| 名稱 | SB-431542 |
| 描述 | SB-431542 is an inhibitor of ALK5/TGF-β type I Receptor (IC50=94 nM) and is selective. SB 431542 also has inhibitory activity against ALK4 and ALK7 but not other proteins. SB 431542 can be used for induced differentiation of stem cells. |
| 細胞實驗 | A498 cells were seeded at 5,000 to 10,000 cells/well in 96-well plates. The cells were serum-deprived for 24 h and then treated with compounds for 48 h to assess the cellular toxicity. Cell viability is determined by incubating cells for 4 h with XTT labeling and electron coupling reagent. Live cells with active mitochondria produce an orange-colored product, formazan, which is detected using a plate reader at between A450 nm and A500 nm with a reference wavelength greater than 600 nm. The absorbance values correlate with the number of viable cells [2]. |
| 激酶實驗 | Kinase assays were performed with 65 nM GSTALK5 and 184 nM GST-Smad3 in 50 mM HEPES, 5 mM MgCl2, 1 mM CaCl2, 1 mM dithiothreitol, and 3 M ATP. Reactions were incubated with 0.5 μCi of [33P]γATP for 3 h at 30°C. Phosphorylated protein was captured on P-81 paper, washed with 0.5% phosphoric acid, and counted by liquid scintillation. Alternatively, Smad3 or Smad1 protein was also coated onto FlashPlate Sterile Basic Microplates. Kinase assays were then performed in FlashPlates with same assay conditions using either the kinase domain of ALK5 with Smad3 as a substrate or the kinase domain of ALK6 (BMP receptor) with Smad1 as substrate. Plates were washed three times with phosphate buffer and counted by TopCount [2]. |
| 動物實驗 | BALB/c mice received intraperitoneal (i.p.) injections of colon-26 tumor cells. Three days after tumor cell inoculation, SB-431542 (1 μM solution, 100 μl/animal) or vehicle alone was directly injected into the peritoneal cavity. CTL activities were measured by a standard 4 h 51Cr release assay after culturing spleen cells with γ-irradiated tumor cells for five days in the absence of added growth factors. In vitro experiments, cell lysate of HLA-A*2402 positive gastric cancer cell line, OCUM-8, was incubated with human DC cultures for 4 h. After washing extensively, PBMCs obtained from the same volunteer as DCs were incubated for 7 days and measured CTL activity by 51Cr release assay. NK activity was tested using 51Cr release assay against K562 [4]. |
| 體外活性 | 方法：人腎細胞 293T、人類結腸癌來源的非致瘤性細胞 FET 和人肺腺癌細胞 A549 用 SB-431542 (2-10 μM) 和 TGF-β1 (12.5 ng/mL) 處理 1 h，使用 Western Blot 方法檢測靶點蛋白表達水平。
結果：TGF-β1 誘導了 Smad2/3 和 Smad4 之間的復合物形成。SB-431542 以劑量依賴的方式，通過阻斷 TGF-β 誘導的磷酸化和Smad2/3的激活，阻斷復合物形成。[1]
方法：人膠質母細胞瘤細胞 D54MG 用 SB-431542 (1-10 μM) 處理 24 h，使用 Transwell 方法檢測細胞遷移情況。
結果： SB-431542抑制 TGF-βRI 信號傳導以濃度依賴的方式阻斷 D54MG 細胞的細胞遷移。[2]
方法：人多能干細胞 hESCs 用含 SB-431542 (10 nM) 和 Noggin (500 ng/mL) 的 KSR 培養(yǎng)基培養(yǎng) 11 天，使用 Immunofluorescence 方法檢測細胞分化情況。
結果：神經(jīng)誘導通過 PAX6 的表達來監(jiān)測，PAX6 是神經(jīng)直腸真皮分化的早期標志物。與單獨使用 Noggin 或 SB-431542 時不到 10% 的 PAX6+ 細胞相比，用 Noggin 和 SB-431542 聯(lián)合處理顯著提高了神經(jīng)誘導的效率，使其超過總細胞的 80%。[3] |
| 體內活性 | 方法：為檢測體內抗腫瘤活性，將 SB-431542 (10 mg/kg in 20% DMSO/80% corn oil) 腹腔注射給攜帶小鼠乳腺癌腫瘤 4T1 的 Balb/c 小鼠，每周三次，持續(xù)四周。
結果：SB-431542 可顯著抑制 4T1 乳腺腫瘤的肺轉移。[4]
方法：為研究肌腱損傷的治療方法，將 SB-431542 (10 mg/kg) 腹腔注射給大面積肩袖撕裂的 C57B/6J 小鼠模型，每天一次，持續(xù)二或六周。
結果：SB-431542 抑制 TGF-β1 信號傳導可減少纖維化、脂肪浸潤和肌肉重量減輕。SB-431542 治療通過促進纖維/脂肪生成祖細胞 (FAP) 的凋亡，減少了損傷肌肉中 FAP 的數(shù)量，F(xiàn)AP 是肩袖肌纖維化和脂肪浸潤的重要細胞來源。[5] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 16.67 mg/mL (43.36 mM)
10% DMSO+90% Saline : 0.1 mg/mL (0.26 mM), Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
Ethanol : 3.8 mg/mL (10 mM)
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| 關鍵字 | Transforming growth factor beta receptors | inhibit | TGF-β Receptor | SB431542 | SB-431542 | Inhibitor |
| 相關產品 | Monocrotaline | Pirfenidone | Ceritinib | A 83-01 | Halofuginone | Chromenone 1 | LY-364947 | BMP signaling agonist sb4 | Alectinib hydrochloride | Galunisertib | Alantolactone | Hydrochlorothiazide |
| 相關庫 | 抑制劑庫 | 血管生成庫 | 抗癌活性化合物庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 高選擇性抑制劑庫 | TGF-β/Smad靶點化合物庫 | 膜蛋白靶向化合物庫 | 酪氨酸激酶分子庫 |