Agerafenib (CEP-32496)

Agerafenib (CEP-32496) is a highly potent inhibitor of BRAF(V600E/WT) and c-Raf with K_d of 14 nM/36 nM and 39 nM, also potent to Abl-1, c-Kit, Ret (c-Ret), PDGFRβ and VEGFR2, respectively; insignificant affinity for MEK-1, MEK-2, ERK-1 and ERK-2. Phase 1/2.

CAS No. 1188910-76-0

サイズ	価格(税別)	在庫状況
10mM (1mL in DMSO)	JPY 94900	国内在庫あり
5mg	JPY 46800	国内在庫あり
10mg	JPY 63400	国内在庫あり

代表番号: 045-509-1970\|電子メール：[email protected] よく尋ねられる質問

文献中Selleckの製品使用例（6）

製品安全説明書

現在のバッチを見る： S801501 DMSO] 9 mg/mL] false] Water] Insoluble] false] Ethanol] Insoluble] false 純度： 99.19%

99.19

Agerafenib (CEP-32496)関連製品

関連ターゲット	C-Raf/Raf-1 B-Raf A-raf	もっと見る
関連化合物ライブラリー	Kinase Inhibitor Library MAPK Inhibitor Library Cell Cycle compound library TGF-beta/Smad compound library Anti-alzheimer Disease Compound Library	もっと見る

シグナル伝達経路

Rafシグナル伝達経路

Raf阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
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
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
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
HCC827	Function assay			Displacement of [3H]-cyclopamine from SMO V404M mutant in gefitinib resistant human HCC827 cells by scintillation counting, Ki=0.1878μM.	28787156
他の多くの細胞株試験データをご覧になる場合はこちらをクリックして下さい

生物活性

製品説明

Agerafenib (CEP-32496) is a highly potent inhibitor of BRAF(V600E/WT) and c-Raf with K_d of 14 nM/36 nM and 39 nM, also potent to Abl-1, c-Kit, Ret (c-Ret), PDGFRβ and VEGFR2, respectively; insignificant affinity for MEK-1, MEK-2, ERK-1 and ERK-2. Phase 1/2.

特性

High binding affinity for both BRAF (V600E) mutation and related c-Raf, but no significant affinity for other kinases of MAPK pathway.

Targets

c-Kit ^[1]	LCK ^[1]	PDGFRβ ^[1]	RET ^[1]	Abl1 ^[1]	もっとクリックする
2 nM(Kd)	2 nM(Kd)	2 nM(Kd)	2 nM(Kd)	3 nM(Kd)	もっとクリックする

In Vitro
In vitro	CEP-32496 inhibits A375 cell (BRAF^V600E) proliferation with EC50 of 78 nM. CEP-32496 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] CEP-32496 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 of 78 nM and 60 nM, respectively. ^[2]
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 CEP-32496. Kd values 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.
細胞実験	細胞株	A375 cell lines
	濃度	78 nM
	反応時間	72 hours
	実験の流れ	Cells are seeded at 10⁴ cells per well in DMEM with 10% fetal calf serum and allowed to attach. The cells are washed with PBS and switched to DMEM with 0.5% of serum and incubated overnight. CEP-32496 is then added at various concentrations with a final DMSO concentration of 0.5% and incubated for 72 h. At the end of incubation, a Cell Titer Blue is added per instructions, and incubation is continued for 3 hours. Remaining viable cells are quantified by measuring the strength of the fluorescence signal using SoftMax Pro (excitation at 560 nm and emission at 590 nm). IC50 values are derived using a 9-point curve fitted with Igor Pro and are presented as mean values from experiments performed in duplicate. Variability between individual values is less than 2-fold.
実験結果図	Methods	Biomarkers	結果図	PMID
	Growth inhibition assay	Cell viability		31695841
	Western blot	p-RET / RET / p-PLCγ / PLCγ / p-ERK / ERK / p-MEK / MEK		28011461

In Vivo
In Vivo	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. CEP-32496 (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. CEP-32496 (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] CEP-32496 is orally bioavailable in multiple preclinical species (>95% in rats, dogs, and monkeys). CEP-32496 (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]
動物実験	動物モデル	Colo-205 xenograft mouse model
	投与量	100 mg/kg
	投与経路	oral gavage

参考
[1] Rowbottom MW, et al. J Med Chem, 2012, 55(3), 1082-1105. [2] James J, et al. Mol Cancer Ther, 2012, 11(4), 930-941.

参考

化学情報

分子量	517.46	化学式	C₂₄H₂₂F₃N₅O₅
CAS No.	1188910-76-0	SDF	Download Agerafenib (CEP-32496) SDFをダウンロードする
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
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1年-80°Cin solvent
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1个月-20°Cin solvent

In vitro
Batch:

DMSO : 9 mg/mL ( (17.39 mM)；吸湿したDMSOは溶解度を減少させます。新しいDMSOをご使用ください。)

Water : Insoluble

Ethanol : Insoluble

モル濃度計算器

in vivo
Batch:

Add solvents to the product individually and in order.

投与溶液組成計算機

実験計算

モル濃度計算器

質量	濃度	体積	分子量
=	×	×

希釈計算器分子量計算器

投与溶液組成計算機(クリア溶液)

ステップ１：実験データを入力してください。（実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します）

投与量 mg/kg 動物平均体重 g 投与体積（動物毎）μL 動物数匹

ステップ２：投与溶媒の組成を入力してください。（ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください）

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

計算結果：

投与溶媒濃度： mg/ml;

DMSOストック溶液調製方法： mg 試薬を μL DMSOに溶解する（濃度 mg/mL, 注：濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法：Take μL DMSOストック溶液に μL PEG300，を加え、完全溶解後μL Tween 80，を加えて完全溶解させた後 μL ddH₂O，を加え完全に溶解させます。

投与溶媒調製方法：μL DMSOストック溶液に μL Corn oil，を加え、完全溶解。

注意：１.ストック溶液に沈殿、混濁などがないことをご確認ください；
２.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):