| 名稱 | Saracatinib |
| 描述 | Saracatinib (AZD0530) is a small molecule inhibitor belonging to the Src family kinase inhibitors (IC50=2.7–11 nM), featuring high selectivity, cell permeability, and oral bioavailability, with anti-fibrotic, anti-inflammatory, and potential anti-tumor activities. |
| 細胞實驗 | Cell proliferation was assessed using a colorimetric 5‐bromo‐2′‐deoxyuridine (BrdU) Cell Proliferation ELISA kit, as described previously. Briefly, cells were plated onto 96‐well plates (1.5×10^4 cells/well), the following day 0.039–20μM AZD0530 in DMSO (at a final concentration of 0.5%) was added and the cells were incubated for 24h. The cells were pulse-labeled with BrdU for 2h and fixed. Cellular DNA was then denatured with the provided solution and incubated with anti-BrdU peroxidase for 90min. Following three washes with phosphate‐buffered saline, tetramethylbenzidine substrate solution was added and the plates were incubated on a plate shaker for 10–30min until the positive control absorbance at 690nm was approximately 1.5 absorbance units [1]. |
| 激酶實驗 | Inhibition of tyrosine kinase activity was examined using an ELISA with recombinant catalytic domains of a panel of receptor and non‐receptor tyrosine kinases (in some cases only part of the catalytic domain was used). This method has been described previously. AZD0530 dose ranges varied depending on the activity versus the particular kinase tested, but were typically 0.001–10μM. Specificity assays against a panel of serine/threonine kinases were performed using a filter capture assay with 32P. Briefly, multidrop 384 plates containing 0.5μL AZD0530 or controls (DMSO alone or pH 3.0 buffer controls) were incubated with 15μL of enzyme plus peptide/protein substrate for 5min before the reaction was initiated by the addition of 10μL of 20mM Mg.ATP. For all enzymes the final concentration was approximated to the Michaelis constant (Km). Assays were carried out for 30min at room temperature before termination by the addition of 5μL orthophosphoric acid. After mixing, the well contents were harvested onto a P81 Unifilter plate, using orthophosphoric acid as the wash buffer. Microcal Origin software was used to interpolate IC50 values by nonlinear regression [1]. |
| 動物實驗 | Female athymic mice (nu/nu: Alpk) and rats (RH‐rnu/rnu) were housed and maintained as previously described. Src3T3 and human tumor lines (as indicated in Table 3) were inoculated subcutaneously in the left flank of animals. Tumor growth was monitored by bi‐dimensional caliper measurements twice weekly. The tumor volume was calculated by the following formula: (length×width)×√(length×width)×(π/6) and supported by excision and weighing of tumors at the end of the studies. Dosing started when the average tumor volume reached 0.2–0.5cm3 (except MDA‐MB‐231 and HT29). Animals were treated once daily by oral gavage with either vehicle alone or AZD0530 6.25–50mg/kg for 10–91 days. Tumor growth inhibition was calculated as described previously. For pharmacokinetic and pharmacodynamic analysis animals were humanely sacrificed and samples (plasma and tumor) were collected. Tumor samples were homogenized with 5 volumes of water and extracted with chloroform. Plasma and tumor samples were analyzed for AZD0530 concentration using high‐performance liquid chromatography with tandem mass spectrometric detection after solid‐phase extraction [1]. |
| 體外活性 | 方法：通過BRE-Luc報告基因?qū)嶒?，在C2C12細胞中評估Saracatinib對caALK2的抑制活性，IC50為14 nM；在MDA-MB-231細胞中，Saracatinib對BMP6誘導(dǎo)的BRE-Luc信號抑制IC50為8.9 nM。
結(jié)果：Western blot顯示，在C2C12細胞中，100 nM Saracatinib可完全抑制BMP7誘導(dǎo)的SMAD1/5磷酸化；在FOP患者原代成纖維細胞中，100 nM Saracatinib有效抑制Activin A誘導(dǎo)的SMAD1/5磷酸化。[1]
方法：在NRK-49F細胞中，采用Western blot實驗，以Src抑制劑Saracatinib預(yù)處理1小時后，再加入Vitronectin刺激。
結(jié)果：Saracatinib可抑制Vitronectin誘導(dǎo)的Src磷酸化及下游纖維化相關(guān)蛋白的表達，證實其通過抑制Src信號阻斷成纖維細胞活化。[2] |
| 體內(nèi)活性 | 方法：在HCC-1954乳腺癌荷瘤裸鼠模型中，Saracatinib以25 mg/kg每日口服灌胃給藥，溶劑為0.25%羧甲基纖維素鈉，連續(xù)治療28天。
結(jié)果：Saracatinib單藥可有效抑制腫瘤生長，與抗ErbB2抗體H2-18聯(lián)用后抗腫瘤效果顯著增強，且未觀察到明顯毒性。[3] |
| 存儲條件 | Store at low temperature,Keep away from moisture
Pure form: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | DMSO : 260 mg/mL (479.68 mM), Sonication is recommended.
Ethanol : 29 mg/mL (53.5 mM), Sonication is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 5 mg/mL (9.22 mM), Sonication is recommended.
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| 關(guān)鍵字 | Src | Saracatinib | Lyn | Lck | Inhibitor | inhibit | Fyn | FGR | EGFR (L861Q) | EGFR (L858R) | c-Yes | c-Src | BLK | AZD-0530 | AZD 0530 | Autophagy |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Aceglutamide | Alginic acid | Hemin | Hydroxychloroquine | Sildenafil citrate | Stavudine | Tamoxifen | Adenosine | Paeonol | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 臨床失敗化合物庫 | 抗衰老化合物庫 | 抗病毒庫 | 免疫/炎癥分子化合物庫 | 膜蛋白靶向化合物庫 | 藥物功能重定位化合物庫 | 疼痛相關(guān)化合物庫 | 抗癌臨床化合物庫 |