Structure-activity Study of hCGRP8-37, a Calcitonin Gene-Related Peptide Receptor Antagonist

J Med Chem. 1992 Jun 12;35(12):2163-8. doi: 10.1021/jm00090a003.

Abstract

A structure-activity study was carried out to determine the importance of the N-terminal amino acids of hCGRP8-37 in binding and antagonistic activity to CGRP receptors. Therefore, fragments of hCGRP8-37 as well as analogs obtained by the replacement of residues 9-12 by L-alanine were synthesized by solid-phase peptide synthesis, using BOP as a coupling reagent. The affinities of the peptides to CGRP receptors were evaluated in the rat brain, guinea pig atrium, and guinea pig vas deferens membrane preparations. Their antagonistic activities were measured in the guinea pig atria and rat vas deferens bioassays. The pharmacological characterization showed that arginine-11 and leucine-12 play a crucial role for the affinity of hCGRP8-37. Interestingly, it was observed that [Ala11]hCGRP8-37 was able to potentiate the twitch response of the electrically stimulated rat vas deferens. On the other hand, the substantial antagonistic potencies of analogs [Ala9]-, [Ala10]-, and [Ala12]hCGRP8-37, as compared to those of the fragments hCGRP10-37, hCGRP11-37, and hCGRP12-37, suggest that the side chains of Thr-9, His-10, and Leu-12 assume mainly a structural role. Accordingly, the conformational characterization of these peptides by circular dichroism spectroscopy revealed that the residues 9-12 are important for the integrity of the amphiphilic alpha-helix of hCGRP8-37.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Brain / metabolism
  • Calcitonin Gene-Related Peptide / chemistry*
  • Calcitonin Gene-Related Peptide / metabolism
  • Calcitonin Gene-Related Peptide / pharmacology*
  • Cell Membrane / metabolism
  • Chromatography, High Pressure Liquid
  • Electric Stimulation
  • Guinea Pigs
  • Heart Atria / metabolism
  • Male
  • Molecular Sequence Data
  • Peptide Fragments / chemical synthesis*
  • Peptide Fragments / chemistry*
  • Peptide Fragments / metabolism
  • Peptide Fragments / pharmacology*
  • Protein Conformation
  • Rats
  • Rats, Inbred Strains
  • Receptors, Calcitonin
  • Receptors, Cell Surface / antagonists & inhibitors*
  • Structure-Activity Relationship
  • Vas Deferens

Substances

  • Peptide Fragments
  • Receptors, Calcitonin
  • Receptors, Cell Surface
  • calcitonin gene-related peptide (8-37)
  • Calcitonin Gene-Related Peptide