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New Gaba-containing analogues of human Growth Hormone-Releasing Hormone (1–30)-amide: I. Synthesis and in vitro biological activity

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Abstract

Analogues of human growth hormone-releasing hormone (1–30)-amide have been developed. All analogues have been modified in position 27 with Nle and with Gaba in position 30. Additional D-amino-acids have been inserted in the GHRH(1–30)-NH2 sequence

A-1741: Nle27, Gaba30-GH-RH (1–30)-NH2 A-495: D-Ala2, Nle27, Gaba30-GH-RH (1–30 )-NH2 A-515: D Ala2, Leu15, Nle27, Gaba30-GH-RH (1–30)-NH2 A-527: D-Ala2, D-Arg11, Leu15, Nle27, Gaba30-GH-RH (1–30)-NH2.

Our analogues were synthesized by solid phase peptide synthesis and were tested is two different in vitro systems and in rat pituitary cell cultures. A-495 and A-1741 were found to be the most active in releasing GH, however they showed different activities in the two different test systems. A-495 exhibited higher potency in the superfusion system (1.63 fold potency of the GHRH (1–29)-amide), while A-1741 evoked higher GH release from cultured pituitary cells (1.5–2.5 times higher than the GH-RH(1–44)-amide). The other analogues (A-515 and A-527) were found to be equipotent to the standard molecule. We can conclude that Nle27 and Gaba30 substitutions appeared to be a good modification in in vitro test systems, and Gaba30 substitution served as a good spacer during the synthesis, since it made the coupling of the C-terminal amino acids easier and produced quantitative coupling. In addition to the advantageous properties in the synthesis these modifications with or without D-Ala at the N-terminus increased the in vitro biological activity to 1.5–2.5 fold of the GHRH molecule. The additional substitution of Gly15 with Leu and Arg11 with D-Arg did not improve the in vitro GH-releasing activity of our analogues. A detailed in vivo investigation, which is essential for the future clinical use, hasbeen performed and written in Part II of this paper.

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Authors and Affiliations

  1. 1st. Institute of Biochemistry, Semmelweis University Medical School, 1444 Budapest 8., P.O. Box 260, Hungary

    Imre Mezö, B. Szöke, E. Z. Szabó & I. Teplán

  2. University Medical School, Pecs, Hungary

    M. Kovács & J. Horváth

  3. Institute of Experimental Medicine, Budapest, Hungary

    G. B. Makara & Gy. Rappay

  4. Hungarian Academy of Sciences, Central Institute for Chemistry, Budapest, Hungary

    J. Tamás

Authors
  1. Imre Mezö
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  2. M. Kovács
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  3. B. Szöke
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  4. E. Z. Szabó
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  5. J. Horváth
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  6. G. B. Makara
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  7. Gy. Rappay
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  8. J. Tamás
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  9. I. Teplán
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Mezö, I., Kovács, M., Szöke, B. et al. New Gaba-containing analogues of human Growth Hormone-Releasing Hormone (1–30)-amide: I. Synthesis and in vitro biological activity. J Endocrinol Invest 16, 793–798 (1993). https://doi.org/10.1007/BF03348929

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  • DOI: https://doi.org/10.1007/BF03348929

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