| 名稱 | Posaconazole |
| 描述 | Posaconazole (POS) is a sterol C14ɑ demethylase inhibitor (IC50: 0.25 nM). |
| 細(xì)胞實(shí)驗(yàn) | The epimastigote form of the parasite is cultivated in liver infusion tryptose medium,supplemented with 10% new born calf serum at 28 °C with strong (120 rpm) agitation.Cultures are initiated at a cell density of 2 × 106 epimastigotes/mL,and Posaconazole is added at a cell density of 0.5−1.0 × 107 epimastigotes/mL.Cell densities are measured by using an electronic particle counter as well as by direct counting with a hemocytometer.Cell viability is followed by Trypan blue exclusion,using light microscopy.Amastigotes are cultured in Vero cells maintained in minimal essential medium supplemented with 1% fetal calf serum in a humidified atmosphere (95% air−5% CO2) at 37 °C.Cells are infected with 10 tissue culture-derived trypomastigotes per cell for 2 hours and then washed three times with phosphate-buffered saline (PBS) to remove nonadherent parasites.Fresh medium with and without Posaconazole is added,and the cells are incubated for 96 hours with a medium change at 48 hours.The percent of infected cells and the numbers of parasites per cell are determined directly using light microscopy,and a statistical analysis of the results is carried out.IC50 values are calculated by nonlinear regression,using the program GraFit.Fractional inhibitory concentrations (FIC) are calculated.Cytoplasmic free Ca2+ concentrations in control and drug-treated extracellular epimastigotes are determined by fluorimetric methods using Fura-2.Subcellular Ca2+ levels and mitochondrial membrane potentials are monitored on individual Vero cells infected with T.cruzi amastigotes by using time-scan confocal microscopy.Briefly,Vero cells heavily infected (72 hours) with T.cruzi amastigotes are plated onto 22 × 40 mm glass coverslips (0.15 mm thickness) and incubated simultaneously with 10 μM cell-permeant Rhod-2 and 10 μg/mL Rhodamine-123 for 50 minutes at 37 ° |
| 激酶實(shí)驗(yàn) | In vitro Aβ reduction assays : Human embryonic kidney cells (American Type Culture Collection CRL-1573), transfected with the gene for APP751 (HEK 293) are used for routine Aβ reduction assays. Cells are plated in 96-well plates and allowed to adhere overnight in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum. DAPT are diluted from stock solutions in dimethylsulfoxide (DMSO) to yield a final concentration equal to 0.1% DMSO in media. Cells are pre-treated for 2 hours at 37 °C with DAPT, media are aspirated off and fresh compound solutions applied. After an additional 2-hour treatment period, conditioned media is drawn off and analyzed by a sandwich ELISA (266–3D6) specific for total Aβ. Reduction of Aβ production is measured relative to control cells treated with 0.1% DMSO and expressed as a percentage inhibition. Data from at least six doses in duplicate are fitted to a four-parameter logistical model using XLfit software in order to determine potency. Human and PDAPP mouse neuronal cultures are grown in serum-free media to enhance their neuronal characteristics, and appeared to be greater than 90% neurons after maturation prior to use. Conditioned media to establish baseline Aβ values are collected by adding fresh media to each well and incubated for 24 hours at 37 °C in the absence of DAPT. Cultures are then treated with fresh media containing DAPT at the desired range of concentrations for an additional 24 hours at 37 °C, and conditioned media collected. For the measurement of total Aβ, samples are analyzed with the same ELISA (266–3D6) as used for the HEK 293 cell assays. Analyses of samples for Aβ42 production are performed by a separate ELISA (21F12–3D6) that utilizes a capture antibody specific for the Aβ42 C-terminus. Inhibition of production for both total Aβ and Aβ42 are determined by the difference between the values for the compound treatment and baseline periods.After plotting percentage inhibition versus DAPT concentration, data are analyzed with XLfit software, as above, to determine potency. |
| 體外活性 | Posaconazole的生物利用度可被食物,尤其是高脂肪膳食顯著增加.與高脂和脫脂食物一起消耗時(shí),全身接觸Posaconazole分別增加其4倍和2.6倍的消耗.Posaconazole（≥15 mg/kg,b.i.d）可使小鼠壽命延長(zhǎng),并減少組織負(fù)擔(dān).在感染的動(dòng)物中,胺碘酮單獨(dú)使用可減少寄生蟲(chóng)血癥,增加60天生存期(未處理的對(duì)照組為0%,胺碘酮處理的動(dòng)物為40%),與Posaconazole聯(lián)用可延緩寄生蟲(chóng)血癥的進(jìn)程.Posaconazole 和胺碘酮可能產(chǎn)生有效的抗T. cruzi治療,且副作用低.在禁食狀態(tài),Posaconazole與Boost Plus同時(shí)服用增加藥物暴露. |
| 體內(nèi)活性 | Posaconazole對(duì)臨床相關(guān)的細(xì)胞內(nèi)無(wú)鞭毛體寄生蟲(chóng)形式的效能更好。Posaconazole的最低抑菌濃度和IC50值分別為3 nM和0.25 nM。Posaconazole對(duì)耐氟康唑、伏立康唑和兩性霉素B的念珠菌和曲霉菌株具有活性,且效果比其它三唑類抗接合菌藥更好。胺碘酮與Posaconazole可產(chǎn)生協(xié)同效果。Posaconazole還會(huì)影響并打亂T. cruzi 中Ca2+內(nèi)穩(wěn)態(tài)。Posaconazole對(duì)前鞭體(細(xì)胞外)階段的增殖具有明顯的劑量依賴性作用,最低抑菌濃度為20 nM,IC50為14 nM。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 93 mg/mL (132.7 mM)
|
| 關(guān)鍵字 | Inhibitor | SCH56592 | inhibit | SCH-56592 | Posaconazole | Fungal |
| 相關(guān)產(chǎn)品 | 2-Butyl-1,2-benzisothiazolin-3-one | Potassium gluconate | Dehydroacetic acid sodium | Sorbic acid | Geraniol | Paclobutrazol | Veratraldehyde | Lauryl betaine | Methyl 2-amino-5-bromobenzoate | Chlorobutanol hemihydrate | Chitosan (MW 150000) | Tebuconazole |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 抗真菌庫(kù) | 已知活性化合物庫(kù) | EMA 上市藥物庫(kù) | 抗COVID-19化合物庫(kù) | FDA 上市藥物庫(kù) | 臨床期小分子藥物庫(kù) | 藥物功能重定位化合物庫(kù) | 兒童藥物庫(kù) |