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. 2013 Aug;102(8):2882-8.
doi: 10.1002/jps.23648. Epub 2013 Jul 12.

In vitro protein binding of liraglutide in human plasma determined by reiterated stepwise equilibrium dialysis

Anne Plum  1 , Lisbeth Bjerring Jensen, Jesper Bøggild Kristensen
Affiliations
Free PMC article

In vitro protein binding of liraglutide in human plasma determined by reiterated stepwise equilibrium dialysis

Anne Plum et al. J Pharm Sci. 2013 Aug.
Free PMC article

Abstract

Liraglutide is a human glucagon-like peptide-1 (GLP-1) analogue approved for the treatment of type 2 diabetes. It is based on human GLP-1 with the addition of a 16-carbon fatty acid, which facilitates binding to plasma proteins, thus prolonging the elimination half-life and allowing once-daily administration. It has not been possible to quantify liraglutide protein binding by ultrafiltration (the usual method of choice), as the lipophilic molecule becomes trapped in the filter membrane. Therefore, the aim of this study was to develop a methodology that could determine the extent of liraglutide binding to plasma proteins in vitro. We report here the details of a novel reiterated stepwise equilibrium dialysis assay that has successfully been used to quantify liraglutide plasma protein binding. The assay allowed quantification of liraglutide binding to proteins in purified plasma protein solutions and human plasma samples and was effective at plasma dilutions as low as 5%. At a clinically relevant liraglutide concentration (10(4) pM), greater than 98.9% of liraglutide was bound to protein. Specific binding to human serum albumin and α1-acid glycoprotein was 99.4% and 99.3%, respectively. The novel methodology described herein could have an application in the quantification of plasma protein binding of other highly lipophilic drug molecules.

Keywords: acylated peptides; equilibrium dialysis; human serum albumin; in vitro method; insulin detemir; liraglutide; plasma proteins; protein binding; type 2 diabetes mellitus.

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Figures

Figure 1
Figure 1
The reiterated stepwise equilibrium dialysis assay. (a) Illustration of the experimental set-up used to perform the assay; (b) schematic representation of the inner and outer chamber buffer solutions at the start of the assay.
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