Purmorphamine

Purmorphamine (Shh Signaling Antagonist VI), which directly binds and activates Smoothened, blocks BODIPY-cyclopamine binding to Smo with IC50 of ~ 1.5 μM in HEK293T cell and also is an inducer of osteoblast differentiation with EC50 of 1 μM. Purmorphamine can reduce both basal and induced autophagy.

CAS No. 483367-10-8

サイズ	価格(税別)	在庫状況
5mg	JPY 22000	国内在庫あり
25mg	JPY 70500	国内在庫あり

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Purmorphamine関連製品

関連ターゲット	Hedgehog Smoothened GLI	もっと見る
関連製品	Vismodegib (GDC-0449) Cyclopamine GANT61 SAG (Smoothened Agonist) HCl Sonidegib Itraconazole SANT-1 SAG (Smoothened Agonist) BMS-833923 HPI-4 (Ciliobrevin A) Taladegib (LY2940680) PF-5274857 Glasdegib Jervine Ciliobrevin D MK-4101 JK184	もっと見る
関連化合物ライブラリー	Kinase Inhibitor Library FDA-approved Drug Library Stem Cell Signaling Compound Library GPCR Compound Library Stem Cell Differentiation Compound Library	もっと見る

シグナル伝達経路

Hedgehog/Smoothenedシグナル伝達経路

Hedgehog/Smoothened阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
HEK293T	Function assay	5 uM	4 hrs	Inhibition of BODIPY-cyclopamine binding to Smo N-terminal cysteine domain expressed in HEK293T cells at 5 uM after 4 hrs by fluorescence microscopy	16408088
HEK293T	Function assay	5 uM	4 hrs	Inhibition of BODIPY-cyclopamine binding to Smo C-terminal cytoplasmic domain expressed in HEK293T cells at 5 uM after 4 hrs by fluorescence microscopy	16408088
Shh Light2	Function assay		30 hrs	Activation of Shh in mouse Shh Light2 cells after 30 hrs by luciferase reporter gene assay, EC50 = 1 μM.	16408088
C3H10T1/2	Function assay		6 days	Activity at Smo in mouse C3H10T1/2 cells assessed as induction of cell differentiation into osteoblast incubated for 6 days by alkaline phosphatase assay, EC50 = 0.8 μM.	27429255
HEK293T	Function assay		1 hr	Inhibition of BODIPY-cyclopamine binding to Smo expressed in HEK293T cells after 1 hr by fluorescence microscopy, IC50 = 1.5 μM.	16408088
Shh Light2	Function assay		30 hrs	Activation of Shh in mouse Shh Light2 cells assessed as beta-galactosidase activity after 30 hrs by luciferase reporter gene assay in presence of 100 nM 3-keto-N-aminoethyl-N'-aminocaproyldihydrocinnamoyl cyclopamine	16408088
C3H10T1/2	Function assay			Induction of osteogenesis in mouse C3H10T1/2 cells assessed as induction of osteoblast specific marker alkaline phosphatase by immunofluorescence method, EC50 = 1 μM.	16408003
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
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生物活性

製品説明

Purmorphamine (Shh Signaling Antagonist VI), which directly binds and activates Smoothened, blocks BODIPY-cyclopamine binding to Smo with IC50 of ~ 1.5 μM in HEK293T cell and also is an inducer of osteoblast differentiation with EC50 of 1 μM. Purmorphamine can reduce both basal and induced autophagy.

Targets

Smoothened ^[1]
(HEK293T cells)

~1.5 μM

In Vitro
In vitro	Purmorphamine activates the Hedgehog pathway by directly binds and activates Smoothened with IC50 of ~ 1.5 μM in compete with cyclopamine, a Smo antagonist. ^[1] Purmorphamine is a potent inducer of osteogenesis in multipotent C3H10T1/2 cells. The EC50 (based on ALP expression) for Purmorphamine is 1 μM in C3H10T1/2 cells. Purmorphamine (1 μM) and BMP-4 (100 ng/mL) together increase ALP activity more than 90-fold in 3T3-L1 cells. ^[2] In contrast to BMP-4, Purmorphamine induces osteogenesis by activating Hedgehog signaling in multipotent mesenchymal progenitor cells. ^[3]
Kinase Assay	Binding assay
Kinase Assay	Smo binding assays are conducted with BODIPY-cyclopamine and Smo-overexpressing cells as previously described4,5, using CMV promoter-based, SV40 origin-containing expression constructs for Smo-Myc3, the deletion mutant SmoCRD (deletion of amino acids 68 to 182), and SmoCT (deletion of amino acids 556 to 793). HEK 293T cells are grown on poly-D-lysine-treated glass coverslips in 12-well plates until 70% confluency and then transfected with the appropriate expression construct (0.5 g/well) using FuGene 6 according the manufacturer
細胞実験	細胞株	C3H10T1/2 cell
	濃度	0.5-10 μM
	反応時間	4 days
	実験の流れ	C3H10T1/2 cells are expanded in T175 flasks; cells at 13th passage are detached by trypsin/EDTA and diluted in the growth media. The resulting cell suspension is then plated into black clear bottom 384-well plates with 2500 cells/well in 100 µL growth medium using a Multi-drop^TM liquid delivery system. After overnight incubation, cells attached to the bottom of the wells. A stock solution of each Purmorphamine in DMSO (500 nL) is delivered into corresponding well using a Mini Trak^TM multiposition dispenser system to make a final concentration of 5μM of Purmorphamine. Cells are then incubated at 37 ℃ with 5% CO₂ in air atmosphere. After 4 days, the medium is removed and 10 μL of passive lysis buffer is added into each well. After 5 min, 10 μL of alkaline phosphatase substrate solution is added to each well. After incubating 15 min at room temperature, the plates are read on an Acquest high-throughput plate reader following the manufacturer's protocol.
実験結果図	Methods	Biomarkers	結果図	PMID
	Western blot	Patch1 / Gli1 / LC3 / p62		26609469
	Immunofluorescence	SOX18		26588701
	Growth inhibition assay	Cell viability		26588701

In Vivo
In Vivo	Purmorphamine up-regulates ALP expression in human mesenchymal stem cell-based constructs on rats. ^[4]
動物実験	動物モデル	Male C57BL/6J mouse pups
	投与量	10 mg/kg
	投与経路	i.p.

参考
[1] Sinha S, et al. Nat Chem Biol, 2006, 2(1), 29-30. [2] Wu X, et al. J Am Chem Soc, 2002, 124(49), 14520-14521. [3] Wu X, et al. Chem Biol, 2004, 11(9), 1229-1238. [4] Faghihi F, Biomed Pharmacother, 2012, 3322(12). [5] Liu D, et al. Front Pharmacol. 2020 Mar 4;11:204. + 展開

参考

+ 展開

化学情報

分子量	520.62	化学式	C₃₁H₃₂N₆O₂
CAS No.	483367-10-8	SDF	Download Purmorphamine SDFをダウンロードする
Smiles	C1CCC(CC1)N2C=NC3=C(N=C(N=C32)OC4=CC=CC5=CC=CC=C54)NC6=CC=C(C=C6)N7CCOCC7
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 : 4 mg/mL ( (7.68 mM)； Warmed with 50℃ water bath; 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):