雷公藤甲素作用機制 - Medchemexpress - MCE中國

1、Product Data SheetTriptolideCat. No.: HY-32735CAS No.: 38748-32-2分式: CHO分量: 360.4作靶點: NF-B; Apoptosis作通路: NF-B; Apoptosis儲存式: 4C, stored under nitrogen* In solvent : -80C, 6 months; -20C, 1 month (stored under nitrogen)溶解性數(shù)據(jù)體外實驗 DMSO : 33 mg/mL (91.56 mM)* means soluble, but saturation unknown.Solve

2、ntMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 2.7747 mL 13.8735 mL 27.7469 mL5 mM 0.5549 mL 2.7747 mL 5.5494 mL10 mM 0.2775 mL 1.3873 mL 2.7747 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month (stored under nitrogen)。-80C 儲存時，請在 6 個內(nèi)使，-20C 儲存時，請在 1 個內(nèi)使。體

3、內(nèi)實驗請根據(jù)您的實驗動物和給藥式選擇適當?shù)娜芙獍?。以下溶解案都請先按?In Vitro 式配制澄清的儲備液，再依次添加助溶劑：為保證實驗結(jié)果的可靠性，澄 的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 1.17 mg/mL (3.25 mM); Clear solution此案可獲得 1.17 mg/mL (3.2

4、5 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 11.7 mg/mL 的澄 DMSO 儲備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 1.17 mg/mL (3.25 mM); Clear solution此案可獲得 1.17 mg/mL (3.25 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 11.7 mg/mL 的澄

5、 DMSO 儲備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合Page 1 of 2 www.MedChemE均勻。3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 1.17 mg/mL (3.25 mM); Clear solution此案可獲得 1.17 mg/mL (3.25 mM，飽和度未知) 的澄 溶液，此案不適于實驗周 期在半個以上的實驗。以 1 mL 作液為例，取 100 L 11.7 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Triptolide抑

6、制劑。從雷公藤根中提取的萜類三環(huán)氧化物，具有免疫抑制，抗炎和抗增殖作。 雷公藤內(nèi)酯是NF-B 活化的IC & Target HSP90 MDM-2/p5347-73 nM (IC50)體外研究 Triptolide induces apoptosis in cultured and primary Chronic Lymphocytic Leukemia (CLL) B-cells. Treatment of CD19+ B cells with Triptolide, induces a dose-dependent increase in apoptosis in cultured and

7、 primary CLL cells. Triptolideis selectively toxic to both high risk (n=5) and low risk CLL (n=12) B cells (10 to 50 nM range) while largely sparingnormal B-cells (n=5). Consistent with the inhibition of heat-shock induced HSP transcription, treatment with Triptolideattenuates heat-shock induced exp

8、ression of HSPs1. Triptolide is a natural product derived from the Chinese plantTripterygium wilfordii, is reported to exhibit antitumor effects in a broad range of cancers. Triptolide inhibits MDM2expression in a dose-dependent manner, even at low concentrations spanning 20-100 nM in acute lymphobl

9、asticleukemia (ALL) cells. Triptolide exhibits strongly cytotoxic activity in all 8 cell lines having native MDM2overexpression, with IC50 values range from 47 to 73 nM. Triptolide exhibits much less cytotoxic effect on EU-4 cellsthat express very low level of MDM2, while it effectively kill these c

10、ells when MDM2 is stably transfected (IC50 values:725 nM vs. 88 nM)2. Differentiated PC12 cells are incubated with different concentrations of Triptolide (0.01, 0.1, and1 nM) in the presence of 10 M A25-35 for 24 hours and MTT assay is used to detect the effect of Triptolide. Theresults show that A2

11、5-35 can decrease the cell viability and when treated with Triptolide the viability of differentiatedPC12 cells is significantly increased. The results indicate that Triptolide can alleviate cellular damage caused by A25-35, which means that Triptolide has a neuroprotective effect3.體內(nèi)研究 The Triptoli

12、de (TP) plasma concentrations are declined rapidly in mice after receive an intravenous dose. After 2h ofinjection, the Triptolide concentrations are dropped below the lower limit of quantification for all three groups. Acomparison of the parameters is made between the control and the treated groups

13、 to assess the effect of P-gpinhibition on the Triptolide exposure and elimination. Treatment with the mdr1a-siRNA can significantly enhance theTriptolide plasma exposure, with the Cmax increases from 41374 to 51094 ng/mL (P0.05) and the AUC from103.59.6 to 154.330.2 ngh/mL (P0.05). In the concomita

14、nt group with Tariquidar, the significantly increased AUCis also noted, from 103.59.6 of the control to 145.924.6 ngh/mL of the Triptolide+Tariquidar group (P0.05).Accordingly, the total body clearance of Triptolide in mice is remarkably decreased, from 95641024.2 mL/min/kg ofthe control to 6576.414

15、38.5 (P0.05) and 5755.41200.1 mL/min/kg (P0.05) for Triptolide+Tariquidar andTriptolide+mdr1a-siRNA groups, respectively4.PROTOCOLCell Assay 3 The viability of differentiated PC12 cells treated with different concentrations of Triptolide. After differentiated PC12cells are cultured on 96-well plates

16、 with RPMI 1640 medium for stabilization, differentiated PC12 cells are incubatedwith different concentrations of Triptolide (0.01, 0.1, and 1 nM) for 24 hours. The concentrations in this study arechosen. Then cell viability is determined by the MTT assay. Each condition and experiment is repeated t

17、hree times3.Page 2 of 3 www.MedChemEMCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice4Administration 4 Male BALB/C mice (weight, 18-22 g) are used. For Triptolide (TP) plasma kinetic study and toxicological evaluation,mice are divided into fou

18、r groups (n=5 each) to collect blood and tissue samples: (1) normal+saline group; (2) 1.0mg/kg Triptolide+15 nmol negative control (NC) siRNA-siRNA group; (3) 1.0 mg/kg Triptolide+15 nmol mdr1a-siRNAgroup; (4) 1.0 mg/kg Triptolide+10 mg/kg Tariquidar group. In order to avoid the complication caused

19、by drugabsorption or possible intestinal first-pass effect, Triptolide and the inhibitor are intravenously administrated to mice.The siRNA group is intravenously injected with NC-siRNA or mdr1a-siRNA 2 days before Triptolide dose. ForTriptolide+Tariquidar group, the mice are received an intravenous

20、Tariquidar dose 20 min prior to the Triptolideinjection. Blood samples are collected at 2, 5, 10, 15, 30, 60 and 120 min after Triptolide dosing. To assess the liverexposure of Triptolide, liver tissue samples are collected from another set of mice at 5, 30, 60 and 120 min afterdosing. Three Triptol

21、ide groups are design for this experiment, including Triptolide+NC-siRNA group,Triptolide+mdr1a-siRNA group and Triptolide+Tariquidar group. The liver tissue samples are weighed and thenhomogenized in 10 volume (w:v) of ice-cold saline. The concentrations of Triptolide in plasma and liver tissue are

22、measured by a validated LC-MS/MS method.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻 Blood. 2020 Apr 14. pii: blood.2019003990. Proc Natl Acad Sci U S A. 2020 Apr 20. pii: 201913633. Clin Cancer Res. 2020 Jan 14. pii: clincanres.2884.2019

23、. Cell Rep. 2018 Sep 4;24(10):2553-2560.e5. Cell Death Dis. 2019 Aug 9;10(8):602.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Ganguly S, et al. Targeting HSF1 disrupts HSP90 chaperone function in chronic lymphocytic leukemia. Oncotarget. 2015 Oct 13;6(31):31767-79.2. Huang M, et al. Triptolide inhibits MDM2 and induces apoptosis in acute lymphoblastic leukemia cells through a p53-independent p