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Synthesis, Stability and Direct Antiproliferative Effect of New Cysteine Modified GnRH Analogs

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Abstract

It is demonstrated that gonadotropin-releasing hormone (GnRH) analogs can directly inhibit the proliferation of reproductive tissue cancer cells, but the poor pharmacokinetic properties still restrict their application in treating hormone-dependent diseases. Modifications in position 6 and 10 of natural GnRH can improve the metabolic stability. In order to study the effect of incorporation Cys6 substitution with C-terminal Pro9-NHEt modification and dimerization of linear peptides on metabolic stability and antiproliferative activity of GnRH analogs, two new GnRH analogs [l-Cys6, desGly10, Pro9-NHEt]-GnRH (1) and [d-Cys6, desGly10, Pro9-NHEt]-GnRH (2), and their corresponding dimer derivatives ([l-Cys6, desGly10, Pro9-NHEt]-GnRH)2 (3) and ([d-Cys6, desGly10, Pro9-NHEt]-GnRH)2 (4) were synthesized. Incubation of these analogs with human serum was carried out to evaluate their metabolic stability, and direct growth inhibitory effect of the two dimer derivatives 3 and 4 on MCF-7 human breast cancer cell line was examined by MTT assay. The metabolic stability of dimer derivatives 3 and 4 was remarkably improved in comparison with natural GnRH. The d-Cys6 substituted dimer derivative 4 exhibited higher inhibitory effect (29.6–39.7% growth reduction) on cell growth than its corresponding counterpart 3 (21.8–26.2% growth reduction) at concentration range of 50, 100 and 200 μΜ. The cell growth inhibition of leuprolide was 16.4–27.2% at the tested concentrations. The dimer derivative 4 was the most stable and active GnRH analog in this study and has the potential for future preclinical investigations as promising antitumor drug candidate.

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Abbreviations

GnRH:

Gonadotropin-releasing hormone

LHRH:

Luteinizing hormone-releasing hormone

LH:

Luteinizing hormone

FSH:

Follicle-stimulating hormone

DIC:

N,N′-Diisopropylcarbodiimide

HOBt:

1-Hydroxy-benzotriazol

PyBOP:

(Benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate

DIPEA:

N,N-Diisopropylethylamine

TFA:

Trifluoroacetic acid

EDT:

1, 2-Ethanedithiol

DCM:

Dichloromethane

HPLC:

High performance liquid chromatography

MALDI-TOF MS:

Matrix-assisted laser desorption/ionization time of flight mass spectrometry

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Acknowledgements

This study was supported by the Scientific Research Projects in Colleges and Universities of Hebei Province (No. BJ2017055), in part by the Natural Science Foundation of Hebei Province (No. H2017406022), the Undergraduate innovation and entrepreneurship training program of Chengde Medical University (No. 201721) and the Key Discipline Construction Projects of Higher School in Hebei Province ([2013]4).

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Authors and Affiliations

  1. Hebei Province Key Laboratory of Research and Development of Traditional Chinese Medicine, Chengde Medical University, Chengde, 067000, Hebei, People’s Republic of China

    Songtao Li, Hongling Zhao, Ruxing Wang, Xiaoxia Mao, Ting Hao, Zhifeng Yin, Shuhua Deng, Yaqi Yang & Huina Wang

  2. Department of Immunology, Chengde Medical University, Chengde, 067000, Hebei, People’s Republic of China

    Jianping Wang & Xiaomin Chang

  3. The Third Department of Surgery, First Affiliated Hospital of Chengde Medical University, Chengde, 067000, Hebei, People’s Republic of China

    Enhong Zhao

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  1. Songtao Li
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  2. Hongling Zhao
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  3. Ruxing Wang
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  4. Jianping Wang
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  5. Xiaoxia Mao
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  7. Xiaomin Chang
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Correspondence to Songtao Li.

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Li, S., Zhao, H., Wang, R. et al. Synthesis, Stability and Direct Antiproliferative Effect of New Cysteine Modified GnRH Analogs. Int J Pept Res Ther 25, 1361–1367 (2019). https://doi.org/10.1007/s10989-018-9781-y

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  • DOI: https://doi.org/10.1007/s10989-018-9781-y

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