Perifosine-KRX-0401-DataSheet-MedChemExpressVIP

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPerifosineCat. No.: HY-50909CAS No.: 157716-52-4Synonyms: KRX-0401; NSC 639966; D21266分式: CHNOP分量: 461.66作靶點: Akt; Autophagy作通路: PI3K/Akt/mTOR; Autophagy儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)

2、體外實驗 H2O : 153.33 mg/mL (332.13 mM)DMSO : 1 mg/mL (insoluble or slightly soluble)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.1661 mL 10.8305 mL 21.6610 mL5 mM 0.4332 mL 2.1661 mL 4.3322 mL10 mM 0.2166 mL 1.0830 mL 2.1661 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

3、，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 Perifosine種有服活性的 Akt 抑制劑，抑制不同腫瘤細胞系增殖的 IC50 值為0.6-8.9 M。IC50 & Target Akt Autophagy1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE體外研究 The IC50 for growth of Ntv-a/LacZ cell lines is determined by MTT assay. When the cells are cultured for 48hours in 10% FCS-s

4、upplemented media, the IC50 for cells with constitutively active PDGF, Ras, or Aktsignaling is similar and found to be 45 M 1.Perifosine, a oral-bioavailable alkylphospholipid (ALK), on thecell cycle kinetics of immortalized keratinocytes (HaCaT) as well as head and neck squamous carcinomacells. Pro

5、liferation is assessed by the incorporation of 3Hthymidine into cellular DNA. Exposure to Perifosine(0.1-30 M) for 24 h results in a dose-dependent inhibition of 3Hthymidine uptake in all cell lines tested. TheIC50s for growth are between 0.6 and 8.9 M, reaching IC80s of 10 M. Perifosine blocks cell

6、 cycleprogression of head and neck squamous carcinoma cells at G1-S and G2-M by inducing p21WAF1,irrespective of p53 function, and may be exploited clinically because the majority of human malignanciesharbor p53 mutations. Perifosine (20 M) induces both G1-S and G2-M cell cycle arrest, together with

7、p21WAF1 expression in both p53 wild-type and p53-/- clones 2.體內(nèi)研究 Mice are identified with tumors by bioluminescence imaging and either treated them with 100 mg/kgTemozolomide, or 30 mg/kg Perifosine, or a combination with 100 mg/kg Temozolomide and 30 mg/kgPerifosine (Temozolomide+Perifosine) for 3

8、 to 5 days. The mice are sacrificed and tumors analyzedhistologically for cell proliferation by Ki-67 immunostaining. Ki-67 staining index is significantly reduced inmice treated with either Temozolomide (Ki-67 staining index=5.51.2%, n=4, P=0.0019) or Perifosine (Ki-67staining index=3.21.1%, n=3, P

9、=0.001) compared with Control, demonstrating the inhibitory effect onproliferation. Most importantly, the tumors treated with Temozolomide+Perifosine have the lowest Ki-67staining index (1.71.2%, n=3, P=0.0005). The additional treatment with Perifosine results in a significantlylower proliferation r

10、ate than Temozolomide alone (P=0.0087) 1. Perifosine markedly decreases p-Akt from10 min to 24 hours and subsequently, moderately decreased p-S6 from 1h to 24 h after injection 3.PROTOCOLKinase Assay 2 Exponentially growing cells (HN12, HN30, and HaCaT) are lysed, and 500 g of total cellular protein

11、 areused to immunoprecipitate active cdc2 and cdk2 complexes. After capturing with gammabind G Sepharoseand subsequent washes, the active immune complexes are assessed for activity in the presence ofincreasing concentrations of Perifosine (0.1-30 M) or flavopiridol (300 nM) in the kinase assay buffe

12、rcontaining -32PATP (3000 Ci/mmol) and 0.2 mg/mL histone H1, 25 M ATP. Reactions are incubated at37C for 30 min and terminated by the addition of SDS-gel loading buffer, resolved in SDS, and driedgels are subjected to autoradiography and phosphorimaging 2.MCE has not independently confirmed the accu

13、racy of these methods. They are for reference only.Cell Assay 2 Cell proliferation studies by measuring the uptake of 3Hthymidine is performed. Briefly, HNSCC and HaCaTcells (1-2104/well) are grown overnight in 24-well plates and exposed to either Perifosine (0.1-30 M) orPBS (control). After treatme

14、nt (24-48 h), cells are pulsed with 3Hthymidine (1 Ci/well) for 4-6 h, fixed (5%trichloroacetic acid), and solubilized (0.5 M NaOH) before scintillation counting. Experiments are performedin triplicates 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.

15、Animal Mice 1Administration 13 Drug treatment of tumor-bearing mice. Image-positive Ef-luc Ntv-a mice are treated daily with i.p.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEadministration of buffer alone as a control, or i.p. administration of 100 mg/kg Temozolomide, or oraladministration of 30

16、 mg/kg Perifosine, or a combination with Perifosine and Temozolomide for 3 to 5 days.The mean doses of the treatments are: Control, 5 (all five); Temozolomide, 3.75 (three to five); Perifosine,3.75 (three to four); and Perifosine+Temozolomide, 3 (all three). Control buffer solution consisted of 5%DM

17、SO and 1% Tween 80 in distilled water.Rats 3To further determine whether the paradoxical effect of rapamycin on S6 phosphorylation is related toupstream signals of Akt-mTOR, rats are treated with Perifosine (20 mg/kg, ip, once), an Akt inhibitor, 30 minbefore rapamycin administration. Rats are sacri

18、ficed 1 h or 6 h after rapamycin injection.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Int J Biol Sci. 2016 Mar 30;12(5):607-16.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Momota H, et al. Perifosine inhibits multiple signaling pathways in glial progenitors and cooperates with temozolomide to arrest cellproliferation in gliomas in vivo. Cancer Res, 2005, 65(16), 7