A8228-S | Purmorphamine [483367-10-8] Clinisciences

Purmorphamine is the first small-molecule agonist developed for the protein Smoothened [1]. Purmorphamine activates the Hedgehog (Hh) signaling pathway, resulting in the up- and downregulation of its downstream target genes, including Gli1 and Patched. Purmorphamine may ultimately be useful in the treatment of bone-related disease andneurodegenerative disease [2].

In vitro: Purmorphamine activated the Hedgehog pathway by directly bound and activated Smoothened with the IC50value of ~ 1.5 μM in competing with cyclopamine, a Smo antagonist [1]. In multi-potent C3H10T1/2 cells, purmorphamine was a potent inducer of osteogenesis. In C3H10T1/2 cells, The EC50 values assessed based on ALP expression for purmorphamine was 1 μM. Purmorphamine (1 μM) in combination with BMP-4 (100 ng/mL) increased ALP activity more than 90-fold in 3T3-L1 cells [3].Administration of purmorphamine at 3 and 5 μM up-regulated the level of osteocalcin on day 14 (P ≤ 0.05)[4].

In vivo: In rats transplanted with stem cell-based constructs subcutaneously, treatment with purmorphamine tended to up-regulate ALP transcripts when compared with those injected by either dexamethasone or injection water (P ≤ 0.05) [4].

References:
[1]. Sinha S, Chen J K. Purmorphamine activates the Hedgehog pathway by targeting Smoothened[J]. Nature chemical biology, 2006, 2(1): 29.
[2]. Wu X, Walker J, Zhang J, et al. Purmorphamine induces osteogenesis by activation of the hedgehog signaling pathway[J]. Chemistry & biology, 2004, 11(9): 1229-1238.
[3]. Wu X, Ding S, Ding Q, et al. A small molecule with osteogenesis-inducing activity in multipotent mesenchymal progenitor cells[J]. Journal of the American Chemical Society, 2002, 124(49): 14520-14521.
[4]. Faghihi F, Eslaminejad M B, Nekookar A, et al. The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells[J]. Biomedicine & Pharmacotherapy, 2013, 67(1): 31-38.

1. Xiu D, Cheng M, et al. "Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling." Exp Biol Med (Maywood). 2020;245(3):213–220. PMID:31903775

Physical Appearance	A solid
Storage	Store at -20°C
M.Wt	520.62
Cas No.	483367-10-8
Formula	C₃₁H₃₂N₆O₂
Solubility	insoluble in H2O; ≥1.82 mg/mL in EtOH with ultrasonic; ≥8.68 mg/mL in DMSO
Chemical Name	9-cyclohexyl-N-(4-morpholin-4-ylphenyl)-2-naphthalen-1-yloxypurin-6-amine
SDF	Download SDF
Canonical SMILES	C1CCC(CC1)N2C=NC3=C2N=C(N=C3NC4=CC=C(C=C4)N5CCOCC5)OC6=CC=CC7=CC=CC=C76
Shipping Condition	Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips	We do not recommend long-term storage for the solution, please use it up soon.

Cell experiment [1]:
Cell lines	Mesenchymal stem cells (MSCs)
Preparation method	The solubility of this compound in DMSO is
Reaction Conditions	14 d; 2 μM
Applications	Modulation of Hh signaling by purmorphamine in hMSCs was evaluated at 7 and 14 days through the gene expression of the membrane receptors SMO and PTCH1, and the transcriptional factors GLI1 and GLI2. Gene expression of SMO was up-regulated at 7 days (P≤0.05) and down-regulated at 14 days (P≤0.05) by purmorphamine. PTCH1 expression was increased by purmorphamine at 7 days (P≤0.05) and not affected at 14 days (P≤0.05). Purmorphamine up-regulated the expression of GLI1 and GLI2 at 7 (P≤ 0.05) and 14 days (P≤0.05).
Animal experiment [2]:
Animal models	Normal male Wistar rats
Dosage form	5 μM; s.c.
Applications	Subcutaneous transplantation of human mesenchymal stem cell-based constructs into rats. According to the histology sections, labeled cells were present inside the scaffolds. Based on real-time PCR results, it has been shown the up-regulation of human osteoblast genes, ALP, osteocalcin, Runx-2, and collagen I in transplanted cell constructs.
Other notes	Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.
References: [1] Oliveira F S, Bellesini L S, Defino H L A, et al. Hedgehog signaling and osteoblast gene expression are regulated by purmorphamine in human mesenchymal stem cells[J]. Journal of cellular biochemistry, 2012, 113(1): 204-208. [2]Faghihi F, Baghaban Eslaminejad M, Nekookar A, et al. The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells[J]. Biomedicine & Pharmacotherapy, 2013, 67(1): 31-38.

Description	Purmorphamine is a blocker of BODIPY-cyclopamine binding to Smo with IC50 of ~ 1.5 μM and also is an inducer of osteoblast differentiation with EC50 of 1 μM.
Targets	Smoothened
IC50	~ 1.5 μM