Radiomitigation and Tissue Repair Activity of Systemically Administered Therapeutic Peptide TP508 Is Enhanced by PEGylation

AAPS J. 2017 May;19(3):743-753. doi: 10.1208/s12248-016-0043-7. Epub 2017 Jan 17.

Abstract

TP508 is a synthetically derived tissue repair peptide that has previously demonstrated safety and potential efficacy in phase I/II clinical trials for the treatment of diabetic foot ulcers. Recent studies show that a single injection of TP508 administered 24 h after irradiation significantly increases survival and delays mortality in murine models of acute radiation mortality. Thus, TP508 is being developed as a potential nuclear countermeasure. Because of the short plasma half-life of TP508, we hypothesize that increasing the peptide bioavailability would increase TP508 efficacy or reduce the dosage required for therapeutic effects. We, therefore, evaluated the covalent attachment of various sizes of polyethylene glycol to TP508 at either its N-terminus or at an internal cysteine. A size-dependent increase in TP508 plasma half-life due to PEGylation was observed in blood samples from male CD-1 mice using fluorescently labeled TP508 and PEGylated TP508 derivatives. Biological activity of PEGylated TP508 derivatives was evaluated using a combination of biologically relevant assays for wound closure, angiogenesis, and DNA repair. PEG5k-TP508 enhanced wound closure after irradiation and enhanced angiogenic sprouting in murine aortic ring segments relative to equimolar dosages of TP508 without enhancing circulating half-life. PEG30k-TP508 extended the plasma half-life by approximately 19-fold while also showing enhanced biological activity. Intermediate-sized PEGylated TP508 derivatives had enhanced plasma half-life but were not active in vivo. Thus, increased half-life does not necessarily correlate with increased biological activity. Nevertheless, these results identify two candidates, PEG5k-TP508 and PEG30k-TP508, for potential development as second-generation TP508 injectable drugs.

Keywords: drug development; peptide conjugation; radiomitigation; tissue repair.

MeSH terms

  • Animals
  • Cells, Cultured
  • DNA Repair / drug effects*
  • Drug Delivery Systems
  • Half-Life
  • Histones
  • Humans
  • Male
  • Mice
  • Neovascularization, Physiologic / drug effects
  • Peptide Fragments / administration & dosage
  • Peptide Fragments / chemistry
  • Peptide Fragments / pharmacology*
  • Polyethylene Glycols / chemistry
  • Radiation-Protective Agents / administration & dosage
  • Radiation-Protective Agents / chemistry
  • Radiation-Protective Agents / pharmacology*
  • Thrombin / administration & dosage
  • Thrombin / chemistry
  • Thrombin / pharmacology*
  • Wound Healing / drug effects*

Substances

  • H2AX protein, human
  • Histones
  • Peptide Fragments
  • Radiation-Protective Agents
  • Polyethylene Glycols
  • rusalatide acetate
  • Thrombin