| 名稱 | Pictilisib |
| 描述 | Pictilisib (GDC-0941) (GDC-0941) is a potent pan inhibitor of class I catalytic subunits of PI3K (IC50s: 3/33/3/75 nM for p110α/β/δ/γ). |
| 細(xì)胞實(shí)驗(yàn) | All drug treatments were tested in quadruplicate during a 4-day incubation period, and the relative number of viable cells was estimated using CellTiter-Glo. Total luminescence was measured on a Wallac Multilabel Reader. Cells were treated simultaneously with docetaxel (dose range = 0.0003–0.020 μmol/L) or GDC-0941 (dose range = 0.083–5 μmol/L) in an 8 × 10 matrix of concentrations chosen to encompass clinically relevant doses (24). The concentration of drug resulting in EC50 was determined using Prism software. Combination synergy of GDC-0941 and docetaxel was determined by Bliss independence analyses. A Bliss expectation for a combined response (C) was calculated by the equation: C = (A + B) ? (A × B) where A and B are the fractional growth inhibitions of drug A and B at a given dose. The difference between the Bliss expectation and the observed growth inhibition of the combination of drugs A and B at the same dose is the 'Delta.Bliss.' Delta.Bliss scores were summed across the dose matrix to generate a Bliss sum. Bliss sum = 0 indicates that the combination treatment is additive (as expected for independent pathway effects); Bliss sum > 0 indicates activity greater than additive (synergy); and Bliss sum < 0 indicates the combination is less than additive (antagonism). Statistical analysis comparing the Bliss sums for each cell line was conducted by the Student t-test [2]. |
| 激酶實(shí)驗(yàn) | Recombinant human PI3Kα, PI3Kβ, and PI3Kδ are coexpressed in a Sf9 baculovirus system with the p85α regulatory subunit and purified as GST-fusion proteins using affinity chromatography on glutathione-sepharose. Recombinant human PI3Kγ is expressed as monomeric GST-fusions and purified similarly. GDC-0941 is dissolved in DMSO and added to 20 mM Tris-HCl (pH 7.5) containing 200 μg yttrium silicate (Ysi) polylysine SPA beads, 4 mM MgCl2, 1 mM dithiothreitol (DTT), 1 μM ATP, 0.125 μCi [γ-33P]-ATP, and 4% (v/v) DMSO in a total volume of 50 μL. The recombinant GST-fusion of PI3Kα (5 ng), PI3Kβ (5 ng), PI3Kδ (5 ng), or PI3Kγ (5 ng) is added to the assay mixture to initiate the kinase reaction. After incubation for 1 hour at room temperature, the kinase reaction is terminated with 150 μL PBS. The mixture is then centrifuged for 2 minutes at 2000 rpm and read using a Wallac Microbeta counter. The reported IC50 values are calculated using a sigmoidal, dose-response curve fit in MDL Assay Explorer [1]. |
| 動(dòng)物實(shí)驗(yàn) | Female nu/nu mice were inoculated subcutaneously with MCF7-neo/HER2 or MX-1 breast cancer cells. When tumors reached a mean volume of 200 to 250 mm3, animals were size-matched and distributed into groups consisting of 10 animals per group. Docetaxel formulated in 3% EtOH, 97% saline was administered intravenously once weekly. GDC-0941, formulated in MCT (0.5% methylcellulose, 0.2% Tween-80) was dosed orally and daily. MAXF1162 is a HER2+/ER+/PR+ patient-derived breast cancer tumor xenograft model established by directly implanting tumors subcutaneously from patient to NMRI nu/nu mice. Tumor volume was calculated as follows: tumor size (mm3) = (longer measurement × shorter measurement2) × 0.5. Tumor sizes were recorded twice weekly over the course of a study. Following data analysis, P values were determined using the Dunnett t test. For pharmacodynamic studies, tumor samples (n = 4) were immediately frozen or fixed in 10% neutral-buffered formalin. Tumors were dissociated in cell extraction buffer, and lysates were analyzed by Western blotting as described above. Immunohistochemistry was conducted using 5-μm paraffin sections of formalin-fixed tissue on a Ventana Benchmark XT instrument by deparaffinization, treatment with antigen retrieval buffer, and incubation with anti-cleaved caspase-3 primary antibody at 37°C. Bound antibody was detected using DABMap technology, and sections were counterstained with hematoxylin [2]. |
| 體外活性 | Pictilisib是這些細(xì)胞系中細(xì)胞增殖的強(qiáng)效抑制劑，具有亞μM級(jí)的IC50。在U87MG、PC3和MDA-MB-361細(xì)胞中觀察到對(duì)Akt (Ser473) 磷酸化的強(qiáng)效抑制，其IC50分別為46、37和28 nM [1]。與單一化合物治療相比，Pictilisib和多西他賽聯(lián)合使用在體外測(cè)試的乳腺癌細(xì)胞系中減少了80%以上的腫瘤細(xì)胞存活率。在MDA-MB-453細(xì)胞系中計(jì)算出的Bliss和為0，表明了添加效應(yīng)的組合效果，而其他腫瘤細(xì)胞系計(jì)算出的Bliss和>0，表明了協(xié)同效應(yīng) [2]。使用250 nM Pictilisib處理2小時(shí)，在所有測(cè)試的細(xì)胞系中pAKT的抑制率為40%-85%。Pictilisib通過(guò)劑量依賴性降低細(xì)胞增殖/存活率來(lái)抑制PI3K/AKT途徑。Pictilisib抑制了曲妥珠單抗敏感和不敏感細(xì)胞的生長(zhǎng)。Pictilisib的IC50值在150到950 nM之間，與曲妥珠單抗的敏感性無(wú)關(guān) [3]。 |
| 體內(nèi)活性 | 給攜帶MCF7-neo/HER2乳腺癌異種移植瘤的動(dòng)物以7.5 mg/kg 的docetaxel或150 mg/kg的Pictilisib治療，分別導(dǎo)致腫瘤生長(zhǎng)延遲和腫瘤停滯。100 mg/kg的Pictilisib與docetaxel的聯(lián)合使用在治療期間導(dǎo)致腫瘤停滯，并在停藥后持續(xù)維持[2]。AZD8055 (20mg/kg)或Pictilisib (75mg/kg)的給藥引起血糖水平的短暫上升。無(wú)論是AZD8055還是Pictilisib的治療均顯著抑制了Akt的活性及其Thr308和Ser473的磷酸化。AZD8055或GDC-941還抑制了Akt底物PRAS40和Foxo-1/3a的磷酸化[4]。 |
| 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 : 41 mg/mL (79.8 mM)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | Autophagy | Inhibitor | Apoptosis | inhibit | Phosphoinositide 3-kinase | RG 7321 | GDC 0941 | Pictilisib | GDC0941 | PI3K | RG-7321 |
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