PIK-75_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:PIK–75 is a p110α inhibitor with IC50 of 5.8 nM (200–fold more potently than p110β), isoform–specific mutants at Ser773, and also potently inhibits DNA–PK with IC50 of 2 nM.IC50 value: 5.8/2 nM(p110α/DNA–PK) [1] [2]Target: p110α; DNA–PKin vitro: PIK–75 shows the impressive potency and isoform selectivity at p110α while the corresponding IC50 values are 1300 nM, 76nM and 510 nM for other PI3K isoforms, p110β, –γ, and –δ, respectively. Furthermore, when binding to purified p110α, PIK–75 is a noncompetitive inhibitor with respect to ATP with Ki of 36 nM and competitive with respect to the substrate PI with Ki of 2.3 nM [1].PIK–75 also shows potent inhibition of DNA–PK [2]. PIK–75 (1 μM) reduces cell survival by significantly decreasing mitochondrial activity in unstimulated nonasthmatic airway smooth muscle (ASM) cells, asthmatic ASM cells, and lung fibroblasts. While inTGFβ–stimulated ASM cells, PIK75 only decreases mitochondrial activity in asthmatic cells without effects in nonasthmatic cells [3].in vivo: In the ErbB3WT tumor model, PIK–75 reduces in vitro chemotactic response to HRGβ1 and lowers pAkt levels by 40%. Besides,PIK–75 significantly reduces tumor cell motility and in vivo invasion in ErbB3WT primary tumors [4]. In the CD1 male mice, PIK–75leads to serious impairments in the insulin tolerance test (ITT) and glucose tolerance test (GTT), and an increase in glucose production during a pyruvate tolerance test (PTT) [5].PROTOCOL (Extracted from published papers and Only for reference)Cell assay [6]A total of 2,000 human pancreatic cancer cells (MIA PaCa–2 or AsPC–1) per well were plated in 96–well flat–bottom plates and then treated with either gemcitabine, PIK–75 alone or in combination of both drugs with indicated concentrations. At the indicated times,20 μl of 1 mg/ml MTT in PBS was added to each well and further incubated for 4 h. After centrifugation and removal of the medium,150 μl of DMSO was added to each well to dissolve the formazan crystals. The absorbance was measured at 562 nm using an ELx808absorbance microplate reader. Absorbance of untreated cells was designated as 100%, and the relative viable cells were expressed as a percentage of this value. The drug interaction was evaluated by using the combination index (CI) according to the method of Chou and Talalay using CompuSyn software.Animal administration [6]Animal use procedures were approved by the Institutional Animal Care and Use Committee of Georgetown University Medical Center.MIA PaCa–2 cells (1.7×106 cells/mouse) mixed with Matrigel were injected subcutaneously into the flank of male athymic nude(Foxn1nu) mice aged 6–weeks. Gemcitabine (50 mg/ml) was dissolved in PBS and PIK–75 (20 mg/ml) was dissolved in DMSO. Injection solution was made as 10% of Cremophor EL and 3% of poly(ethylene glycol) 400 in sterile water. Before administration of compounds,gemcitabine was further diluted in PBS and DMSO or PIK–75 was further diluted in the injection solution and sterilized by 0.2 μm filterProduct Name:PIK–75Cat. No.:HY-13281CAS No.:372196-77-5Molecular Formula:C 16H 15BrClN 5O 4S Molecular Weight:488.74Target:DNA–PK; DNA–PK Pathway:PI3K/Akt/mTOR; Cell Cycle/DNA Damage Solubility:10 mM in DMSOunit. These diluents were mixed with 1:1 ratio and administered into peritoneal cavity of the mouse. Gemcitabine (20 mg/kg) or gemcitabine (20 mg/kg)/PIK–75 (2 mg/kg) combination was administered twice per week and vehicle control and PIK–75 (2 mg/kg) were administered 5 times per week. The body weights and tumor sizes were measured 3 times per week. Tumor volumes were calculated as width (mm) × length (mm) × height (mm)/2.References:[1]. Zheng Z, et al. Isoform–selective inhibition of phosphoinositide 3–kinase: identification of a new region of nonconserved amino acids critical for p110αinhibition. Mol Pharmacol. 2011, 80(4), 657–664.[2]. WO/2003/072557, 09/04/2003[3]. Moir LM, et al. Phosphatidylinositol 3–kinase isoform–specific effects in airway mesenchymal cell function. J Pharmacol Exp Ther. 2011, 337(2), 557–566.[4]. Smirnova T, et al. Phosphoinositide 3–kinase signaling is critical for ErbB3–driven breast cancer cell motility and metastasis. Oncogene. 2012, 31(6), 706–715.[5]. Smith GC, et al. Effects of acutely inhibiting PI3K isoforms and mTOR on regulation of glucose metabolism in vivo. Biochem J. 2012, 442(1), 161–169.[6]. Duong HQ, et al. Inhibition of NRF2 by PIK–75 augments sensitivity of pancreatic cancer cells to gemcitabine. Int J Oncol. 2014 Mar;44(3):959–69.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。