| 名稱 | Idelalisib |
| 描述 | Idelalisib (GS-1101) is a small molecule inhibitor of the PI3K catalytic subunit p110δ (IC50: 2.5 nM). The selectivity for p110δ is 40- to 300-fold than p110α/β/γ. |
| 細(xì)胞實(shí)驗 | MTT assays were performed to determine cytotoxicity. Briefly, 1 × 10^5 cells (CLL B cells or healthy volunteer T cells or NK cells) were incubated for 48 hours with different concentrations of CAL-101, 25μM LY294002, or vehicle control. MTT reagent was then added, and plates were incubated for an additional 20 hours before washing with protamine sulfate in phosphate-buffered saline. Dimethyl sulfoxide was added, and absorbance was measured by spectrophotometry at 540 nm in a Labsystems plate reader. Cell viability was also measured at various time points with the use of annexin/PI flow cytometry. Data were analyzed with Expo-ADC32 software package. At least 10 000 cells were counted for each sample. Results were expressed as the percentage of total positive cells over untreated control. Experiments examining caspase-dependent apoptosis included the addition of 100μM Z-VAD. Experiments examining survival signals included the addition of 1 μg/mL CD40L, 800 U/mL IL-4, 50 ng/mL BAFF, 20 ng/mL TNF-α, or coculturing on fibronectin or stromal (HS-5 cell line) coated plates. Stromal coculture was done by plating a 75-cm2 flask (80%-100% confluent) per 6-well plate 24 hours before the addition of CLL cells [2]. |
| 激酶實(shí)驗 | PI3K assay was performed on whole-cell lysates from CLL or normal B cells. A PI3K ELISA assay was performed according to the manufacturer's instructions. Briefly, whole-cell extracts were added to a mixture of PI(4,5)P2 substrate and reaction buffer containing adenosine triphosphate (ATP) and allowed to incubate at room temperature. The reaction was stopped by adding PI(3,4,5)P3 detector mixed with EDTA (ethylenediaminetetraacetic acid) and allowed to incubate at room temperature for 1 hour. After this time, the mixture was transferred from each well to a PI3K ELISA plate and allowed to incubate 1 hour. Plates were washed and then incubated with a secondary detector for 30 minutes. Plates were washed again, and 3,3′,5,5′-tetramethylbenzidine solution was added for 5 minutes at which time H2SO4 was added to stop all reactions. Plates were read at 450 nm on a Labsystems 96-well plate reader [2]. |
| 動物實(shí)驗 | For Idelalisib (CAL-101) treatment, wild-type C57BL/6 mice were administered either 40 mg kg 1 CAL-101 or vehicle DMSO, by 25 ml infusion into the femoral vein, 15 min before I/R (pre-treatment), or 3 and 6 h after initiation of reperfusion (post-treatment). Controls and animals treated with CAL-101 underwent cerebral blood flow (CBF) measurements using a laser Doppler perfusion monitor. The CBF measurements obtained immediately before and after MCAO and again at 3 h after reperfusion showed a B90–95% reduction in the blood flow to the MCAO infarct region, which did not differ between groups [4]. |
| 體外活性 | Idelalisib 是一種口服的 p110δ 抑制劑，目前正在對 B 細(xì)胞惡性腫瘤患者進(jìn)行臨床評估。Idelalisib 對 p110δ 的選擇性比對其他 PI3K 類 I 酶（IC50 p110δ = 2.5nM; p110α、p110β 和 p110γ 的 IC50 分別為 820、565 和 89nM）高出 40 至 300 倍。相對于 C2β、hVPS34、DNA-PK 和 mTOR 等相關(guān)激酶，其選擇性更高（400 至 4000 倍），而在 10μM 濃度下對一個包含 402 種多樣激酶的測試組未顯示任何活性[1]。Idelalisib 能夠在劑量和時間依賴的方式下促進(jìn)原發(fā)性 CLL 細(xì)胞的凋亡，這一過程與常見的預(yù)后標(biāo)記物無關(guān)。Idelalisib 引發(fā)的細(xì)胞毒性依賴于半胱天冬酶，并且通過在基質(zhì)細(xì)胞上共培養(yǎng)并未減弱[2]。CAL-101 抑制 CLL 細(xì)胞向 CXCL12 和 CXCL13 的趨化以及在基質(zhì)細(xì)胞下的遷移（偽包裹現(xiàn)象）。Idelalisib 還能下調(diào)在基質(zhì)共培養(yǎng)和 BCR 觸發(fā)后的趨化因子的分泌。Idelalisib 減少了來自 BCR 或像護(hù)士一樣的細(xì)胞的生存信號，并抑制 BCR- 和趨化因子受體誘導(dǎo)的 AKT 和 MAP 激酶（ERK）活化[3]。 |
| 體內(nèi)活性 | 單次靜脈注射40 mg/kg的Idelalisib，于缺血前15分鐘給藥（預(yù)處理），在野生型小鼠中顯著降低了72小時后的梗塞面積。然而，較低劑量elalisib（20、10和1 mg/kg）未能實(shí)現(xiàn)顯著的保護(hù)效果。重要的是，即便在再灌注開始后3小時給藥（治療后），每千克體重40 mg的Idelalisib劑量仍能有效減少梗塞體積，與對照組相比平均降低了44%[4]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : Insoluble
Ethanol : 22 mg/mL (53 mM)
DMSO : 50 mg/mL (120.36 mM)
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| 關(guān)鍵字 | GS 1101 | PI3K | GS1101 | CAL101 | Idelalisib | Phosphoinositide 3-kinase | Autophagy | Inhibitor | inhibit | CAL 101 |
| 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 | 抗癌活性化合物庫 | 已知活性化合物庫 | EMA 上市藥物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | FDA 上市藥物庫 | FDA 上市激酶抑制劑庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |