Evaluation of the Biological Potency of New Agmatine Analogs of Growth Hormone-Releasing Hormone in the Bovine

Abstract

Four new growth hormone-releasing hormone (GHRH) analogs with C-terminal agmatine were compared with the parent human GHRH(1–29)NH₂ fragment to assess their abilities to increase serum concentrations of growth hormone (GH) in the bovine. The four analogs were: [D-Ala², Nle²⁷] GHRH(1–28)Agm (JG-73); [desNH₂-Tyr¹, Ala¹⁵, Nle²⁷] GHRH(1–28)Agm (MZ-2–51); [desNH₂-Tyr¹, Ala¹⁵, D-Lys²¹, Nle²⁷] GHRH(1–28)Agm (M2–2–75); and [desNH₂-Tyr¹, D-Lys^12,21, Ala¹⁵, Nle²⁷] GHRH(1–28)Agm (MZ-2–87). The special characteristic of all four GHRH analogs is that arginine was replaced by agmatine (Agm) in Position 29. Five pregnant Holstein cows received these peptides subcutaneously at the following doses: 0.0156, 0.0625, 0.25, 1, and 4 μg/kg body wt. Each cow received each analog-dose combination according to a 5 × 5 Greco-Latin square design repeated for the 5-week treatment. Each cow also received saline vehicle only at the end of the 5-week treatment. Blood samples were collected from 30 min before until 360 min after treatment injection. Total area under the GH response curves for the 6-hr sampling period for each dose of each GHRH analog was compared. There was a linear dose-dependent GH release in response to hGHRH(1–29)NH₂ and its four GHRH(1–28)Agm analogs. At the dose of 0.25 μg/kg, two GHRH analogs, JG-73 and MZ-2–75, stimulated greater GH release than hGHRH(1–29)NH₂ (P < 0.05). No differences were seen at the two lowest doses, 0.0625 and 0.156 μg/kg. When both total area under the GH response curves and GH peak amplitudes for each treatment were averaged for all doses, JG-73 and MZ-2–75 stimulated greater GH release than hGHRH(1–29)NH₂ (P < 0.05). In summary, three GHRH(1–28)Agm analogs, JG-73, MZ-2–75, and MZ-2–51, were found to be 11.8, 11.3, and 6.5 times more potent, respectively, on a weight basis, than hGHRH(1–29)NH₂ in stimulating the release of GH in cows.