New functional assay of P-glycoprotein activity using Hoechst 33342

被引：59

作者：

Mueller, Henrik ^{[1
]}

Klinkhammer, Werner ^{[1
]}

Globisch, Christoph ^{[1
]}

Kassack, Matthias U. ^{[1
]}

Pajeva, Ilza K. ^{[1
,2
]}

Wiese, Michael ^{[1
]}

机构：

[1] Univ Bonn, Inst Pharm, D-53121 Bonn, Germany

[2] Bulgarian Acad Sci, Ctr Biomed Engn, BU-1113 Sofia, Bulgaria

来源：

BIOORGANIC & MEDICINAL CHEMISTRY | 2007年 / 15卷 / 23期

关键词：

P-glycoprotein; multidrug resistance; Hoechst; 33342; 3rd generation modulators;

D O I：

10.1016/j.bmc.2007.07.024

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In this study we describe a simplified, HTS-capable functional assay for the multidrug resistance (MDR) transporter P-glycoprotein (P-gp) based on its substrate Hoechst 33342. The physicochemical properties of Hoechst 33342 and the enormous milieu dependency of its fluorescence intensity allowed performing the assay in a homogeneous manner. This new assay served as an effective tool to estimate the potency of 10 well recognized P-gp substrates and modulators. Further, the potency of these compounds was also estimated in the calcein AM assay. The Hoechst 33342 and calcein AM assays yielded significantly comparable results for all compounds tested. Principal component analysis (PCA) applied to literature data on inhibition of P-gp activity and our results obtained in the Hoechst 33342 and calcein AM assay indicated similarity of compared functional transport assays. However, no correlation could be detected between these functional assays and the ATPase activity assay. (C) 2007 Elsevier Ltd. All rights reserved.

引用

页码：7470 / 7479

页数：10

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