ChemicalBook Product Catalog Biochemical Engineering Inhibitors Mitogen-activated protein kinase (MAPK) Raf inhibitors CEP-32496 (free base)

CEP-32496 (free base)

Product NameCEP-32496 (free base)
CAS1188910-76-0
MFC24H22F3N5O5
MW517.46
EINECS
MOL File1188910-76-0.mol

Chemical Properties

Boiling point	544.839±50.00 °C(Press: 760.00 Torr)(predicted)
Density	1.406±0.06 g/cm3(Temp: 25 °C; Press: 760 Torr)(predicted)
storage temp.	Store at -20°C
solubility	≥25.85 mg/mL in DMSO; insoluble in H2O; insoluble in EtOH
form	A crystalline solid
pka	12.337±0.70(predicted)
color	White to off-white
CAS DataBase Reference	1188910-76-0

Usage And Synthesis

Description

B-Raf is a MAP kinase kinase kinase, which functions downstream of Ras family GTPases to activate MEK1/2 and ERK1/2 signaling. Mutations of B-Raf, particularly at Val⁶⁰⁰, are common in melanomas and melanocytic nevi. CEP-32496 is a potent inhibitor of B-Raf^V600E (K_d = 14 nM in an in vitro binding assay). It blocks B-Raf^V600E-dependent phosphorylation of MEK in human melanoma A375 and colorectal cancer COLO 205 cells (IC₅₀s = 78 and 60 nM, respectively). CEP-32496 binds kinases other than B-Raf but displays selective cytotoxicity for cells expressing B-Raf^V600E. It displays good oral bioavailability in rats, dogs, and monkeys and has single oral dose pharmacodynamics inhibition of both pMEK and pERK in B-Raf^V600E colon carcinoma xenografts in nude mice.

Uses

CEP 32496 is an orally active BRAFV600E inhibitor with selective cellular and in vivo antitumor activity.

Synthesis

3-[(6,7-dimethoxy-4-quinazolinyl)oxy]aniline

1188908-37-3

[5-(2,2,2-TRIFLUORO-1,1-DIMETHYL-ETHYL)-ISOXAZOL-3-YL]-CARBAMIC ACID PHENYL ESTER

1188911-77-4

1188910-76-0

A 10L Chemglass jacketed reactor, equipped with an N2 inlet/outlet, was charged with 3-[(6,7-dimethoxy-4-quinazolinyl)oxy]aniline (Compound 2, 200.0 g, 637 mmol) and phenyl (5-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazol-3-yl)carbamate (Compound 3, 177.0 g, 596 mmol). equipped with N2 inlet/outlet, 4-dimethylaminopyridine (DMAP, 2.88 g) and 4.0 L of isopropylacetic acid were added. The internal temperature of the reaction mixture was raised to 70 °C and maintained at this temperature for 9 hours. The slurry was maintained during the reaction and HPLC monitoring showed that compound 2 was completely consumed after a period of heating. Subsequently, 2.0 L of heptane was added at 70 °C and the reaction mixture was cooled to 20 °C. Stirring was continued for 1 h to precipitate the solid, filtered, and the filter cake was washed with 2.0 L of a 1:1 (v/v) isopropyl acetate/heptane solvent mixture. The resulting white solid was dried under vacuum at 55 °C and 75 mbar by N2 exhaust to give 295 g (96% yield) of form A0 product with 99.3% HPLC purity.

in vivo

Oral administration of Agerafenib (CEP-32496) to Colo-205 tumor xenograft-bearing mice results in significant inhibition of pMEK in tumor cell lysates. For instance, a single 30 mg/kg (po) dose of Agerafenib leads to a 50 and 75% inhibition of normalized pMEK in tumor lysates at the 2 and 6 h postdose time point, respectively (p<0.03), while a 55 mg/kg (po) dose resulted in a 75% to 57% (p<0.03) inhibition of pMEK at 2 through 10 h post administration, with normalization to baseline by 24 h. Agerafenib exhibits an exceptional PK profile in mouse, dog, and cynomolgus monkey. Administration of Agerafenib to beagle dogs (single dose of 1 mg/kg iv and 10 mg/kg po) results in low clearance (CL=5.0 (mL/min)/kg) and excellent bioavailability (%F=100). Similarly, in cynomolgus monkey, the administration of Agerafenib (single dose of 1 mg/kg iv and 10 mg/kg po) leads to high oral exposure due to low clearance (CL=6.7 mL/min/kg) and excellent bioavailability (%F=100)^[1].

IC 50

BRaf^V600E: 14 nM (Kd); Braf: 36 nM (Kd); CRAF: 39 nM (Kd); c-Kit: 2 nM (Kd); Ret: 2 nM (Kd); LCK: 2 nM (Kd); Abl-1: 3 nM (Kd); VEGFR-2: 8 nM (Kd); CSF-1R: 9 nM (Kd); EPHA2: 14 nM (Kd); EGFR: 22 nM (Kd); c-Met: 513 nM (Kd); JAK-2: 4700 nM (Kd); MEK-1: 7100 nM (Kd); MEK-2: 8300 nM (Kd)

References

[1] rowbottom mw1, faraoni r, chao q, campbell bt, lai ag, setti e, ezawa m, sprankle kg, abraham s, tran l, struss b, gibney m, armstrong rc,gunawardane rn, nepomuceno rr, valenta i, hua h, gardner mf, cramer md, gitnick d, insko de, apuy jl, jones-bolin s, ghose ak, herbertz t, ator ma,dorsey bd, ruggeri b, williams m, bhagwat s, james j, holladay mw. identification of 1-(3-(6,7-dimethoxyquinazolin-4-yloxy)phenyl)-3-(5-(1,1,1-trifluoro-2 -methylpropan- 2-yl) isoxazol-3-yl) urea hydrochloride (cep-32496), a highly potent and orally efficacious inhibitor of v-raf murine sarcoma viral oncogene homologue b1 (braf) v600e. j med chem. 2012 feb 9;55(3):1082-105. doi: 10.1021/jm2009925. epub 2012 jan 23.

CEP-32496 (free base)

Chemical Properties

Usage And Synthesis

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