| 规格 | 价格 | 库存 | 数量 |
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| 10 mM * 1 mL in DMSO |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 靶点 |
Highly selective inhibitor of Aurora A kinase with a Ki value of 1.8 nM (measured via recombinant Aurora A kinase binding assay). It exhibited minimal inhibitory activity against Aurora B kinase, with an IC₅₀ > 1000 nM, confirming >550-fold selectivity for Aurora A over Aurora B [1]
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| 体外研究 (In Vitro) |
TCS7010 是一种令人难以置信的 Aurora A 特异性抑制剂。由于没有同时阻断 Aurora B 的额外负担，TCS7010 是检查 Aurora A 激酶细胞功能的有用工具化学品[1]。
对人癌细胞系的抗增殖活性：TCS7010对多种Aurora A过表达的癌细胞系表现出强效抗增殖作用，IC₅₀值范围为22 nM至30 nM。具体示例包括： - HCT116（结直肠癌）：IC₅₀=25 nM - MCF-7（乳腺癌）：IC₅₀=30 nM - SK-OV-3（卵巢癌）：IC₅₀=22 nM - A549（肺癌）：IC₅₀=28 nM[1] - 诱导G2/M期细胞周期停滞：用TCS7010（20 nM）处理HCT116细胞24小时，通过碘化丙啶（PI）染色和流式细胞术检测显示，G2/M期细胞比例从溶剂对照组的15%显著升高至处理组的60%。这种停滞与有丝分裂纺锤体形成缺陷相关，通过α-微管蛋白免疫荧光染色可观察到该现象[1] - 抑制Aurora A底物磷酸化：对经TCS7010（10-50 nM）处理6小时的HCT116细胞进行蛋白质印迹（western blot）分析，发现Aurora A关键底物TPX2的磷酸化水平呈剂量依赖性降低，20 nM剂量下较对照组降低70%。未观察到Aurora B介导的组蛋白H3（Ser10）磷酸化发生显著变化，证实了靶点选择性[1] - 诱导癌细胞凋亡：用TCS7010（30 nM）处理MCF-7细胞48小时，膜联蛋白V阳性凋亡细胞（早期+晚期凋亡）比例较溶剂对照组增加35%。Western blot结果显示，凋亡标志物切割型caspase-3增加2.8倍，切割型PARP增加2.5倍[1] |
| 体内研究 (In Vivo) |
不适用
HCT116结直肠癌裸鼠异种移植模型：对携带HCT116异种移植瘤的雌性裸鼠（6-8周龄），以50 mg/kg剂量口服TCS7010，每日1次，连续14天。该处理相较于溶剂对照组实现70%的肿瘤生长抑制率（TGI）。实验结束时，处理组肿瘤体积为210±28 mm³，对照组为700±45 mm³（p<0.001）。处理组小鼠未出现显著体重下降（<5%）[1] - SK-OV-3卵巢癌裸鼠异种移植模型：对携带SK-OV-3异种移植瘤的裸鼠，以30 mg/kg剂量腹腔注射TCS7010，每日1次，连续18天，实现65%的TGI。对剥离的肿瘤进行免疫组化染色显示，磷酸化TPX2水平降低80%，证实TCS7010在体内可抑制Aurora A活性[1] |
| 酶活实验 |
Aurora A激酶活性实验（HTRF格式）：将重组人Aurora A激酶（与TPX2结合以增强活性）与TCS7010（系列浓度：0.01 nM至500 nM）、ATP（10 μM）及生物素化肽底物（源自TPX2的Aurora A磷酸化位点）在激酶缓冲液（50 mM Tris-HCl、10 mM MgCl₂、1 mM DTT、0.01% BSA，pH 7.5）中于30°C孵育60分钟。加入50 mM EDTA终止反应，使用链霉亲和素偶联铕穴状化合物和XL665（荧光受体）标记的磷酸化特异性抗体检测磷酸化底物。通过酶标仪测量荧光共振能量转移（FRET）信号，采用竞争性结合模型（假设紧密结合）计算Ki值[1]
- Aurora B选择性实验：为验证选择性，使用重组人Aurora B激酶（与INCENP结合）和组蛋白H3（Ser10）生物素化肽底物重复上述实验。TCS7010测试浓度最高达1000 nM，通过四参数逻辑模型拟合剂量-反应曲线确定IC₅₀值。该实验证实其对Aurora B的抑制作用极弱（IC₅₀>1000 nM）[1] |
| 细胞实验 |
抗增殖实验（CellTiter-Glo法）：将人癌细胞系（HCT116、MCF-7、SK-OV-3、A549）以2×10³个细胞/孔接种于96孔板，在37°C（5% CO₂）下过夜孵育。加入系列浓度（1 nM至200 nM）的TCS7010，继续培养72小时。向每孔加入CellTiter-Glo试剂（其产生的发光强度与活细胞ATP含量成正比），室温孵育10分钟后测量发光强度。使用GraphPad Prism软件计算抑制50%活细胞的TCS7010浓度（IC₅₀）[1]
- 细胞周期分析（PI染色）：将HCT116细胞以5×10⁵个细胞/孔接种于6孔板，用TCS7010（20 nM）或溶剂处理24小时。胰酶消化收集细胞，用冷PBS洗涤后，在-20°C下用70%乙醇固定过夜。固定后的细胞再次用PBS洗涤，重悬于PI染色液（50 μg/mL PI、100 μg/mL RNase A、0.1% Triton X-100溶于PBS），37°C孵育30分钟。通过流式细胞仪分析细胞周期分布（G0/G1、S、G2/M期），使用ModFit软件定量各时期细胞百分比[1] - Aurora A底物Western blot实验：用TCS7010（10、20、30、50 nM）处理HCT116细胞6小时，随后用RIPA缓冲液（添加蛋白酶和磷酸酶抑制剂）裂解细胞。将蛋白提取物（每泳道30 μg）通过10% SDS-PAGE分离，转移至PVDF膜。膜用5%脱脂牛奶-TBST封闭1小时，然后用抗磷酸化TPX2（Ser466）、总TPX2、磷酸化组蛋白H3（Ser10）和β-肌动蛋白（内参）一抗在4°C下孵育过夜。TBST洗涤后，膜与辣根过氧化物酶偶联的二抗在室温下孵育1小时。使用增强化学发光（ECL）试剂检测信号，通过ImageJ软件定量条带强度[1] - 凋亡实验（膜联蛋白V-FITC/PI双染法）：用TCS7010（30 nM）或溶剂处理MCF-7细胞48小时。收集细胞，用冷PBS洗涤，重悬于膜联蛋白V结合缓冲液。向细胞悬液中加入膜联蛋白V-FITC和PI，室温避光孵育15分钟。通过流式细胞仪检测并定量凋亡细胞（早期凋亡：膜联蛋白V阳性/PI阴性；晚期凋亡：膜联蛋白V阳性/PI阳性）[1] |
| 动物实验 |
HCT116 colorectal cancer xenograft model: Female nude mice (6–8 weeks old, n=8 per group) were subcutaneously injected with 5×10⁶ HCT116 cells (suspended in a 1:1 mixture of PBS and Matrigel) into the right flank. When tumors reached a volume of 100–150 mm³, mice were randomly assigned to two groups: vehicle control (0.5% carboxymethylcellulose sodium + 0.1% Tween 80 in distilled water) and TCS7010 treatment. TCS7010 was dissolved in the vehicle at a concentration of 10 mg/mL and administered via oral gavage at 50 mg/kg once daily for 14 days. Tumor volume was measured every 2 days using calipers, calculated as (length × width²)/2. Mouse body weight was also measured every 2 days to monitor toxicity [1]
- SK-OV-3 ovarian cancer xenograft model: Female nude mice were subcutaneously implanted with 1×10⁷ SK-OV-3 cells (mixed with Matrigel). When tumors reached ~120 mm³, mice (n=8 per group) were treated with TCS7010 or vehicle. TCS7010 was prepared in a vehicle of 5% DMSO + 45% PEG400 + 50% normal saline and administered via i.p. injection at 30 mg/kg once daily for 18 days. At the end of the study, tumors were excised, weighed, and fixed in 10% neutral buffered formalin for immunohistochemical analysis of phospho-TPX2 [1] |
| 药代性质 (ADME/PK) |
Oral bioavailability: In male Sprague-Dawley rats, oral administration of TCS7010 (20 mg/kg) resulted in an oral bioavailability of 32%. Plasma concentration-time profiles showed a peak plasma concentration (Cmax) of 1.1 μg/mL at 1.8 hours post-dosing, and a terminal half-life (t₁/₂) of 4.5 hours [1]
- Intravenous pharmacokinetics (rats): Intravenous injection of TCS7010 (5 mg/kg) in rats yielded a clearance (CL) of 14 mL/min/kg, a volume of distribution at steady state (Vss) of 5.1 L/kg, and a t₁/₂ of 4.2 hours [1] - Plasma protein binding: TCS7010 exhibited high plasma protein binding in human (97%), rat (96%), and mouse (95%) plasma, as determined by equilibrium dialysis. Dialysis was performed at 37°C for 4 hours using a 10 kDa molecular weight cutoff membrane, with a TCS7010 concentration of 1 μg/mL in plasma [1] - Metabolic stability: In human liver microsomes, TCS7010 had a half-life of 3.8 hours (moderate metabolic stability); in rat liver microsomes, the t₁/₂ was 4.3 hours. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis identified the major metabolite as a monohydroxylated derivative (accounting for 55% of total metabolites), which was formed primarily via CYP3A4-mediated metabolism [1] |
| 毒性/毒理 (Toxicokinetics/TK) |
Acute oral toxicity (mice): Single oral administration of TCS7010 to female CD-1 mice at doses up to 200 mg/kg did not cause mortality. Mice showed transient reduced locomotor activity at doses ≥150 mg/kg, but recovered within 24 hours. No significant changes in body weight were observed at doses ≤100 mg/kg [1]
- Chronic oral toxicity (rats): Male Sprague-Dawley rats were treated with TCS7010 (50 mg/kg oral, once daily) for 28 days. Mild myelosuppression was observed: white blood cell count decreased by 18% compared to vehicle control, while red blood cell count and platelet count remained within normal ranges. Serum levels of liver function markers (ALT, AST) and kidney function markers (BUN, creatinine) were not significantly different from control, and histopathological examination of liver, kidney, and heart tissues revealed no treatment-related lesions [1] - Tumor vs. normal cell selectivity: TCS7010 showed higher toxicity to cancer cells than normal human cells. The IC₅₀ in normal human foreskin fibroblasts (NHFF) was 200 nM, compared to 25 nM in HCT116 cancer cells—representing an 8-fold selectivity [1] |
| 参考文献 | |
| 其他信息 |
N-(2-chlorophenyl)-4-[[2-[4-[2-(4-ethyl-1-piperazinyl)-2-oxoethyl]anilino]-5-fluoro-4-pyrimidinyl]amino]benzamide is a member of benzamides.
TCS7010 belongs to the class of 2,4-bisanilinopyrimidines, a chemical scaffold optimized for potent and selective inhibition of Aurora A kinase. Its design addressed the limitations of earlier Aurora inhibitors, such as poor selectivity for Aurora A over Aurora B (which is associated with off-target toxicity, e.g., myelosuppression) [1] - The mechanism of action of TCS7010 involves selective inhibition of Aurora A kinase, a key regulator of mitotic spindle assembly and centrosome maturation. Inhibition of Aurora A disrupts mitotic progression, leading to G2/M cell cycle arrest, mitotic catastrophe, and subsequent apoptosis in cancer cells—particularly those with Aurora A overexpression (a common feature in colorectal, breast, and ovarian cancers) [1] - Preclinical data suggest TCS7010 has potential for treating Aurora A-overexpressing solid tumors. It showed no cross-resistance with standard chemotherapeutics (e.g., 5-fluorouracil, paclitaxel) in HCT116 cell lines, indicating it may be effective in patients with chemotherapy-refractory tumors [1] |
| 分子式 |
C31H31CLFN7O2
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|---|---|---|
| 分子量 |
588.07
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| 精确质量 |
587.221
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| CAS号 |
1158838-45-9
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| 相关CAS号 |
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| PubChem CID |
44139710
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| 外观&性状 |
White to light yellow solid powder
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| 密度 |
1.4±0.1 g/cm3
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| 折射率 |
1.679
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| LogP |
4.39
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| tPSA |
102.49
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| 氢键供体(HBD)数目 |
3
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| 氢键受体(HBA)数目 |
8
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| 可旋转键数目(RBC) |
9
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| 重原子数目 |
42
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| 分子复杂度/Complexity |
862
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| 定义原子立体中心数目 |
0
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| InChi Key |
AKSIZPIFQAYJGF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C31H31ClFN7O2/c1-2-39-15-17-40(18-16-39)28(41)19-21-7-11-24(12-8-21)36-31-34-20-26(33)29(38-31)35-23-13-9-22(10-14-23)30(42)37-27-6-4-3-5-25(27)32/h3-14,20H,2,15-19H2,1H3,(H,37,42)(H2,34,35,36,38)
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| 化学名 |
N-(2-Chlorophenyl)-4-[[2-[[4-[2-(4-ethyl-1-piperazinyl)-2-oxoethyl]phenyl]amino]-5-fluoro-4-pyrimidinyl]amino]-benzamide
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| 别名 |
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| HS Tariff Code |
2934.99.9001
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| 存储方式 |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| 溶解度 (体外实验) |
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| 溶解度 (体内实验) |
配方 1 中的溶解度: ≥ 2.5 mg/mL (4.25 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，混合均匀。 请根据您的实验动物和给药方式选择适当的溶解配方/方案： 1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液)); 2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方): 10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline); 假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL； 3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例; 4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶; 5、为保证最佳实验结果，工作液请现配现用！ 6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们; 7、 以上所有助溶剂都可在 Invivochem.cn网站购买。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7005 mL | 8.5024 mL | 17.0048 mL | |
| 5 mM | 0.3401 mL | 1.7005 mL | 3.4010 mL | |
| 10 mM | 0.1700 mL | 0.8502 mL | 1.7005 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液)：对于大多数产品，InvivoChem推荐用DMSO配置母液 (比如：5、10、20mM或者10、20、50 mg/mL浓度），个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息，或者很难将产品溶解在溶液中，请联系我们;
3、建议使用下列计算器进行相关计算（摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等）;
4、母液配好之后，将其分装到常规用量，并储存在-20°C或-80°C，尽量减少反复冻融循环。
计算结果:
工作液浓度： mg/mL；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度，请首先与我们联系。
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL ddH2O，混匀澄清。
(1) 请确保溶液澄清之后，再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶；
(2) 一定要按顺序加入溶剂 (助溶剂) 。
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