Rineterkib (LTT-462; ERK-IN-1; ERK-IN1), the compound B extracted from WO2018051306A1, is a novel and potent RAF and ERK1/2 inhibitor with anticancer activity. It has the potential to treat a proliferative disease marked by mutations that activate the MAPK pathway. The focus of the activity is on the treatment of KRAS-mutant non-small cell lung cancer (NSCLC), BRAF-mutant NSCLC, KRAS-mutant pancreatic cancer, KRAS-mutant colorectal cancer (CRC), and KRAS-mutant ovarian cancer.

Physicochemical Properties

Molecular Formula	C26H27BRF3N5O2
Molecular Weight	578.4241
Exact Mass	577.13
Elemental Analysis	C, 53.99; H, 4.71; Br, 13.81; F, 9.85; N, 12.11; O, 5.53
CAS #	1715025-32-3
Related CAS #	Rineterkib hydrochloride;1715025-34-5
PubChem CID	118045847
Appearance	White to light yellow solid powder
LogP	3.1
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	7
Heavy Atom Count	37
Complexity	757
Defined Atom Stereocenter Count	4
SMILES	CNC[C@H](C1=CC(=CC(=C1)Br)F)NC(=O)C2=C(C=C(C=C2)C3=NC(=CN=C3N)[C@H]4CC[C@@H]([C@H](C4)F)O)F
InChi Key	YFCIFWOJYYFDQP-PTWZRHHISA-N
InChi Code	InChI=1S/C26H27BrF3N5O2/c1-32-11-21(15-6-16(27)10-17(28)7-15)35-26(37)18-4-2-14(9-19(18)29)24-25(31)33-12-22(34-24)13-3-5-23(36)20(30)8-13/h2,4,6-7,9-10,12-13,20-21,23,32,36H,3,5,8,11H2,1H3,(H2,31,33)(H,35,37)/t13-,20-,21+,23-/m0/s1
Chemical Name	4-[3-amino-6-[(1S,3S,4S)-3-fluoro-4-hydroxycyclohexyl]pyrazin-2-yl]-N-[(1S)-1-(3-bromo-5-fluorophenyl)-2-(methylamino)ethyl]-2-fluorobenzamide
Synonyms	ERK-IN 1; ERK IN-1; ERK-IN-1
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 and light.
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	RAF; ERK1; ERK2
ln Vivo	ERK-IN-1 (compound B) (50, 75 mg/kg, p.o., qd/q2d, 27 days), in the Calu-6 human NSCLC subcutaneous tumor xenograft model in mice, treatment significantly reduces the tumor volume.
Animal Protocol	Calu-6 NSCLC xenograft tumor models in mice[1]. 50, 75 mg/kg. Orally either daily (qd) or every other day (q2d) for 27 days.
References	[1]. THERAPEUTIC COMBINATIONS COMPRISING A RAF INHIBITOR AND A ERK INHIBITOR. WO2018051306A1. [2]. Targeting RAS-RAF-MEK-ERK signaling pathway in human cancer: current status in clinical trials. Genes & Diseases, 2022.
Additional Infomation	Rineterkib is an inhibitor of extracellular signal-regulated kinase (ERK) being investigated in NCT04097821 (A Randomized, Open-label, Phase I/II Open Platform Study Evaluating Safety and Efficacy of Novel Ruxolitinib Combinations in Myelofibrosis Patients). Rineterkib is an orally available inhibitor of extracellular signal-regulated kinase (ERK), with potential antineoplastic activity. Upon oral administration, rineterkib binds to and inhibits ERK, thereby preventing the activation of ERK-mediated signal transduction pathways. This results in the inhibition of ERK-dependent tumor cell proliferation and survival. The mitogen-activated protein kinase (MAPK)/ERK pathway is upregulated in numerous tumor cell types and plays a key role in tumor cell proliferation, differentiation and survival.

Solubility Data

Solubility (In Vitro)

DMSO: ~200 mg/mL (~345.8 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 5 mg/mL (8.64 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 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7288 mL	8.6442 mL	17.2885 mL
	5 mM	0.3458 mL	1.7288 mL	3.4577 mL
	10 mM	0.1729 mL	0.8644 mL	1.7288 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.