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
Gianni L, Grasselli G, Cresta S, Locatelli A, Vigano L, Minotti G (2003) Anthracyclines. Cancer Chemother Biol Response Modif 21:29–40
Dorr RT, Von Hoff DD (1994) Cancer chemotherapy handbook, 2nd edn. Appleton and Lange, Norwalk
Cattel L, Ceruti M, Dosio F (2003) From conventional to stealth liposomes: a new frontier in cancer chemotherapy. Tumori 89:237–249
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
Kratz F, Warnecke A, Schmid B, Chung DE, Gitzel M (2006) Prodrugs of anthracyclines in cancer chemotherapy. Curr Med Chem 13:763–771
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
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
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
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
Kratz F (2008) Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. J Control Release 132:171–183
Kratz F, Beyer U (1998) Serum proteins as drug carriers of anticancer agents: a review. Drug Deliv 5:281–299
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
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
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
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
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
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
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
Carl PL, Chakravarty PK, Katzenellenbogen JA (1981) A novel connector linkage applicable in prodrug design. J Med Chem 24:479–480
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
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
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
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
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
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
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
Walker MA, Dubowchik GM, Hofstead SJ, Trail PA, Firestone RA (2002) Synthesis of an immunoconjugate of camptothecin. Bioorg Med Chem Lett 12:217–219
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
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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