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Purmorphamine Smoothened Receptor Agonist

Name: Purmorphamine
Brand: Selleck Chemicals
SKU: S3042
Availability: InStock
Rating: 5 (61 reviews)

Cat.No.S3042

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.

Chemical Structure

Molecular Weight: 520.62

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Quality Control

Batch: Purity: 99.97%

99.97

Related Targets	JAK TGF-beta/Smad Wnt/beta-catenin ERK GSK-3 ROCK PKA Secretase STAT Casein Kinase
Other Hedgehog/Smoothened Inhibitors	SAG (Smoothened Agonist) Hydrochloride Cyclopamine (11-Deoxojervine) GANT61 SAG (Smoothened Agonist) SANT-1 HPI-4 (Ciliobrevin A) BMS-833923 Taladegib (LY2940680) Ciliobrevin D Jervine

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
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
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			Induction of osteogenesis in mouse C3H10T1/2 cells assessed as induction of osteoblast specific marker alkaline phosphatase by immunofluorescence method, EC50 = 1 μM.	16408003
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
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
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
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 4 mg/mL (7.68 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	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	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.
Clear solution	2%DMSO 1 30%PEG300 2 5%Tween80 3 63%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	1.000mg/ml (1.92mM)	Taking the 1 mL working solution as an example, add 20 μL of clarified DMSO stock solution of 50 mg/ml to 300 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify it; then continue to add 630 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

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Average weight of animals: g

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Chemical Information, Storage & Stability

Molecular Weight	520.62	Formula	C₃₁H₃₂N₆O₂	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	483367-10-8	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	Shh Signaling Antagonist VI			Smiles	C1CCC(CC1)N2C=NC3=C(N=C(N=C32)OC4=CC=CC5=CC=CC=C54)NC6=CC=C(C=C6)N7CCOCC7

Mechanism of Action

Targets/IC₅₀/Ki	Smoothened (HEK293T cells) ~1.5 μM
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. This compound is a potent inducer of osteogenesis in multipotent C3H10T1/2 cells. The EC50 (based on ALP expression) for this chemical is 1 μM in C3H10T1/2 cells. It (1 μM) and BMP-4 (100 ng/mL) together increase ALP activity more than 90-fold in 3T3-L1 cells. In contrast to BMP-4, this compound induces osteogenesis by activating Hedgehog signaling in multipotent mesenchymal progenitor cells.
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
In vivo	Purmorphamine up-regulates ALP expression in human mesenchymal stem cell-based constructs on rats.
References	[1] http://www.ncbi.nlm.nih.gov/pubmed?term=16408088 [2] https://pubmed.ncbi.nlm.nih.gov/12465946/ [3] http://www.ncbi.nlm.nih.gov/pubmed?term=15380183 [4] http://www.ncbi.nlm.nih.gov/pubmed?term=23228449 [5] https://pubmed.ncbi.nlm.nih.gov/32194421/