Purmorphamine (Hedgehog/Smoothened agonist),Beyotime_specification/price/image

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Cat.No.: SF6822-10mM
Package: 10mM×0.2ml

Description

Cat. No.	Product Name	Package	Price (CNY)
SF6822-10mM	Purmorphamine (Hedgehog/Smoothened agonist)	10mM×0.2ml	252.00
SF6822-5mg	Purmorphamine (Hedgehog/Smoothened agonist)	5mg	807.00
SF6822-25mg	Purmorphamine (Hedgehog/Smoothened agonist)	25mg	2540.00

Chemical information:

Chemical Name	9-cyclohexyl-N-(4-morpholin-4-ylphenyl)-2-naphthalen-1yloxypurin-6-amine
Abbreviations	Purmorphamine
Alias	UNII-PB12M2F8KY, Shh Signaling Antagonist VI, PB12M2F8KY
Chemical Formula	C31H32N6O2
Molecular Weight	520.62
CAS No.	483367-10-8
Purity	98%
Solvent/Solubility	Water <1mg/ml; DMSO 4mg/ml warming; Ethanol <1mg/ml
Solution Preparation	Add 0.96ml of DMSO for 5mg, or 1ml of DMSO for every 5.21mg to prepare a 10mM solution. SF6822-10mM is formulated with DMSO.

Biological information:

Description	Purmorphamine directly binds and activates Smoothened and blocks BODIPY-cyclopamine binding to Smo with an IC50 of approximately 1.5μM in HEK293T cells and also induces osteoblast differentiation with an EC50 of 1μM.
Signaling Pathways	Stem Cells & Wnt; GPCR & G Protein
Targets	Smoothened	-	-	-	-
IC50	~1.5μM	-	-	-	-
In vitro Studies	Purmorphamine competes with Cyclopamine (a Smo antagonist) for direct binding and activation of Smoothened, while activating the Hedgehog pathway with an IC50 of 1.5μM. Purmorphamine acts on allosteric C3H10T1/2 cells and is a potent inducer of osteogenesis. Purmorphamine acts on C3H10T1/2 cells with an EC50 (based on ALP expression) of 1 μM. Purmorphamine (1μM) in combination with BMP4 (100ng/ml) acts on 3T3-L1 cells, resulting in a more than 90-fold enhancement of ALP activity. In contrast to BMP-4, Purmorphamine acts on pluripotent mesenchymal progenitor cells and induces osteogenesis, by activating Hedgehog signaling.
In vivo Studies	Purmorphamine acts on rat structure-based human mesenchymal stem cells to upregulate ALP expression.
Clinical Trials	N/A
Characterization	N/A

Relevant experimental data (this data is from published literature and Beyotime does not guarantee its validity):

Enzyme Activity Assay Experiment
Method	Smo binding experiments were performed using BODIPY-Cyclopamine and cells overexpressing Smo to obtain Smo-Myc3, deletion mutants Smo CRD (amino acid deletion at positions 68 to 182) and Smo CT (amino acid deletion at positions 556 to 793) using a CMV promoter, SV40 starting point-containing expression vector. HEK 293T cells were grown on poly-D-lysine-treated glass coverslips in 12-well plates until 70% confluence, and then transfected by appropriate expression vectors (0.5g per well) using FuGene 6. Two days after transfection, HEK 293T cells were incubated with DMEM medium containing 0.5% calf serum, 5nM BODIPY-Cyclopamine and different concentrations of Purmorphamine (0, 1.5 or 5M) (1ml per well) for 1h at 37℃. Then cells overexpressing Smo were washed using 1×PBS buffer (1 ml per well), placed in DAPI-containing medium and observed using a Leica DM4500B fluorescence microscope. For binding assays using fixed cells, HEK293T cells overexpressing Smo were fixed with 3% paraformaldehyde dissolved in 1× PBS buffer (1ml per well) for 10 min at room temperature, then treated with 1× PBS buffer (1ml per well) containing 10mM glycine and 0.2% sodium azide for 5 min and washed with 1X PBS buffer (1ml per well). The medium containing Purmorphamine was then used again for 4h at room temperature.
Cell Experiment
Cell Line	C3H10T1/2 cells
Concentration	0.5-10μM
Treatment Time	4 days
Method	C3H10T1/2 cells were amplified in T175 flasks and passage 13 cells were isolated by trypsin/EDTA and then diluted in growth medium. Using the Multi-dropTM Liquid Transfer System, the resulting cell suspension was inoculated at 2500 cells per well into black clear bottom 384-well plates with 100µl of growth medium in the wells. After incubation overnight, cells are attached to the bottom of the wells. Using the Mini TrakTM Multi-Dispensing System, each group of Purmorphamine stock solution dissolved in DMSO (500nl) was delivered to the corresponding wells, ensuring a final concentration of Purmorphamine of 5µM. Cells were then incubated at 37℃ in air with 5% CO2. After 4 days, the medium was removed and 10μl of passive lysis buffer was added to each well. 5 min later, 10μl of alkaline phosphatase substrate solution was added to each well. After incubation at room temperature for 15 min, the experimental plates were read on an Acquest high-throughput plate reader.
Animal Experiment
Animal Models	N/A
Formulation	N/A
Dosage	N/A
Administration Method	N/A

References：
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).

Packing List：

Item	Component	Quantity
SF6822-10mM	Purmorphamine (Hedgehog/Smoothened Agonist)	10mM×0.2ml
SF6822-5mg	Purmorphamine (Hedgehog/Smoothened Agonist)	5mg
SF6822-25mg	Purmorphamine (Hedgehog/Smoothened Agonist)	25mg
Manual	-	1 copy

Storage Conditions：
Store at -20℃, valid for at least 1 year. SF6822-5mg and SF6822-25mg can also be stored at room temperature for at least 6 months. If dissolved in non-DMSO solvents, it is recommended to store aliquots at -80℃, valid for 6 months.

Precautions：
This product is for R&D only. Not for drug, household, or other uses.
For your safety and health, please wear a lab coat and disposable gloves during the operation.

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