in plasma and tumors are measured.
References	[1]. Discovery of (S)-1-(1-(4-Chloro-3-fluorophenyl)-2-hydroxyethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one (GDC-0994), an Extracellular Signal-Regulated Kinase 1/2 (ERK1/2) Inhibitor in Early Clinical Developme [2]. Abstract DDT02-03: Discovery of GDC-0994, a potent and selective ERK1/2 inhibitor in early clinical development. Proceedings: AACR Annual Meeting 2014; April 5-9, 2014. [3]. Opportunity for Pharmaceutical Intervention in Lung Cancer: Selective Inhibition of JAK1/2 to Eliminate EMT-Derived Mesenchymal Cells.

Solubility Data

Solubility (In Vitro)

DMSO: > 100 mg/mL Water: N/A Ethanol: N/A

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.24 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 (5.24 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 25.0 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 3: 2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 30mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.0950 mL	10.4752 mL	20.9503 mL
	5 mM	0.4190 mL	2.0950 mL	4.1901 mL
	10 mM	0.2095 mL	1.0475 mL	2.0950 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.