Characterization of binding by repaglinide and nateglinide with glycated human serum albumin using high-performance affinity microcolumns

Susan T. Ovbude, Pingyang Tao, Zhao Li, David S. Hage

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

High-performance affinity microcolumns were used to characterize binding by the anti-diabetic drugs repaglinide and nateglinide with normal and glycated forms of human serum albumin. The microcolumns contained only nmol amounts of protein and provided a detailed analysis of these drug interactions with good precision and in a matter of minutes per experiment. The overall binding by repaglinide to normal and glycated albumin fits a model with two types of binding sites: a set of one or two moderate-to-high affinity regions and a larger set of weaker regions with association equilibrium constants of ∼105 and 103 M−1, respectively, at pH 7.4 and 37°C. Competition studies gave site-specific association constants for repaglinide and nateglinide at Sudlow site I of 4.2 × 104 and 5.0 × 104 M−1 for normal albumin, with a decrease of 26%–30% being seen for nateglinide with glycated albumin and no significant change being noted for repaglinide. At Sudlow site II, repaglinide and nateglinide had association constants for normal albumin of 6.1 × 104 and 7.1 × 105 M−1, with glycated albumin giving an increase in the association constant at this site for repaglinide of 1.6- to 1.8-fold and a decrease for nateglinide of 51%–58%.

Original languageEnglish (US)
Pages (from-to)4176-4186
Number of pages11
JournalJournal of Separation Science
Volume45
Issue number23
DOIs
StatePublished - Dec 2022

Keywords

  • drug-protein binding
  • glycation
  • high-performance affinity microcolumn
  • human serum albumin
  • meglitinide

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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