| 名稱 | Ruxolitinib |
| 描述 | Ruxolitinib (INCB018424) is a JAK1/2 inhibitor (IC50=3.3/2.8 nM) that is potent and selective. Rixolitinib exhibits antitumor activity and induces autophagy and apoptosis. |
| 細(xì)胞實(shí)驗(yàn) | Cells were seeded at 2000/well of white bottom 96-well plates, treated with compounds from DMSO stocks (0.2% final DMSO concentration), and incubated for 48 hours at 37°C with 5% CO2. Viability was measured by cellular ATP determination using the Cell-Titer Glo luciferase reagent or viable cell counting. Values were transformed to percent inhibition relative to vehicle control, and IC50 curves were fitted according to nonlinear regression analysis of the data using PRISM GraphPad [1]. |
| 激酶實(shí)驗(yàn) | The kinase domains of human JAK1 (837-1142), JAK2 (828-1132), JAK3 (781-1124), and Tyk2 (873-1187) were cloned by PCR with N-terminal epitope tags. Recombinant proteins were expressed using Sf21 cells and baculovirus vectors and purified with affinity chromatography. JAK kinase assays used a homogeneous time-resolved fluorescence assay with the peptide substrate (-EQEDEPEGDYFEWLE). Each enzyme reaction was carried out with test compound or control, JAK enzyme, 500nM peptide, adenosine triphosphate (ATP; 1mM), and 2.0% dimethyl sulfoxide (DMSO) for 1 hour. The 50% inhibitory concentration (IC50) was calculated as the compound concentration required for inhibition of 50% of the fluorescent signal. Biochemical assays for CHK2 and c-MET enzymes were performed using standard conditions (Michaelis constant [Km] ATP) with recombinantly expressed catalytic domains from each protein and synthetic peptide substrates. An additional panel of kinase assays (Abl, Akt1, AurA, AurB, CDC2, CDK2, CDK4, CHK2, c-kit, c-Met, EGFR, EphB4, ERK1, ERK2, FLT-1, HER2, IGF1R, IKKα, IKKβ, JAK2, JAK3, JNK1, Lck, MEK1, p38α, p70S6K, PKA, PKCα, Src, and ZAP70) was performed using standard conditions using 200nM INCB018424. Significant inhibition was defined as more than or equal to 30% (average of duplicate assays) compared with control values [1]. |
| 動(dòng)物實(shí)驗(yàn) | All of the procedures were conducted in accordance with the US Public Health Service Policy on Humane Care and Use of Laboratory Animals. Mice were fed standard rodent chow and provided with water ad libitum. Ba/F3-JAK2V617F cells (10^5 per mouse) were inoculated intravenously into 6- to 8-week-old female BALB/c mice. Survival was monitored daily, and moribund mice were humanely killed and considered deceased at time of death. Treatment with vehicle (5% dimethylacetamide, 0.5% methocellulose) or INCB018424 began within 24 hours of cell inoculation, twice daily by oral gavage. Hematologic parameters were measured using a Bayer Advia120 analyzed, and statistical significance was determined using Dunnett testing [1]. |
| 體外活性 | 方法：Ba/F3-EpoR-JAK2V617F 細(xì)胞用 Ruxolitinib (0-10 μM) 處理 48 h，使用 Cell-Titer Glo 檢測(cè)的細(xì)胞活力。
結(jié)果：Ruxolitinib 劑量依賴性降低細(xì)胞活力，IC50 為 126 nM。[1]
方法：霍奇金淋巴瘤細(xì)胞 HDLM-2 用 Ruxolitinib (10-100 nM) 處理 24 h，使用 Western Blot 方法檢測(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：Ruxolitinib 顯著抑制下游活性 p-STAT3 和 p-STAT5，且呈劑量依賴性，而總 STAT3 和 STAT5 水平保持不變。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Ruxolitinib (3-30 mg/kg，5% dimethyl acetamide, 0.5% methocellulose) 灌胃給藥給攜帶腫瘤 Ba/F3-JAK2V617F 的 BALB/c 小鼠，每天兩次，持續(xù)三周。
結(jié)果：Ruxolitinib 顯著降低了脾腫大和炎癥細(xì)胞因子的循環(huán)水平，并優(yōu)先消除了腫瘤細(xì)胞，從而顯著延長(zhǎng)了生存期，而沒有骨髓抑制或免疫抑制作用。[1]
方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Ruxolitinib (150 mg/kg) 口服給藥給攜帶人結(jié)直腸腫瘤 LS411N 的 BALB/c nude 小鼠，每?jī)商煲淮?，持續(xù)兩周。
結(jié)果：口服 Ruxolitinib 可顯著抑制人結(jié)直腸腫瘤的體內(nèi)生長(zhǎng)，而不會(huì)引起肝毒性。[3] |
| 存儲(chǔ)條件 | Store at low temperature
Pure form: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 242 mg/mL (789.92 mM), Sonication is recommended.
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 5.7 mg/mL (18.61 mM), Solution.
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| 關(guān)鍵字 | TyrosineKinases | Tyrosine Kinases | Tyk2 | Ruxolitinib | Mitophagy | Mitochondrial Autophagy | Janus kinase | JAK2 | JAK1 | JAK | Inhibitor | inhibit | INCB-18424 | INCB18424 | INCB-018424 | INCB 18424 | INCB 018424 | Autophagy | Apoptosis |
| 相關(guān)產(chǎn)品 | Urea | Formamide | Guanidine hydrochloride | Naringin | Aceglutamide | Alginic acid | Dextran sulfate sodium salt (MW 5000) | Hemin | Hydroxychloroquine | Stavudine | Tamoxifen | Paeonol |
| 相關(guān)庫 | 抑制劑庫 | 抗癌上市藥物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | EMA 上市藥物庫 | 抗衰老化合物庫 | FDA 上市藥物庫 | 膜蛋白靶向化合物庫 | 免疫/炎癥分子化合物庫 | 藥物功能重定位化合物庫 | FDA 上市激酶抑制劑庫 | 抗癌臨床化合物庫 |