| 名稱 | Enzastaurin |
| 描述 | Enzastaurin (LY317615) (LY317615) is an effective PKCβ selective inhibitor (IC50: 6 nM), 6- to 20-fold selectivity against PKCα/γ/ε. |
| 細(xì)胞實(shí)驗(yàn) | Induction of apoptosis by enzastaurin is measured by nucleosomal fragmentation and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) and staining in HCT116 and U87 mg cell lines. Briefly, 5 × 103 cells are plated per well in 96-well plates (1% FBS-supplemented media conditions), incubated with or without Enzastaurin for 48 to 72 hours. The absorbance values are normalized to those from control-treated cells to derive a nucleosomal enrichment factor at all concentrations as per the manufacturer's protocol. The concentrations studied ranges from 0.1 to 10 μM. In situ TUNEL staining is assayed with the In situ Cell Death Detection, Fluorescein kit. Cells (7.5 ×104) are plated per well in 6-well plates and incubated 72 hours in 1% FBS-supplemented media Enzastaurin. Fluorescein-labeled DNA strand breaks are detected with the BD epics flow cytometer. Ten thousand, single-cell, FITC-staining events are collected for each test. (Only for Reference) |
| 激酶實(shí)驗(yàn) | Kinase inhibition assays: The inhibition of PKCβII, PKCα, PKCε, or PKCγ activity by enzastaurin is determined using a filter plate assay format measuring 33P incorporation into myelin basic protein substrate. Reactions are done in 100 μL reaction volumes in 96-well polystyrene plates with final conditions as follows: 90 mM HEPES (pH 7.5), 0.001% Triton X-100, 4% DMSO, 5 mM MgCl2, 100 μM CaCl2, 0.1 mg/mL phosphatidylserine, 5 μg/mL diacetyl glyerol, 30 μM ATP, 0.005 μCi/μL 33ATP, 0.25 mg/mL myelin basic protein, serial dilutions of enzastaurin (1-2,000 nM), and recombinant human PKCβII, PKCα, PKCε, or PKCγ enzymes (390, 169, 719, or 128 pM, respectively). Reactions are started by addition of the enzyme and incubated at room temperature for 60 minutes. They are then quenched with 10% H3PO4, transferred to multiscreen anionic phosphocellulose 96-well filter plates, incubated for 30 to 90 minutes, filtered and washed with 4 volumes of 0.5% H3PO4 on a vacuum manifold. Scintillation cocktail is added and plates are read on a Microbeta scintillation counter. IC50 values are determined by fitting a three-variable logistic equation to the 10-point dose-response data using ActivityBase 4.0. |
| 體外活性 | Enzastaurin 在所有研究的MM細(xì)胞系中，包括MM.1S、MM.1R、RPMI 8226 (RPMI)、RPMI-Dox40 (Dox40)、NCI-H929、KMS-11、OPM-2和U266，均顯示出顯著的劑量依賴性生長(zhǎng)抑制作用，IC50范圍為0.6-1.6 μM。Enzastaurin 直接作用于人類腫瘤細(xì)胞，誘導(dǎo)凋亡并抑制培養(yǎng)腫瘤細(xì)胞的增殖。Enzastaurin 還抑制了GSK3βser9、核糖體蛋白S6S240/244和AKTThr308的磷酸化，但對(duì)VEGFR磷酸化無(wú)直接影響。[1] Enzastaurin 提高了CTCL惡性淋巴細(xì)胞的凋亡率。與GSK3抑制劑聯(lián)合使用時(shí)，enzastaurin 顯示出提高的細(xì)胞毒性水平。使用enzastaurin與GSK3抑制劑AR-A014418的組合處理，增加了β-catenin總蛋白水平和β-catenin介導(dǎo)的轉(zhuǎn)錄。阻斷β-catenin介導(dǎo)的轉(zhuǎn)錄或小發(fā)夾RNA (shRNA)敲減β-catenin產(chǎn)生了與enzastaurin加AR-A014418相同的細(xì)胞毒作用。此外，enzastaurin和AR-A014418的治療降低了CD44的mRNA水平和表面表達(dá)。[2] |
| 體內(nèi)活性 | 將異種移植瘤用Enzastaurin和放射線聯(lián)合處理，比單獨(dú)使用任一治療能更大幅度降低微血管密度。微血管密度的減少與腫瘤生長(zhǎng)延遲相對(duì)應(yīng)。[3] |
| 存儲(chǔ)條件 | store at low temperature | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 10.3 mg/mL (20 mM)
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| 關(guān)鍵字 | Apoptosis | LY-317615 | Autophagy | PKC | Protein kinase C | LY 317615 | inhibit | Enzastaurin | Inhibitor |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | L-Glutamic acid | Gefitinib | Hydroxychloroquine | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | L-Ascorbic acid | Paeonol | Sodium 4-phenylbutyrate |
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