生物活性
Prexasertib dihydrochloride (LY2606368 HCl) 是一种有效的,ATP竞争性的选择性Chk1抑制剂,Ki值为0.9 nM,对CHK1和CHK2的IC50值分别为 <1 nM 和 8 nM。LY2606368处理将导致处于S期的细胞很快出现TUNEL和pH2AXy阳性的双链DNA断裂。在含p53缺陷的HeLa细胞中,LY2606368可有效取消doxorubicin激活的G2-M检查点,EC50为9 nM。LY2606368可广泛地在多种细胞中发挥抗增殖作用,在多数敏感细胞系中,IC50s < 50 nM;仅有少数细胞系对LY2606368具有抗性,IC50s > 1000 nM。
在体内研究中,LY2606368能有效地抑制肿瘤异种移植模型中的肿瘤生长,发挥抗肿瘤活性。在SKOV3卵巢癌模型中,LY2606368可抑制原发肿瘤的生长并显著地减少转移率和腹水积累。在SW1990原位胰脏癌模型中,LY2606368可抑制92%的原发肿瘤生长,阻止其向淋巴结结核、脾脏和肠道的转移。
化学数据
分子量 | 438.31 |
分子式 | C18H19N7O2.2HCl |
CAS号 | 1234015-54-3 |
纯度 | >99% |
溶解性(25°C) | DMSO 2 mg/mL |
储存和运输条件 | 固体粉末: -20°C 冷藏长期储存 常温运输及临时存放 |
实验操作 来自于公开的文献,仅供相同实验参考(如实验材料、目的不同,请参考其他文献)
细胞实验 |
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细胞系 | HeLa cells |
方法 | HeLa cells were plated onto T25 flasks and allowed to recover for 24 hours. LY2606368 was then added to give final concentrations of 33 or 100 nmol/L. In some experiments, 20μmol/L Z-VAD-FMK was included during the drug treatment. Cells were treated for 12 hours, and during the last 2 hours, colchicine was added to 1 μg/mL. Fixation of nuclei for metaphase spreads was done following the method of Bayani and Squire. Chromosome spreads were done. A 12-μL volume of cell suspension in 3:1 methanol/acetic acid fixative was dropped from a height of 3 cm onto dry glass slides or coverslips. The slides were then heated for 45 seconds on a 43°C metal block, before being removed to allow drying to complete at room temperature. Coverslips were mounted on slides with Vectashield Hard Set mounting medium with DAPI. Slides were examined with a Leica DMR fluorescence microscope and images were captured using a SPOT RT3 Slider camera. |
浓度 | 33 or 100 nmol/L |
处理时间 | 12 h |
动物实验 |
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动物模型 | Female CD-1 nu-/nu- mice |
配制 | 20% Captisol, pH4 |
剂量 | 15 mg/kg |
给药处理 | s.c. |
不同实验动物依据体表面积的等效剂量转换表(数据来源于FDA指南)
| 小鼠 | 大鼠 | 兔 | 豚鼠 | 仓鼠 | 狗 |
重量 (kg) | 0.02 | 0.15 | 1.8 | 0.4 | 0.08 | 10 |
体表面积 (m2) | 0.007 | 0.025 | 0.15 | 0.05 | 0.02 | 0.5 |
Km系数 | 3 | 6 | 12 | 8 | 5 | 20 |
动物 A (mg/kg) = 动物 B (mg/kg) × | 动物 B的Km系数 |
动物 A的Km系数 |
例如,依据体表面积折算法,将化合物用于小鼠的剂量20 mg/kg 换算成大鼠的剂量,需要将20 mg/kg 乘以小鼠的Km系数(3),再除以大鼠的Km系数(6),得到化合物用于大鼠的等效剂量为10 mg/kg。
储备液配制
以下数据基于产品分子量,对于特殊产品,请参照COA中的储备液配制条件和说明进行操作。
Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
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1 mM | 2.2815 mL | 11.4075 mL | 22.8149 mL |
5 mM | 0.4563 mL | 2.2815 mL | 4.563 mL |
10 mM | 0.2281 mL | 1.1407 mL | 2.2815 mL |