Antibodies
PP2
- 貨號 Cat. No.
- S7008
- 規格 Size
- 5~200mg
- 品牌 Brand
- Selleckchem
Description
PP2
Catalog No.S7008
PP2 is Src family kinase inhibitor, potently inhibits Lck/Fyn with IC50 of 4 nM/5 nM, ~100-fold less potent to EGFR, inactive for ZAP-70, JAK2 and PKA.
| Size | Price | Stock | Quantity | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ||||||||||||
Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw
Biological Activity
| Description | PP2 is Src family kinase inhibitor, potently inhibits Lck/Fyn with IC50 of 4 nM/5 nM, ~100-fold less potent to EGFR, inactive for ZAP-70, JAK2 and PKA. | |||||
|---|---|---|---|---|---|---|
| Targets | Lck | Fyn | ||||
| IC50 | 4 nM | 5 nM | ||||
| In vitro | PP2 (5 μM) blocks in vivo phosphorylation and signaling of the RET/PTC1 oncoprotein in NIH3T3 and NIH-RET/PTC3 cell lysates. PP2 (5 μM) prevents serum-independent growth of RET/PTC1-transformed NIH3T3 fibroblasts and of TPC1 and FB2, two human papillary thyroid carcinoma cell lines that carry spontaneous RET/PTC1 rearrangements. PP2 (5 μM) blocks invasion of type I collagen matrix by TPC1 cells. [1] PP2 inhibits Src by binding to an area of the molecule that does not overlap with the ATP binding domain. [2] PP2 (20 μM) induces 40-50% growth inhibition of HT29 cells, this concentration reduces the Src activity as early as 1 hour and maintains a 35% inhibition of Src activity for 2 days. PP2 (100 mM) decreases the Src activity of HT29 cells in a dose-dependent manner. PP2 (1 mM-100 mM) causes a dose-dependent growth inhibition of human colon cancer cells (HT29, SW480, and PMCO1), liver cancer cells (PLC/PRF/5, KYN-2, Li7, and HepG2), and breast cancer cells (MCF-7, MDA-MB-468, and BT-474). PP2 (20 μM) significantly increases aggregation in most of the cancer cells (HT29, SW480, PMCO1, PLC/PRF/5, KYN-2, Li7, MCF-7, and MDA-MB-468) in E-cadherin dependent manner. PP2 (20 μM) enhances E-cadherin expression and also strongly increases E-cadherin’s association with the actin cytoskeleton in cancer cells. PP2 (20 μM) increases the expression of α-catenin, β-catenin, and γ-catenin in HT29 cells, whereas in PLC/PRF/5 and MCF-7 cells, the total protein level of α-catenin does not change, but the levels of β- catenin and γ-catenin increases slightly. [3] PP2 inhibits proliferation of two cervical cancer cells (HeLa and SiHa) in a time- and dose-dependent manner. PP2 (10 μM) down-regulates pSrc-Y416, pEGFR-Y845, and -Y1173 expression levels in HeLa and SiHa cells. PP2 (10 μM) could modulate cell cycle arrest by up-regulating p21(Cip1) and p27(Kip1) in both HeLa and SiHa cells and down-regulating expression of cyclin A, and cyclin dependent kinase-2, -4 (Cdk-2, -4) in HeLa and of cyclin B and Cdk-2 in SiHa.[4] | |||||
| In vivo | PP2 (5 mg/kg/day) induces some slowing in the growth rate of the primary tumors relative to the control treated with vehicle in SCID mice inoculated HT29 cells in the spleen. PP2 (5 mg/kg/day) induces some slowing in the growth rate of the primary tumors relative to the control treated with vehicle in SCID mice inoculated HT29 cells in the spleen. PP2 (5 mg/kg/day) significantly reduces the relative liver weight and liver metastasis volume compared with the controls in SCID mice inoculated HT29 cells in the spleen. [3] PP2 (1.5 mg/kg i.p.) treated rats show approximately 50% reduction of infarct size on T2-weighted MRI and in TTC staining compared with controls in rats with focal ischemic brain injury. PP2 (1.5 mg/kg i.p.) results in better the neurological score than controls in rats with focal ischemic brain injury. [5] | |||||
| Features | ||||||
Protocol(Only for Reference)
Kinase Assay: [1]
| In vitro kinase assays | Subconfluent HEK293 cells are transiently transfected with the appropriate plasmids and solubilized in lysis buffer. Proteins are immunoprecipitated with the required antibodies; immunocomplexes are recovered with protein A Sepharose beads, washed five times with kinase buffer and incubated (20 min at room temperature) in kinase buffer containing 200 μM poly(L-glutamic acid-L-tyrosine) (poly-GT), 2.5 μCi [γ-32P]ATP and unlabeled ATP to a final concentration of 20 μM in the presence of PP2 or vehicle. Samples are spotted on Whatman 3MM paper and washed five times with 1% orthophosphoric acid. 32P incorporation is measured with a β-counter scintillator. Glutathione-S-transferase (GST)-RET/tyrosine kinase (TK) is purified from HEK293 cell lysates using glutathione Sepharose according to standard procedures. |
|---|
Cell Assay: [3]
| Cell lines | HT29, SW480, PMCO1, PLC/PRF/5, KYN-2, Li7, HepG2, MCF-7, MDA-MB-468 and BT-474 cell lines |
|---|---|
| Concentrations | ~100 μM |
| Incubation Time | 2 days |
| Method | Cell viability is determined using an in vitro toxicology assay kit following the manufacturer’s instructions. Cells are seeded in 96-well plates at day 0. Starting at day 1, cells are treated for 2 days with each of a series of increasing concentrations of PP2 (1 μM, 10 μM, and 100 μM). At the end of this period, cell proliferation is evaluated by a colorimetric assay based on the cleavage of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide by mitochondria dehydrogenase in viable cells, leading to formazan formation. This experiment is repeated three times with 10 determinations/tested concentration. |
Animal Study: [3]
| Animal Models | SCID mice inoculated HT29 cells in the spleen |
|---|---|
| Dosages | 5 mg/kg/day |
| Administration | intraperitoneal injection |
| Solubility | 1% DMSO/30% polyethylene glycol/1% Tween 80, 30 mg/mL |
References
Clinical Trial Information( data from http://clinicaltrials.gov)
| NCT Number | Recruitment | Conditions | Sponsor /Collaborators | Start Date | Phases |
|---|---|---|---|---|---|
| NCT00695409 | Recruiting | Lymphoma | City of Hope Medical Center|National Cancer Institute (NCI) | 2008-03 | Phase 2 |
| NCT00797797 | Completed | Fibromyalgia | Forest Laboratories | 2008-11 | Phase 3 |
| NCT01047605 | Completed | Healthy | Pascoe Pharmazeutische Praeparate GmbH | 2010-01 | Phase 1 |
| NCT01434472 | Recruiting | Post-transplant Lymphoproliferative Disorder|Recurrent Adult Burkitt Lymphoma|Recurrent Adult Diffuse Large Cell Lymphoma|Recurrent Adult Diffuse Mixed Cell Lymphoma|Recurrent Adult Grade III Lymphomatoid Granulomatosis etc. | Fred Hutchinson Cancer Research Center|Fred Hutchinson Cancer Research Center/University of Washington Cancer Consortium|National Cancer Institute (NCI) | 2011-11 | Phase 2 |
| NCT01686165 | Recruiting | Anaplastic Large Cell Lymphoma|Recurrent Adult Diffuse Large Cell Lymphoma|Recurrent Mantle Cell Lymphoma | University of Arizona|Thomas P. Miller|National Cancer Institute (NCI) | 2012-09 | Phase 2 |
Chemical Information
| Molecular Weight (MW) | 301.77 |
|---|---|
| Formula | C15H16ClN5 |
| CAS No. | 172889-27-9 |
| Storage | 3 years -20℃Powder |
|---|---|
| 6 months-80℃in DMSO | |
| Syonnyms | AG 1879 |
| Solubility (25°C) * | In vitro | DMSO | 60 mg/mL (198 mM) |
|---|---|---|---|
| Water | <1 mg/mL (<1 mM) | ||
| Ethanol | 2 mg/mL (6 mM) | ||
| In vivo | 1% DMSO/30% polyethylene glycol/1% Tween 80, | 30 mg/mL | |
| * <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. | |||
| Chemical Name | 1H-Pyrazolo[3,4-d]pyrimidin-4-amine, 3-(4-chlorophenyl)-1-(1,1-dimethylethyl)- |
|---|
Preparing Stock Solutions
| Stock Solution (1ml DMSO) | 1mM | 10mM | 20mM | 30mM |
|---|---|---|---|---|
| Mass(mg) | 0.30177 | 3.0177 | 6.0354 | 9.0531 |
