In vitro stability of growth hormone releasing factor (GRF) analogs in porcine plasma

• Published in: Hormone and metabolic research Vol. 23; no. 1; p. 15
• Main Authors: Su, C M, Jensen, L R, Heimer, E P, Felix, A M, Pan, Y C, Mowles, T F
• Format: Journal Article
• Language: English
• Published: Germany 01-01-1991
• Subjects: Amino Acid Sequence; Animals; Chromatography, High Pressure Liquid; Drug Stability; Growth Hormone-Releasing Hormone - analogs & derivatives; Growth Hormone-Releasing Hormone - blood; Half-Life; Molecular Sequence Data; Peptide Fragments - blood; Sermorelin; Swine - blood
• ISSN: 0018-5043
• DOI: 10.1055/s-2007-1003601

The stability of growth-hormone releasing factor (growth regulating factor; GRF) analogs in porcine plasma was examined. GRF analogs were incubated in porcine plasma at 37 degrees C, extracted and subsequently analyzed using high performance liquid chromatography (HPLC). GRF(1-29)-NH2 was rapidly broken down in the plasma with a degradation rate of t1/2 = 13 min. The primary degradation product was identified as GRF(3-29)-NH2. Substitution of Gly15 by Ala15 slightly prolonged the plasma half-life (t1/2 = 17 min) and the major degradative fragment was found to be [Ala15]GRF(3-29)-NH2. The cleavage between the 2 and 3 position of the peptide was not inhibited by trasylol at a concentration of 1,000 KIU/ml but was dramatically reduced by the combined use of diprotin A and trasylol. Absence of the free amino group at the N-terminus and/or substitution of a D-amino acid residue at the penultimate position completely prevented cleavage between the 2 and 3 position in the structural linear GRF analogs. Side-chain to side-chain cyclization between Asp8 and Lys12 amino acid residues significantly improved the stability of GRF in plasma with t1/2 greater than 2 hr. An additional stability was provided by substitution of D-Ala2 for Ala2 in the structural cyclic analog. Cyclization between Lys21 and Asp25 also improved the stability of the GRF peptide in the plasma. Stability was further enhanced by the presence of D-Ala2 or by forming a dicyclic analog through an additional linkage between Asp8 and Lys12.