| 名稱 | Tofacitinib Citrate |
| 描述 | Tofacitinib Citrate (CP-690550 citrate) is a a potent, cell-permeable inhibitor of JAK1/2/3 (IC50s: 1/20/112 nM). |
| 細胞實驗 | Apoptotic cells were detected by flow cytometry using recombinant human Annexin-V conjugated with allophycocyanin. Briefly, after exposure to CP-690,550 for different periods of time, cells were washed in Ca2+-free PBS and resuspended in 100 μL of binding buffer (10 mM HEPES pH 7.4; 0.15 M NaC1; 5 mM KCl; 1 mM MgCl2; 1.8 mM CaCl2) to which Annexin-V-APC had been previously added. Cells were incubated for 20 min in the dark at room temperature, washed and resuspended in 0.3 mL binding buffer. Cells were analyzed on a FACSCalibur flow cytometer equipped with the Cell Quest Pro software [2]. |
| 激酶實驗 | The JAK1, JAK2, and JAK3 kinase assays utilize a protein expressed in baculovirus-infected SF9 cells (a fusion protein of GST and the catalytic domain of human JAK enzyme) purified by affinity chromatography on glutathione sepharose. The substrate for the reaction was polyglutamic acid-tyrosine [PGT (4:1)], coated onto Nunc Maxi Sorp plates at 100 μg/mL overnight at 37 °C. The plates were washed three times, and JAK enzyme was added to the wells, which contained 100 μL of kinase buffer (50 mM HEPES, pH 7.3, 125 mM NaCl, 24 mM MgCl2) + ATP + 1 mM sodium orthovanadate). After incubation at room temperature for 30 min, the plates were washed three times. The level of phosphorylated tyrosine in a given well was determined by standard ELISA assay utilizing an anti-phosphotyrosine antibody [5]. |
| 動物實驗 | Mice received tofacitinib in PEG300 (100 mg/ml) or vehicle alone (PEG300) by osmotic pump infusion (Alzet Model 2004, 0.25 μl/hour, 28 days). Four days prior to immunization, mice were anesthetized and their dorsal surface was shaved. A one cm incision was made on the back to create a subcutaneous pocket and insert the pump. The incision site was closed with wound clips. Mice were injected weekly (i.p.) with SS1P recombinant immunotoxin (RIT; 5 μg/mouse) beginning on day 0; control mice received injections of saline alone. Every week before SS1P or vehicle immunization, ~50 μl of blood was drawn to obtain serum samples. Sera were stored at ?80°C until analyzed [4]. |
| 體外活性 | 雖然Tofacitinib (CP-690,550) 對JAK3的抑制作用極為強大（酶抑制效力為1 nM），但其對JAK2和JAK1的抑制作用分別低20至100倍。CP-690,550能以比對GM-CSF誘導(dǎo)的增殖30倍更強的效力抑制IL-2誘導(dǎo)的增殖。CP-690,550在使用鼠、猴或人細胞的混合淋巴細胞反應(yīng)中展現(xiàn)出強大的抑制效果。與其作用機制一致，這些細胞活性與CP-690,550阻斷IL-2誘導(dǎo)的JAK3及其關(guān)鍵底物STAT5的磷酸化能力相關(guān)[1]。在治療含有人類野生型或V617F JAK2的鼠類因子依賴細胞Patersen-erythropoietin receptor (FDCP-EpoR)時，CP-690,550能抑制細胞增殖，其IC50分別為2.1 μg/ml和0.25 μg/ml。對JAK2(V617F)-陽性PV患者體外擴增的紅細胞祖細胞進行CP-690,550治療，表現(xiàn)出特異的抗增殖（IC50：0.2 μg/ml）和促凋亡活性[2]。Tofacitinib對JAK3的藥理抑制作用與IMA在CML細胞中的抗腫瘤效果協(xié)同增強[3]。 |
| 體內(nèi)活性 | CP-690,550治療顯著延長了與對照組相比的移植物存活時間。12只動物中有4只接受CP-690,550治療（每個劑量組各兩只）存活至研究結(jié)束，且腎功能正常，根據(jù)組織病理學(xué)判斷只有輕度排斥反應(yīng)[1]。用tofacitinib單藥治療小鼠能夠抑制對由細菌蛋白質(zhì)Pseudomonas exotoxin A衍生的免疫毒素以及模型抗原鑰孔血藍蛋白的抗體（Ab）反應(yīng)。實驗顯示，在免疫后21天觀察到對兩種抗原IgG1滴度的千倍降低。Tofacitinib治療還導(dǎo)致CD127+前B細胞數(shù)量減少[4]。 |
| 存儲條件 | store at low temperature,keep away from moisture,keep away from direct sunlight | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 55 mg/mL (109.02 mM)
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| 關(guān)鍵字 | inhibit | CP-690550 Citrate | Influenza Virus | Tofacitinib Citrate | Apoptosis | CP-690550 | CP690550 Citrate | JAK | CP690550 | Janus kinase | CP 690550 | Fungal | CP 690550 Citrate | Tasocitinib Citrate | Tofacitinib | Inhibitor | Tasocitinib | Bacterial |
| 相關(guān)產(chǎn)品 | Dehydroacetic acid sodium | Doxycycline | Methyl anthranilate | Neomycin sulfate | Geraniol | Dimethyl sulfoxide | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Ampicillin sodium | Sulfamethoxazole sodium | Kanamycin sulfate | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 抗真菌庫 | EMA 上市藥物庫 | 激酶抑制劑庫 | FDA 上市藥物庫 | 藥物功能重定位化合物庫 | 酪氨酸激酶分子庫 | 抗癌臨床化合物庫 |