Name: Agerafenib (CEP-32496)
Brand: Selleck Chemicals
SKU: S8015
Availability: InStock
Rating: 5 (6 reviews)

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

Cited in Nature Medicine for its top-tier quality
COA
NMR
HPLC
SDS
Datasheet

99.19

Related Targets	ERK p38 MAPK K-Ras JNK MEK Ras S6 Kinase MAP4K TAK1 Mixed Lineage Kinase
Other Raf Inhibitors	LY3009120 GDC-0879 PLX-4720 AZ 628 SB590885 TAK-632 RAF265 (CHIR-265) GW5074 Avutometinib (Ro 5126766) PLX7904

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	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

Solubility

In vitro
Batch:

DMSO : 9 mg/mL (17.39 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	15% Captisol Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	10.000mg/ml (19.33mM)	Taking the 1 mL working solution as an example, take 10 mg of this product, add it to 1 ml of 15% Captisol clear solution, and mix evenly to form a uniform suspension. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

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Chemical Information, Storage & Stability

Molecular Weight	517.46	Formula	C₂₄H₂₂F₃N₅O₅	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	1188910-76-0	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
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

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 2 nM(Kd) LCK 2 nM(Kd) PDGFRβ 2 nM(Kd) RET 2 nM(Kd) Abl1 3 nM(Kd) VEGFR2 8 nM(Kd) CSF-1R 9 nM(Kd) B-Raf (V600E) 14 nM(Kd) EphA2 14 nM(Kd) EGFR 22 nM(Kd) B-Raf 36 nM(Kd) C-Raf 39 nM(Kd) c-Met 513 nM(Kd)
In vitro	Agerafenib (CEP-32496) inhibits A375 cell (BRAF^V600E) 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). 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.
Kinase Assay	Binding assay
Kinase 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 t_1/2 > 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. 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.
References	[1] https://pubmed.ncbi.nlm.nih.gov/22168626/ [2] https://pubmed.ncbi.nlm.nih.gov/22319199/

Applications

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

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Agerafenib (CEP-32496) Raf inhibitor

Chemical Structure

Molecular Weight: 517.46