| 名稱 | 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. |
| 細(xì)胞實(shí)驗(yàn) | 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) |
| 激酶實(shí)驗(yàn) | 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). |
| 體外活性 | 在H1975(非小細(xì)胞肺癌, 突變 EGFR [L858R, T790M])移植瘤模型中,以PKI-587（25 mg/kg）處理7天,實(shí)驗(yàn)處理組存活率可達(dá)90%.而PKI-587（25 mg/kg,i.v.）會(huì)使裸鼠產(chǎn)生高容量分布(7.2 L/kg),低血漿清除力(7(mL/min)/kg)和較長(zhǎng)半衰期(14.4 h).在MDA-361移植瘤模型中,PKI-587具有較好的抗腫瘤效果,最低有效劑量為3 mg/kg,最大耐受劑量為30 mg/kg. |
| 體內(nèi)活性 | 在MDA-361和PC3-MM2 細(xì)胞系中,PKI-587對(duì)腫瘤細(xì)胞生長(zhǎng)有抑制作用（IC50：4/13.1 nM）。PKI-587對(duì)PI3Kα突變形式也有效,尤其是H1047R和E545K（IC50：0.6/0.6 nM）。 |
| 存儲(chǔ)條件 | 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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| 關(guān)鍵字 | mTOR | PKI587 | Gedatolisib | Mammalian target of Rapamycin | PKI 587 | Inhibitor | PI3K | Phosphoinositide 3-kinase | PF05212384 | inhibit | PF 05212384 |
| 相關(guān)產(chǎn)品 | (2S,3R,4S)-4-Hydroxyisoleucine | Quercetin | Capivasertib | Rapamycin | Quercetin Dihydrate | Apilimod | L-Leucine | Myricetin | Duvelisib (R enantiomer) hydrochloride | Isoprenaline hydrochloride | Erucic acid | Sapanisertib |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 抗癌活性化合物庫(kù) | 經(jīng)典已知活性庫(kù) | 已知活性化合物庫(kù) | 激酶抑制劑庫(kù) | 神經(jīng)再生化合物庫(kù) | 抗衰老化合物庫(kù) | 藥物功能重定位化合物庫(kù) | 抗癌臨床化合物庫(kù) | 抗癌藥物庫(kù) |