PIK-75-PIK-75-Hydrochloride-DataSheet-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPIK-75Cat. No.: HY-13281CAS No.: 372196-77-5Synonyms: PIK-75 Hydrochloride分式: CHBrClNOS分量: 488.74作靶點: DNA-PK; PI3K作通路: Cell Cycle/DNA Damage; PI3K/Akt/mTOR儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性

2、數(shù)據(jù)體外實驗 DMSO : 30 mg/mL (61.38 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.0461 mL 10.2304 mL 20.4608 mL5 mM 0.4092 mL 2.0461 mL 4.0922 mL10 mM 0.2046 mL 1.0230 mL 2.0461 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 PIK-75種 D

3、NA-PK 和 PI3K 抑制劑，抑制 DNA-PK，p110 和 p110，IC50 分別為 2, 5.8 和 76 nM。PIK-75 抑制 p110 效果抑制 p110 (IC50=1.3 M) 200 多倍。IC50 & Target DNA-PK p110 p110 p1102 nM (IC50) 5.8 nM (IC50) 76 nM (IC50) 510 nM (IC50)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEp110 hsVPS34 PI3KC2 PI3KC21.3 M (IC50) 2.6 M (IC50) 1 M

4、(IC50) 10 M (IC50)mTORC1 mTORC2 ATM ATR1 M (IC50) 10 M (IC50) 2.3 M (IC50) 21 M (IC50)PI4KIII50 M (IC50)體外研究 PIK-75 also inhibits p110, PI3KC2, mTORC1, ATM, hsVPS34, PI3KC2, mTORC2, ATR and PI4KIII withIC50s of 510 nM, 1 M, 1 M, 2.3 M, 2.6 M, 10 M, 10 M, 21 M, 50 M, respectively. PIK-75alone blocks

5、Thr 308 phosphorylation in L6 myotubes and 3T3-L1 adipocytes with IC50 values of 1.2 and 1.3M, respectively 1. PIK-75 is a competitive p110 inhibitor with respect to a substrate, phosphatidylinositol(PI) in contrast to most other PI3K inhibitors, which bind at or near the ATP site. Using sequence an

6、alysisand the existing crystal structures of inhibitor complexes with the p110 and p110 isoforms, a new region ofnonconserved amino acids (region 2) is identified that is postulated to be involved in PIK-75 p110selectivity. Analysis of region 2, using in vitro mutation of identified nonconserved ami

7、no acids to alanine,shows that Ser773 is a critical amino acid involved in PIK-75 binding, with an 8-fold-increase in the IC50compared with wild-type. Further kinetic experiments are undertaken to determine the effect of PIK-75 on thekinetics of binding of ATP and PI to the p11 S773D mutant. Activit

8、y is estimated using a range of PIconcentrations at the concentrations of 0, 50, 100 and 200 nM PIK-75. The Km for PI is 11.2 M comparedwith 7.0 M for the wild-type enzyme. The Ki for PIK-75 is estimated to be 146 nM, a 64-fold increase on thevalue estimated for the wild-type enzyme (2.3 nM) 2. MIA

9、PaCa-2 and AsPC-1 cells are treated withincreasing concentration of PIK-75 for 48 h and the cell viability is determined by MTT assay. PIK-75 inhibitsthe proliferation of pancreatic cancer cells via apoptotic cell death. Submicromolar concentration of PIK-75 issufficient to inhibit the proliferation

10、 of pancreatic cancer, MIA PaCa-2 and AsPC-1 cells after 48-h treatment.PIK-75 also reduces the colony formation of pancreatic cancer MIA PaCa-2 and AsPC-1 cells 3體內(nèi)研究 PIK-75 enhances the antitumor effect of Gemcitabine in vivo. The effect of PIK-75/Gemcitabine combinationis further demonstrated by

11、in vivo mouse xenograft model. Mice bearing tumors of MIA PaCa-2 areadministered with Gemcitabine (20 mg/kg), PIK-75 (2 mg/kg), or combination of both drugs. Since PIK-75 is areversible inhibitor, PIK-75 is administered 5 times per week to ensure maintaining sufficient inhibitoryeffects. Gemcitabine

12、 is administered twice per week. Gemcitabine or PIK-75 reduces the tumor growth tosimilar degree 3PROTOCOLKinase Assay 2 PI3K enzyme activity is determined in 50 L of 20 mM HEPES, pH 7.5, and 5 mM MgCl2 containing 180 Mphosphatidyl inositol, with the reaction starts by the addition of 100 M ATP (con

13、taining 2.5 Ci of -32PATP). After a 30-min incubation at room temperature, the enzyme reaction is stopped by the addition of 50L of 1 M HCl. Phospholipids are then extracted with 100 L of chloroform/methanol 1:1 (v/v) and 250 L of2 M KCl followed by liquid scintillation counting. Inhibitors (e.g., P

14、IK75) ) are diluted in 20% (v/v) DMSO togenerate a concentration versus inhibition of enzyme activity curve, which is then analyzed with the use of2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEPrism version 5.00 for Windows to calculate the IC50 2.MCE has not independently confirmed the accuracy

15、of these methods. They are for reference only.Cell Assay 3 MIA PaCa-2 cells are maintained in Dulbeccos modified Eagles medium (DMEM) containing 10% heat-inactivated fetal bovine serum (HI-FBS), 2.5% horse serum (HS) and 100 U/mL Penicillin/Streptomycin.AsPC-1 cells are cultured in RPMI-1640 media s

16、upplemented with 20% HI-FBS, 100 U/mLPenicillin/Streptomycin and 1 mM sodium pyruvate. A total of 2,000 human pancreatic cancer cells (MIAPaCa-2 or AsPC-1) per well are plated in 96-well flat-bottom plates and then treated with either Gemcitabine,PIK-75 alone (0.1 M, 0.3 M and 1 M) or in combination

17、 of both drugs with indicated concentrations. At theindicated times, 20 L of 1 mg/mL MTT in PBS is added to each well and further incubated for 4 h. Aftercentrifugation and removal of the medium, 150 L of DMSO is added to each well to dissolve the formazancrystals. The absorbance is measured at 562

18、nm using an ELx808 absorbance microplate reader.Absorbance of untreated cells is designated as 100%, and the relative viable cells are expressed as apercentage of this value 3MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 3Administration 3

19、MIA PaCa-2 cells (1.7106 cells/mouse) mixed with Matrigel are injected subcutaneously into the flank ofmale athymic nude (Foxn1nu) mice aged 6-weeks. Gemcitabine (50 mg/mL) is dissolved in PBS and PIK-75(20 mg/mL) is dissolved in DMSO. Injection solution is made as 10% of Cremophor EL and 3% ofpoly(

20、ethylene glycol) 400 in sterile water. Before administration of compounds, Gemcitabine is further dilutedin PBS and DMSO or PIK-75 is further diluted in the injection solution and sterilized by 0.2 m filter unit.These diluents are mixed with 1:1 ratio and administered into peritoneal cavity of the m

21、ouse. Gemcitabine(20 mg/kg) or Gemcitabine (20 mg/kg)/PIK-75 (2 mg/kg) combination is administered twice per week andvehicle control and PIK-75 (2 mg/kg) are administered 5 times per week. The body weights and tumor sizesare measured 3 times per week. Tumor volumes are calculated.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENC