Purmorphamine is a smoothened/Smo receptor agonist with an EC₅₀ of 1 μM.

IC50: 1.5 μM (Smoothened)

Purmorphamine (10, 20 μM) in combination with sirolimus significantly decreases cell numbers according to the MTT assay. Purmorphamine induces up-regulation of alkaline phosphatase activity and expression of RUNX-2 at day 14. Up-regulation of osteocalcin is detected at the 3 and 5 μM doses of purmorphamine on day 14 post-induction. Matrix mineralization remains unchanged in the presence or absence of purmorphamine^[1]. Purmorphamine induces STAT3 phosphorylation in mouse ES cell line ES14 and mesenchymal stem cell line C3H10T1/2^[2]. Purmorphamine up-regulates the expressionof markers of the osteoblast phenotype-ALP activity and bone-like nodule formationd-in human bonemarrow mesenchymal cells^[3].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

520.62

C₃₁H₃₂N₆O₂

483367-10-8

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : 50 mg/mL (96.04 mM; Need ultrasonic)

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (4.80 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (4.80 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. F. Faghihia, et al. The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. Biomedicine & Pharmacotherapy. 2013, 67(1): 31-38.

  [2]. Gu D, et al. A role for transcription factor STAT3 signaling in oncogene smoothened-driven carcinogenesis. J Biol Chem. 2012 Nov 2;287(45):38356-66.

  [3]. Beloti MM, et al. Purmorphamine enhances osteogenic activity of human osteoblasts derived from bone marrow mesenchymal cells. Cell Biol Int. 2005 Jul;29(7):537-41.

To determine non-toxic doses of the small molecules, 5×10⁵ passaged-3 human MSCs are seeded in each well of a six-well culture plate and incubated in expansion medium (as mentioned above) at 37°Cand 5% CO₂. Two days later, the medium is exchanged with osteogenic medium (OM) that consisted of α-MEM supplemented with 10% FBS, 10 nM dexamethasone, 50 μg/mL ascorbic acid 2-phosphate, and 10 mM beta-glycerol phosphate. This OM is supplemented with different concentrations of purmorphamine (1, 3, 5, 10, and 20 μM) and sirolimus (0.1, 1, 10, 100, and 200 nM). The cultures are maintained for an additional two days and then assessed for the presence of viable cells with the MTT assay, by the addition of MTT solution (5 mg/mL in PBS) to the medium at a ratio of 1:5. Cells are then incubated at 37°C and 5% CO₂. Two hours later, the medium is removed and 500 μL of DMSO is added to the treated cells in order to dissolve the formazone precipitate. The optical absorption rate is read at 570 nm. Cell viability is calculated as percent value relative to the control group which is only treated with OM.

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. F. Faghihia, et al. The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. Biomedicine & Pharmacotherapy. 2013, 67(1): 31-38.

  [2]. Gu D, et al. A role for transcription factor STAT3 signaling in oncogene smoothened-driven carcinogenesis. J Biol Chem. 2012 Nov 2;287(45):38356-66.

  [3]. Beloti MM, et al. Purmorphamine enhances osteogenic activity of human osteoblasts derived from bone marrow mesenchymal cells. Cell Biol Int. 2005 Jul;29(7):537-41.