Development of a Novel Human Serum Albumin-Based Tool for Effective Drug Discovery: The Investigation of Protein Quality and Immobilization

Balázs Kenéz; Gábor Koplányi; Balázs Decsi; Zsófia Molnár; Péter Horváth; Gábor Katona; György T Balogh; Diána Balogh-Weiser

doi:10.1021/acs.jmedchem.4c02136

Development of a Novel Human Serum Albumin-Based Tool for Effective Drug Discovery: The Investigation of Protein Quality and Immobilization

J Med Chem. 2025 Jan 15. doi: 10.1021/acs.jmedchem.4c02136. Online ahead of print.

Authors

Balázs Kenéz¹, Gábor Koplányi¹, Balázs Decsi², Zsófia Molnár¹, Péter Horváth³, Gábor Katona⁴, György T Balogh^{2

3

5}, Diána Balogh-Weiser^{1

6}

Affiliations

¹ Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, H-1111Budapest, Hungary.
² Department of Chemical and Environmental Process Engineering, Budapest University of Technology and Economics, Műegyetem rkp. 3, H-1111Budapest, Hungary.
³ Department of Pharmaceutical Chemistry, Semmelweis University, Hőgyes E. Street 7-9, H-1092 Budapest, Hungary.
⁴ Institute of Pharmaceutical Technology and Regulatory Affairs, Faculty of Pharmacy, University of Szeged, Eötvös u. 6, H-6720 Szeged, Hungary.
⁵ Center for Pharmacology and Drug Research & Development, Semmelweis University, Üllői Street 26, H-1085Budapest, Hungary.
⁶ Department of Physical Chemistry and Materials Science, Budapest University of Technology and Economics, Műegyetem rkp. 3, H-1111Budapest, Hungary.

PMID: 39813641
DOI: 10.1021/acs.jmedchem.4c02136

Abstract

The binding ability of human serum albumin (HSA) on active pharmaceutical ingredients (APIs) is one of the most important parameters in the early stages of drug discovery. In this study, an immobilized HSA-based tool was developed for the rapid and easy in vitro screening of API binding. The work explored the serious incompleteness in the identification of HSA used for in vitro screening published in the last five years. To mitigate this problem, a comprehensive analysis and immobilization studies were performed on the most used HSA types. Serious differences in the colloidal stability of HSAs and their API binding ability on a selected set of APIs were observed. HSAs were immobilized on magnetic nanoparticles with glutardialdehyde (GDA) or cyclohexyl-diglycidyl ether (CDGE) linkers, which have never been used for HSA immobilization before. The HSA-MNP-CDGE complexes achieved a higher immobilization yield and preserved API binding ability; however, the esterase-like enzymatic activity of HSA reduced significantly.