环孢霉素A作用机制 - Medchemexpress - MCE中国

1、Product Data SheetCyclosporin ACat. No.: HY-B0579CAS No.: 59865-13-3分式: CHNO分量: 1202.61作靶点: Complement System作通路: Immunology/Inflammation储存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性数据体外实验 DMSO : 62.5 mg/mL (51.97 mM; Need ultrasonic)H2O : 0.1 mg/mL

2、(insoluble)SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 0.8315 mL 4.1576 mL 8.3152 mL5 mM 0.1663 mL 0.8315 mL 1.6630 mL10 mM 0.0832 mL 0.4158 mL 0.8315 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；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 (protect from light)。-80C 储存时，请在 6 个内使，-20C 储存时

3、，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. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.08 mg/mL (1.73 mM); Suspended solution; Need

4、 ultrasonic此案可获得 2.08 mg/mL (1.73 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 20.8 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)Solubility: 2.08 mg/mL (1.73 mM); Suspended solution; Need ultrasonic此案可获得 2.08 m

5、g/mL (1.73 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 20.8 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合Page 1 of 2 www.MedChemE均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.08 mg/mL (1.73 mM); Clear solution此案可获得 2.08 mg/mL (1.73 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 20.8 m

6、g/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Cyclosporin A种免疫抑制剂，能与亲环素结合，抑制 calcineurin 活性的IC50 值为 7 nM3。Cyclosporin A 也抑制CD11a/CD18 粘附分8。IC & Target IC50: 7 nM (calcineurin)体外研究 Cyclosporin A is able to bind with the cyclophilin in T cells1. Cyclosporin A works by forming a Cyclophilin-C

7、yclosporin A complex to inhibit calcineurin2. Cyclosporin A inhibits calcineurin in stimulated cells with an IC50 valueof 7 nM3. Cyclosporin A suppresses the nuclear translocation of NF-AT4. Cyclosporin A shows an effect onmitochondria via preventing the MTP from opening with an IC50 of 39 nM5.体内研究

8、Cyclosporin A has immunosuppressive activity, and is active via parenteral and p.o. administration in mice, rat andguinea pigs6. Cyclosporin A can be used in organ transplantation to prevent rejection7.PROTOCOLKinase Assay 3 Reaction mixtures with purified enzyme contains 100 nM calcineurin, 100 nM

9、calmodulin, and 5 M 32P-labeledphosphopeptide, in 60 L (total volume) of assay buffer containing 20 mM Tris (pH 8), 100 mM NaCl, 6 mM MgCl2,0.5 mM dithiothreitol, 0.1 mg of bovine serum albumin per mL, and either 0.1 mM CaCl2 or 5 mM EGTA. Reactionmixtures with cell lysates contains 20 L of undilute

10、d lysate, 5 M 32P-labeled phosphopeptide, and 40 L of assaybuffer. Reaction mixtures contains 50 M peptide 412 or 413 and/or 500 nM okadaic acid, a specific inhibitor ofphosphatases 1 and 2A; 500 nM okadaic acid is sufficient for inhibition of Ca2+-independent phosphatases, whereashigher concentrati

11、ons partially inhibit Ca2+-dependent activity as well. After 15 min at 30C, reactions are terminatedby the addition of 0.5 mL of 100 mM potassium phosphate buffer (pH 7.0) containing 5% trichloroacetic acid. Freeinorganic phosphate is isolated by Dowex cation-exchange chromatography and quantitated

12、by scintillation countingas described.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 3 Immunosuppressive agents are dissolved in ethanol at concentrations 1000-fold more than the concentration desiredfor cell treatments. Cells (106) are susp

13、ended in 1 mL of complete medium in microcentrifuge tubes; 1 L of ethanolor of the ethanolic solution of Cyclosporin A is added, and the cells are incubated at 37C for 1 hr. Cells are washedtwice with 1 mL of PBS on ice and lysed in 50 L of hypotonic buffer containing 50 mM Tris (pH 7.5); 0.1 mM EGT

14、A; 1mM EDTA; 0.5 mM dithiothreitol; and 50 g of phenylmethylsulfonyl fluoride, 50 g of soybean trypsin inhibitor, 5 gof leupeptin, and 5 g of aprotinin per mL. Lysates are subjected to three cycles of freezing in liquid nitrogenfollowed by thawing at 30C and then are centrifuged at 4C for 10 min at

15、12,000g.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Rats are immunized on day 0 i.p. with 0 5 mL and mice i.v. with 0 2 mL of a 10% suspension of washed sheepAdministration 6 erythrocytes (SE). To elicit a secondary response, mice are boosted

16、 8-11 weeks after the primary immunization withan i.v. injection of 0-2 mL of 0 25% washed SE (107 cells). For prolonged treatment the animals receive on the averagePage 2 of 3 www.MedChemE45 mg/kg cyclosporin A daily in the food during 13 weeks. These rats are immunized 5 days before killing.MCE ha

17、s not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Environ Sci Technol. 2017 Dec 5;51(23):13938-13948. Cell Death Dis. 2018 May 22;9(6):598. Neurobiol Dis. 2019 Sep;129:44-55. Int J Biol Macromol. 2020 Feb 7. pii: S0141-8130(20)30191-4. J Immunol. 20

18、19 Dec 23. pii: ji1901065.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Handschumacher RE, et al. Cyclophilin: a specific cytosolic binding protein for cyclosporin A. Science. 1984 Nov 2;226(4674):544-7.2. Liu J, et al. Calcineurin is a common target of cyclophilin

19、-cyclosporin A and FKBP-FK506 complexes. Cell. 1991 Aug 23;66(4):807-15.3. Fruman DA, et al. Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. Proc Natl Acad Sci U S A. 1992 May1;89(9):3686-90.4. Flanagan WM, et al. Nuclear association of a T-cell transcript

20、ion factor blocked by FK-506 and cyclosporin A. Nature. 1991 Aug 29;352(6338):803-7.5. Nicolli A, et al. Interactions of cyclophilin with the mitochondrial inner membrane and regulation of the permeability transition pore, and cyclosporin A-sensitive channel. J Biol Chem. 1996 Jan 26;271(4):2185-92.6. Borel JF, et al. Effects of the new anti-lymphocytic peptide cyclosporin A in animals. Immunology. 1977