PHA-767491 HCl

別名：CAY10572, NMS 1116354

製品コード：S2742 純度：99.96%

PHA-767491 (CAY10572, NMS 1116354) HCl is a potent ATP-competitive dual Cdc7/CDK9 inhibitor with IC50 of 10 nM and 34 nM in cell-free assays, respectively.It displays ~20-fold selectivity against CDK1/2 and GSK3-β, 50-fold selectivity against MK2 and CDK5, 100-fold selectivity against PLK1 and CHK2.

CAS No. 942425-68-5

サイズ	価格(税別)	在庫状況
10mM (1mL in DMSO)	JPY 29500	国内在庫あり
10mg	JPY 22000	国内在庫あり
50mg	JPY 55500	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

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製品安全説明書

現在のバッチを見る：純度： 99.96%

99.96

PHA-767491 HCl関連製品

関連ターゲット	CDK1 CDK2 CDK3 CDK4 CDK5 CDK6 CDK7 CDK9 CLK Cdc CDK8 CDK12 CDK13 CDK19 CDK11 CDK/cyclin complexes	もっと見る
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関連化合物ライブラリー	Kinase Inhibitor Library PI3K/Akt Inhibitor Library MAPK Inhibitor Library DNA Damage/DNA Repair compound Library Cell Cycle compound library	もっと見る

シグナル伝達経路

CDKシグナル伝達経路

CDK阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
human HeLa cells	Function assay	5 μM	24 h	Induction of apoptosis in human HeLa cells assessed as appearance of PARP at 5 uM after 24 hrs	18469809
NHDF	Function assay	5 μM	16 h	Induction of cell cycle arrest in thymidine deficient NHDF assessed as DNA synthesis in S-phase at 5 uM after 16hrs FACS analysis in presence of serum	18469809
human SF268 cells	Proliferation assay		72 h	Antiproliferative activity against p53 deficient human SF268 cells after 72 hrs, IC50=0.86 μM	18469809
human HCT116 cells	Proliferation assay		72 h	Antiproliferative activity against human HCT116 cells expressing p53 gene after 72 hrs by proliferative assay, IC50=0.97 μM	18469809
human HCT16 cells	Proliferation assay		72 h	Antiproliferative activity against human HCT16 cells after 72 hrs by luciferase based assay, IC50=1 μM	19115845
human SW403 cells	Proliferation assay		72 h	Antiproliferative activity against human SW403 cells after 72 hrs by luciferase based assay, IC50=1 μM	19115845
human A2780 cells	Proliferation assay		72 h	Antiproliferative activity against human A2780 cells expressing p53 gene after 72 hrs by proliferative assay, IC50=1.07 μM	18469809
human SW48 cells	Proliferation assay		72 h	Antiproliferative activity against human SW48 cells after 72 hrs by luciferase based assay, IC50=1.2 μM	19115845
human MCF7 cells	Proliferation assay		72 h	Antiproliferative activity against human MCF7 cells after 72 hrs by luciferase based assay, IC50=1.3 μM	19115845
human U2OS cells	Proliferation assay		72 h	Antiproliferative activity against human U2OS cells expressing p53 gene after 72 hrs by proliferative assay, IC50=1.49 μM	18469809
human COLO205 cells	Proliferation assay		72 h	Antiproliferative activity against human COLO205 cells after 72 hrs by luciferase based assay, IC50=1.5 μM	19115845
human OVCAR8 cells	Proliferation assay		72 h	Antiproliferative activity against p53 deficient human OVCAR8 cells after 72 hrs by proliferative assay, IC50=1.56 μM	18469809
human L363 cells	Proliferation assay		72 h	Antiproliferative activity against human L363 cells after 72 hrs by luciferase based assay, IC50=1.6 μM	19115845
human NHDF cells	Proliferation assay		72 h	Antiproliferative activity against human NHDF cells after 72 hrs by luciferase based assay, IC50=1.6 μM	19115845
human NCI-H929 cells	Proliferation assay		72 h	Antiproliferative activity against human NCI-H929 cells after 72 hrs by luciferase based assay, IC50=1.8 μM	19115845
human SF539 cells	Proliferation assay		72 h	Antiproliferative activity against human SF539 cells expressing p53 gene after 72 hrs by proliferative assay, IC50=2.34 μM	18469809
human SW480 cells	Proliferation assay		72 h	Antiproliferative activity against p53 deficient human SW480 cells after 72 hrs by proliferative assay, IC50=2.67 μM	18469809
human Jurkat cells	Proliferation assay		72 h	Antiproliferative activity against p53 deficient human Jurkat cells after 72 hrs by proliferative assay, IC50=3.2 μM	18469809
human HCT15 cells	Proliferation assay		72 h	Antiproliferative activity against human HCT15 cells expressing p53 gene after 72 hrs by proliferative assay, IC50=3.81 μM	18469809
human OPM2 cells	Proliferation assay		72 h	Antiproliferative activity against human OPM2 cells after 72 hrs by luciferase based assay, IC50=4.5 μM	19115845
human HT-29 cells	Proliferation assay		72 h	Antiproliferative activity against human HT-29 cells after 72 hrs by luciferase based assay, IC50=5 μM	19115845
human K562 cells	Proliferation assay		72 h	Antiproliferative activity against p53 deficient human K562 cells after 72 hrs, IC50=5.87 μM	18469809
human NCI60 cells	Proliferation assay			Antiproliferative activity against human NCI60 cells, IC50=3.1 μM	18469809
U937 cells	Function assay			Inhibition of TNFalpha production in U937 cells, IC50=19 μM	17480064
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

特性

The first inhibitor that directly affects the mechanisms controlling initiation as opposed to elongation in DNA replication.

Targets

Cdc7 ^[1] (Cell-free assay)	CDK9 ^[1] (Cell-free assay)	GSK-3β ^[1] (Cell-free assay)	CDK2 ^[1] (Cell-free assay)	CDK1 ^[1] (Cell-free assay)	もっとクリックする
10 nM	34 nM	220 nM	240 nM	250 nM	もっとクリックする

In Vitro
In vitro	PHA-767491 displays approximately 20-fold selectivity for Cdk1, Cdk2 and GSK3-β, 50-fold selectivity for MK2 and Cdk5 and 100-fold selectivity for PLK1 and CHK2. PHA-767491 inhibits cell proliferation in a variety of human cell lines with IC50 of 0.86 μM for SF-268 to 5.87 μM for K562, and significantly induces apoptosis in a p53-independent manner in almost all cell lines in contrast with 5-FU which only works in a few of cell lines. Unlike current DNA synthesis inhibitors, PHA-767491 treatment at 5 μM blocks the initiation of DNA replication but not replication fork progression, due to specific inhibition of Cdc7 kinase and Mcm2 phosphorylation at the Cdc7-dependent Ser40 site. ^[1] The up-regulated Mcl-1 levels in ABT-737-resistant OCI-LY1 and SU-DHL-4 cells can be significantly decreased by PHA-767491 treatment at 3 μM possibly due to the inhibition of Cdk9, leading to the restoration of the sensitivity to ABT-737. ^[2] The direct mitochondrial dependent pro-apoptosis effect of PHA-767491 is also observed when applied at 1 μM in quiescent chronic lymphocytic leukemia (CLL) cells through the similar mechanism with EC50 of 0.34-0.97 μM. While in proliferating CLL cells stimulated by CD154 and interleukin-4, PHA-767491 treatment at 5 μM abolishes DNA synthesis by inhibiting Cdc7 rather than triggering cell death. ^[3]
Kinase Assay	In vitro kinase assays
Kinase Assay	The inhibition of Cdc7 and Cdk9 by PHA-767491 (IC50) is determined using the strong anion exchanger (Dowex 1-X8 resin, formate form)-based assay. For each enzyme, the absolute K_m values for ATP and the specific substrate are initially determined, and each assay is then run at optimized ATP/³³P-γ-ATP mix (2K_m) and substrate (5K_m) concentrations. Cdc7 kinase assay is performed in a buffer containing 50 mM Hepes pH 7.9, 15 mM MgCl₂, 2 mM β- glycerylphosphate, 0.2 mg/mL BSA, 1 mM DTT, 3 μM Na₃VO₄, 2K_m ATP/³³P-γ-ATP mix, 5K_m Mcm2 (aa 10-294), 37 nM of recombinant Cdc7/Dbf4 and increasing concentration of PHA-767491 in a final volume of 30 μL, and incubated for 1 hour at 25 °C. Cdk9 kinase assay is performed using 50 nM of recombinant Cdk9/cyclin T in 50 mM HEPES pH 7.5, 10 mM MgCl₂, 1 mM DTT, 3 μM Na₃VO₄, 2K_m ATP/³³P-γ-ATP mix, 5K_m RNA polymerase CDT peptide and increasing concentration of PHA-767491 in a final volume of 30 μL, and incubated for 1 hour at 25 °C. After incubation, an amount of 150 μL of resin/formate (pH 3.0) is added to stop the reaction and capture unreacted ³³P-γ-ATP, separating it from the phosphorylated substrate in solution. After 1 hour of rest, a volume of 50 μL supernatant is transferred to Optiplate 96-well plates. After the additon of 150 μL of Microscint 40, the radioactivity is counted in the TopCount.
細胞実験	細胞株	HeLa, MCF7, HCT-116, U2OS, A2780, K562, SF-539, SF-268, Ovcar8, SW480, COLO205, HCT-15, Jurkat, PC3, and NHDF
	濃度	Dissolved in DMSO, final concentrations ~ 20 μM
	反応時間	24 or 72 hours
	実験の流れ	Cells are exposed to PHA-767491 for 24 or 72 hours. Cells are lysed and the ATP content in the well, used as a measure of viable cells, is determined using a thermostable firefly luciferase–based assay. Activation of caspase-3 and caspase-7 is measured as a ratio between treated sample and untreated control with a luciferase-based assay, containing a specific proluminescent substrate. DNA replication is measured as incorporation of nucleotide analog BrdU into DNA by flow cytometry.
実験結果図	Methods	Biomarkers	結果図	PMID
実験結果図	Western blot	p-MCM2 / CDC7 RNA Pol II / p-RNA Pol II / Caspase-3 / PARP / Mcl-1 / XIAP / Bcl-xL / Bcl-2 / NOXA		24902048

In Vivo
In Vivo	Administration of PHA-767491 twice a day for 5 days significantly inhibits the growth of HL60 xenograft in a dose-dependent manner with TGI of 50% and 92% at dose of 20 mg/kg and 30 mg/kg, respectively, the effect of which is also marked in A2780, Mx-1, and HCT-116 xenograft models as well as the mammary carcinomas, and correlates with Cdc7 inhibition and subsequently decreased phosphorylation of Mcm2 at the Cdc7-dependent site Ser40 ^[1]
動物実験	動物モデル	Female SCID mice subcutaneously implanted with HL60 cells, male Hsd, athymic nu-nu mice subcutaneously implanted with HCT116 cells, A2780 or Mx-1 cells, and female Sprague-Dawley rats with mammary carcinomas
	投与量	~50 mg/kg
	投与経路	Intravenous or oral administration twice a day

参考
[1] Montagnoli A, et al. Nat Chem Biol, 2008, 4(6), 357-365. [2] Yecies D, et al. Blood, 2010, 115(16), 3304-3313. [3] Natoni A, et al. Mol Cancer Ther, 2011, 10(9), 1624-1634.

参考

化学情報

分子量	249.7	化学式	C₁₂H₁₁N₃O.HCl
CAS No.	942425-68-5	SDF	Download PHA-767491 HCl SDFをダウンロードする
Smiles	C1CNC(=O)C2=C1NC(=C2)C3=CC=NC=C3.Cl
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ケ月-20°Cin solvent

In vitro
Batch:

DMSO : 24 mg/mL ( (96.11 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。

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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):