Purmorphamine|483367-10-8|安捷凯

Background

Purmorphamine is the first small molecule agonist developed for Smoothened protein. Purmorphamine directly binds and activates Smoothened, blocking the binding of BODIPY-cyclopamine to Smo. Purmorphamine activates the Hedgehog(Hh) signaling pathway, resulting in up-regulation and down-regulation of its downstream target genes, including Gli1 and Patched Purmorphamine can be used to treat bone-related diseases and neurodegenerative diseases[1-2].

Purmorphamine(2 μM;4 days) induces osteogenesis in pluripotent mesenchymal progenitor cells[3]. Purmorphamine enhances osteogenic activity of human osteoblasts derived from bone marrow mesenchymal cells, Purmorphamine(1, 2 or 3 μM;21days) did not affect cell proliferation or viability, but increased ALP activity and bone-like nodule formation[4]. Purmorphamine(2 μM ) had a promotive effect on adhesion, proliferation and osteogenic differentiation of human dental pulp stem cells (hDPSCs) cultured on β-TCP[5]. Purmorphamine substantially suppressed proliferation, enhanced and accelerated alkaline phosphatase activity and calcium sedimentation and increased the expression of osteopontin and osteocalcin in rat mesenchymal stem cells[6].

Purmorphamine(10 mg/kg; i.p.) prevents Hypoxic-Ischemic(HI)-induced brain injury. Purmorphamine exerted long-term protective effects upon tissue loss and improved neurobehavioral outcomes as determined at 14 and 28 days post-HI insult[7].Purmorphamine(1mg/kg; i.p.) activated Sonic hedgehog (SHH) signal to attenuate LPS-induced inflammatory response in mice[8].

参考文献:
[1]. Sinha S, Chen JK. Purmorphamine activates the Hedgehog pathway by targeting Smoothened. Nat Chem Biol. 2006 Jan;2(1):29-30. doi: 10.1038/nchembio753. Epub 2005 Nov 20. PMID: 16408088.
[2]. Faghihi F, Baghaban Eslaminejad M, et,al.The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. Biomed Pharmacother. 2013 Feb;67(1):31-8. doi: 10.1016/j.biopha.2012.10.004. Epub 2012 Nov 16. PMID: 23228449.
[3]. Wu X, Walker J, et,al.Purmorphamine induces osteogenesis by activation of the hedgehog signaling pathway. Chem Biol. 2004 Sep;11(9):1229-38. doi: 10.1016/j.chembiol.2004.06.010. PMID: 15380183.
[4]. Beloti MM, Bellesini LS, et,al. Purmorphamine enhances osteogenic activity of human osteoblasts derived from bone marrow mesenchymal cells. Cell Biol Int. 2005 Jul;29(7):537-41. doi: 10.1016/j.cellbi.2005.02.007. PMID: 15979909.
[5]. Rezia Rad M, Khojaste M, et,al. Purmorphamine increased adhesion, proliferation and expression of osteoblast phenotype markers of human dental pulp stem cells cultured on beta-tricalcium phosphate. Biomed Pharmacother. 2016 Aug;82:432-8. doi: 10.1016/j.biopha.2016.05.016. Epub 2016 Jun 1. PMID: 27470382.
[6]. W?ltje M, B?bel M, et,al. Purmorphamine and oxysterols accelerate and promote osteogenic differentiation of mesenchymal stem cells in vitro. In Vivo. 2015 Mar-Apr;29(2):247-54. PMID: 25792653.
[7]. Liu D, Bai X, et,al. Purmorphamine Attenuates Neuro-Inflammation and Synaptic Impairments After Hypoxic-Ischemic Injury in Neonatal Mice via Shh Signaling. Front Pharmacol. 2020 Mar 4;11:204. doi: 10.3389/fphar.2020.00204. PMID: 32194421; PMCID: PMC7064623.
[8]. Shao S, Wang GL, et,al.Activation of Sonic hedgehog signal by Purmorphamine, in a mouse model of Parkinson's disease, protects dopaminergic neurons and attenuates inflammatory response by mediating PI3K/AKt signaling pathway. Mol Med Rep. 2017 Aug;16(2):1269-1277. doi: 10.3892/mmr.2017.6751. Epub 2017 Jun 9. PMID: 28627590; PMCID: PMC5562000.

Purmorphamine是第一个针对Smoothened蛋白开发的小分子激动剂, 能够直接结合并激活Smo,阻断BODIPY-cyclopamine与Smo结合。 Purmorphamine激活Hedgehog(Hh)信号通路,导致其下游靶基因Gli1和Patched上调或下调,Purmorphamine可用于治疗骨相关疾病和神经变性疾病[1-2]。

Purmorphamine(2 μM;4 days) 可以在多能性间充质祖细胞中诱导成骨作用[3]。Purmorphamine可以增强来源于骨髓间充质细胞的人类成骨细胞的成骨活性,Purmorphamine(1, 2 or 3 μM;21days)不影响细胞增殖或存活,但可以增加碱性磷酸酶(ALP)活性和骨瘤结节形成 [4]。Purmorphamine(2 μM) 可以促进人类牙髓干细胞(hDPSCs)在β-TCP上的附着、增殖和成骨分化作用[5]。Purmorphamine可以大幅抑制大鼠间充质干细胞的增殖,同时增强并加速碱性磷酸酶活性和钙沉淀,并增加骨蛋白和骨钙素的表达[6]。

Purmorphamine(10 mg/kg;i.p.) 可以预防缺氧缺血性脑损伤(HI)。Purmorphamine对组织损失产生长期保护效应,并在HI刺激后的14天和28天改善了神经行为结果[7]。 Purmorphamine(1mg/kg;i.p.) 激活Sonic Hedgehog (SHH)信号,在小鼠中减轻了LPS引起的炎症反应[8]。

Protocol

Kinase experiment^[1]:

Preparation method

Smo binding experiments were performed using BODIPY-Cyclopamine and cells overexpressing Smo.

Two days after transfection of HEK 293T cells, the cells were incubated at 37 ℃for 1 hour with DMEM medium containing BODIPY-Cyclopamine and varying concentrations of Purmorphamine (0, 1.5, or 5μM) (1 mL per well).

The overexpressed Smo cells were then washed with 1xPBS buffer (1 mL per well), placed in a medium containing DAPI, observed using fluorescence microscopy, and binding detection was performed using fixed cells.

Smo-overexpressing HEK293T cells were fixed at room temperature with 3% paraformaldehyde dissolved in 1 xPBS buffer (1 mL per well) for 10 minutes, then treated with 1 xPBS buffer (1 mL per well) containing 10 mm of glycerine and 0.2% sodium azide for 5 minutes with 1 xPBS buffer (1 ml per well).

Cells were washed and then treated with a medium containing Purmorphamine at room temperature for 4 hours.

Reaction Conditions

0, 1.5, or 5μM Purmorphamine

Applications

Purmorphamine directly binds and activates Smoothened, blocking BODIPY-cyclopamine binding to Smo, with an IC₅₀ of about 1.5 μM in HEK293T cells.

Cell experiment ^[2]:

Cell lines

C3H10T1/2 cells

Preparation method

Cells were treated with 2 μM of purmorphamine or 0.5% DMSO control for 4 days.

Reaction Conditions

2 μM purmorphamine;4 days

Applications

Purmorphamine induces osteogenesis in pluripotent mesenchymal progenitor cells.

Animal experiment ^[3]:

Animal models

Male C57BL/6J mouse ( HI injury)

Dosage form

10 mg/kg purmorphamine, i.p.

Applications

Purmorphamine prevents Hypoxic-Ischemic(HI)-induced brain injury.

参考文献:

[1]. Sinha S, Chen JK. Purmorphamine activates the Hedgehog pathway by targeting Smoothened. Nat Chem Biol. 2006 Jan;2(1):29-30. doi: 10.1038/nchembio753. Epub 2005 Nov 20. PMID: 16408088.

[2].Wu X, Walker J,et,al. Purmorphamine induces osteogenesis by activation of the hedgehog signaling pathway. Chem Biol. 2004 Sep;11(9):1229-38. doi: 10.1016/j.chembiol.2004.06.010. PMID: 15380183.

[3]. Liu D, Bai X, et,al. Purmorphamine Attenuates Neuro-Inflammation and Synaptic Impairments After Hypoxic-Ischemic Injury in Neonatal Mice via Shh Signaling. Front Pharmacol. 2020 Mar 4;11:204. doi: 10.3389/fphar.2020.00204. PMID: 32194421; PMCID: PMC7064623.

安捷凯在线客服

扫一扫加微信

信号通路

生命科学

有机化学

无机化学

材料科学

Purmorphamine

Purmorphamine详情

Background

Protocol

安捷凯在线客服