PF-477736
(Synonyms: PF 00477736) 目录号 : GC14234
A selective inhibitor of checkpoint kinase 1
Cas No.:952021-60-2
Sample solution is provided at 25 µL, 10mM.
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PF-477736 is a potent, selective ATP-competitive and small-molecule inhibitor of Chk1 with a Ki of 0.49±0.29nM for the in vitro kinase activity of Chk1 [1].
PF-477736 has shown a selective inhibition of Chk1 with the IC50 values of 0.49nM, 9.9μM and 47nM for Chk1, CDK1 and Chk2, respectively. In a dot-blot assay, PF-477736 has been reported to inhibit camptothecin-induced G2 arrest with the EC50 values of 45nM, 38nM and 42nM in P53-mutated human lymphoma CA46 cells, HeLa cells and HT29 cells. Apart from these, PF-477736 has been found to selectively target p53-defective cancer cells while having minimal cytotoxic effects on normal (p53-competent) cells. In addition, PF-00477736 has been revealed to dose-dependently enhance the antitumor activity of a MTD of gemcitabine with no apparent exacerbation of systemic toxicity as assessed by monitoring body weight in the Colo205 xenograft [1].
References:
[1] Blasina A1, Hallin J, Chen E, Arango ME, Kraynov E, Register J, Grant S, Ninkovic S, Chen P, Nichols T, O'Connor P, Anderes K. Breaching the DNA damage checkpoint via PF-00477736, a novel small-molecule inhibitor of checkpoint kinase 1. Mol Cancer Ther. 2008 Aug;7(8):2394-404.
| Kinase experiment [1]: | |
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Kinase assays |
CHK1 activity was measured by a pyruvate kinase-lactate dehydrogenase coupled, continuous spectrophotometric assay method where the phosphorylation of a CHK1 peptide substrate (Syntide-2, PLARTLSVAGLPGKK) was coupled to the oxidation of NADH and the corresponding change in absorbance intensity was measured at 340 nm. The assay was performed in a 96-well plate for 20 minutes at 30°C in 0.1 mL of assay buffer containing 50 mM TRIS pH 7.5, 0.4 M NaCl, 4 mM PEP, 0.15 mM NADH, 28 units of lactate dehydrogenase/mL, 16 units of pyruvate kinase/mL, 3 mM DTT, 0.125 mM Syntide-2, 0.15 mM ATP and 25 mM magnesium chloride. Assays were initiated with 1nM of CHK1 kinase domain. The inhibition of CHK1 activity was determined by measuring initial velocities in the presence of varying concentrations of PF-477736. |
| Cell experiment [2]: | |
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Cell lines |
HT29 cell lines |
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Preparation method |
The solubility of this compound in DMSO is >5.2mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
|
Reacting condition |
69 nM, 24h |
|
Applications |
The gemcitabine and PF-477736 combination induced significant potentiation of the cytotoxic activity of gemcitabine, indicating that the combination treatment caused a permanent cellular damage that cannot be overcome even after drug removal. The cell kill induced by the combination treatment is time and dose dependent. |
| Animal experiment [2]: | |
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Animal models |
Colo205 Xenograft Models in Athymic Nude Mice |
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Dosage form |
i.p., 4-60 mg/kg，q3d × 4 |
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Application |
In xenografts, PF-477736 enhanced the antitumor activity of gemcitabine in a dose-dependent manner. |
|
Other notes |
Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal. |
|
References: [1] Blasina A1, Hallin J, Chen E, Arango ME, Kraynov E, Register J, Grant S, Ninkovic S, Chen P, Nichols T, O'Connor P, Anderes K. Breaching the DNA damage checkpoint via PF-00477736, a novel small-molecule inhibitor of checkpoint kinase 1. Mol Cancer Ther. 2008 Aug;7(8):2394-404. | |
| Cas No. | 952021-60-2 | SDF | |
| 别名 | PF 00477736 | ||
| 化学名 | (R)-2-amino-2-cyclohexyl-N-(2-(1-methyl-1H-pyrazol-4-yl)-6-oxo-5,6-dihydro-4H-[1,2]diazepino[4,5,6-cd]indol-8-yl)acetamide | ||
| Canonical SMILES | O=C(C(C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C1([H])[H])([H])N([H])[H])N([H])C2=C([H])C3=C4C(N=C(C5=C([H])N(C([H])([H])[H])N=C5[H])C4=C([H])N([H])N([H])C3=O)=C2[H] | ||
| 分子式 | C22H25N7O2 | 分子量 | 419.48 |
| 溶解度 | ≥ 5.2mg/mL in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
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 |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
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，
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 saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet