Home Stem Cells & Wnt Hedgehog/Smoothened agonist - Autophagy inhibitor Purmorphamine For research use only.

Purmorphamine

Synonyms: Shh Signaling Antagonist VI

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.

Purmorphamine Chemical Structure

Purmorphamine Chemical Structure

CAS No. 483367-10-8

Purity & Quality Control

Purmorphamine Related Products

Signaling Pathway

Hedgehog/Smoothened Signaling Pathways

Hedgehog/Smoothened Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation 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

Biological Activity

Description 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
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
Cell Research Cell lines C3H10T1/2 cell
Concentrations 0.5-10 μM
Incubation Time 4 days
Method

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-dropTM 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 TrakTM 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.
Experimental Result Images Methods Biomarkers Images 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]
Animal Research Animal Models Male C57BL/6J mouse pups
Dosages 10 mg/kg
Administration i.p.

Chemical Information & Solubility

Molecular Weight 520.62 Formula

C31H32N6O2
CAS No. 483367-10-8 SDF Download Purmorphamine SDF
Smiles C1CCC(CC1)N2C=NC3=C(N=C(N=C32)OC4=CC=CC5=CC=CC=C54)NC6=CC=C(C=C6)N7CCOCC7
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 4 mg/mL ( (7.68 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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