کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9918646 | 1557552 | 2005 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A combined cell based approach to identify P-glycoprotein substrates and inhibitors in a single assay
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کلمات کلیدی
PAMPATEERBcrpP-gpMRPHBSSNCEADMELRPabsorption, distribution, metabolism, eliminationHEPESLSCbasolateral to apicalN-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid - N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acidP-glycoprotein - P-گلیکوپروتئینHigh-throughput - با توان بالاInhibitor - بازدارندهParallel artificial membrane permeability assay - تست نفوذپذیری غشایی موازیCaco-2 cell - سلول Caco-2Liquid scintillation counter - ضد شمعدان مایعImmobilized artificial membrane - غشای مصنوعی ImmobilizedEfflux - فلوکسSubstrate - لایهHank's balanced salt solution - محلول نمک متعادل هانکTrans-epithelial electrical resistance - مقاومت الکتریکی Trans-epithelialIAM - من هستمnew chemical entity - موجودی شیمیایی جدیدPermeability - نفوذپذیریMulti-drug resistance protein - پروتئین مقاوم در برابر چند داروbreast cancer resistance protein - پروتئین مقاومت به سرطان سینه
موضوعات مرتبط
علوم پزشکی و سلامت
داروسازی، سم شناسی و علوم دارویی
علوم دارویی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The objective of this project was to develop a cell based in vitro experimental procedure that can differentiate P-glycoprotein (P-gp) substrates from inhibitors in a single assay. Caco-2 cells grown to confluency on 12-well Transwell® were used for this study. The efflux permeability (B to A) of P-gp specific probe (viz., digoxin) in the presence of test compounds (e.g. substrates, inhibitors and non-substrates of P-gp) was monitored, and the influx permeability (A to B) of test compounds was evaluated after complete P-gp blockade. Radiolabelled digoxin was added on the basolateral side with buffer on the apical side. The digoxin concentration appearing on the apical side represents digoxin efflux permeability during the control phase (0-1 h period). After 1 h, a test compound (10 uM) was added on the apical side. The reduced efflux permeability of digoxin suggests that the added test compound is an inhibitor. The influx permeability of test compound is also determined during the 1-2 h study period by measuring the concentration of the test compound in the basolateral side. At the end of 2 h, a potent P-gp inhibitor (GF120918) was added. The increased influx permeability of test compound during the 2-3 h incubation period indicates that the added test compound is a substrate. Samples were taken from both sides at the end of 1-3 h and the concentrations of the test compounds and digoxin were quantitated. Digoxin efflux permeability remained unchanged when incubated with P-gp substrates (e.g., etoposide, rhodamine123, taxol). However, when a P-gp inhibitor was added to the apical side, the digoxin efflux (B to A permeability) was significantly reduced (ketoconazole = 51% reduction) as expected. The influx permeability of substrates increased significantly (rhodamine123 = 70%, taxol = 220%, digoxin = 290%) after the P-gp inhibitor (GF120918) was introduced, whereas the influx permeability of P-gp inhibitor and non-substrates was not affected by GF120918. Thus, this combined assay provides an efficient cell based in vitro screening tool to simultaneously distinguish compounds that are P-gp substrates from P-gp inhibitors.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Pharmaceutics - Volume 301, Issues 1â2, 14 September 2005, Pages 80-88
Journal: International Journal of Pharmaceutics - Volume 301, Issues 1â2, 14 September 2005, Pages 80-88
نویسندگان
Praveen V. Balimane, Saeho Chong,