Heparin-binding motif mutations of human diamine oxidase allow the development of a first-in-class histamine-degrading biopharmaceutical.

Abstract

Background:

Excessive plasma histamine concentrations cause symptoms in mast cell activation syndrome, mastocytosis, or anaphylaxis. Anti-histamines are often insufficiently efficacious. Human diamine oxidase (hDAO) can rapidly degrade histamine and therefore represents a promising new treatment strategy for conditions with pathological histamine concentrations.

Methods:

Positively charged amino acids of the heparin-binding motif of hDAO were replaced with polar serine or threonine residues. Binding to heparin and heparan sulfate, cellular internalization and clearance in rodents were examined.

Results:

Recombinant hDAO is rapidly cleared from the circulation in rats and mice. After mutation of the heparin-binding motif, binding to heparin and heparan sulfate was strongly reduced. The double mutant rhDAO-R568S/R571T showed minimal cellular uptake. The short α-distribution half-life of the wildtype protein was eliminated, and the clearance was significantly reduced in rodents.

Conclusions:

The successful decrease in plasma clearance of rhDAO by mutations of the heparin-binding motif with unchanged histamine-degrading activity represents the first step towards the development of rhDAO as a first-in-class biopharmaceutical to effectively treat diseases characterized by excessive histamine concentrations in plasma and tissues.

Funding:

Austrian Science Fund (FWF) Hertha Firnberg program grant T1135 (EG); Sigrid Juselius Foundation, Medicinska Understödsförening Liv och Hälsa rft (TAS and SeV).

• Authors: Elisabeth Gludovacz, Kornelia Schuetzenberger, Marlene Resch, Katharina Tillmann, Karin Petroczi, Markus Schosserer, Sigrid Vondra, Serhii Vakal, Gerald Klanert, Jürgen Pollheimer, Tiina A Salminen, Bernd Jilma, Nicole Borth, Thomas Boehm
• Journal: eLife (Elife) Vol. 10 (09 03 2021) ISSN: 2050-084X [Electronic] England
• PMID: 34477104 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2021, Gludovacz et al.

Chemical References

• Biological Products
• Histamine Antagonists
• Recombinant Proteins
• Heparin
• Amine Oxidase (Copper-Containing)

Topics

• Amine Oxidase (Copper-Containing) (chemistry, genetics, metabolism)
• Amino Acid Motifs (genetics)
• Animals
• Biological Products (chemistry, metabolism)
• Heparin (metabolism)
• Histamine Antagonists (chemistry, metabolism)
• Humans
• Mice
• Mutation (genetics)
• Protein Binding (genetics)
• Rats
• Recombinant Proteins (chemistry, genetics, metabolism)