GnRH-III based multifunctional drug delivery systems containing daunorubicin and methotrexate

Ulrike Leurs, Eszter Lajkó, G. Mező, Erika Orbán, Peter Öhlschläger, Andreas Marquardt, L. Kőhidai, Marilena Manea

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Abstract

Here we report on the design, synthesis and biochemical characterization of multifunctional bioconjugates containing two chemotherapeutic agents, daunorubicin and methotrexate, coupled to the GnRH-III decapeptide, which served as a targeting moiety. This represents a possible approach to increase the receptor mediated tumor targeting and consequently the cytostatic effect of anticancer drug-peptide bioconjugates. The multifunctional bioconjugates were prepared according to two drug design approaches recently developed by our group. Both bifunctional GnRH-III derivatives, [ 4Lys]-GnRH-III (Glp-His-Trp-Lys-His-Asp-Trp-Lys-Pro-Gly-NH 2) and [ 8Lys(Lys)]-GnRH-III (Glp-His-Trp-Ser-His-Asp-Trp-Lys(Lys)-Pro-Gly- NH 2), contain two free amino groups suitable for the attachment of two anticancer drugs, such as methotrexate and daunorubicin. The drugs were chosen with respect to their different mechanisms of action, with the goal of increasing the antitumor effect of the bioconjugates. The in vitro cytostatic effect of the bioconjugates was determined on MCF-7 human breast, HT-29 human colon and LNCaP human prostate cancer cells by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay. Their in vitro stability/degradation in human serum and in the presence of rat liver lysosomal homogenate was investigated by liquid chromatography in combination with mass spectrometry. The influence of the multifunctional bioconjugates on the cell adhesion and cell proliferation was studied on Mono Mac 6 human leukemic monocytes. It was found that (1) all synthesized bioconjugates had in vitro cytostatic effect; (2) they were stable in human serum for at least 24 h; (3) they were hydrolyzed in the presence of lysosomal homogenate and (4) they exerted a moderate cell-cell adhesion inducing effect. These results demonstrate that multifunctional bioconjugates containing two different anticancer drugs attached to the same GnRH-III targeting moiety could be successfully prepared and resulted in higher in vitro cytostatic effect than the monofunctional bioconjugates containing either methotrexate or daunorubicin, in particular on HT-29 human colon cancer cells.

Original languageEnglish
Pages (from-to)173-183
Number of pages11
JournalEuropean Journal of Medicinal Chemistry
Volume52
DOIs
Publication statusPublished - Jun 2012

Fingerprint

Daunorubicin
Drug Delivery Systems
Methotrexate
Cytostatic Agents
Pharmaceutical Preparations
Cell adhesion
Cells
Cell Adhesion
prolylglycine
Liquid chromatography
Cell proliferation
Liver
Mass spectrometry
Rats
Tumors
Assays
Drug Design
gonadotropin-releasing hormone-III
Serum
Liquid Chromatography

Keywords

  • Cytostatic effect
  • Daunorubicin
  • GnRH-III
  • Methotrexate
  • Multifunctional drug delivery systems
  • Targeted cancer chemotherapy

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Pharmacology

Cite this

GnRH-III based multifunctional drug delivery systems containing daunorubicin and methotrexate. / Leurs, Ulrike; Lajkó, Eszter; Mező, G.; Orbán, Erika; Öhlschläger, Peter; Marquardt, Andreas; Kőhidai, L.; Manea, Marilena.

In: European Journal of Medicinal Chemistry, Vol. 52, 06.2012, p. 173-183.

Research output: Contribution to journalArticle

Leurs, Ulrike ; Lajkó, Eszter ; Mező, G. ; Orbán, Erika ; Öhlschläger, Peter ; Marquardt, Andreas ; Kőhidai, L. ; Manea, Marilena. / GnRH-III based multifunctional drug delivery systems containing daunorubicin and methotrexate. In: European Journal of Medicinal Chemistry. 2012 ; Vol. 52. pp. 173-183.
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