title: The application of crystal soaking technique to study the effect of zinc and cresol on insulinotropin crystals grown from a saline solution.
abstract: OBJECTIVE
  The purpose of this study is to investigate the effect of zinc and cresol on the structure of insulinotropin crystals.
  
  
  METHODS
  Insulinotropin crystals grown from a saline solution were treated with zinc and/or m-cresol using a crystal soaking technique. The effects of these additives on the crystal structure were investigated with powder X-ray diffraction, photomicrography, and differential scanning calorimetry. The molecular interaction between insulinotropin and m-trifluorocresol in solution was also studied by 19F NMR:
  
  
  RESULTS
  The data suggest that the original crystals grown from a saline solution have relatively weak lattice forces. After the addition of m-cresol to the suspension of the insulinotropin crystals, the crystals were immediately rendered amorphous. The m-cresol molecules which diffused into the crystals through solvent channels may have disturbed the lattice interactions that maintain the integrity of the crystal. In contrast, the zinc added to the suspension stabilized the crystal lattice so that the subsequent addition of m-cresol did not alter the integrity of the crystals. A marked increase in melting point (206 degrees versus 184 degrees) and heat of fusion (24.6 J/g versus 1.4 J/g) of the crystals was observed after the treatment with zinc. The solubility of the zinc treated crystals in a pH 7.1 phosphate buffered saline was 1/20 of that of the original crystals.
  
  
  CONCLUSIONS
  When the insulinotropin crystals were treated with the additives using a crystal soaking method, the crystals underwent structural changes. Zinc stabilized the crystal lattice, and reduced the solubility of the peptide.
authors.affiliation: Central Research Division, Pharmaceutical Research and Development, Pfizer Inc., Groton, Connecticut 06340, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Kim
authors.forename: A M
authors.initials: AM
authors.lastname: Haren
mesh_terms: D000595:Amino Acid Sequence; D002152:Calorimetry, Differential Scanning; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003408:Cresols / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D005934:Glucagon; D052216:Glucagon-Like Peptide 1; D004763:Glucagon-Like Peptides; D006863:Hydrogen-Ion Concentration; D007279:Injections, Subcutaneous; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D010455:Peptides / Q000737:chemistry*; D010784:Photomicrography; D012965:Sodium Chloride; D012995:Solubility; D015032:Zinc / Q000737:chemistry*
chemical_list: D003408:Cresols; D010446:Peptide Fragments; D010455:Peptides; D012965:Sodium Chloride; D004763:Glucagon-Like Peptides; D052216:Glucagon-Like Peptide 1; D005934:Glucagon; D015032:Zinc; C051322:glucagon-like peptide 1 (7-37)