| 名称 | 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. |
| 细胞实验 | 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]. |
| 激酶实验 | 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]. |
| 动物实验 | 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]. |
| 体外活性 | 方法：通过BRE-Luc报告基因实验，在C2C12细胞中评估Saracatinib对caALK2的抑制活性，IC50为14 nM；在MDA-MB-231细胞中，Saracatinib对BMP6诱导的BRE-Luc信号抑制IC50为8.9 nM。
结果：Western blot显示，在C2C12细胞中，100 nM Saracatinib可完全抑制BMP7诱导的SMAD1/5磷酸化；在FOP患者原代成纤维细胞中，100 nM Saracatinib有效抑制Activin A诱导的SMAD1/5磷酸化。[1]
方法：在NRK-49F细胞中，采用Western blot实验，以Src抑制剂Saracatinib预处理1小时后，再加入Vitronectin刺激。
结果：Saracatinib可抑制Vitronectin诱导的Src磷酸化及下游纤维化相关蛋白的表达，证实其通过抑制Src信号阻断成纤维细胞活化。[2] |
| 体内活性 | 方法：在HCC-1954乳腺癌荷瘤裸鼠模型中，Saracatinib以25 mg/kg每日口服灌胃给药，溶剂为0.25%羧甲基纤维素钠，连续治疗28天。
结果：Saracatinib单药可有效抑制肿瘤生长，与抗ErbB2抗体H2-18联用后抗肿瘤效果显著增强，且未观察到明显毒性。[3] |
| 存储条件 | Store at low temperature,Keep away from moisture
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 : 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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| 关键字 | Src | Saracatinib | Lyn | Lck | Inhibitor | inhibit | Fyn | FGR | EGFR (L861Q) | EGFR (L858R) | c-Yes | c-Src | BLK | AZD-0530 | AZD 0530 | Autophagy |
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| 相关库 | 抑制剂库 | 已知活性化合物库 | 激酶抑制剂库 | 临床失败化合物库 | 抗病毒库 | 神经退行性疾病化合物库 | 膜蛋白靶向化合物库 | 免疫/炎症分子化合物库 | 药物功能重定位化合物库 | 疼痛相关化合物库 | 酪氨酸激酶分子库 | 抗癌药物库 |