| 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(xì)胞實(shí)驗(yàn) | 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(shí)驗(yàn) | 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(dòng)物實(shí)驗(yàn) | 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(xì)胞 293T、人類結(jié)腸癌來(lái)源的非致瘤性細(xì)胞 FET 和人肺腺癌細(xì)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(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：TGF-β1 誘導(dǎo)了 Smad2/3 和 Smad4 之間的復(fù)合物形成。SB-431542 以劑量依賴的方式，通過(guò)阻斷 TGF-β 誘導(dǎo)的磷酸化和Smad2/3的激活，阻斷復(fù)合物形成。[1]
方法：人膠質(zhì)母細(xì)胞瘤細(xì)胞 D54MG 用 SB-431542 (1-10 μM) 處理 24 h，使用 Transwell 方法檢測(cè)細(xì)胞遷移情況。
結(jié)果： SB-431542抑制 TGF-βRI 信號(hào)傳導(dǎo)以濃度依賴的方式阻斷 D54MG 細(xì)胞的細(xì)胞遷移。[2]
方法：人多能干細(xì)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(cè)細(xì)胞分化情況。
結(jié)果：神經(jīng)誘導(dǎo)通過(guò) PAX6 的表達(dá)來(lái)監(jiān)測(cè)，PAX6 是神經(jīng)直腸真皮分化的早期標(biāo)志物。與單獨(dú)使用 Noggin 或 SB-431542 時(shí)不到 10% 的 PAX6+ 細(xì)胞相比，用 Noggin 和 SB-431542 聯(lián)合處理顯著提高了神經(jīng)誘導(dǎo)的效率，使其超過(guò)總細(xì)胞的 80%。[3] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)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(jié)果：SB-431542 可顯著抑制 4T1 乳腺腫瘤的肺轉(zhuǎn)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(jié)果：SB-431542 抑制 TGF-β1 信號(hào)傳導(dǎo)可減少纖維化、脂肪浸潤(rùn)和肌肉重量減輕。SB-431542 治療通過(guò)促進(jìn)纖維/脂肪生成祖細(xì)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(rùn)的重要細(xì)胞來(lái)源。[5] |
| 存儲(chǔ)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(guān)鍵字 | Transforming growth factor beta receptors | inhibit | TGF-β Receptor | SB431542 | SB-431542 | Inhibitor |
| 相關(guān)產(chǎn)品 | Monocrotaline | Pirfenidone | Ceritinib | A 83-01 | Halofuginone | Chromenone 1 | LY-364947 | BMP signaling agonist sb4 | Alectinib hydrochloride | Galunisertib | Alantolactone | Hydrochlorothiazide |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 血管生成庫(kù) | 抗癌活性化合物庫(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) | 高選擇性抑制劑庫(kù) | TGF-β/Smad靶點(diǎn)化合物庫(kù) | 膜蛋白靶向化合物庫(kù) | 酪氨酸激酶分子庫(kù) |