For research use only.

Catalog No.S3042 Synonyms: Shh Signaling Antagonist VI

33 publications

Purmorphamine Chemical Structure

CAS No. 483367-10-8

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.

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Selleck's Purmorphamine has been cited by 33 publications

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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.
Smoothened [1]
(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. [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]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
C3H10T1/2 NIixOppHfW6ldHnvckBie3OjeR?= M4TTeFYh\GG7cx?= MlL3RYN1cX[rdImgZZQhW22xIHnuJI1wfXOnIFOzTFExXDFxMjDj[YxteyCjc4Pld5Nm\CCjczDpcoR2[3Srb36gc4Yh[2WubDDkbYZn\XKnboTpZZRqd25iaX70c{Bwe3Snb3LsZZN1KGmwY4XiZZRm\CCob4KgOkBl[Xm|IHL5JIFtc2GuaX7lJJBpd3OyaHH0ZZNmKGG|c3H5MEBGSzVyIE2gNE45KM7:TT6= M3XY[FxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ5NEK5NlU2Lz5{N{SyPVI2PTxxYU6=
Shh Light2 MXnGeY5kfGmxbjDhd5NigQ>? Mlr1N|AhcHK| M4X5XWFkfGm4YYTpc44hd2ZiU3joJIlvKG2xdYPlJHNpcCCOaXfoeFIh[2WubIOgZYZ1\XJiM{CgbJJ{KGK7IHz1Z4ln\XKjc3WgdoVxd3K2ZYKg[4Vv\SCjc4PhfUwhTUN3MDC9JFEh|ryPLh?= M3TGb|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF4NEC4NFg5Lz5zNkSwPFA5QDxxYU6=
C3H10T1/2 MoixSpVv[3Srb36gZZN{[Xl? MUTJcoR2[3Srb36gc4Yhd3O2ZX;n[Y5me2m|IHnuJI1wfXOnIFOzTFExXDFxMjDj[YxteyCjc4Pld5Nm\CCjczDpcoR2[3Srb36gc4Yhd3O2ZX;icIF{fCC|cHXjbYZq[yCvYYLr[ZIh[WytYXzpcoUheGixc4DoZZRie2ViYomgbY1ufW6xZnz1c5Jme2OnbnPlJI1mfGixZDygSWM2OCB;IEGg{txONg>? NYrtfZFZRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMU[0NFgxODNpPkG2OFA5ODB|PD;hQi=>
HEK293T M{XqfWZ2dmO2aX;uJIF{e2G7 NYjtWFVROSCqch?= NWDwUlhqUW6qaXLpeIlwdiCxZjDCU2RKWFlvY4njcI9x[W2rbnWgZolv\GmwZzD0c{BUdW9iZYjwdoV{e2WmIHnuJGhGUzJ7M2SgZ4VtdHNiYX\0[ZIhOSCqcjDifUBndHWxcnXzZ4Vv[2VibXnjdo9{[2:yeTygTWM2OCB;IEGuOUDPxE1w M1n5fFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF4NEC4NFg5Lz5zNkSwPFA5QDxxYU6=
Shh Light2 NWHFUFJ3TnWwY4Tpc44h[XO|YYm= MVyzNEBpenN? M1;kb2FkfGm4YYTpc44hd2ZiU3joJIlvKG2xdYPlJHNpcCCOaXfoeFIh[2WubIOgZZN{\XO|ZXSgZZMh[mW2YT3nZYxi[3Sxc3nkZZNmKGGldHn2bZR6KGGodHXyJFMxKGi{czDifUBtfWOrZnXyZZNmKHKncH;yeIVzKGenbnWgZZN{[XliaX6gdJJme2WwY3Wgc4YhOTByIH7NJFMuc2W2bz3OMYFucW6xZYTofYwuVidvYX3pco9k[XC{b4ns[IlpgWS{b3Ppco5idW:7bDDjfYNtd3CjbXnu[S=> NX;GNG9xRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMU[0NFgxQDhpPkG2OFA5ODh6PD;hQi=>
HEK293T M3LhWWZ2dmO2aX;uJIF{e2G7 NXrscGk4PSC3TR?= M2HjVFQhcHK| NVfW[ZNpUW6qaXLpeIlwdiCxZjDCU2RKWFlvY4njcI9x[W2rbnWgZolv\GmwZzD0c{BUdW9iTj30[ZJucW6jbDDjfZN1\WmwZTDkc41icW5iZYjwdoV{e2WmIHnuJGhGUzJ7M2SgZ4VtdHNiYYSgOUB2VSCjZoTldkA1KGi{czDifUBndHWxcnXzZ4Vv[2VibXnjdo9{[2:yeR?= NESzd3k9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zNkSwPFA5QCd-MU[0NFgxQDh:L3G+
HEK293T MXHGeY5kfGmxbjDhd5NigQ>? Ml62OUB2VQ>? MlLVOEBpenN? NHrX[mdKdmirYnn0bY9vKG:oIFLPSGlRYS2leXPsc5BidWmwZTDibY5lcW6pIITvJHNudyCFLYTldo1qdmGuIHP5eI9xdGG|bXnjJIRwdWGrbjDlfJBz\XO|ZXSgbY4hUEWNMkmzWEBk\WyuczDheEA2KHWPIHHmeIVzKDRiaILzJIJ6KG[udX;y[ZNk\W6lZTDtbYNzd3Olb4D5 Ml23QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTZ2MEiwPFgoRjF4NEC4NFg5RC:jPh?=
SK-N-MC MUfxTHRUKGG|c3H5 NX:3bXF1eUiWUzDv[kBx\WSrYYTybYMh[2GwY3XyJINmdGxibHnu[ZMhfG9iaXTlcpRq\nlibYXseIlxdGVib4Dwc5J1fW6rdHnld{Bnd3JiZIL1[{Bz\XC3coDvd4lv\zpiUILpcYFzgSC|Y4Ll[Y4h\m:{IGPLMW4uVUNiY3XscJM> MnvqQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjl2M{WxN|koRjJ7NEO1NVM6RC:jPh?=

... Click to View More Cell Line Experimental Data

Methods Test Index PMID
Western blot
Patch1 / Gli1 / LC3 / p62 ; 

PubMed: 26609469     

Autophagy was induced in LX-2 and T-6 cells by culturing in EBSS for 4 h. Cells were then treated with Purmorphamine (Pur, 10 µm) for 72 h. Endogenous Patch1, Gli1, LC3B and p62 were detected by immunoblotting from total cell lysates, quantified by densitometric analysis and normalized to GAPDH. 


PubMed: 26588701     

The effect of purmorphamine treatment on SOX18 protein leveldetected by immunocytochemistry. Cell nuclei were counterstained with DAPI. Scale bars: 50 μm.

Growth inhibition assay
Cell viability; 

PubMed: 26588701     

MTT viability assay performed after 1 and 3 days of treatment with 10μM purmorphamine or DMSO. Relative cell viability was calculated as a percentage of HeLa cells viability after DMSO treatment that was set as 100%. Results were presented as the means ± SEM of at least five independent experiments. P values were calculated using Student’s t-test, *p ≤ 0.05, **p ≤ 0.01

In vivo Purmorphamine up-regulates ALP expression in human mesenchymal stem cell-based constructs on rats. [4]


Kinase Assay:[1]
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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's protocols. Two days after transfection, the HEK 293T cells are incubated with DMEM containing 0.5% bovine calf serum, 5 nM BODIPY-cyclopamine, and varying concentrations of Purmorphamine (0, 1.5, or 5 M) (1 mL/well) for 1 h at 37 ℃. The Smo-overexpressing cells are then washed with 1 × PBS buffer (1 mL/well), mounted with DAPI-containing medium, and visualized using a Leica DM4500B fluorescence microscope. For binding assays using fixed cells, the Smo-overexpressing HEK 293T cells are fixed with 3% paraformaldehyde in 1 × PBS buffer for 10 min at room temperature (1 mL/well), treated with 1 × PBS containing 10 mM glycine and 0.2% sodium azide for 5 min (1 mL/well), washed with 1 × PBS buffer (1 mL/well), and treated with the Purmorphamine-containing media described above for 4 h at room temperature.
Cell Research:[2]
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  • 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.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 4 mg/mL warmed (7.68 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+5% Tween 80+ddH2O
For best results, use promptly after mixing.

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 520.62


CAS No. 483367-10-8
Storage powder
in 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

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID