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Morath, V.* ; Maurer, S.* ; Feuchtinger, A. ; Walser, R.* ; Schlapschy, M.* ; Bolze, F.* ; Metzler, T.* ; Bruder, J.* ; Steiger, K.* ; Walch, A.K. ; Klingenspor, M.* ; Skerra, A.*

Long-acting human PASylated leptin reaches the murine central nervous system and offers potential for optimized replacement therapy.

Mol. Pharm., DOI: 10.1021/acs.molpharmaceut.4c01503 (2025)
Verlagsversion DOI PMC
Open Access Gold (Paid Option)
Creative Commons Lizenzvertrag
Despite the multifaceted role of leptin for energy homeostasis and its broad therapeutic potential, the FDA/EMA-approved metreleptin constitutes the only leptin drug to date. To translate the promising results from previous studies on murine PASylated leptin with improved solubility and extended plasma half-life using PASylation technology─a biological alternative to PEGylation─we have developed a second-generation human leptin drug candidate and tested it rigorously in vitro and in vivo. To this end, the exposed hydrophobic Trp residue at position 100 in human leptin was replaced by Gln, which, together with the genetic fusion with a 600-residue PAS polypeptide, yielded a protein with high solubility, folding stability and receptor-stimulatory activity. In a pharmacokinetic (PK) study with wild-type mice, this modified human leptin showed an extended plasma half-life of 18.8 ± 3.6 h after subcutaneous (s.c.) injection. Furthermore, leptin-deficient mice were dosed s.c. with the modified human leptin carrying two different PAS fusion tags, PAS#1 or P/A#1, each comprising 600 residues. After only four doses, the disease phenotype, including morbid adiposity, hyperphagia, and hepatic steatosis, was completely reversed by both PASylated leptin versions, but not by the non-PASylated leptin if administered at the same dose. To assess its tissue distribution, P/A(200)-huLeptinW100Q was doubly labeled with two fluorescent dyes, which were specifically attached to the leptin and the PAS moiety, respectively. Analysis of relevant mouse organs by light sheet fluorescence microscopy after clearance revealed colocalized signals in the kidney and liver, thus indicating general stability of the PAS-leptin fusion protein in vivo. However, discrete signals were observed in the hypothalamic region, only with leptin detectable in the choroid plexus, which implies cleavage of the PAS tag during transcytosis across the physiological barriers. This study should pave the way toward a second-generation leptin drug enabling prolonged dosing intervals.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Pasylation ; Adipokine ; Blood-brain Barrier ; Light Sheet Fluorescence Microscopy ; Obesity ; Plasma Half-life; Plasma Half-life; High-fat Diet; Escherichia-coli; Gene-expression; C57bl/6j Mice; Blood-volume; Protein; Induction; Metreleptin; Sensitivity
ISSN (print) / ISBN 1543-8384
e-ISSN 1543-8392
Zeitschrift Molecular Pharmaceutics
Verlag American Chemical Society (ACS)
Verlagsort 1155 16th St, Nw, Washington, Dc 20036 Usa
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Research Unit Analytical Pathology (AAP)
Förderungen DFG
Federal State of Bavaria
Else Kroner-Fresenius-Stiftung
Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) in frame of the Collaborative Research Centre 824
Animal Research Center (School of Life Sciences, TU Munich)
Deutsche Forschungsgemeinschaft