| 名稱 | Cyclosporin A |
| 描述 | Cyclosporin A is a naturally occurring cyclic polypeptide that is the active metabolite of a fungus. Cyclosporin A is an immunosuppressant that binds to procyclins and inhibits calcineurin (IC50=7 nM). |
| 細(xì)胞實(shí)驗 | Immunosuppressive agents are dissolved in ethanol at concentrations 1000-fold more than the concentration desired for cell treatments. Cells (106) are suspended in 1 ml of complete medium in microcentrifuge tubes; 1 μl of ethanol or of the ethanolic solution of Cyclosporin A is added, and the cells are incubated at 37°C for 1 hr. Cells are washed twice with 1 ml of PBS on ice and lysed in 50μl of hypotonic buffer containing 50 mM Tris (pH 7.5); 0.1 mM EGTA; 1 mM EDTA; 0.5 mM dithiothreitol; and 50 μg of phenylmethylsulfonyl fluoride, 50 μg of soybean trypsin inhibitor, 5 μg of leupeptin, and 5 μg of aprotinin per ml. Lysates are subjected to three cycles of freezing in liquid nitrogen followed by thawing at 30°C and then are centrifuged at 4°C for 10 min at 12,000×g.(Only for Reference) |
| 激酶實(shí)驗 | Phosphatase Assay: Purified bovine brain calcineurin and calmodulin are purchased. Reaction mixtures with purified enzyme contains 100 nM calcineurin, 100 nM calmodulin, and 5 μM 32P-labeled phosphopeptide, in 60 μl (total volume) of assay buffer containing 20 mM Tris (pH 8), 100 mM NaCI, 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. Reaction mixtures with cell lysates contains 20 μl of undiluted lysate, 5 μM 32P-labeled phosphopeptide, and 40 μl of assay buffer. Where indicated, reaction mixtures contains 50 μM peptide 412 or 413 and/or 500 nM okadaic acid, a specific inhibitor of phosphatases 1 and 2A; 500 nM okadaic acid is sufficient for inhibition of Ca2+-independent phosphatases, whereas higher concentrations partially inhibit Ca2+-dependent activity as well. After 15 min at 30°C, reactions are terminated by the addition of 0.5 ml of 100 mM potassium phosphate buffer (pH 7.0) containing 5% trichloroacetic acid. Free inorganic phosphate is isolated by Dowex cation-exchange chromatography and quantitated by scintillation counting as described. |
| 體外活性 | 方法：膠質(zhì)瘤細(xì)胞 C6 用 Cyclosporin A (10 μM) 在缺氧條件下處理 4 h，使用 Western Blot 方法檢測靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：4 h 缺氧誘導(dǎo)了內(nèi)源性 HIF-1α 蛋白的大量積累。Cyclosporin A 阻止了缺氧誘導(dǎo)的 HIF-1α 的大部分穩(wěn)定。[1]
方法：人結(jié)腸癌細(xì)胞 CACO-2 用 Cyclosporin A (2 μM) 處理 24-72 h，使用 Flow Cytometry 檢測細(xì)胞周期情況。
結(jié)果：在用 Cyclosporin A 處理后的 G0/G1 階段檢測到細(xì)胞的積聚。[2] |
| 體內(nèi)活性 | 方法：為檢測體內(nèi)對肌肉疾病的活性，將 Cyclosporin A (5 mg/kg in olive oil) 腹腔注射給肌病 Col6a1-/- 小鼠，每天兩次，持續(xù)十天。
結(jié)果：Cyclosporin A 影響 Col6a1-/- 小鼠的衛(wèi)星細(xì)胞活性并觸發(fā)再生纖維的形成。[3]
方法：為檢測對缺血再灌注損傷 (IRI) 的作用，將 Cyclosporin A (3 mg/kg，在缺血前 1 h 或 10 min；10 mg/kg，在缺血前 10 min) 腹腔注射給缺血再灌注的 C57BL/6J 小鼠。
結(jié)果：Cyclosporin A 10 mg/kg-10 min 和 Cyclosporin A 3 mg/kg-1 h 組的死亡率和腎功能顯著高于 Cyclosporin A 3 mg/kg-10 min 組。[4] |
| 存儲條件 | keep away from moisture | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 60 mg/mL (49.89 mM)
Ethanol : 60.1 mg/mL (50 mM)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 12.03 mg/mL (10 mM), Suspension. Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
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| 關(guān)鍵字 | Phosphatase | CsA | Ciclosporin A | Complement System | Antibiotic | Inhibitor | inhibit | Cyclosporin A |
| 相關(guān)產(chǎn)品 | Cefaclor monohydrate | Stearic acid | Tartaric acid disodium dihydrate | G-418 disulfate | Doxycycline | Neomycin sulfate | Metronidazole | EDTA copper(II) disodium salt | Ampicillin sodium | 3-Phenoxybenzaldehyde | Sulfamethoxazole sodium | Kanamycin sulfate |
| 相關(guān)庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 環(huán)肽庫 | 天然產(chǎn)物庫 | FDA 上市藥物庫 | 藥物功能重定位化合物庫 | 高通量篩選天然產(chǎn)物庫 |