Tripterin-Celastrol-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemETripterinCat. No.: HY-13067CAS No.: 34157-83-0Synonyms: Celastrol分式: CHO分量: 450.61作靶点: Proteasome; Autophagy; Mitophagy作通路: Metabolic Enzyme/Protease; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 mon

2、th溶解性数据体外实验 DMSO : 50 mg/mL (110.96 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.2192 mL 11.0961 mL 22.1921 mL5 mM 0.4438 mL 2.2192 mL 4.4384 mL10 mM 0.2219 mL 1.1096 mL 2.2192 mL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，并请注意储备液的保存式和期限。体内实验 请根据您的实验动物和给药式选择适当的溶解案

3、，配制前请先配制澄的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，当天使；澄的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 5 mg/mL (11.10 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 5 mg/mL (11.10 mM); Clear solution1/3 Master of Sma

4、ll Molecules 您边的抑制剂师www.MedChemEBIOLOGICAL ACTIVITY物活性 Tripterin (Celastrol)种蛋酶体抑制剂，有效且优先抑制20S 蛋酶体的胰凝乳蛋酶样活性，IC50 为2.5 M。IC50 & Target IC50: 2.5 M (20S proteasome) 1体外研究 Tripterin (Celastrol) significantly inhibits the proteasomal chymotrypsin activity in PC-3 cells in aconcentration-dependent manner

5、; at 2.5 M it reaches 55% inhibition, comparable to its potency to apurified 20S proteasome (IC50=2.5 M). Furthermore, increased levels of IB-, Bax, and p27 are observed,three well known target proteins of the proteasome in PC-3 cells treated with Celastrol 1.体内研究 Treatment of PC-3 tumor-bearing nud

6、e mice with Tripterin (Celastrol) (1-3 mg/kg/d, i.p., 1-31 days) results insignificant inhibition (65-93%) of the tumor growth 1. Following treatment with 3 and 6 mg/kg Tripterin(Celastrol), the levels of malondialdehyde (MDA) are significantly decreased by 35.2 and 36.7% (P 2.PROTOCOLKinase Assay 1

7、 A purified rabbit 20S proteasome (0.1 g) is incubated with 40 M of various fluorogenic peptide substratesin 100 L assay buffer (20 mM Tris-HCl ,pH 7.5), in the presence of Celastrol or Oridonin at differentconcentrations or in the solvent DMSO for 2 hours at 37C, followed by measurement of inhibiti

8、on of eachproteasomal activity 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Prostate cancer cells (5,000-8,000) are plated in each well of a 96-well plate and then treated with eitherDMSO, Tripterin (Celastrol), or Oridonin at differen

9、t concentrations for 12 to 16 hours, followed by anadditional 2-hour incubation with Z-Gly-Gly-Leu-AMC (at 40 M). After that, the proteasome activity ismeasured using the whole plate 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 1Adminis

10、tration 12 Male nude immunodeficient mice NCRNU-M, aged 5 weeks, are used. On day 0, human prostate cancer PC-3 or C4-2B cells (5-10106) suspended in 0.1 mL of serum-free RPMI 1640 are inoculated s.c. in the rightflank of each mouse (four mice per group). For the first experiment using PC-3 cells, o

11、n day 14 afterinoculation, the animals started daily i.p. injection with either 50 to 100 L of a vehicle 10% DMSO, 70%Cremophor/ethanol (3:1), and 20% PBS, and 1.0 or 3.0 mg/kg of Tripterin (Celastrol) . Tumor sizes aremeasured daily using calipers and their volumes are calculated using a standard f

12、ormula: width2length/2.Body weight is measured weekly. To study whether the proteasome is inhibited in an early phase of theexperiment, after 3 days of treatment, one control and one 3.0 mg/kg Tripterin (Celastrol) -treated mouse issacrificed. The rest are sacrificed after 16 days of treatment when

13、control tumors reach 1,400 mm3. For thesecond PC-3 tumor experiment, 12 days after inoculation, mice are randomly divided into three groups andtreated with either control, Tripterin (Celastrol) , or Oridonin at 1.5 mg/kg daily for the duration of the study(31 days). In another experiment, to study t

14、he effects of Tripterin (Celastrol) on AR expression, nude mice2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEbearing C4-2B tumors receive daily i.p. injection of the vehicle or 3.0 mg/kg Tripterin (Celastrol) .Rats 2Male Sprague-Dawley (SD) rats (n=90, 6 weeks old), weighing 1619 g, are randomly

15、divided into the control(NC) and the high energy diet (HED) groups. In the control group, the animals receive a standard chow diet,while the rats in the HED group are fed with an additional high energy emulsion. After 8 weeks on theirrespective diets, Streptozotocin (STZ; 45 mg/kg) dissolved in 0.1

16、mol/l citrate buffer (pH 4.5) is injected intothe caudal vein of the rats in the HED group to establish a model of T2DM, while the rats in the control groupare injected with sodium citrate buffer. The rats with blood glucose levels 16.7 mM at 7 days after the STZinjection are selected as the model o

17、f diabetes. On average, 80% of the rats injected with STZ met thesecriteria. At 1 week following the injection of STZ, the rats with successfully-induced diabetes are randomlydivided into the diabetes model (DM) group, the Tripterin (Celastrol) low-dose group (1 mg/kg/day), theTripterin (Celastrol)

18、middle-dose group (3 mg/kg/day) and the Tripterin (Celastrol) high-dose group (6mg/kg/day) (n=15 rats per group). The rats in the treatment groups are administered Tripterin (Celastrol) bygavage, whereas the rats in the NC and DM groups are administered an equal amount of distilled water (2mL). Foll

19、owing 8 weeks of the respective treatments, rats are anesthetized with an intraperitoneal injection ofsodium pentobarbital (30 mg/kg body weight) and tissue samples are collected for analysis. Theparavertebral muscle is excised from the rat bodies, and is cut perpendicularly along the longitudinal a

20、xis andfixed in phosphate-buffered 20% formaldehyde. Histological paraffin-embedded sections (5 m) are thenprepared for H&E staining. The sections of paravertebral muscle are snap-frozen in liquid nitrogen and storedat 80C until further analysis.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Cell Death Dis. 2018 May 22;9(6):601. Life Sci. 2018 Jul 15;205:136-144. RSC Adv. 2016,6, 42537-42544. Mol Biosyst. 2016 Dec 20;13(1):83-91. Harvard Medical School LIN