H 89 2HCl

H 89 2HCl is a potent PKA inhibitor with K_i of 48 nM in a cell-free assay, 10-fold selective for PKA than PKG,500-fold greater selectivity than PKC, MLCK, calmodulin kinase II and casein kinase I/II. H 89 2HCl induces autophagy.

CAS No. 130964-39-5

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
200mg	JPY 132000	国内在庫あり
1g	JPY 295500	国内在庫なし(納期7~10日)

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製品安全説明書

現在のバッチを見る：純度： 99.48%

99.48

H 89 2HCl関連製品

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PKA阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
SK-N-MC	Function Assay	30 μM	24 h	reduces the down-regulation of β1-AR by isoproterenol by 50%	11705454
CHO	Growth Inhibition Assay	10 μM	1 h	inhibits 5-HT induced cAMP production decrease	12569069
LS174T	Function assay	20 uM	6 hrs	Inhibition of PGE2-stimulated TCF/LEF transactivation in human LS174T cells at 20 uM measured after 6 hrs by dual luciferase reporter gene assay	27736063
OVCAR8	Cytotoxicity assay		72 hrs	Cytotoxicity against human OVCAR8 cells assessed as growth inhibition after 72 hrs by MTT assay, IC50 = 10.5 μM.	24508830
OVCAR8	Antiproliferative assay		72 hrs	Antiproliferative activity against human OVCAR8 cells after 72 hrs by MTT assay, IC50 = 10.5 μM.	24389511
HL60	Antiproliferative assay		72 hrs	Antiproliferative activity against human HL60 cells after 72 hrs by MTT assay, IC50 = 11.4 μM.	24389511
U87MG	Antiproliferative assay		96 hrs	Antiproliferative activity against human U87MG cells after 96 hrs by SRB assay, IC50 = 15 μM.	20151677
HCT116	Cytotoxicity assay		72 hrs	Cytotoxicity against human HCT116 cells assessed as growth inhibition after 72 hrs by MTT assay, IC50 = 15.7 μM.	24508830
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay, IC50 = 15.7 μM.	24389511
PC3M	Function assay		96 hrs	Inhibition of PC3M cells by SRB assay after 96 hrs, GI50 = 18 μM.	17451235
Sf9	Function assay			Inhibition of His-tagged human MSK1 expressed in Sf9 cells, IC50 = 0.12 μM.	10998351
Sf9	Function assay			Inhibition of rat ROCK2 expressed in Sf9 cells, IC50 = 0.27 μM.	10998351
SF9	Function assay			Inhibition of human PKBalpha expressed in SF9 cells, IC50 = 2.6 μM.	10998351
PC3M	Function assay			Inhibition of GSK3-beta in human PC3M cells by ELISA, IC50 = 15 μM.	18345609
U87MG	Growth inhibition assay			Growth inhibition of human U87MG cells by SRB assay, IC50 = 15 μM.	18345609
PC3M	Growth inhibition assay			Growth inhibition of human PC3M cells by SRB assay, IC50 = 18 μM.	18345609
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生物活性

製品説明

H 89 2HCl is a potent PKA inhibitor with K_i of 48 nM in a cell-free assay, 10-fold selective for PKA than PKG,500-fold greater selectivity than PKC, MLCK, calmodulin kinase II and casein kinase I/II. H 89 2HCl induces autophagy.

Targets

PKA ^[1] (Cell-free assay)	S6K1 ^[2] (Cell-free assay)	PKG ^[1] (Cell-free assay)
48 nM(Ki)	80 nM	0.48 μM(Ki)

In Vitro
In vitro	H89 2HCl is a potent PKA (cAMP-dependent) protein kinase A inhibitor with K_i of 48 nM, exhibits 10-fold selectivity over PKG, exhibits >500-fold selectivity over PKC, MLCK, calmodulin kinase II and casein kinase I/II. Pretreatment of the cells with H-89 (30 μM) 1 h before the addition of forskolin markedly inhibits the forskolin-induced protein phosphorylation in a dose-dependent manner. ^[1] H89 also inhibits several other kinases with IC50 of 80, 120, 135, 270, 2600 and 2800 nM for S6K1, MSK1, PKA, ROCKII, PKBα and MAPKAP-K1b, respectively. ^[2]^[3] H89 also has activity at some cellular receptors and ion channels, including Kv1.3 K⁺ channels,β1AR and β2AR. ^[4]
Kinase Assay	PKA enzyme activity
Kinase Assay	cAMP-dependent protein kinase activity is assayed in a reaction mixture containing, in a final volume of 0.2 mL, 50 mM Tris-HC1 (pH 7.0), 10 mM magnesium acetate, 2 mM EGTA, 1 μM cAMP or absence of cAMP, 3.3-20 μM [γ-³²P]ATP (4 × 10⁵ cpm), 0.5 μg of the enzyme, 100 μg of histone H2B, and each compound, as indicated.
細胞実験	細胞株	PC12D
	濃度	~30 μM
	反応時間	1 h
	実験の流れ	Levels of intracellular cAMP are determined. After 48 h in culture, PC12D cells are cultured in test medium containing 30 μM H-89 for 1 h and then exposed to fresh medium that contained both 10 μM forskolin and 30 μM H-89. Cells are scraped off with a rubber policeman and sonicated in the presence of 0.5 ml of 6% trichloroacetic acid. To extract trichloroacetic acid, 2 ml of petroleum ether is added, the preparation mixed and centrifuged at 3000 rpm for 10 min. After aspiration of the upper layer, the residue sample solution is used for determination.
実験結果図	Methods	Biomarkers	結果図	PMID
	Western blot	p-eNOS / T-eNOS / p-PKA p-GPIbβ / GPIbβ / p-BAD p-S6K1 / p-rpS6 / p-GSK3β		31035633
	Immunofluorescence	p-eNOS / eNOS iNOS / NFκB p65 myopodin		30513637

In Vivo
In Vivo	H89 causes distinct modifications of protein phosphorylation, with the most robust changes in phosphorylation are fructose-1,6-biphosphatase, heterogeneous nuclear ribonucleoprotein (hnRNP), NSFL1 cofactor p47, all which have potentially regulatory connections to cAMP/PKA. ^[5]
動物実験	動物モデル	rat; mice
	投与量	20 or 200 mg/kg (Rat); 0-5 mg/kg (Mice)
	投与経路	s.c. (Rat); i.p. (Mice)

参考
[1] Chijiwa T, et al. J Biol Chem., 1990, 265(9), 5267-5272. [2] Davies SP, et al. Biochem J, 2000, 351(Pt 1), 95-105. [3] Lochner A, et al. Cardiovasc Drug Rev, 2006, 24(3-4), 261-274. [4] Murray AJ. Sci Signal, 2008, 1(22), re4 [5] Davis MA, et al. J Pharmacol Exp Ther, 2006, 318(2), 589-595. + 展開

参考

+ 展開

化学情報

分子量	519.28	化学式	C₂₀H₂₀BrN₃O₂S.2HCl
CAS No.	130964-39-5	SDF	Download H 89 2HCl SDFをダウンロードする
Smiles	C1=CC2=C(C=CN=C2)C(=C1)S(=O)(=O)NCCNCC=CC3=CC=C(C=C3)Br.Cl.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个月-20°Cin solvent

In vitro
Batch:

DMSO : 100 mg/mL ( (192.57 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 : 10 mg/mL

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):