Synthesis and structure-activity studies on novel analogs of human growth hormone releasing hormone (GHRH) with enhanced inhibitory activities on tumor growth

Marta Zarandi, Renzhi Cai, Magdolna Kovacs, Petra Popovics, Luca Szalontay, Tengjiao Cui, Wei Sha, Miklos Jaszberenyi, Jozsef Varga, Xian Yang Zhang, Norman L Block, Ferenc G. Rick, Gabor Halmos, Andrew V Schally

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The syntheses and biological evaluations of new GHRH analogs of Miami (MIA) series with greatly increased anticancer activity are described. In the design and synthesis of these analogs, the following previous substitutions were conserved: D-Arg2, Har9, Abu15, and Nle27. Most new analogs had Ala at position 8. Since replacements of both Lys12 and Lys21 with Orn increased resistance against enzymatic degradation, these modifications were kept. The substitutions of Arg at both positions 11 and 20 by His were also conserved. We kept D-Arg28, Har29 –NH2 at the C-terminus or inserted Agm or 12-amino dodecanoic acid amide at position 30. We incorporated pentafluoro-Phe (Fpa5), instead of Cpa, at position 6 and Tyr(Me) at position 10 and ω-amino acids at N-terminus of some analogs. These GHRH analogs were prepared by solid-phase methodology and purified by HPLC. The evaluation of the activity of the analogs on GH release was carried out in vitro on rat pituitaries and in vivo in male rats. Receptor binding affinities were measured in vitro by the competitive binding analysis. The inhibitory activity of the analogs on tumor proliferation in vitro was tested in several human cancer cell lines such as HEC-1A endometrial adenocarcinoma, HCT-15 colorectal adenocarcinoma, and LNCaP prostatic carcinoma. For in vivo tests, various cell lines including PC-3 prostate cancer, HEC-1A endometrial adenocarcinoma, HT diffuse mixed β cell lymphoma, and ACHN renal cell carcinoma cell lines were xenografted into nude mice and treated subcutaneously with GHRH antagonists at doses of 1–5 μg/day. Analogs MIA-602, MIA-604, MIA-610, and MIA-640 showed the highest binding affinities, 30, 58, 48, and 73 times higher respectively, than GHRH (1-29) NH2. Treatment of LNCaP and HCT-15 cells with 5 μM MIA-602 or MIA-690 decreased proliferation by 40%–80%. In accord with previous tests in various human cancer lines, analog MIA-602 showed high inhibitory activity in vivo on growth of PC-3 prostate cancer, HT-mixed β cell lymphoma, HEC-1A endometrial adenocarcinoma and ACHN renal cell carcinoma. Thus, GHRH analogs of the Miami series powerfully suppress tumor growth, but have only a weak endocrine GH inhibitory activity. The suppression of tumor growth could be induced in part by the downregulation of GHRH receptors levels.

Original languageEnglish (US)
Pages (from-to)60-70
Number of pages11
JournalPeptides
Volume89
DOIs
StatePublished - Mar 1 2017

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Growth Hormone-Releasing Hormone
Human Growth Hormone
Tumors
Cells
Adenocarcinoma
Growth
lauric acid
Neoplasms
Renal Cell Carcinoma
Cell Line
Non-Hodgkin's Lymphoma
Rats
Prostatic Neoplasms
Substitution reactions
Hormone Antagonists
Competitive Binding
Nude Mice
Amides
Down-Regulation
High Pressure Liquid Chromatography

Keywords

  • Cancer inhibition
  • Growth hormone releasing hormone
  • hGHRH antagonist
  • Hormone antagonist
  • Hypothalamic hormones
  • SAR studies

ASJC Scopus subject areas

  • Physiology
  • Biochemistry
  • Endocrinology
  • Cellular and Molecular Neuroscience

Cite this

Synthesis and structure-activity studies on novel analogs of human growth hormone releasing hormone (GHRH) with enhanced inhibitory activities on tumor growth. / Zarandi, Marta; Cai, Renzhi; Kovacs, Magdolna; Popovics, Petra; Szalontay, Luca; Cui, Tengjiao; Sha, Wei; Jaszberenyi, Miklos; Varga, Jozsef; Zhang, Xian Yang; Block, Norman L; Rick, Ferenc G.; Halmos, Gabor; Schally, Andrew V.

In: Peptides, Vol. 89, 01.03.2017, p. 60-70.

Research output: Contribution to journalArticle

Zarandi, M, Cai, R, Kovacs, M, Popovics, P, Szalontay, L, Cui, T, Sha, W, Jaszberenyi, M, Varga, J, Zhang, XY, Block, NL, Rick, FG, Halmos, G & Schally, AV 2017, 'Synthesis and structure-activity studies on novel analogs of human growth hormone releasing hormone (GHRH) with enhanced inhibitory activities on tumor growth', Peptides, vol. 89, pp. 60-70. https://doi.org/10.1016/j.peptides.2017.01.009
Zarandi, Marta ; Cai, Renzhi ; Kovacs, Magdolna ; Popovics, Petra ; Szalontay, Luca ; Cui, Tengjiao ; Sha, Wei ; Jaszberenyi, Miklos ; Varga, Jozsef ; Zhang, Xian Yang ; Block, Norman L ; Rick, Ferenc G. ; Halmos, Gabor ; Schally, Andrew V. / Synthesis and structure-activity studies on novel analogs of human growth hormone releasing hormone (GHRH) with enhanced inhibitory activities on tumor growth. In: Peptides. 2017 ; Vol. 89. pp. 60-70.
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AU - Zarandi, Marta

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AU - Popovics, Petra

AU - Szalontay, Luca

AU - Cui, Tengjiao

AU - Sha, Wei

AU - Jaszberenyi, Miklos

AU - Varga, Jozsef

AU - Zhang, Xian Yang

AU - Block, Norman L

AU - Rick, Ferenc G.

AU - Halmos, Gabor

AU - Schally, Andrew V

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N2 - The syntheses and biological evaluations of new GHRH analogs of Miami (MIA) series with greatly increased anticancer activity are described. In the design and synthesis of these analogs, the following previous substitutions were conserved: D-Arg2, Har9, Abu15, and Nle27. Most new analogs had Ala at position 8. Since replacements of both Lys12 and Lys21 with Orn increased resistance against enzymatic degradation, these modifications were kept. The substitutions of Arg at both positions 11 and 20 by His were also conserved. We kept D-Arg28, Har29 –NH2 at the C-terminus or inserted Agm or 12-amino dodecanoic acid amide at position 30. We incorporated pentafluoro-Phe (Fpa5), instead of Cpa, at position 6 and Tyr(Me) at position 10 and ω-amino acids at N-terminus of some analogs. These GHRH analogs were prepared by solid-phase methodology and purified by HPLC. The evaluation of the activity of the analogs on GH release was carried out in vitro on rat pituitaries and in vivo in male rats. Receptor binding affinities were measured in vitro by the competitive binding analysis. The inhibitory activity of the analogs on tumor proliferation in vitro was tested in several human cancer cell lines such as HEC-1A endometrial adenocarcinoma, HCT-15 colorectal adenocarcinoma, and LNCaP prostatic carcinoma. For in vivo tests, various cell lines including PC-3 prostate cancer, HEC-1A endometrial adenocarcinoma, HT diffuse mixed β cell lymphoma, and ACHN renal cell carcinoma cell lines were xenografted into nude mice and treated subcutaneously with GHRH antagonists at doses of 1–5 μg/day. Analogs MIA-602, MIA-604, MIA-610, and MIA-640 showed the highest binding affinities, 30, 58, 48, and 73 times higher respectively, than GHRH (1-29) NH2. Treatment of LNCaP and HCT-15 cells with 5 μM MIA-602 or MIA-690 decreased proliferation by 40%–80%. In accord with previous tests in various human cancer lines, analog MIA-602 showed high inhibitory activity in vivo on growth of PC-3 prostate cancer, HT-mixed β cell lymphoma, HEC-1A endometrial adenocarcinoma and ACHN renal cell carcinoma. Thus, GHRH analogs of the Miami series powerfully suppress tumor growth, but have only a weak endocrine GH inhibitory activity. The suppression of tumor growth could be induced in part by the downregulation of GHRH receptors levels.

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