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In vitro and in vivo study of an albumin-binding prodrug of doxorubicin that is cleaved by cathepsin B

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Abstract

Purpose

This study was designed to evaluate the in vitro and in vivo antitumor activity of an albumin-binding prodrug of doxorubicin 1 which incorporates a maleimide moiety and a para-aminobenzyloxycarbonyl (PABC) spacer coupled to the dipeptide Phe-Lys that is cleaved by cathepsin B.

Methods

Cleavage of the albumin conjugate was studied with cathepsin B and in homogenates of MDA-MB 435 tumors. For in vivo studies, nude mice were injected with (a) glucose buffer, (b) doxorubicin (2 × 8 mg/kg, i.v, on days 10 and 17), or (c) compound 1 (3 × 24 mg/kg doxorubicin equivalent, on days 10, 17 and 24).

Results

Prodrug 1 once bound to albumin was effectively cleaved by cathepsin B and in tumor homogenates releasing doxorubicin. A cytotoxicity assay of the albumin conjugate of 1 in two human tumor cell lines showed that doxorubicin was ~6 times more active than the conjugate. In contrast, in an in vivo study, the prodrug exhibited superior antitumor activity (T/C 15%) compared to doxorubicin (T/C 49%) in an equitoxic comparison.

Conclusions

The cathepsin B cleavable spacer Phe-Lys-PABC incorporated in an albumin-binding prodrug is an effective way to increase the therapeutic index of doxorubicin.

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References

  1. Gianni L, Grasselli G, Cresta S, Locatelli A, Vigano L, Minotti G (2003) Anthracyclines. Cancer Chemother Biol Response Modif 21:29–40

    PubMed  CAS  Google Scholar 

  2. Dorr RT, Von Hoff DD (1994) Cancer chemotherapy handbook, 2nd edn. Appleton and Lange, Norwalk

    Google Scholar 

  3. Cattel L, Ceruti M, Dosio F (2003) From conventional to stealth liposomes: a new frontier in cancer chemotherapy. Tumori 89:237–249

    PubMed  CAS  Google Scholar 

  4. Li J, Wu C, Dai Y, Zhang R, Wang X, Fu D, Chen B (2007) Doxorubicin-CdS nanoparticles: a potential anticancer agent for enhancing the drug uptake of cancer cells. J Nanosci Nanotechnol 7:435–439

    PubMed  CAS  Google Scholar 

  5. Kratz F, Warnecke A, Schmid B, Chung DE, Gitzel M (2006) Prodrugs of anthracyclines in cancer chemotherapy. Curr Med Chem 13:763–771

    Article  Google Scholar 

  6. Kratz F, Mueller-Driver R, Hofmann I, Drevs J, Unger C (2000) A novel macromolecular prodrug concept exploiting endogenous serum albumin as a drug carrier for cancer chemotherapy. J Med Chem 43:1253–1256

    Article  PubMed  CAS  Google Scholar 

  7. Kratz F, Warnecke A, Scheuermann K, Stockmar C, Schwab J, Lazar P, Drückes P, Esser N, Drevs J, Rognan D, Bissantz C, Hinderling C, Folkers G, Fichtner I, Unger C (2002) Probing the cysteine-34 position of endogenous serum albumin with thiol-binding doxorubicin derivatives: improved efficacy of an acid-sensitive doxorubicin derivative with specific albumin-binding properties compared to that of the parent compound. J Med Chem 45:5523–5533

    Article  PubMed  CAS  Google Scholar 

  8. Mansour AM, Drevs J, Esser N, Hamada FM, Badary OA, Unger C, Fichtner I, Kratz F (2003) A new approach for the treatment of malignant melanoma: enhanced antitumor efficacy of an albumin-binding doxorubicin prodrug that is cleaved by matrix metalloproteinase 2. Cancer Res 63:4062–4066

    PubMed  CAS  Google Scholar 

  9. Kratz F (2007) DOXO-EMCH (INNO–206): the first albumin-binding prodrug of doxorubicin to enter clinical trials. Expert Opin Investig Drugs 16:855–866

    Article  PubMed  CAS  Google Scholar 

  10. Kratz F (2008) Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. J Control Release 132:171–183

    Article  CAS  PubMed  Google Scholar 

  11. Kratz F, Beyer U (1998) Serum proteins as drug carriers of anticancer agents: a review. Drug Deliv 5:281–299

    Article  CAS  PubMed  Google Scholar 

  12. Maeda H, Wu J, Sawa T, Matsumura Y, Hori K (2000) Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. J Control Release 65:271–284

    Article  PubMed  CAS  Google Scholar 

  13. Graeser R, Chung DE, Esser N, Moor S, Schachtele C, Unger C, Kratz F (2008) Synthesis and biological evaluation of an albumin-binding prodrug of doxorubicin that is cleaved by prostate-specific antigen (PSA) in a PSA-positive orthotopic prostate carcinoma model (LNCaP). Int J Cancer 122:1145–1154

    Article  PubMed  CAS  Google Scholar 

  14. Kratz F, Mansour A, Soltau J, Warnecke A, Fichtner I, Unger C, Drevs J (2005) Development of albumin-binding doxorubicin prodrugs that are cleaved by prostate-specific antigen. Arch Pharm (Weinheim) 338:462–472

    Article  CAS  Google Scholar 

  15. Chung DE, Kratz F (2006) Development of a novel albumin-binding prodrug that is cleaved by urokinase-type-plasminogen activator (uPA). Bioorg Med Chem Lett 16:5157–5163

    Article  PubMed  CAS  Google Scholar 

  16. Schmid B, Chung DE, Warnecke A, Fichtner I, Kratz F (2007) Albumin-binding prodrugs of camptothecin and doxorubicin with an Ala-Leu-Ala-Leu-linker that are cleaved by cathepsin B: synthesis and antitumor efficacy. Bioconjug Chem 18:702–716

    Article  PubMed  CAS  Google Scholar 

  17. Duncan R, Ulbrich K (1993) Development of N-(2-hydroxypropyl)methacrylamide copolymer conjugates for delivery of cancer chemotherapy. Makromol Chem Macromol Symp 70(71):157–162

    Google Scholar 

  18. Dubowchik GM, Firestone RA, Padilla L, Willner D, Hofstead SJ, Mosure K, Knipe JO, Lasch SJ, Trail PA (2002) Cathepsin B-labile dipeptide linkers for lysosomal release of doxorubicin from internalizing immunoconjugates: model studies of enzymatic drug release and antigen-specific in vitro anticancer activity. Bioconjug Chem 13:855–869

    Article  PubMed  CAS  Google Scholar 

  19. Carl PL, Chakravarty PK, Katzenellenbogen JA (1981) A novel connector linkage applicable in prodrug design. J Med Chem 24:479–480

    Article  PubMed  CAS  Google Scholar 

  20. King HD, Dubowchik GM, Mastalerz H, Willner D, Hofstead SJ, Firestone RA, Lasch SJ, Trail PA (2002) Monoclonal antibody conjugates of doxorubicin prepared with branched peptide linkers: inhibition of aggregation by methoxytriethyleneglycol chains. J Med Chem 45:4336–4343

    Article  PubMed  CAS  Google Scholar 

  21. Abu Ajaj K, Kratz F (2009) Development of dual-acting prodrugs for circumventing multidrug-resistance in breast cancer. Bioorg Med Chem Lett 19(3):995–1000

    Article  PubMed  CAS  Google Scholar 

  22. Abu Ajaj K, Biniossek ML, Kratz F (2009) Development of protein-binding bifunctional spacers for the development of dual-acting prodrugs. Bioconjug Chem. doi:10.1021/bc800429q

    Google Scholar 

  23. Graeser R, Bornmann C, Esser N, Ziroli V, Jantscheff P, Unger C, Hopt UT, Schaechtele C, von Dobschuetz E, Massing U (2009) Antimetastatic effects of liposomal gemcitabine and empty liposomes in an orthotopic mouse model of pancreatic cancer. Pancreas. doi:10.1097/MPA.0b013e31819436e6

    PubMed  Google Scholar 

  24. Kratz F, Roth T, Fichtner I, Schumacher P, Fiebig HH, Unger C (2000) In vitro and in vivo efficacy of acid-sensitive transferrin and albumin doxorubicin conjugates in a human xenograft panel and in the MDA-MB-435 mamma carcinoma model. J Drug Target 8:305–318

    Article  PubMed  CAS  Google Scholar 

  25. Trail PA, Willner D, Lasch SJ, Henderson AJ, Greenfield RS, King D, Zoeckler ME, Braslawsky GR (1992) Antigen-specific activity of carcinoma-reactive BR64-doxorubicin conjugates evaluated in vitro and in human tumor xenograft models. Cancer Res 52:5693–5700

    PubMed  CAS  Google Scholar 

  26. Moon SJ, Govindan SV, Cardillo TM, D’Souza CA, Hansen HJ, Goldenberg DM (2008) Antibody conjugates of 7-ethyl-10-hydroxycamptothecin (SN-38) for targeted cancer chemotherapy. J Med Chem 51:6916–6926

    Article  PubMed  CAS  Google Scholar 

  27. Walker MA, Dubowchik GM, Hofstead SJ, Trail PA, Firestone RA (2002) Synthesis of an immunoconjugate of camptothecin. Bioorg Med Chem Lett 12:217–219

    Article  PubMed  CAS  Google Scholar 

  28. Dubowchik GM, Mosure K, Knipe JO, Firestone RA (1998) Cathepsin B-sensitive dipeptide prodrugs. 2. Models of anticancer drugs paclitaxel (Taxol), mitomycin C and doxorubicin. Bioorg Med Chem Lett 8:3347–3352

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Tumor Biology Center, Breisacher Strasse 117, 79106, Freiburg, Germany

    Khalid Abu Ajaj & Felix Kratz

  2. ProQinase GmbH, Breisacher Strasse 117, Freiburg, Germany

    Ralph Graeser

  3. Max-Delbrück Center, Robert-Rössle-Strasse 10, 13122, Berlin, Germany

    Iduna Fichtner

Authors
  1. Khalid Abu Ajaj
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  2. Ralph Graeser
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  3. Iduna Fichtner
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  4. Felix Kratz
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Corresponding author

Correspondence to Felix Kratz.

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Abu Ajaj, K., Graeser, R., Fichtner, I. et al. In vitro and in vivo study of an albumin-binding prodrug of doxorubicin that is cleaved by cathepsin B. Cancer Chemother Pharmacol 64, 413–418 (2009). https://doi.org/10.1007/s00280-009-0942-8

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  • DOI: https://doi.org/10.1007/s00280-009-0942-8

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