Rodríguez-Alfonso, A.; Heck, A.; Ruiz-Blanco, Y.B.; Gilg, A.; Ständker, L.; Kuan, S.L.; Weil, T.; Sanchez-Garcia, E.; Wiese, S.; Münch, J.; et al. Advanced EPI-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability. International Journal of Molecular Sciences 2022, 23, 15029, doi:10.3390/ijms232315029.
Rodríguez-Alfonso, A.; Heck, A.; Ruiz-Blanco, Y.B.; Gilg, A.; Ständker, L.; Kuan, S.L.; Weil, T.; Sanchez-Garcia, E.; Wiese, S.; Münch, J.; et al. Advanced EPI-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability. International Journal of Molecular Sciences 2022, 23, 15029, doi:10.3390/ijms232315029.
Rodríguez-Alfonso, A.; Heck, A.; Ruiz-Blanco, Y.B.; Gilg, A.; Ständker, L.; Kuan, S.L.; Weil, T.; Sanchez-Garcia, E.; Wiese, S.; Münch, J.; et al. Advanced EPI-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability. International Journal of Molecular Sciences 2022, 23, 15029, doi:10.3390/ijms232315029.
Rodríguez-Alfonso, A.; Heck, A.; Ruiz-Blanco, Y.B.; Gilg, A.; Ständker, L.; Kuan, S.L.; Weil, T.; Sanchez-Garcia, E.; Wiese, S.; Münch, J.; et al. Advanced EPI-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability. International Journal of Molecular Sciences 2022, 23, 15029, doi:10.3390/ijms232315029.
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:
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