Synthesis and in vitro testing of new potent polyacridine-melittin gene delivery peptides

Bioconjug Chem. 2010 Jan;21(1):74-83. doi: 10.1021/bc9003124.

Abstract

The combination of a polyacridine peptide modified with a melittin fusogenic peptide results in a potent gene transfer agent. Polyacridine peptides of the general formula (Acr-X)(n)-Cys were prepared by solid-phase peptide synthesis, where Acr is Lys modified on its epsilon-amine with acridine, X is Arg, Leu, or Lys and n is 2, 3, or 4 repeats. The Cys residue was modified by either a maleimide-melittin or a thiolpyridine-Cys-melittin fusogenic peptide resulting in reducible or non-reducible polyacridine-melittin peptides. Hemolysis assays established that polyacridine-melittin peptides retained their membrane lytic potency relative to melittin at pH 7.4 and 5. When combined with plasmid DNA, the membrane lytic potency of polyacridine-melittin peptides was neutralized. Gene transfer experiments in multiple cell lines established that polyacridine-melittin peptides mediate expression as efficiently as PEI. The expression was very dependent upon a disulfide bond linking polyacridine to melittin. The gene transfer was most efficient when X is Arg and n is 3 or 4 repeats. These studies establish polyacridine peptides as a novel DNA binding anchor peptide.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acridines / chemical synthesis*
  • Acridines / chemistry
  • Acridines / metabolism*
  • Acridines / pharmacology
  • Amino Acid Sequence
  • Animals
  • Arginine / chemistry
  • Arginine / metabolism
  • Cell Line
  • Cysteine / chemistry
  • Cysteine / metabolism
  • DNA / administration & dosage
  • DNA / chemistry
  • DNA / metabolism
  • Gene Transfer Techniques*
  • Hemolysis / drug effects
  • Hemolysis / physiology
  • Hydrogen-Ion Concentration
  • Melitten / chemical synthesis*
  • Melitten / chemistry
  • Melitten / metabolism*
  • Melitten / pharmacology
  • Mice
  • Mice, Inbred ICR
  • Molecular Sequence Data
  • Plasmids / administration & dosage
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Polymers / chemical synthesis*
  • Polymers / chemistry
  • Polymers / metabolism*
  • Polymers / pharmacology
  • Sulfhydryl Compounds / chemistry
  • Sulfhydryl Compounds / metabolism

Substances

  • Acridines
  • Polymers
  • Sulfhydryl Compounds
  • Melitten
  • DNA
  • Arginine
  • Cysteine