Intramolecular azo-bridge as a cystine disulfide bond surrogate: Somatostatin-14 and brain natriuretic peptide (BNP) analogs

Bioorg Med Chem. 2011 Jan 15;19(2):798-806. doi: 10.1016/j.bmc.2010.12.014. Epub 2010 Dec 13.

Abstract

Cystine disulfide bond is a common feature in numerous biologically active peptides and proteins and accordingly its replacement by various surrogates presents a potential route to obtain analogs with improved pharmacokinetic characteristics. The purpose of the present study was to assess whether an azo-bridge can serve as such a surrogate. In view of the marked clinical significance of somatostatin and the brain natriuretic peptide (BNP) we choose these peptides as a model. Three cyclic-azo somatostatin analogs and three cyclic-azo BNP analogs were effectively prepared in solution through azo bond formation between p-amino phenylalanine and His or Tyr residues that were positioned in the peptide sequences in place of the native Cys residues. The peptides binding affinities to the sst₂ and ANP-receptor (NPR-A) expressed on rat acinar pancreating carcinoma AR4-2J cell membranes and HeLa cells, respectively, were examined. The somatostatin analogs displayed good to moderate affinities to the rat sst₂ in the nM range with best results obtained with peptide 2, that is, IC₅₀ = 8.1 nM. Molecular dynamics simulations on these peptides suggests on a correlation between the observed binding potencies and the degree of conformational space overlapping with that of somatostatin. The BNP analogs exhibited binding affinities to the NPR-A in the nM range with best results obtained with BNP-1, that is, IC₅₀ = 60 nM.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Azo Compounds / chemistry*
  • Cell Line, Tumor
  • Cystine / chemistry*
  • Disulfides / chemistry*
  • Humans
  • Hydrogen-Ion Concentration
  • Molecular Dynamics Simulation
  • Natriuretic Peptide, Brain / analogs & derivatives*
  • Natriuretic Peptide, Brain / chemical synthesis
  • Natriuretic Peptide, Brain / metabolism
  • Protein Binding
  • Rats
  • Somatostatin / analogs & derivatives*
  • Somatostatin / chemical synthesis
  • Somatostatin / metabolism

Substances

  • Azo Compounds
  • Disulfides
  • Natriuretic Peptide, Brain
  • Cystine
  • Somatostatin