New functional assay of P-glycoprotein activity using Hoechst 33342.

Article Details

Citation

Muller H, Klinkhammer W, Globisch C, Kassack MU, Pajeva IK, Wiese M

New functional assay of P-glycoprotein activity using Hoechst 33342.

Bioorg Med Chem. 2007 Dec 1;15(23):7470-9. Epub 2007 Aug 21.

PubMed ID
17890094 [ View in PubMed
]
Abstract

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.

DrugBank Data that Cites this Article

Binding Properties
DrugTargetPropertyMeasurementpHTemperature (°C)
CyclosporineP-glycoprotein 1IC 50 (nM)1413N/AN/ADetails
CyclosporineP-glycoprotein 1IC 50 (nM)234423N/AN/ADetails
DiltiazemP-glycoprotein 1IC 50 (nM)70794578N/AN/ADetails
DiltiazemP-glycoprotein 1IC 50 (nM)42658N/AN/ADetails
DiltiazemP-glycoprotein 1IC 50 (nM)61660N/AN/ADetails
ImatinibP-glycoprotein 1IC 50 (nM)66069345N/AN/ADetails
ImatinibP-glycoprotein 1IC 50 (nM)7244N/AN/ADetails
ImatinibP-glycoprotein 1IC 50 (nM)12589N/AN/ADetails
NicardipineP-glycoprotein 1IC 50 (nM)4786N/AN/ADetails
NicardipineP-glycoprotein 1IC 50 (nM)5129N/AN/ADetails
NicardipineP-glycoprotein 1IC 50 (nM)3630781N/AN/ADetails
ProgesteroneP-glycoprotein 1IC 50 (nM)48977882N/AN/ADetails
ProgesteroneP-glycoprotein 1IC 50 (nM)47863N/AN/ADetails
ProgesteroneP-glycoprotein 1IC 50 (nM)48978N/AN/ADetails
ReserpineP-glycoprotein 1IC 50 (nM)13182567N/AN/ADetails
ReserpineP-glycoprotein 1IC 50 (nM)3020N/AN/ADetails
ReserpineP-glycoprotein 1IC 50 (nM)3236N/AN/ADetails
VerapamilP-glycoprotein 1IC 50 (nM)4571N/AN/ADetails
VerapamilP-glycoprotein 1IC 50 (nM)6607N/AN/ADetails
VerapamilP-glycoprotein 1IC 50 (nM)134896288N/AN/ADetails
VinblastineP-glycoprotein 1IC 50 (nM)10471N/AN/ADetails
VinblastineP-glycoprotein 1IC 50 (nM)10000N/AN/ADetails
VinblastineP-glycoprotein 1IC 50 (nM)8709636N/AN/ADetails