Home MAPK Raf inhibitor - CSF-1R inhibitor - c-Kit inhibitor - PDGFR inhibitor - c-RET inhibitor Agerafenib (CEP-32496)

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Agerafenib (CEP-32496) Raf inhibitor

Cat.No.S8015

Agerafenib (CEP-32496) is a highly potent inhibitor of BRAF(V600E/WT) and c-Raf with Kd of 14 nM/36 nM and 39 nM, respectively. It also shows potent activity against Abl-1, c-Kit, Ret (c-Ret), PDGFRβ and VEGFR2, but has insignificant affinity for MEK-1, MEK-2, ERK-1 and ERK-2. This compound is currently in Phase 1/2 clinical trials.
Agerafenib (CEP-32496) Raf inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 517.46

Jump to Mechanism of Action Cell Culture & Working Concentration Solubility Chemical Information, Storage & Stability Specificity

Quality Control

Batch: S801501 DMSO]9 mg/mL]false]Water]Insoluble]false]Ethanol]Insoluble]false Purity: 99.19%
99.19

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HEK293 Function assay 1 hr Inhibition of LCK in human HEK293 cells after 1 hr by competition binding assay, Kd=0.002μM. 22168626
HEK293 Function assay 1 hr Inhibition of PDGFRbeta in human HEK293 cells after 1 hr by competition binding assay, Kd=0.002μM. 22168626
HEK293 Function assay 1 hr Inhibition of cKit in human HEK293 cells after 1 hr by competition binding assay, Kd=0.002μM. 22168626
HEK293 Function assay 1 hr Inhibition of Ret in human HEK293 cells after 1 hr by competition binding assay, Kd=0.002μM. 22168626
HEK293 Function assay 1 hr Inhibition of Abl1 in human HEK293 cells after 1 hr by competition binding assay, Kd=0.003μM. 22168626
HEK293 Function assay 1 hr Inhibition of VEGFR2 in human HEK293 cells after 1 hr by competition binding assay, Kd=0.008μM. 22168626
HEK293 Function assay 1 hr Inhibition of CSF1R in human HEK293 cells after 1 hr by competition binding assay, Kd=0.009μM. 22168626
HEK293 Function assay 1 hr Inhibition of EPHA2 in human HEK293 cells after 1 hr by competition binding assay, Kd=0.014μM. 22168626
HEK293 Function assay 1 hr Inhibition of BRAF V600E mutant in human HEK293 cells after 1 hr by competition binding assay, Kd=0.014μM. 22168626
HEK293 Function assay 1 hr Inhibition of EGFR in human HEK293 cells after 1 hr by competition binding assay, Kd=0.022μM. 22168626
HEK293 Function assay 1 hr Inhibition of wild type BRAF in human HEK293 cells after 1 hr by competition binding assay, Kd=0.036μM. 22168626
COLO205 Cytotoxicity assay 72 hrs Cytotoxicity against human COLO205 cells expressing BRAF V600E mutant after 72 hrs by cell titer blue assay, EC50=0.036μM. 22168626
HEK293 Function assay 1 hr Inhibition of CRAF in human HEK293 cells after 1 hr by competition binding assay, Kd=0.039μM. 22168626
A375 Cytotoxicity assay 72 hrs Cytotoxicity against human A375 cells expressing BRAF V600E mutant after 72 hrs by cell titer blue assay, IC50=0.078μM. 22168626
A375 Function assay 2 hrs Inhibition of BRAF V600E mutant-mediated MEK phosphorylation in human A375 cells after 2 hrs, IC50=0.082μM. 22168626
HCC827 Function assay Displacement of [3H]-cyclopamine from SMO V404M mutant in gefitinib resistant human HCC827 cells by scintillation counting, Ki=0.1878μM. 28787156
COLO679 Cytotoxicity assay 72 hrs Cytotoxicity against human COLO679 cells expressing BRAF V600E mutant after 72 hrs by cell titer blue assay, EC50=0.211μM. 22168626
HT144 Cytotoxicity assay 72 hrs Cytotoxicity against human HT144 cells expressing BRAF V600E mutant after 72 hrs by cell titer blue assay, EC50=0.228μM. 22168626
SK-MEL-28 Cytotoxicity assay 72 hrs Cytotoxicity against human SK-MEL-28 cells expressing BRAF V600E mutant after 72 hrs by cell titer blue assay, EC50=0.454μM. 22168626
HEK293 Function assay 1 hr Inhibition of cMET in human HEK293 cells after 1 hr by competition binding assay, Kd=0.513μM. 22168626
HCT116 Cytotoxicity assay 72 hrs Cytotoxicity against human HCT116 cells expressing wild type BRAF after 72 hrs by cell titer blue assay, EC50=0.669μM. 22168626
Hs578T Cytotoxicity assay 72 hrs Cytotoxicity against human Hs578T cells expressing wild type BRAF after 72 hrs by cell titer blue assay, EC50=2.736μM. 22168626
DU145 Cytotoxicity assay 72 hrs Cytotoxicity against human DU145 cells expressing wild type BRAF after 72 hrs by cell titer blue assay, EC50=2.911μM. 22168626
HEK293 Function assay 1 hr Inhibition of JAK2 in human HEK293 cells after 1 hr by competition binding assay, Kd=4.7μM. 22168626
PC3 Cytotoxicity assay 72 hrs Cytotoxicity against human PC3 cells expressing wild type BRAF after 72 hrs by cell titer blue assay, EC50=6.257μM. 22168626
LNCAP Cytotoxicity assay 72 hrs Cytotoxicity against human LNCAP cells expressing wild type BRAF after 72 hrs by cell titer blue assay, EC50=6.631μM. 22168626
HEK293 Function assay 1 hr Inhibition of MEK1 in human HEK293 cells after 1 hr by competition binding assay, Kd=7.1μM. 22168626
HEK293 Function assay 1 hr Inhibition of MEK2 in human HEK293 cells after 1 hr by competition binding assay, Kd=8.3μM. 22168626
COLO205 Antitumor assay 10 mg/kg 14 days Antitumor activity against human COLO205 cells xenografted in athymic nude mouse at 10 mg/kg, po bid for 14 days 22168626
COLO205 Antitumor assay 30 mg/kg 14 days Antitumor activity against human COLO205 cells xenografted in athymic nude mouse at 30 mg/kg, po bid for 14 days 22168626
COLO205 Antitumor assay 100 mg/kg 14 days Antitumor activity against human COLO205 cells xenografted in athymic nude mouse at 100 mg/kg, po bid for 14 days 22168626
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight 517.46 Formula

C24H22F3N5O5

 Storage (From the date of receipt)
CAS No. 1188910-76-0 Download SDF Storage of Stock Solutions

Synonyms RXDX-105 Smiles CC(C)(C1=CC(=NO1)NC(=O)NC2=CC(=CC=C2)OC3=NC=NC4=CC(=C(C=C43)OC)OC)C(F)(F)F

Solubility

In vitro
Batch:

DMSO : 9 mg/mL ( (17.39 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo
Batch:

In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Mechanism of Action

Features
High binding affinity for both BRAF (V600E) mutation and related c-Raf, but no significant affinity for other kinases of MAPK pathway.
Targets/IC50/Ki
c-Kit [1]
2 nM(Kd)
LCK [1]
2 nM(Kd)
PDGFRβ [1]
2 nM(Kd)
RET [1]
2 nM(Kd)
Abl1 [1]
3 nM(Kd)
VEGFR2 [1]
8 nM(Kd)
CSF-1R [1]
9 nM(Kd)
B-Raf (V600E) [1]
14 nM(Kd)
EphA2 [1]
14 nM(Kd)
EGFR [1]
22 nM(Kd)
B-Raf [1]
36 nM(Kd)
C-Raf [1]
39 nM(Kd)
c-Met [1]
513 nM(Kd)
In vitro
Agerafenib (CEP-32496) inhibits A375 cell (BRAFV600E) proliferation with an EC50 of 78 nM. It exhibits more sensitive cytotoxicity for tumor cell lines (A375, SK-MEL-28, Colo-205, Colo-679, and HT-144) expressing mutant BRAF than those expressing wild-type BRAF (HCT116, Hs578T, LNCaP, DU145, and PC-3). [1] This compound inhibits mitogen-activated protein (MAP)/extracellular signal-regulated (ER) kinase (MEK) phosphorylation (pMEK) in human melanoma (A375) and colorectal cancer (Colo-205) cell lines with IC50 values of 78 nM and 60 nM, respectively. [2]
Kinase Assay
Binding assay
Kinases are produced displayed on T7 phage or by expression in HEK-293 cells and tagged with DNA. Binding reactions are performed at room temperature for 1 hour, and the fraction of kinase not bound to test compound is determined by capture with an immobilized affinity ligand and quantitation by quantitative PCR. Each kinase is tested individually against Agerafenib (CEP-32496). Kd values for this compound are determined using eleven serial 3-fold dilutions and presented as mean values from experiments performed in duplicate. Variability between individual values is less than 2-fold.
In vivo
Agerafenib (CEP-32496) exhibits good stability in mouse, dog, monkey, and human liver microsomal preparations with measured intrinsic clearance values of <23 (μL/min)/mg and t1/2 > 60 min in all assays. This compound (30 mg/kg, orally, BID) exhibits tumor stasis and a 40% incidence of partial tumor regressions (PRs) in Colo-205 xenograft mouse model, whereas the 100 mg/kg dose group exhibits both tumor stasis and an 80% incidence of PRs. It (30 mg/kg, orally, BID) leads to a 50% and 75% inhibition of normalized pMEK in tumor lysates at the 2 hours and 6 hours postdose time point, respectively, while a 55 mg/kg dose results in a 75% to 57% inhibition of pMEK at 2 hours through 10 hours post administration in Colo-205 xenograft mouse model. [1] It is orally bioavailable in multiple preclinical species (>95% in rats, dogs, and monkeys). This compound (100 mg/kg) results in inhibition of pMEK and pERK and sustained tumor stasis and regressions in BRAF(V600E) colon carcinoma xenografts in nude mice. [2]
References

Applications

Methods Biomarkers Images PMID
Growth inhibition assay Cell viability S8015-viability1 31695841
Western blot p-RET / RET / p-PLCγ / PLCγ / p-ERK / ERK / p-MEK / MEK S8015-WB1 28011461

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