| 名称 | Gedatolisib |
| 描述 | Gedatolisib (PF-05212384) is a highly effective dual inhibitor targeting the PI3Kα/γ (IC50: 0.4/5.4 nM)and mTOR (IC50: 1.6 nM) in the PI3K/mTOR signaling pathway. |
| 细胞实验 | Cells are plated in 96-well culture plates at about 3000 cells per well. One day following plating, PKI-587 is added to cells. Three days after PKI-587 treatment, viable cell densities are determined by measuring metabolic conversion (by viable cells) of the dye MTS, a previously established cell proliferation assay. For each assay, MTS and PMS stocks are freshly thawed and mixed (MTS/PMS, 20:1). The MTS/PMS mixture is then added to 96-well cell plates at 20 μL/well, and plates are incubated for 1 hour–2 hours in cell culture incubator. MTS assay results are read in a 96-well format plate reader by measuring absorbance at 490 nm. The effect of each PKI-587 treatment is calculated as a percentage of control cell growth obtained from vehicle-treated cells grown in the same culture plate.(Only for Reference) |
| 激酶实验 | PI3K and mTOR kinase assay : Enzyme assays are done in fluorescent polarization (FP) format, adapted from the Echelon K-1100 PI3K FP assay kit protocol. Human class I PI3Ks and PI3K-α mutants (E545K and H1047R) are produced in Sf9 or purchased from Upstate Biotech. GST-GRP1 (murine) is produced in Escherichia coli and isolated by GST-Sepharose. Assay buffers are reaction buffer [20 mM HEPES (pH 7.1), 2 mM MgCl2, 0.05% CHAPS, and 0.01% β-mercaptoethanol] and stop/detection buffer [100 mM HEPES (pH 7.5), 4 mM EDTA, 0.05% CHAPS]. FP reaction is run for 30 minutes at room temperature in 20 μL of reaction buffer containing 20 μM phosphatidylinositol 4,5-bisphosphate (PIP2), 25 μM ATP, and <4% DMSO. FP reaction is stopped with 20 μL of stop/detection buffer (10 nM probe and 40 nM GST-GRP), and after 2 hours, data are collected using an Envision plate reader. The routine assays with purified FLAG-TOR (FL and 3.5) are performed in 96-well plates as follows. Enzymes are first diluted in kinase assay buffer (10 mM Hepes (pH 7.4), 50 mM NaCl, 50 mM β-glycerophosphate, 10 mM MnCl2, 0.5 mM DTT, 0.25 μM microcystin LR, and 100 μg/mL BSA). To each well, 12 μL of the diluted enzyme is mixed briefly with 0.5 μL test inhibitor or control vehicle dimethyl sulfoxide (DMSO). The kinase reaction is initiated by adding 12.5 μL kinase assay buffer containing ATP and His6-S6K to give a final reaction volume of 25 μL containing 800 ng/mL FLAG-TOR, 100 μM ATP, and 1.25 μM His6-S6K. The reaction plate is incubated for 2 hours (linear at 1–6 hours) at room temperature with gentle shaking and then terminated by adding 25 μL Stop buffer (20 mM Hepes (pH 7.4), 20 mM EDTA, and 20 mM EGTA). |
| 体外活性 | 在H1975(非小细胞肺癌, 突变 EGFR [L858R, T790M])移植瘤模型中,以PKI-587（25 mg/kg）处理7天,实验处理组存活率可达90%.而PKI-587（25 mg/kg,i.v.）会使裸鼠产生高容量分布(7.2 L/kg),低血浆清除力(7(mL/min)/kg)和较长半衰期(14.4 h).在MDA-361移植瘤模型中,PKI-587具有较好的抗肿瘤效果,最低有效剂量为3 mg/kg,最大耐受剂量为30 mg/kg. |
| 体内活性 | 在MDA-361和PC3-MM2 细胞系中,PKI-587对肿瘤细胞生长有抑制作用（IC50：4/13.1 nM）。PKI-587对PI3Kα突变形式也有效,尤其是H1047R和E545K（IC50：0.6/0.6 nM）。 |
| 存储条件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : < 1 mg/mL (insoluble）
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| 关键字 | mTOR | PKI587 | Gedatolisib | Mammalian target of Rapamycin | PKI 587 | Inhibitor | PI3K | Phosphoinositide 3-kinase | PF05212384 | inhibit | PF 05212384 |
| 相关产品 | (2S,3R,4S)-4-Hydroxyisoleucine | Quercetin | Capivasertib | Rapamycin | Quercetin Dihydrate | Apilimod | L-Leucine | Myricetin | Duvelisib (R enantiomer) hydrochloride | Isoprenaline hydrochloride | Erucic acid | Sapanisertib |
| 相关库 | 抑制剂库 | 抗癌活性化合物库 | 经典已知活性库 | 已知活性化合物库 | 激酶抑制剂库 | 神经再生化合物库 | 抗衰老化合物库 | 药物功能重定位化合物库 | 抗癌临床化合物库 | 抗癌药物库 |