| 名稱 | AZ6102 |
| 描述 | AZ6102 is a potent TNKS1/2 inhibitor that has 100-fold selectivity against other PARP family enzymes and shows IC50 of 5 nM for Wnt pathway inhibition in DLD-1 cells. |
| 激酶實(shí)驗(yàn) | Sphingosine Kinase Assays: The IC50 values for ABC294640 and DMS are determined by a newly developed HPLC-based SK activity assay. In brief, the test compounds are incubated with recombinant SK1 or SK2 and NBD-Sph in the isozyme-selective assay buffers detailed below with 1 mg/ml fatty acid-free bovine serum albumin, 100 μM ATP, and 400 μM MgCl2. The product, i.e., NBD-S1P, is separated from NBD-Sph by HPLC as follows: Waters 2795 HPLC system with a Waters 2495 fluorescence detector, C8 Chromolith RP-8e column (100 × 4.6 mm), 1 ml/min mobile phase (acetonitrile/20 mM sodium phosphate buffer, pH2.5, at 45:55). Fluorescence is monitored with excitation at 465 nm and emission at 531 nm. The ratio of NBD-S1P/(NBD-Sph + NBD-S1P) is used as a measure of SK activity. SK-isozyme selective assay buffers each contained 20 mM Tris, pH7.4, 5 mM EDTA, 5 mM EGTA, 3 mM β-mercaptoethanol, 5% glycerol, 1× protease inhibitors and 1× phosphatase inhibitors. For the SK1 assay buffer, 0.25% (final) Triton X-100 is added; and for the SK2 buffer, 1 M (final) KCl is added. Assays are run for 2 h at room temperature, and then a 1.5 volume of methanol is added to terminate the kinase reaction. After 10 min, the samples are centrifuged at 20,000 g to pellet the precipitated protein, and the supernatants are analyzed by HPLC. In experiments to determine the Ki for inhibition of SK2 by ABC294640, the ADP Quest assay system is used to measure kinase activity in the presence of varying concentrations of sphingosine and ABC294640. To determine the effects of ABC294640 on cellular SK activity, near-confluent MDA-MB-231 cells are serum-starved overnight, and then treated with varying concentrations of ABC294640. The cells are then incubated with [3H]sphingosine at a final concentration of 1 μM. The cells take up the exogenous sphingosine, which is converted to S1P via SK activity, and [3H]S1P is separated from [3H]sphingosine by extraction and quantified by scintillation counting. |
| 體外活性 | AZ6102在酶促實(shí)驗(yàn)和TCF4報(bào)告基因?qū)嶒?yàn)中（<5 nM）抑制TNKS1和TNKS2。在Colo320DM細(xì)胞中，AZ6102能抑制增殖（GI50約40 nM），但在含有β-catenin突變的HCT-116細(xì)胞系和BRCA突變的MDA-MB-436細(xì)胞系中不顯示抗增殖活性。在Colo320DM中，AZ6102穩(wěn)定了axin2蛋白并以劑量和時(shí)間依賴的方式在體內(nèi)外調(diào)控Wnt目標(biāo)基因[1]。 |
| 體內(nèi)活性 | 裸鼠每千克體重被給予25mg的AZ-6102。該化合物半衰期為4小時(shí)，清除率(Clearance, CL)為24 mL/min.kg。進(jìn)一步在小鼠和大鼠的分析顯示，AZ-6102的生物利用度分別為12%和18%，表現(xiàn)出中等水平。通過對(duì)DLD-1細(xì)胞處理后的Western blot 分析發(fā)現(xiàn)，AZ-6102在較低濃度(at 24, 48 和 72h)時(shí)，相比XAV-939對(duì)TNKS1、TNSK2 和Axin2的穩(wěn)定作用更強(qiáng)且持續(xù)時(shí)間更長。在臨床前物種中，AZ-6102表現(xiàn)出良好的藥代動(dòng)力學(xué)特性，具有低Caco2外排作用（以避免可能的腫瘤抗性機(jī)制）。此外，這種化合物可以使用SBECD作為輔料，在pH4時(shí)配制成每毫升20mg的臨床相關(guān)靜脈注射溶液。利用AZ-6102作為靜脈(i.v.)探針化合物探索其對(duì)腫瘤異種移植物和正常組織中TNKS1及TNSK2抑制作用的體內(nèi)效果的結(jié)果即將公布[2]。 |
| 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 | DMSO : 21.4 mg/mL (49.94 mM), Sonication is recommended.
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| 關(guān)鍵字 | βcatenin | Wnt/β-catenin | Wnt/betacatenin | Wnt/b-catenin | Wnt | TNKS2 | TNKS1 | poly ADP ribose polymerase | PARP | Inhibitor | inhibit | beta-catenin | betacatenin | bcatenin | AZ-6102 | AZ6102 | AZ 6102 |
| 相關(guān)產(chǎn)品 | Urea | Bisdemethoxycurcumin | Niraparib | Nefopam hydrochloride | XAV-939 | Olaparib | 3-Aminobenzamide | Benzamide | OUL35 | TNIK-IN-3 | CHIR-99021 | Wnt pathway activator 1 |
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