Development and Analytical Characterization of Pegunigalsidase Alfa, a Chemically Cross-Linked Plant Recombinant Human α-Galactosidase-A for Treatment of Fabry Disease

Bioconjug Chem. 2018 May 16;29(5):1630-1639. doi: 10.1021/acs.bioconjchem.8b00133. Epub 2018 May 3.

Abstract

The current treatment of Fabry disease by enzyme replacement therapy with commercially available recombinant human α-Galactosidase A shows a continuous deterioration of the disease patients. Human recombinant α-Galactosidase A is a homodimer with noncovalently bound subunits and is expressed in the ProCellEx plant cell-based protein expression platform to produce pegunigalsidase alfa. The effect of covalent bonding between two α-Galactosidase A subunits by PEG-based cross-linkers of various lengths was evaluated in this study. The results show that cross-linking by a bifunctional PEG polymer of 2000 Da produces a more stable protein with improved pharmacokinetic and biodistribution properties. The chemical modification did not influence the tertiary protein structure but led to an increased thermal stability and showed partial masking of immune epitopes. The developed pegunigalsidase alfa is currently tested in phase III clinical trials and has a potential to show superior efficacy versus the currently used enzyme replacement therapies in the treatment of Fabry disease patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cross-Linking Reagents / chemistry*
  • Enzyme Stability
  • Fabry Disease / drug therapy
  • Humans
  • Mice
  • Models, Molecular
  • Polyethylene Glycols / chemistry*
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / pharmacokinetics
  • Recombinant Proteins / therapeutic use
  • Tissue Distribution
  • Tobacco / genetics
  • alpha-Galactosidase / chemistry*
  • alpha-Galactosidase / genetics
  • alpha-Galactosidase / pharmacokinetics
  • alpha-Galactosidase / therapeutic use

Substances

  • Cross-Linking Reagents
  • Recombinant Proteins
  • Polyethylene Glycols
  • alpha-Galactosidase
  • alpha-galactosidase A, human