米替福新作用機制 - Medchemexpress - MCE中國

1、Product Data SheetMiltefosineCat. No.: HY-13685CAS No.: 58066-85-6分式: CHNOP分量: 407.57作靶點: Akt; HIV作通路: PI3K/Akt/mTOR; Anti-infection儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 H2O : 50 mg/mL (122.68 mM; Need ultrasonic)DMSO : 5 mg/mL (12.27 mM; Need ultrasonic)Sol

2、ventMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 2.4536 mL 12.2678 mL 24.5357 mL5 mM 0.4907 mL 2.4536 mL 4.9071 mL10 mM 0.2454 mL 1.2268 mL 2.4536 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；旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲存時，請在 6 個內使，-20C 儲存時，請在 1 個內使。BIOLOGICAL ACTIVITY物活性

3、Miltefosine 種譜抗微物，抗利什曼原，磷脂劑，通過抑制 PI3K/Akt 活性起作。Miltefosine是CTP 磷酸膽 堿胞苷基轉移酶 (CCT) 的抑制劑IC & Target Akt HIV-1體外研究Treatment of HIV-1 infected macrophages with Miltefosine inhibits the recruitment of PH-AktGFP to the plasmamembrane. Since Miltefosine inhibits Akt through mimicry of the PH domain, it is l

4、ikely that Miltefosine binds to PIP3,blocking the recruitment of PH-Akt to the membrane1. Miltefosine (HePC) inhibits protein kinase C (PKC) fromNIH3T3 cells in cell-free extracts with a IC50 of about 7 M. Inhibition is competitive with regard tophosphatidylserine with a Ki of 0.59 M2. Miltefosine i

5、s an alkylphospholipid that inhibit activation of Akt.Miltefosine is a direct inhibitor of Akt, and induces dose-dependent inhibition of primary effusion lymphoma (PEL) inPage 1 of 2 www.MedChemEculture and also inhibits the downstream targets of Akt, such as mTOR, leading to reduced phosphorylation

6、 andactivation of S6K and S6. Importantly, Miltefosine also inhibits Akt targets that are not part of the mTOR pathway, eg,FOXO1, and are therefore expected to have a greater therapeutic impact than mTORC1 inhibitors alone3.體內研究 Mice are randomized into groups of 5 and injected intraperitoneally 5 d

7、ays a week with 50 mg/kg of eitherMiltefosine or Perifosine dissolved in PBS, or equivalent volume of vehicle (PBS). Both Miltefosine and Perifosineinhibit the growth rate of tumors compared with vehicle-treated mice. By day 14 after treatment, there is anapproximately 50% decrease in average tumor

8、volume in Perifosine- and Miltefosine-treated mice, compared withvehicle-treated mice (P0.04). Tumor growth is also significantly retarded (P0.04 for Perifosine and P0.055 forMiltefosine by linear mixed-effects model analysis). Immunohistochemical analyses display an overall reduction instaining for

9、 phosphorylated ribosomal S6 protein in tumor sections from Miltefosine- and Perifosine-treated micecompared with the PBS-treated mice. This reduced phosphorylation correlated with the delay in tumor progression indrug-treated animals3.PROTOCOLKinase Assay 3 Levels of enzymatically active caspase-3

10、are quantified using the ApoAlert Caspase Fluorescent assay kit. Briefly, 1106 BC-1 PEL cells are treated with 50 M Miltefosine, 50 M Perifosine, or 20 nM NVP-BEZ235, as well as the respectivevehicle controls. Cells are harvested and lysed 12 hours later. Equivalent micrograms of cell lysate for all

11、 samples areincubated with a fluorogenic caspase-3 substrate (DEVD-AFC). Cleavage of DEVD by caspase-3 releases AFC, thefluorescence of which is measured using a FLUOstar OPTIMA fluorometer, with excitation and emission filterwavelengths set to 400 and 505 nm, respectively3.MCE has not independently

12、 confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 NIH3T3 cells are grown in DMEM supplemented with 10% FCS in a humidified atmosphere of 95% air with 5% CO2.Cells are plated on 35-mm culture dishes (6-well plates) at 0.5-0.8105 cells/well. Growth is established for 1

13、8-24 hand the cell number of representative wells is determined (time 0). The experiments are started by addition of freshprepared solution of Miltefosine at given concentrations to the cells or equal volumes of Tris-HCI to control cells.After incubation for 60 h, cells are counted with an electroni

14、c counter. Cellular multiplication is calculated2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice3Administration 34 PEL cells are washed in ice-cold phosphate buffered saline, counted, and diluted in 100 L of PBS mixed with 100 Lof growth fa

15、ctor-depleted Matrigel. A total of 1105 to 7.5105 BC-1 cells are injected subcutaneously into the rightflank of NOD.CB17-Prkdcscid/J or CB17-Prkdcscid/J mice. The mice are monitored on alternate days for developmentof palpable tumors (2 mm3), at which point drug or vehicle treatments are initiated,

16、and are administered eitherintraperitoneally (Perifosine) or by oral gavage (Rosiglitazone, NVP-BEZ235) 5 days a week. Groups of 5 to 7 mice areused to generate PEL tumors and treated with either vehicle or drug cocktail. Each biologic experiment is repeatedmultiple times. For Rosiglitazone, 0.25% m

17、ethylcellulose is used as vehicle, and 30 mg/kg or 60 mg/kg Rosiglitazoneis suspended in methylcellulose. For Perifosine and Miltefosine, PBS is used as a vehicle and 50 mg/kg Perifosine orMiltefosine is dissolved in PBS. For NVP-BEZ235, the compound is dissolved in a 1:9 vol/vol mixture of 1-methyl

18、-2-pyrrolidone and polyethylene glycol 300. A dose of 40 mg/kg NVP-BEZ235 or equal volume of the vehicle isadministered. Tumor diameters are measured using digital calipers, and tumor volume is calculated. The tumors areexcised and fixed in formalin. Statistical analyses are performed using linear m

19、odel fit by maximum likelihood withindividual animals treated as random effect.Rats4Male Sprague-Dawley rats (weight 270-290 g) are divided into five groups (n=5). Rats in the treatment groups areadministered a single 10 mg/kg oral dose of Miltefosine (MFS) either as an aqueous solution or MFS-LNCs

20、dispersionby gastric gavage. This dose is equivalent to the 20 mg/kg Miltefosine dose administered to mice in the preclinicalstudy after correction for rats. Following administration, blood samples are collected via the orbital plexus underPage 2 of 3 www.MedChemEanesthesia at time intervals of 0.5,

21、 1, 2, 4, 7, 10, 24, 48, 72 and 216 h in Eppendorf tubes containing EDTA. Bloodsamples are then centrifuged immediately at 4000 rpm for 10 min. Plasma samples are frozen and maintained at -80C pending analysis.MCE has not independently confirmed the accuracy of these methods. They are for reference

22、only.戶使本產(chǎn)品發(fā)表的科研獻 J Cell Mol Med. 2019 May 26. Chem Biol Interact. 2019 Jun 29;310:108731. Pathogens. 2020 May 20;9(5):E393. Molecules. 2020 Apr 23;25(8). pii: E1980. Acta Trop. 2018 May 4;185:69-76.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Chugh P, et al. Akt i

23、nhibitors as an HIV-1 infected macrophage-specific anti-viral therapy. Retrovirology. 2008 Jan 31;5:112. Uberall F, et al. Hexadecylphosphocholine inhibits inositol phosphate formation and protein kinase C activity. Cancer Res. 1991 Feb 1;51(3):807-12.3. Bhatt AP, et al. Dual inhibition of PI3K and mTOR inhib