生物活性
PF-477736是一种有效的，选择性，ATP竞争性Chk1抑制剂，Ki为0.49 nM，也抑制VEGFR2， Aurora-A， FGFR3， Flt3， Fms (CSF1R)， Ret和Yes。作用于Chk1比作用于Chk2选择性高100倍左右。PF 477736(128 nM)作用于CA46 和 HeLa细胞，废除Camptothecin诱导的DNA损伤检查点，这种作用具有剂量依赖性。PF 477736作用于HT29细胞，有效废除Gemcitabine诱导的S期停滞，使凋亡细胞群相应增加。PF 477736(540 nM)作用于HT29细胞，增强Gemcitabine诱导的细胞毒性,这种作用具有时间和剂量依赖性。
化学数据
| 分子量 | 419.48 |
| 分子式 | C22H25N7O2 |
| CAS号 | 952021-60-2 |
| 纯度 | 99.21% |
| 溶解性（25°C） | DMSO 40 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 |
|---|
| 细胞系 | p53-defective human cancer cell lines HT29 cells |
| 方法 | IC50 MTT.
The proliferation assays were done using an MTT assay. The IC50 assay measures the antiproliferative effects of PF-00477736 on p53-defective human cancer cell lines. Cells in each line were seeded in complete medium at an exponentially growing density in 96-well assay plate and allowed to attach for 16 h. Serial dilutions of PF-00477736 were then done, and appropriate controls were added to each plate. Cells were incubated with drug for 96 h. After incubation, MTT working stock diluted in complete medium was added to each well, and cells were incubated for 4 h. After centrifugation and supernatant removal, DMSO was added to each well and plates were read on SpectraMax plate reader at 540 nm. IC50 and PF50 results were calculated by concentration-response curve fitting using a Microsoft Excel–based four-variable analysis method. |
| 浓度 | 0, 180, 360 and 540 n M |
| 处理时间 | 96 h |
| 动物实验 |
|---|
| 动物模型 | Colo205 cells tumour Xenograft Models in Athymic Nude Mice |
| 配制 | 50 nM sodium acetate buffer and 4% dextrose (pH 4) |
| 剂量 | 4-60 mg/kg q3d × 4 |
| 给药处理 | i.p. |
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 |
|---|
| 1 mM | 2.3839 mL | 11.9195 mL | 23.839 mL |
| 5 mM | 0.4768 mL | 2.3839 mL | 4.7678 mL |
| 10 mM | 0.2384 mL | 1.192 mL | 2.3839 mL |
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