辛伐他汀作用机制 - Medchemexpress - MCE中国

1、Product Data SheetSimvastatinCat. No.: HY-17502CAS No.: 79902-63-9分式: CHO分量: 418.57作靶点: HMG-CoA Reductase (HMGCR); Autophagy; Mitophagy; Apoptosis; Ferroptosis作通路: Metabolic Enzyme/Protease; Autophagy; Apoptosis储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 Ethanol :

2、 100 mg/mL (238.91 mM; Need ultrasonic)DMSO : 50 mg/mL (119.45 mM)H2O : 0.1 mg/mL (insoluble)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 2.3891 mL 11.9454 mL 23.8909 mL5 mM 0.4778 mL 2.3891 mL 4.7782 mL10 mM 0.2389 mL 1.1945 mL 2.3891 mL请根据产品在不同溶剂中的溶解度选择合

3、适的溶剂配制储备液；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。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。以下溶解案都请先按照 In Vitro 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 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶

4、剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.97 mM); Clear solution此案可获得 2.5 mg/mL (5.97 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Page 1 o

5、f 2 www.MedChemESolubility: 2.5 mg/mL (5.97 mM); Suspended solution; Need ultrasonic此案可获得 2.5 mg/mL (5.97 mM) 的均匀悬浊液，悬浊液可于服和腹腔注射。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄均匀。DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.97 mM); Clear solution此案可获得 2.5 mg/mL

6、 (5.97 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。4. 请依序添加每种溶剂： 10% EtOH 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (5.97 mM); Clear solution此案可获得 2.5 mg/mL (5.97 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 EtOH 储备液加到 900 L 20% 的 SBE-CD 理

7、盐溶液中，混合均匀。5. 请依序添加每种溶剂： 10% EtOH 90% corn oilSolubility: 2.5 mg/mL (5.97 mM); Clear solution此案可获得 2.5 mg/mL (5.97 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 EtOH 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Simvastatin种竞争性的 HMG-CoA reductase 抑制剂，Ki 值为 0.2 nM。IC & Target Ki: 0

8、.2 nM (HMG-CoA reductase)1体外研究 Simvastatin needs to be activated by NaOH in EtOH treatment before use for cell assay (Activation of Simvastatin: 2mg of Simvastatin in 50 L of ethanol and 75 L of 0.1 N NaOH, incubated at 50C for 2 hours. The pH is adjusted to7.2 with HCl). Simvastatin suppresses chol

9、esterol synthesis in mouse L-M cell, rat H4II E cell, and human Hep G2 cellwith IC50s of 19.3 nM, 13.3 nM and 15.6 nM, respectively1. Simvastatin causes a dose-dependent increase in serine473 phosphorylation of Akt within 30 minutes, with maximal phosphorylation occurring at 1.0 M. Simvastatin (1.0

10、M) enhances phosphorylation of the endogenous Akt substrate endothelial nitric oxide synthase (eNOS), inhibitsserum-free media undergo apoptosis and accelerates vascular structure formation2. Simvastatin shows anti-inammatory effects, reduces anti-CD3/anti-CD28 antibody-stimulated proliferation of P

11、B-derived mononuclear cellsand synovial uid cells from rheumatoid arthritis blood, as well as IFN- release at 10 M. Simvastatin (10 M) alsoblocks cell-mediated macrophage TNF- release induced via cognate interactions by appr 30%3. Simvastatin (5 M)significantly reduces the expression of ABCA1 in ast

12、rocytes and neuroblastoma cells, the expression of apolipoproteinE in astrocytes, and increases cyclin-dependent kinase 5 and glycogen synthase kinase 3 expression in SK-N-SH cells7.体内研究 Simvastatin suppresses the conversion of radiolabeled acetate to cholesterol with an IC50 of 0.2 mg/kg via p.o.ad

13、ministration1. Simvastatin (4 mg/day, p.o. for 13 weeks) returns the cholesterol-induced increases in totalcholesterol, LDL-cholesterol and HDL-cholesterol to normal level in rabbits fed an atherogenci cholesterol-rich diet4.Simvastatin (6 mg/kg) increases LDL receptor-dependent binding and the numb

14、er of hepatic LDL receptors in rabbitsfed a diet containing 0.25% cholesterol5. Simvastatin affects inflammation independent of its effect on plasmacholesterol level. Simvastatin (20 mg/kg/day) causes a 1.3-fold less macrophage content in lesions, and 2-fold lessvascular cell adhesion molecule-1, in

15、terleukin-1beta, and tissue factor expression, companied by a 2.1-fold increasesin lesional smooth muscle cell and collagen content in cynomolgus monkeys fed an atherogenic diet6.Page 2 of 3 www.MedChemEPROTOCOLKinase Assay 3 For assessment of Akt protein kinase activity in vitro, substrate (2 g his

16、tone H2B or 25 g eNOS peptide) isincubated with Akt immunoprecipitated from cell lysate using goat polyclonal anti-Akt1 antibody. Kinase reactionsare initiated following the addition of Simvastatin to a final concentration of ATP (50 M) containing 10 Ci of 32P-ATP, dithiotreitol (1 mM), HEPES buffer

17、 (20 mM, pH 7.4), MnCl2 (10 mM), MgCl2 (10 mM). After incubation for 30 minat 30C, phosphorylated histone H2B is visualized after SDS(15%) and autoradiography. To estimate the extentof 32P incorporation into eNOS peptides, each reaction mixture is measured by spotting onto phosphocellulose discfilte

18、r and the amount of phosphate incorporated is measured by Cerenkov counting. The wild-type peptide sequenceis 1174-RIRTQSFSLQERHLRGAVPWA-1194, and the mutant eNOS peptide is identical except that serine 1179 issubstituted by alanine3.MCE has not independently confirmed the accuracy of these methods.

19、 They are for reference only.Cell Assay 7 Before use, Simvastatin needs to be converted to open acid form by reconstitution of 50mg in 1mL ethanol, additionof 0.813 mL 1N NaOH, and pH adjustment to 7.2 with 1N HCl9. Primary human astrocytes from four different donorsare prepared from tissue obtained

20、 from legally aborted fetuses. Primary human astrocytes and SK-N-SHneuroblastoma cell line (ATCC) are plated on 6-well plates and grown in DMEM supplemented with 5% or 8% fetalcalf serum, respectively, at 37C, 5% CO2 until 80% confluent. For measurement of gene expression levels at baseline,cells ar

21、e just washed and RNA is prepared and assayed as outlined below. Baseline gene expression levels inastrocytes are measured in primary human astrocytes obtained from two donors. For experimental purposes, cells areincubated under serum-free conditions. Primary human astrocytes are obtained from four

22、donors. Based onpreliminary time-dependent studies, 48-h incubation is used for all of experiments. Based on the dose-responsestudies, a majority of our experiments are conducted using the following concentrations of active compounds:Simvastatin at 5 M, pravastatin at 10 M, mevalonate at 50 M, and G

23、GPP and FPP at 10 M. Following incubation,cells are extensively washed to remove dead cells and cell debris and prepared for further analyses7.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Monkeys1Administration 1 Thirty-nine adult male cynomol

24、gus monkeys are initially fed a moderately atherogenic diet containing 0.28 mgcholesterol per calorie of diet, with 16.7% from protein, 45.1% from lipids, and 38.1% from carbohydrates. Afterconsuming the atherogenic diet for 3 months, the monkeys are divided into 3 groups (n=13 each) that are equiva

25、lentin their total plasma cholesterol (TPC), LDL-C, and HDL cholesterol (HDL-C) concentrations; these groups consumethe atherogenic diet and receive statin (or control) treatment for an additional 15 months. Control monkeys are fedthe atherogenic diet with no added statins. Prava-treated monkeys hav

26、e 40 mg Prava/kg body wt per day added tothe atherogenic diet. Simvastatin-treated monkeys consumed 20 mg Simvastatin/kg body wt per day1.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 Biomaterials. 2020 Apr 17;250:119963. Clin Cancer Res.

27、2020 Jan 14. pii: clincanres.2884.2019. Oxid Med Cell Longev. 2017;2017:3861914. J Biol Chem. 2018 Sep 14;293(37):14328-14341. Cell Cycle. 2019 Oct 10:1-14.See more customer validations on HYPERLINK www.MedChemE www.MedChemEPage 3 of 4 www.MedChemEREFERENCES1. Slater, E.E., et al. Mechanism of actio

28、n and biological profile of HMG CoA reductase inhibitors. A new therapeutic alternative. Drugs, 1988. 36 Suppl 3: p.72-82.2. Kureishi, Y., et al. The HMG-CoA reductase inhibitor simvastatin activates the protein kinase Akt and promotes angiogenesis in normocholesterolemicanimals. Nat Med, 2000. 6(9)

29、: p. 1004-10.3. Leung BP, et al. A novel anti-inflammatory role for simvastatin in inflammatory arthritis. J Immunol. 2003 Feb 1;170(3):1524-30.4. Kobayashi M, et al. Preventive effect of MK-733 (simvastatin), an inhibitor of HMG-CoA reductase, on hypercholesterolemia and atherosclerosis inducedby cholesterol feeding in rabbits. Jpn J Pharmacol. 1989 Jan;49(1):125-33.5. Ishida F, et al. Comparative effects of simvastatin (MK-733) and pravastatin (CS-514) on hypercholesterolemia induced by cholesterol feeding in rabbits.Biochim Biophys Acta. 1990 Feb 23;1042(3):365-73.6. S