| 名稱 | SB-505124 |
| 描述 | SB505124 is a selective inhibitor of TGFβR for ALK4, ALK5. |
| 細(xì)胞實(shí)驗(yàn) | Cell viability is measured as described by Laping et al., 2002 or by using the modified tetrazolium salt WST-1. XTT assay: The cells are serum-deprived for 24 hours and then treated with SB505124 for 48 hours to assess the cellular toxicity. Cell viability is determined by incubating cells for 4 hours with XTT labeling and electron coupling reagent according to the manufacturer's directions. 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. Modified tetrazolium salt WST-1: Approximately 2000 cells are seeded in 96-well dishes in 100 μL of 0.2% FBS phenol red-free media overnight. The cells are treated with 50 μL of SB505124 (to achieve the final concentrations indicated) for 30 minutes before being treated with or without TGF-β1 and TNF-α to a final volume of 200 μL. Cell growth is measured at the indicated time points by incubating each well with 10 μL of WST-1 for 3 hours at 37 °C. Metabolically active cells cleave WST-1 to water-soluble formazan, which is directly quantitated with an enzyme-linked immunosorbent assay plate reader. Each experiment is done at least twice, and treatment for each cell line is done in triplicate.(Only for Reference) |
| 激酶實(shí)驗(yàn) | In Vitro Protein Kinase Assay : Kinase assays are performed as described by Laping et al., 2002 using the kinase domain of ALK5 and full-length N-terminal fused GST-Smad3. Kinase assays are performed with 65 nM GST-ALK5 and 184 nM GST-Smad3 in 50 mM HEPES, 5 mM MgCl2, 1 mM CaCl2, 1 mM dithiothreitol, and 3 μM ATP. Reactions are incubated with 0.5 μCi of [33P]γATP for 3 hours at 30 °C. Phosphorylated protein is captured on P-81 paper , washed with 0.5% phosphoric acid, and counted by liquid scintillation. Alternatively, Smad3 or Smad1 protein is also coated onto FlashPlate Sterile Basic Microplates. Kinase assays are then performed in FlashPlates with same assay conditions using either the kinase domain of ALK5 with Smad3 as substrate or the kinase domain of ALK6 (BMP receptor) with Smad1 as substrate. Plates are washed three times with phosphate buffer and counted by TopCount. |
| 體外活性 | 在兔GFS模型中,SB505124降低了GFS手術(shù)部位的眼內(nèi)壓（IOP）水平并減少了結(jié)膜下細(xì)胞浸潤和瘢痕形成.在他克莫司（TAC）治療的小鼠和FK12EC KO小鼠中,SB505124抑制內(nèi)皮細(xì)胞TGF-β受體的活化和腎小動(dòng)脈玻璃樣變性的誘導(dǎo). |
| 體內(nèi)活性 | SB505124被鑒定為ALK4和ALK5的可逆ATP競爭性和選擇性ALK抑制劑。SB-505124抑制緊密相關(guān)的ALK4,IC50為129 nM,選擇性比作用于ALK5低2.5倍。SB-505124濃度高達(dá)10 μM時(shí)也不抑制ALK2。SB-505124抑制COS-1細(xì)胞內(nèi)源性Smad2磷酸化。ALK4和ALK5激活Smad1和Smad2,加入SB-505124抑制激活。SB-505124作用于HepG2人類肝癌細(xì)胞,C2C12小鼠成肌細(xì)胞和Mv1Lu水貂肺細(xì)胞,濃度依賴性抑制轉(zhuǎn)化生長因子-β誘導(dǎo)的Smad2磷酸化。SB-505124有效抑制轉(zhuǎn)化生長因子-β和活化素誘導(dǎo)CAGA12-熒光素酶和ARE-熒光素酶受體結(jié)構(gòu)的活力,這種作用存在濃度依賴性。在人臍靜脈內(nèi)皮細(xì)胞（HUVEC）中,SB505124（500 nM）阻斷TGF-β1對F-actin組裝的影響,并阻止TGF-β誘導(dǎo)的ROS產(chǎn)生。 |
| 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 : 50 mg/mL (149.08 mM), Sonication is recommended.
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| 關(guān)鍵字 | Transforming growth factor beta receptors | TGF-β/Smad | TGF-β Receptor | TGFβ | TGFbeta/Smad | TGF-beta | TGFbeta | TGF-b/Smad | TGFb | Smad | SB-505124 | SB505124 | SB 505124 | Inhibitor | inhibit | ALK5 | ALK4 |
| 相關(guān)產(chǎn)品 | Entrectinib | Pirfenidone | Ceritinib | A 83-01 | Chebulinic acid | Chromenone 1 | LY-364947 | BMP signaling agonist sb4 | Alectinib hydrochloride | Alantolactone | Galunisertib | Hydrochlorothiazide |
| 相關(guān)庫 | 抑制劑庫 | 抗乳腺癌化合物庫 | 血管生成庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 抗癌化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 臨床前化合物庫 | TGF-β/Smad靶點(diǎn)化合物庫 | 膜蛋白靶向化合物庫 | 酪氨酸激酶分子庫 |