A66

生物活性产品描述 A66是一种有效的，特异性的p110α抑制剂，IC50为32 nM，作用于p110α比作用于其他I型PI3K亚型选择性高100倍以上。 靶点 p110α [1] PI4Kβ [1] C2β [1] p110δ [1] p110γ [1] View More IC50 32 nM 236 nM 462 nM >1.25 μM 3.48 μM 体外研究 In addition to the wild-type p110α, A66 also potently inhibits the oncogenic forms of p110α such as p110α E545K and p110α H1047R with IC50 of 30 nM and 43 nM, respectively. Unlike PIK-75, A66 displays >100 fold selectivity for p110α over other class-I PI3K isoforms. Among the class-II PI3Ks, class-III PI3K and PI4Ks, A66 only exhibits limited cross-reactivity with the class-II PI3K PI3KC2β and the PI4Kβ isoform of PI4K with IC50 of 462 nM and 236 nM, respectively. A66 exhibits no inhibitory activity against other lipid kinases or the related kinases DNA-PK and mTOR. A66 has a higher degree of specificity compared with PIK-75 when tested at 10 μM against two large panels of 110 protein kinases and 318 kinases. Inhibition of p110α alone by A66 treatment is sufficient to block insulin signalling to Akt/PKB in certain cell lines that harbor H1047R mutations in PIK3CA and have high levels of p110α and class-Ia PI3K activity. [1] A66 treatment at 0.7 μM induces a 75-80% reduction in focus formation by the highly transforming p85α iSH2 mutants KS459delN, DKRMN-S560del, and K379E, and reduces the phosphorylation of Akt on T308 by all p85 mutants. [2] 体内研究 A single dose of A66 at 100 mg/kg induces a profound reduction in the phosphorylation of Akt/PKB and p70 S6 kinase, but not of ERK, in SK-OV-3 tumour tissue in vivo at both 1 hour and 6 hours after dosing. A66 dosed at 100 mg/kg once daily (QD) for 21 days or 75 mg/kg twice daily (BID) for 16 days induces a significant delay in growth of SK-OV-3 xenografted tumors with average TGI of 45.9% and 29.9%, respectively, which is even greater than that induced by the well-established pan-PI3K inhibitor BEZ-235. QD dosing of A66 in the HCT-116 xenograft model also induces a significant reduction in tumour volume with TGI of 77.2%, but causes a non-significant reduction in tumor volume in the U87MG xenograft model. [1] Administration of A66 at 10 mg/kg in male CD1 mice induces significant impairments in the ITT (insulin tolerance test) and GTT (glucose tolerance test), and an increase in glucose production during a PTT (pyruvate tolerance test), almost to the same level as the pan-PI3K inhibitors. [3] 特征 A66 is highly selective for p110α isoform. 推荐的实验操作(此推荐来自于公开的文献所以Selleck并不保证其有效性) 激酶实验: [1] PI3K (human) HTRF Assay The p110α isoform is produced in-house by co-expressing full-length human p85α with the indicated human full-length catalytic subunit containing a histidine tag at the N-terminus to allow purification. The p110α is titrated and used at a concentration between EC65-EC80 values. IC50 value is evaluated using the PI3K (human) HTRF Assay. 动物实验: [1] 动物模型 Age-matched specific pathogen-free Rag1-/- or NIH-III mice inoculated subcutaneously with U87MG, SK-OV-3 or HCT-116 cells 配制 Formulated in 20% 2-hydroxypropyl-β-cyclodextrin in water 剂量 100 mg/kg once daily (QD) or 75 mg/kg twice daily (BID) 给药处理 Intraperitoneal injection 溶解度 15% Captisol 8 mg/mL * 溶解度检测是由Selleck技术部门检测的，可能会和文献中提供的溶解度有所差异，这是由于生产工艺和批次不同产生的正常现象。 不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南） 小鼠 大鼠 兔 豚鼠 仓鼠 狗 猴 狒狒 重量 (kg) 0.02 0.15 1.8 0.4 0.08 10 3 12 体表面积 (m2) 0.007 0.025 0.15 0.05 0.02 0.5 0.24 0.6 Km 系数 3 6 12 8 5 20 12 20 动物A (mg/kg) = 动物B (mg/kg) × 动物B的Km系数 动物A的Km系数 例如，依据体表面积折算法，将白藜芦醇用于小鼠的剂量22.4 mg/kg 换算成大鼠的剂量，需要将22.4 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到白藜芦醇用于大鼠的等效剂量为11.2 mg/kg。 大鼠剂量 (mg/kg) = 小鼠剂量 (22.4 mg/kg) × 小鼠的Km系数(3) = 11.2 mg/kg 大鼠的Km系数(6) 1 参考文献 [1] Jamieson S, et al. Biochem J, 2011, 438(1), 53-62. [2] Sun M, et al. Proc Natl Acad Sci U S A, 2010, 107(35), 15547-15552. view more [3] Smith GC, et al. Biochem J, 2012, 442(1), 161-169. 化学数据点击下载A66 (SDF格式文件) 分子量 393.53 化学式 C17H23N5O2S2 CAS号 1166227-08-2 稳定性 3年 -20℃粉状 6个月-80℃溶于溶剂 别名 N/A 溶解性 (25°C) * 体外 DMSO 79 mg/mL (200.74 mM) 乙醇 1 mg/mL (2.54 mM) 水 <1 mg/mL (<1 mM) 体内 15% Captisol 8 mg/mL * <1 mg/ml 指产品微溶或不溶 * 溶解度检测是由Selleck技术部门检测的，可能会和文献中提供的溶解度有所差异，这是由于生产工艺和批次不同产生的正常现象。 . Chemical Name (2S)-N1-(5-(2-tert-butylthiazol-4-yl)-4-methylthiazol-2-yl)pyrrolidine-1,2-dicarboxamide 制备储备液 1 mg 5 mg 10 mg 1 mM 2.5411 mL 12.7055 mL 25.4110 mL 5 mM 0.5082 mL 2.5411 mL 5.0822 mL 10 mM 0.2541 mL 1.2706 mL 2.5411 mL 50 mM 0.0508 mL 0.2541 mL 0.5082 mL 摩尔浓度计算器 稀释计算器 分子量计算器 Research Area 客户使用Selleck产品的实验数据(3) 点击放大 Effects of PI3K inhibitors on lipid kinase activity and pAkt abundance. (C) Western blot analyses of pAkt in MMTV-MT tumorderived cells treated with various PI3K inhibitors (in micromolar) as indicated following growth factor starvation. The bar graphs show the relative amounts of pAkt(T308) 6 SD (n = 6). (D) Western blot analyses of pAkt in MMTV-NeuT tumor-derived cells treated with various PI3K inhibitors (in micromolar) as indicated. The bar graphs show the relative amounts of pAkt(T308) 6 SD (n = 6). (*) P < 0.01; (**) P < 0.001 (Student’s t-test). 评级 数据来源于 Genes Dev, 2012, 26, 1573–1586. A66购于Selleck 方法 Western Blot 细胞系 MMEC cells 浓度 0.1-1uM 处理时间 结果 As expected, both pan-PI3K and p110a-selective inhibitors dramatically reduced lipid kinase activity and pAkt in MT and NeuT tumor cells (Fig. C–D).