| 名稱(chēng) | BX795 |
| 描述 | BX795 is an effective and selective PDK1 inhibitor (IC50: 6 nM), and its selectivity is 140- and 1600-fold for PDK1 over PKA and PKC in cell-free assays, respectively. Meanwhile, the selectivity for PDK1 is 100-fold than GSK3β. |
| 細(xì)胞實(shí)驗(yàn) | Cells seeded at a low density (1,500–3,000 cells/well, 0.1 mL/well, 96-well plates) are incubated overnight. Compound treatments are made by adding 10 μL/well of the compound in 1% dimethyl sulfoxide and growth medium (final concentration of dimethyl sulfoxide, 0.1%), followed by brief shaking. Treated cells are incubated for 72 hours, and viability is measured by the addition of 10 μL of the metabolic dye WST-1. The WST-1 signal is read in a plate reader at 450 nm, and a no cell, or zero time cell, background is subtracted to calculate the net signal. Results are reported as the average ± S.E. of two or more replicates.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Kinase assays: PDK1 is assayed in a direct kinase assay and a coupled assay format measuring PDK1- and PtdIns-3,4-P2-mediated activation of AKT2. For the coupled assay, the final assay mixture (60 μL) contained: 15 mM MOPS, pH 7.2, 1 mg/mL bovine serum albumin, 18 mM β-glycerol phosphate, 0.7 mM dithiothreitol, 3 mM EGTA, 10 mM MgOAc, 7.5 μM ATP, 0.2 μCi of [γ-33P]ATP, 7.5 μM biotinylated peptide substrate (biotin-ARRRDGGGAQPFRPRAATF), 0.5 μL of PtdIns-3,4-P2-containing phospholipid vesicles, 60 pg of purified recombinant human PDK1, and 172 ng of purified recombinant human AKT2. After incubation for 2 h at room temperature, the biotin-labeled peptide is captured from 10 μl of the assay mixture on streptavidin-coated SPA beads, and product formation is measured by scintillation proximity in a Wallac MicroBeta counter. The product formed is proportional to the time of incubation and to the amount of PDK1 and inactive AKT2 added. PDK1 is added at suboptimal levels so that the assay could sensitively detect inhibitors of AKT2 activation as well as direct inhibitors of PDK1 or AKT2. To measure PDK1 activity directly, the final assay mixture (60 μL) contained 50 mM Tris-HCl, pH 7.5, 0.1 mM EGTA, 0.1 mM EDTA, 0.1% β-mercaptoethanol, 1 mg/mL bovine serum albumin, 10 mM MgOAc, 10 μM ATP, 0.2 μCi of [γ-33P]ATP, 7.5 μM substrate peptide (H2N-ARRRGVTTKTFCGT), and 60 ng of purified recombinant human PDK1. After 4 h at room temperature, we add 25 mM EDTA and spotted a portion of the reaction mixture on Whatman P81 phosphocellulose paper. The filter paper is washed three times with 0.75% phosphoric acid and once with acetone. After drying, the filter-bound labeled peptide is quantified using a Fuji phosphorimager. |
| 體內(nèi)活性 | BX795（1 μM）對(duì)以下酪氨酸蛋白激酶無(wú)抑制作用: 肝配蛋白受體A2和B3,Bruton's酪氨酸激酶,Syk和FGFR1。但BX795對(duì)抑制血管內(nèi)皮生長(zhǎng)因子受體的抑制效果低于TBK1。BX795對(duì)IRF3 Ser396位點(diǎn)磷酸化（TBK1-催化）的抑制作用隨著ATP濃度上升而下降,意味著B(niǎo)X795是ATP競(jìng)爭(zhēng)性抑制劑。BX795也抑制NUAK1,MARK1/2/4,VEGFR和MLK1/2/3,IC50分別為5,55,53,19,157,50,46和42 nM。BX795對(duì)IRF3依賴的基因轉(zhuǎn)錄具有抑制作用。BX795對(duì)巨噬細(xì)胞的IFN-β分泌也有抑制作用。BX795阻斷IKKε和TBK1調(diào)節(jié)的IRF3激活,并對(duì)IFN-β的產(chǎn)量有抑制效果。BX795對(duì)TBK1/IKKε的抑制不會(huì)引起p38α MAPK和JNK1/2的激活。BX795可抑制聚(I:C)處理后IRF3在核中的積累。BX795不影響脂多糖刺激的p70核糖體S6激酶1Thr229位點(diǎn)的磷酸化,該位點(diǎn)是PDK1的作用靶點(diǎn)。BX795不影響NFκB依賴的基因轉(zhuǎn)錄激活（由IKKα/β復(fù)合體或脂多糖、聚(I:C)、IL-1α或TNFα促進(jìn)）。BX795對(duì)IL-1α或TNFα刺激的MEFs有效,并阻斷p38α MAPKJ和NK1/2磷酸化。 |
| 存儲(chǔ)條件 | Pure form: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | DMSO : 250 mg/mL (422.68 mM), Sonication is recommended.
Ethanol : 59.2 mg/mL (100.09 mM), Sonication is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 5 mg/mL (8.45 mM), Sonication is recommended.
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| 關(guān)鍵字 | PDK-1 | PDK1 | IκB kinase | IκB | Inhibitor | inhibit | IKK | IkB/IKK | IkB | I kappa B kinase | c-Kit | cKit | Chk1 | CDK2/CyclinE | BX-795 | BX795 | BX 795 | Autophagy |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | 2-Chloropyrazine | Aceglutamide | Alginic acid | Hemin | Hydroxychloroquine | Sildenafil citrate | Stavudine | Tamoxifen | Adenosine | Paeonol |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 細(xì)胞焦亡化合物庫(kù) | 經(jīng)典已知活性庫(kù) | 抗癌活性化合物庫(kù) | 已知活性化合物庫(kù) | 激酶抑制劑庫(kù) | 高選擇性抑制劑庫(kù) | 抗衰老化合物庫(kù) | 膜蛋白靶向化合物庫(kù) | 免疫/炎癥分子化合物庫(kù) | 酪氨酸激酶分子庫(kù) | 抗前列腺癌化合物庫(kù) |