PF-477736 (Synonyms: PF 00477736) |
| Catalog No.GC14234 |
Le PF-477736 (PF 00477736) est un inhibiteur puissant, sélectif et compétitif pour l'ATP de Chk1, avec un Ki de 0,49 nM, c'est aussi un inhibiteur de Chk2, avec un Ki de 47 nM. PF-477736 montre une sélectivité \u003c100 fois pour Chk1 sur VEGFR2, Fms, Yes, Aurora-A, FGFR3, Flt3 et Ret (IC50 \u003d 8 (Ki), 10, 14, 23, 23, 25 et 39 nM, respectivement). Le PF-477736 peut renforcer l'activité antitumorale de la gemcitabine in vitro et in vivo.
Products are for research use only. Not for human use. We do not sell to patients.
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]: | |
|
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]: | |
|
Cell lines |
HT29 cell lines |
|
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]: | |
|
Animal models |
Colo205 Xenograft Models in Athymic Nude Mice |
|
Dosage form |
i.p., 4-60 mg/kg，q3d × 4 |
|
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 | |
| Synonymes | PF 00477736 | ||
| Chemical Name | (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] | ||
| Formula | C22H25N7O2 | M.Wt | 419.48 |
| Solubility | ≥ 5.2mg/mL in DMSO | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
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 |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
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Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 25 reference(s) in Google Scholar.)
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