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In vitro and in vivo toxicity of 5-FdU-alendronate, a novel cytotoxic bone-seeking duplex drug against bone metastasis

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Summary

Background Bone remains one of the most common anatomic sites for cancer metastases, and the limited therapeutic options aggravate cancer-related morbidity and mortality in multiple malignancies. The covalent conjugation of the amino-bisphosphonate alendronate (ale) with the antimetabolite 5-fluoro-2′-desoxyuridine (5-FdU) results in N4-(butyl-(4-hydroxy-4-phosphono)phosphate)-5-fluoro-2′-desoxyuridine (5-FdU-alendronat, 5-FdU-ale), an effective, novel bone-targeting duplex drug directed against skeletal cancer manifestations. Methods In vitro cytotoxicity of ale, 5-FdU or 5-FdU-ale was measured with Alamar Blue and MUH cell viability assays in 14 malignant melanoma, multiple myeloma, bone marrow-derived stromal cell and osteoblast-like cell lines. In vivo toxicity was evaluated using the chicken embryo assay and evaluation of nephrotoxicity and the systemic toxicity in Balb/c nude mice. The effect of 5-FdU-ale on osteoclast was evaluated with Balb/c nude mice in a metastatic breast cancer mouse model. Results A cell line-specific, dose-related cytotoxicity was observed for 5-FdU-ale in all cancer cell lines tested, which was significantly less toxic than 5-FdU alone when compared to the benign osteoblasts or stromal cells. The embryotoxicity of 5-FdU-ale was significantly less than that of the parental drugs alendronate or 5-FdU. 5-FdU-ale showed no signs of unwanted side effects, weight loss or nephrotoxicity in mice. In a bone metastasis mouse model, 5-FdU-ale reduced the number of tumor-associated osteoclasts. Conclusion The coupling of an amino-bisphosphonate with an antimetabolite via an N-alkyl-bonding offers a new strategy for the preparation of amino-bisphosphonates conjugates with a cancer cell-specific, efficacious cytotoxic bone-targeting potential along with a reduced systemic toxicity. The innovative duplex drug 5-FdU-ale therefore warrants further clinical investigation.

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Acknowledgments

The research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. This work was supported by grants from the DFG SFB 773: “Understanding and overcoming drug resistance of solid tumors” to CB. SS received a research scholarship from the University of Heidelberg. SS and SV received a research grant from the “Stiftung für Krebs und Scharlachforschung Mannheim”. The sponsors had no involvement in the study design, the collection, analysis and interpretation of data, the writing of the manuscript or in the decision to submit the manuscript for publication.

Conflict of interest

All authors claim no conflicts of interest

Ethical standards

The Ethics Committee for Animal Experiments at the Christian-Albrechts-Universität zu Kiel, Germany (V312-72241.121-10) approved this study. Animal experiments and care were performed in accordance with the guidelines of institutional authorities.

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Authors and Affiliations

  1. Department of Gynecology and Obstetrics, University of Heidelberg, Im Neuenheimer Feld 440, 69120, Heidelberg, Germany

    Sarah Schott

  2. Department. of Medical Oncology, National Center for Tumor Diseases Heidelberg, Heidelberg University Hospital, Im Neuenheimer Feld 460, 69120, Heidelberg, Germany

    Sarah Schott & Sonia Vallet

  3. Section Biomedical Imaging, Department of Radiology and Neuroradiology, University Hospital Schleswig-Holstein, Campus Kiel, Am Botanischen Garten 14, 24118, Kiel, Germany

    Robert J. Tower & Sanjay Tiwari

  4. Section of Dermato-Oncology, Department of Dermatology and Allergology, University of Tuebingen, Liebermeisterstr.25, 72076, Tuebingen, Germany

    Seema Noor & Christian Busch

  5. Department of Gynecology and Obstetrics, University Medical Centre Schleswig-Holstein, Campus Kiel, Arnold-Heller-Str. 3, Haus 24, 24105, Kiel, Germany

    Christian Schem

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  1. Sarah Schott
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  2. Sonia Vallet
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Correspondence to Sarah Schott.

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Schott, S., Vallet, S., Tower, R.J. et al. In vitro and in vivo toxicity of 5-FdU-alendronate, a novel cytotoxic bone-seeking duplex drug against bone metastasis. Invest New Drugs 33, 816–826 (2015). https://doi.org/10.1007/s10637-015-0253-3

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  • DOI: https://doi.org/10.1007/s10637-015-0253-3

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