| 名稱 | AZ960 |
| 描述 | AZ960 is an effective ATP competitive JAK2 inhibitor (IC50/Ki: <3 nM and0.45 nM). |
| 細胞實驗 | Cellular proliferation is evaluated using the fluorometric/colorimetric BIOSOURCE AlamarBlue Assay and read in the Spectra Max Gemini EM microplate reader. SET-2 cells are plated at 20,000 cells/well, TEL-JAK2 Ba/F3 cells at 2000 cells/well, and all other TEL-JAKs at 5000 cells/well in 96-well plates. Cells are treated with AZ 960 24 hours after plating and grown for 72 hours for SET-2 and 48 hours for TEL-JAK Ba/F3 cells. Following the indicated growth period Alamar Blue (10 μL/well) is added, cells are incubated at 37 °C in 5% CO2 for 2 hours, and fluorescence is measured at 545 (excitation) and 600 nm (emission). Data are normalized to percent of the control, and GI50 values (the concentration that causes 50% growth inhibition) are calculated using Xlfit4 version 4.2.2 for Microsoft Excel. (Only for Reference) |
| 激酶實驗 | Enzyme Biochemical Assay and Kinase Profiling: Inhibition studies of AZ 960 are performed using a recombinant JAK2 kinase (amino acids 808–1132) at a peptide (Tyk2 peptide) concentration of 100 nM and an ATP concentration of 15 μM. Concentrations of AZ 960 ranging from 0.003 μM to 30 μM are used. The mode of inhibition and inhibition constant (Ki) of AZ960 against JAK2 kinase are further evaluated by inhibition kinetics. Specifically, a series of JAK2-catalyzed reactions are set up in HEPES buffer (75 mM, pH 7.3) with a fixed concentration of peptide (FL-Ahx-IPTSPITTTYFFFKKK-COOH), and varied concentrations of ATP and AZ 960. The progress of each reaction is subsequently monitored by the Caliper LC3000 system, and the initial velocity of each reaction is extracted from the corresponding reaction time course. To define the mode of inhibition, initial velocities are plotted against corresponding ATP concentrations using Lineweaver-Burk plots and the characteristic convergence of the lines on the y axis demonstrates the competitiveness of AZ 960 to ATP. Initial inspection of Ki using the Michealis-Menten equation revealed that AZ960 is a tight-binding inhibitor of JAK2. AZ960 is profiled against 83 kinases at three inhibitor concentrations (0.01 μM, 0.10 μM, and 1.0 μM). |
| 體內活性 | AZ960（0.1 μM）對83 種激酶中的31種也有抑制作用,如 JAK3(IC50：9 nM),Aurora,TrkA,和ARK5。AZ 960明顯抑制 TEL-JAK2,-JAK1,-JAK3和-Tyk2細胞增殖（GI50：25/230/279/214 nM）。AZ 960還可降低 STAT3/5磷酸化水平,從而有效抑制SET-2 細胞增殖（GI50：~33 nM）。AZ 960可抑制TEL-JAK2細胞中STAT5磷酸化（IC50：15 nM）,對TEL-JAK2驅動的STAT5磷酸化比對其他JAK激酶家族成員(TEL-JAK1, -JAK3和 -TYK2)驅動的細胞系選擇性高15到30倍。AZ 960還使人類 T-細胞淋巴嗜淋巴細胞病毒1型感染的T細胞生長停滯和凋亡,且通過小干擾RNA增強AZ 960對MT-1細胞的抗增殖效果,從而抑制Bcl-xL。AZ 960顯著抑制患者新分離的AML細胞克隆性生長,并誘導細胞凋亡。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | 10% DMSO+40% PEG300+5% Tween 80+45% Saline : 10 mg/mL (28.22 mM), Suspension.
Ethanol : 3 mg/mL (8.47 mM), Sonication is recommended.
10% DMSO+90% Saline : < 10 mg/mL (28.22 mM), Lower concentrations may be soluble, but exact solubility limit is unknown.
H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 250 mg/mL (705.5 mM), Sonication is recommended.
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| 關鍵字 | VirusProtease | Virus Protease | Parasite | Janus kinase | JAK2 | JAK | Inhibitor | inhibit | AZ-960 | AZ960 | Apoptosis |
| 相關產品 | Formamide | 2-Amino-2-methyl-1-propanol | Urea | Aceglutamide | Fenpyroximate | Alginic acid | Doxycycline | Metronidazole | Hydroxychloroquine | Stavudine | Tamoxifen | Thymidine |
| 相關庫 | 抑制劑庫 | 血管生成庫 | 經典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 抗病毒庫 | 抗寄生蟲庫 | 免疫/炎癥分子化合物庫 | 膜蛋白靶向化合物庫 | 酪氨酸激酶分子庫 |