| 名稱(chēng) | AZD1080 |
| 描述 | AZD1080 is a selective, orally active, brain permeable GSK3 inhibitor. |
| 細(xì)胞實(shí)驗(yàn) | 3T3 fibroblasts are engineered to stably express four-repeat tau protein. These cells have high endogenous levels of GSK3 that is able to phosphorylate tau protein constitutively. This phosphorylation is inhibited by LiCl. After treatment with different compounds, cultures are washed twice with 5 mM MgCl2-PBS. Extracts for Western blot analysis are prepared by homogenizing cells in ice-cold extraction buffer consisting of 20 mM HEPES, pH 7.4, 100 mM NaCl, 10 mM NaF, 1% Triton X-100, 1 mM sodium orthovanadate, 10 mM EDTA, and protease inhibitors (2 mM phenylmethylsulfonyl fluoride, 10 μg/ml aprotinin, 10 μg/ml leupeptin, and 10 μg/ml pepstatin). The samples are homogenized at 4 °C, and protein content is determined by Bradford method. Total protein (25 μg) is electrophoresed on 10% SDS-PAGE gel and transferred to a nitrocellulose membrane. The experiments are performed using the following primary antibodies: tau Ser(P)-396, 1:1000; Tau5, 1:1000; and anti-GSK3β, 1:1000. The filters are incubated with the antibody at 4 °C overnight in 5% nonfat dried milk. A secondary antibody (1:5000), followed by ECL detection reagents are used for immunodetection. Quantitation of immunoreactivity is performed by densitometric scanning.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Kinase Assay: GSK3 scintillation proximity assay is done. The competition experiments are carried out in duplicate with 10 concentrations of the inhibitor in clear-bottomed microtiter plates. The biotinylated peptide substrate biotin-AAEELDSRAGS(PO3H2)PQL, is added at a final concentration of 2 μM in an assay buffer containing 6 milliunits of recombinant human GSK3 (equal mix of both α and β), 12 mM MOPS, pH 7.0, 0.3 mM EDTA, 0.01% β-mercaptoethanol, 0.004% Brij 35, 0.5% glycerol, and 0.5 μg of bovine serum albumin/25 μl and preincubated for 10–15 min. The reaction is initiated by the addition of 0.04 μCi of [γ-33P]ATP and unlabeled ATP in 50 mM Mg(Ac)2 to a final concentration of 1 μM ATP and assay volume of 25 μl. Blank controls without peptide substrate are used. After incubation for 20 min at room temperature, each reaction is terminated by the addition of 25 μl of stop solution containing 5 mM EDTA, 50 μM ATP, 0.1% Triton X-100, and 0.25 mg of streptavidin-coated SPA beads corresponding to 35 pmol of binding capacity. After 6 h the radioactivity is determined in a liquid scintillation counter. |
| 體外活性 | AZD1080逆轉(zhuǎn)小鼠的認(rèn)知缺陷并拯救其功能失調(diào)的突觸.口服AZD1080抑制大鼠腦中tau蛋白磷酸化,在峰濃度下腦/血漿暴露比率為0.5-0.8.急性口服AZD1080抑制外圍GSK3的活性,劑量依賴(lài)性降低磷酸化糖原合成酶和總糖原合成酶的比率,在最高劑量為10mol/kg時(shí)達(dá)到平均最大抑制效果49％. |
| 體內(nèi)活性 | AZD1080抑制表達(dá)人類(lèi)tau的細(xì)胞中的tau磷酸化,IC50為324 nM。AZD1080抑制人GSK3α和GSK3β,Ki 值分別為6.9 nM和31 nM,對(duì)cdk2、cdk5、cdk1和Erk2表現(xiàn)出> 14倍的選擇性。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 5.43 mg/mL (16.24 mM), Sonication is recommended.
10% DMSO+90% Saline : 0.54 mg/mL (1.61 mM), Solution.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 2 mg/mL (5.98 mM), Sonication is recommended.
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| 關(guān)鍵字 | Inhibitor | inhibit | GSK-3β | GSK-3α | GSK-3 | GSK3 | Glycogen synthase kinase-3 | Glycogen synthase kinase 3 | AZD-1080 | AZD1080 | AZD 1080 |
| 相關(guān)產(chǎn)品 | Cromolyn sodium | ROCK-IN-5 | 4-Methyl-6-Methoxycoumarin | Ginsenoside Rg2 | 4-Chloro-2'-bromoacetophenone | AT7519 | Ceftriaxone sodium hydrate | 5-Bromoindole | GSK-3 Inhibitor 5 | GSK 3 Inhibitor IX | Indazole | Ethyl linoleate |
| 相關(guān)庫(kù) | 神經(jīng)保護(hù)化合物庫(kù) | 抑制劑庫(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) | 抗衰老化合物庫(kù) | 免疫/炎癥分子化合物庫(kù) | 神經(jīng)退行性疾病化合物庫(kù) | 膜蛋白靶向化合物庫(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) |