| 名稱 | AZD 6482 |
| 描述 | AZD 6482 (KIN-193) is a potent and selective inhibitor of p110β and PI3Kβ with IC50 values ??of 0.69nM and 10nM, respectively. AZD 6482 is particularly effective in blocking AKT signaling and tumor growth dependent on p110β activation or PTEN loss. |
| 細(xì)胞實(shí)驗(yàn) | For assay of washed platelet aggregation (WPA), the platelet pellets are isolated from human blood and re-suspended to 2 × 1015/L in Tyrodes buffer (TB) containing 1 μM hirudin and 0.02 U/mL apyrase. Then, the platelet suspension is left to rest at room temperature for 30 min. Just prior to time for assay, CaCl2 is added to a final concentration of 2 mM. AZD6482, dissolved in DMSO, is added to a 96-well plate prior to the addition of the washed platelet suspension. The platelet suspension is preincubated with AZD6482 for 5 min. Light absorption at 650 nm is recorded before and after a 5 min plate shake and referred to as recording 0 (R0) and Rl. A mouse anti-human CD9 antibody is added (at a donor specific concentration) to each well prior to next 10 min plate shake and light absorption recording; R2. For data analysis, light absorbance in wells with TB are subtracted from all readings before percent aggregation is calculated according the formula: [(R1-R2)/R1] × 100 = % aggregation. Spontaneous aggregation or pro-aggregatory effect of the inhibitor is evaluated by the same formula, [(R0-Rl)/R0] × 100 = % aggregation. IC50 values are calculated according to the equation, y = {a+[(b-a)/(l+(x/IC50)s)]}, where y = % inhibition; a = 0%; b = 100%; s = the slope of the concentration-response curve; x = AZD6482 concentration. (Only for Reference) |
| 激酶實(shí)驗(yàn) | Assay of PI3K enzyme inhibition: The inhibition of PI3Kβ, PI3Kα, PI3Kγ, and PI3Kδ is evaluated in an AlphaScreen based enzyme activity assay using human recombinant enzymes. The assay measures PI3K-mediated conversion of PIP2 to PIP3. Biotinylated PIP3, a GST-tagged pleckstrin homology (PH) domain and the two AlphaScreen beads form a complex that elicits a signal upon laser excitation at 680 nm. The PIP3 formed in the enzyme reaction competes with the biotinylated PIP3 for binding to the PH domain thus reducing the signal with increasing enzyme product. The AZD6482 is dissolved in DMSO and added to 384 well plates. PBKβ, PBKα, PBKγ, or PBKδ is added in a Tris buffer (50 mM Tris pH 7.6, 0.05% CHAPS, 5 mM DTT, and 24 mM MgCl2) and allowed to preincubate with AZD6482 for 20 minutes prior to the addition of substrate solution containing PIP2 and ATP. The enzyme reaction is stopped after 20 minutes by addition of stop solution containing EDTA and biotin-PIP3, followed by addition of detection solution containing GST-grpl PH and AlphaScreen beads. Plates are left for a minimum of 5 hours in the dark prior to analysis. The final concentration of DMSO, ATP and PIP2 in the assay are, 0.8%, 4 μM, and 40 μM, respectively. IC50 values are calculated according to the equation, y = {a+[(b-a)/(l+(x/IC50)s)]}, where y = % inhibition; a = 0%; b = 100%; s = the slope of the concentration-response curve; x = AZD6482 concentration. |
| 體外活性 | 方法：AZD 6482 (KIN-193) （0.625，1.25，2.5，5，10，20 ， 40 μM，48小時）測定處理 U87 和 U118細(xì)胞，CCK8測定其對 U87 和 U118 細(xì)胞增殖的抑制作用。
結(jié)果：AZD 6482以劑量依賴式方式抑制兩種細(xì)胞系的活力，其中U118 細(xì)胞比 U87 細(xì)胞更敏感，IC50值分別為 7.989 和 9.061μM。[4] |
| 體內(nèi)活性 | 方法：AZD 6482 (KIN-193) （20 mg/kg，腹腔注射，每天兩次）或 GDC-0941（150 mg/kg，口服，每天一次）處理 HCC70、PC3 和 HCC1954 的異種移植腫瘤小鼠，研究KIN-193 對 PTEN 缺陷腫瘤的體內(nèi)效應(yīng)。
結(jié)果：AZD 6482 顯著抑制 HCC70 和 PC3 腫瘤的腫瘤生長，但未能阻斷 HCC1954 腫瘤的生長。[2] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 45 mg/mL (110.17 mM)
1eq. HCl : 4.1 mg/mL (10 mM)
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| 關(guān)鍵字 | Inhibitor | AZD-6482 | inhibit | Autophagy | PI3K | KIN-193 | KIN193 | AZD 6482 | Phosphoinositide 3-kinase |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | Taurine | Gefitinib | Aceglutamide | Hydroxychloroquine | Curcumin | Stavudine | Salicylic acid | Paeonol | 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 | 代謝化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 糖酵解化合物庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |