Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Advanced Epi-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability

Armando Rodríguez-Alfonso
,
Astrid Heck
,
Yasser Ruiz-Blanco
,
Andrea Gilg
,
Ludger Ständker
ORCID logo ,
Seah Ling Kuan
,
Tanja Weil
,
Elsa Sanchez-Garcia
,
Sebastian Wiese
,
Jan Münch
ORCID logo ,
Mirja Harms
* ORCID logo
Version 1 : Received: 5 October 2022 / Approved: 6 October 2022 / Online: 6 October 2022 (08:50:49 CEST)

How to cite: Rodríguez-Alfonso, A.; Heck, A.; Ruiz-Blanco, Y.; Gilg, A.; Ständker, L.; Kuan, S.L.; Weil, T.; Sanchez-Garcia, E.; Wiese, S.; Münch, J.; Harms, M. Advanced Epi-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability. Preprints 2022, 2022100058. https://doi.org/10.20944/preprints202210.0058.v1 Rodríguez-Alfonso, A.; Heck, A.; Ruiz-Blanco, Y.; Gilg, A.; Ständker, L.; Kuan, S.L.; Weil, T.; Sanchez-Garcia, E.; Wiese, S.; Münch, J.; Harms, M. Advanced Epi-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability. Preprints 2022, 2022100058. https://doi.org/10.20944/preprints202210.0058.v1

Abstract

Advanced derivatives of the CXCR4 antagonist EPI-X4 have shown therapeutic efficacy upon topical administration in animal models of asthma and dermatitis. Here, we studied plasma stability of the EPI-X4 lead compounds WSC02 and JM#21, using mass spectrometry to monitor the chemical integrity of the peptides and a functional fluorescence-based assay that determines peptide function in a CXCR4-antibody competition assay. Although mass spectrometry revealed a very rapid disappearance of both peptides in human plasma within seconds, the functional assay revealed a significantly higher half-life of 9 minutes for WSC02 and 6 minutes for JM#21, respectively. Further analyses demonstrated that WSC02 and JM#21 interact with low molecular weight plasma components and serum albumin. Albumin binding is mediated by the formation of a disulfide bridge between Cys10 in the EPI-X4 peptides and Cys34 in albumin. These covalently linked albumin-peptide complexes have a higher stability in plasma as compared to the non-bound peptides and retain the ability to bind and antagonize CXCR4. Remarkably, chemically synthesized albumin-EPI-X4 conjugates coupled by non-breakable bonds have a drastically increased plasma stability of over 2 hours. Thus, covalent coupling of EPI-X4 to albumin in vitro before administration or in vivo post administration may significantly increase the pharmacokinetic properties of the new class of CXCR4 antagonists.

Keywords

EPI-X4; albumin carrier; peptide stability; CXCR4 antagonist;

Subject

Medicine and Pharmacology, Pharmacology and Toxicology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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