| 名稱 | SB 525334 |
| 描述 | SB-525334 is a potent and selective inhibitor of the TGF-β1R and ALK5 (IC50: 14.3 nM). |
| 細(xì)胞實(shí)驗(yàn) | RPTE cells were seeded on microscope slides. The following day, the cells were starved by removal of epidermal growth factor and serum for 24 h prior to dosing. Cells were dosed with 10 ng/ml TGF- 1 or 1 M SB-525334 or a combination of both. Slides were pretreated with SB-525334 or starve media for 3 h prior to a 1-h incubation at 37°C with TGF- 1 or starve media. The cells were then fixed for 15 min in 4% ice-cold paraformaldehyde. The cells were permeabilized for 10 min in 0.3% Triton X-100/PBS at room temperature. The slides were incubated for 30 min in a blocking solution containing 0.3% bovine serum albumin, 10% FBS, 0.3% Triton X-100/PBS, and 5% milk in PBS. A 1:200 dilution of primary mouse anti-Smad2/3 antibody was applied to each slide for overnight incubation. A 1:200 dilution of anti-mouse IgG fluorescein secondary antibody was applied to each slide for 30 min at room temperature. The slides were then viewed using an argon blue 488 nM laser in a confocal microscope. Nuclear signal intensity was analyzed using 1D Image Analysis software. The relative intensity was determined by the mean intensity of the nucleus and expressed as percent control [1]. |
| 激酶實(shí)驗(yàn) | To determine the potency of the ALK5 inhibitor SB-525334 at the enzyme level, purified GST-tagged kinase domain of ALK5 was incubated with purified GST-tagged full-length Smad3 in the presence of 33P-γATP and different concentrations of SB525334. The readout is radioactively labeled Smad3. To determine the selectivity of SB-525334, purified GST-tagged kinase domain of ALK2 and ALK4 were incubated with GST-tagged full-length Smad1 and Smad3, respectively, in the presence of different concentrations of SB-525334 (n=3). IC50 value determinations were calculated with GraphPad software using a sigmoidal dose-response curve [1]. |
| 動(dòng)物實(shí)驗(yàn) | To identify the optimal treatment length for puromycin aminonucleoside's effect on extracellular matrix in the kidney, 18 Sprague-Dawley (SD) rats (200 –250 g) were injected with 15 mg/100 g of puromycin aminonucleoside in 0.9% saline or sham 0.9% saline only intraperitoneally. Animals were sacrificed at 24 h (n = 3+2 control), day 4 (n=3), day 8 (n = 3), day 10 (n = 3), day 15 (n = 2), and day 20 (n = 2). A 24-h urine collection and plasma sample were taken at 9:00 AM everyday. Urine and plasma chemistry were measured at GlaxoSmithKline Laboratories Animal Science using an Olympus clinical analyzer. Proteinuria was measured as a concentration (mg/deciliter) and then converted to total protein excreted over a 24-h period using urine flow (mL/24 h). The creatinine clearance was calculated by multiplying urine creatinine levels (mg/mL) by urine flow (mg/mL/100 g b.wt.) and then dividing that product by plasma creatinine (mg/mL). To determine the effect of SB-525334 on renal disease in the PAN model, SD rats were pretreated by oral gavage with 1, 3, or 10 mg/kg/day of SB-525334 once a day. The following day, PAN was injected at 15 mg/100 g to the appropriate rats. Treatment groups continued to receive SB-525334. Ten days after PAN injection the rats were sacrificed, and blood, urine, and kidneys were collected at the termination point for analysis [1]. |
| 體外活性 | SB-525334（1 μM）在腎小管近端細(xì)胞中阻斷了TGF-beta1誘導(dǎo)的Smad2/3的磷酸化和核內(nèi)轉(zhuǎn)移，并抑制了TGF-beta1在A498腎上皮癌細(xì)胞中誘導(dǎo)的纖溶酶原激活物抑制劑-1（PAI-1）及前膠原α1(I) mRNA表達(dá)的增加[1]。與SB525334的聯(lián)合應(yīng)用顯著增強(qiáng)了在親本細(xì)胞和耐古美替尼的胰腺癌細(xì)胞中古美替尼的細(xì)胞毒性。SB525334在耐古美替尼的細(xì)胞中顯著增加了凋亡細(xì)胞死亡[2]。 |
| 體內(nèi)活性 | 口服給予1、3或10 mg/kg/day的SB 525334，連續(xù)11天，能顯著降低腎臟PAI-1 mRNA表達(dá)[1]。SB 525334（10 mg/kg或30 mg/kg）每日兩次口服。在Bleomycin（BLM）處理后的第5、7、9和14天，分離肺部進(jìn)行研究。BLM處理導(dǎo)致顯著的肺纖維化變化，伴隨ECM mRNA表達(dá)顯著上調(diào)、Smad2/3核轉(zhuǎn)位、CTGF表達(dá)增加、肌成纖維細(xì)胞增殖以及I型膠原沉積。SB 525334處理減輕了肺部的組織病理學(xué)改變，并顯著降低了I型和III型前膠原及纖維連接蛋白mRNA表達(dá)[3]。 |
| 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 : ＜1 mg/mL (insoluble or slightly soluble)
1eq. HCl : 34.3 mg/mL (100 mM)
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| 關(guān)鍵字 | inhibit | Inhibitor | TGF-β Receptor | Transforming growth factor beta receptors | SB-525334 | SB 525334 |
| 相關(guān)產(chǎn)品 | Monocrotaline | Pirfenidone | Ceritinib | A 83-01 | Chromenone 1 | LY-364947 | BMP signaling agonist sb4 | Alectinib hydrochloride | SB-431542 | Galunisertib | Alantolactone | Hydrochlorothiazide |
| 相關(guān)庫 | 抑制劑庫 | 腫瘤免疫治療小分子化合物庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 激酶抑制劑庫 | NO PAINS 化合物庫 | 癌細(xì)胞分化化合物庫 | 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 |