| 名稱 | BX-912 |
| 描述 | BX-912 is a specific inhibitor of 3-Phosphoinositide-dependent Kinase-1 (PDK1, IC50: 12 nM). The selectivity of BX-912 is more 10 fold than C-Kit, EGFR, PKA, PKC etc. |
| 細(xì)胞實(shí)驗(yàn) | Cells such as MDA-468, MDA-453 are seeded at a low density (1.5-3 × 103 cells/well, 0.1 mL/well, 96-well plates) and are incubated overnight. BX912 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. (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 hours 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 inhibitor BX912 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 hours at room temperature, 25 mM EDTA is added and a portion of the reaction mixture on P81 phosphocellulose paper is spotted. 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 phosphorimager. |
| 體外活性 | BX-912對ChcK1（IC50：0.83 μM）,PKA（IC50：0.11 μM）,c-kit（IC50：0.85 μM）和KDR（IC50：0.41 μM）有抑制作用。在腫瘤細(xì)胞中,BX-912可阻斷PDK1/Akt信號,能抑制多種腫瘤細(xì)胞的貼壁依賴型生長(比如PC-3細(xì)胞)或者誘導(dǎo)凋亡。許多Akt活性升高的癌細(xì)胞株在軟瓊脂中對BX-912引起的生長抑制的敏感性比在組織培養(yǎng)塑料中高30倍,與PDK1/Akt信號通路的細(xì)胞生存功能一致,這對未貼壁細(xì)胞尤為重要。BX-912對PDK1酶活有顯著的抑制效果, 但不影響AKT2活性（IC50>10 μM）,故BX-912是PDK1的直接抑制劑。BX-912使含4 NDNA的MDA-468細(xì)胞數(shù)顯著增多,并使細(xì)胞停在G2/M期。BX-912有效抑制細(xì)胞生長（IC50：0.32 μM）。BX-912（1 μM）對HCT-116細(xì)胞生長的抑制也十分有效（抑制率達(dá)96%）。 |
| 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 : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 87 mg/mL (184.6 mM)
Ethanol : 87 mg/mL (184.6 mM)
|
| 關(guān)鍵字 | BX-912 | Apoptosis | PDK-1 | BX 912 | inhibit | BX912 | Inhibitor |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | Sorafenib | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 抗癌細(xì)胞代謝庫 | 激酶抑制劑庫 | 高選擇性抑制劑庫 | 抗衰老化合物庫 | 膜蛋白靶向化合物庫 | 酪氨酸激酶分子庫 |