Summary
The two monoclonal antibodies (mAb), L6 (anti-carcinoma), and 1F5 [anti-(B-cell-lymphoma)], were chemically linked to the enzyme penicillin-V amidase (PVA), which hydrolyzes phenoxyacetamides, to explore the potential of using mAb-enzyme conjugates for the localizaton of chemotherapeutic drugs at tumor cells. The phenoxyacetamide derivatives of doxorubicin and melphalan were prepared, yielding the less toxic amides, doxorubicin-N-p-hydroxyphenoxyacetamide (DPO) and melphalan-N-p-hydroxyphenoxyacetamide (MelPO). These were hydrolyzed by PVA to doxorubicin and melphalan respectively.In vitro studies with the L6-positive lung carcinoma cell line, H2981, and the 1F5-positive B-cell lymphoma line, Daudi, showed that DPO was 80-fold less toxic to H2981 cells and 20-fold less toxic to Daudi cells than doxorubicin, and its toxicity was substantially increased when the H2981 cells were pretreated with L6-PVA or the Daudi cells were pretreated with 1F5-PVA. The cytotoxic effect was antigen-specific, since only the binding mAb-enzyme conjugate increased the cytotoxicity of the prodrug. MelPO was more than 1000-fold less toxic than melphalan to H2981 cells and more than 100-fold less toxic than melphalan to Daudi cells. Pretreatment with the mAb-PVA conjugates did not enhance the toxicity of MelPO in either cell line, because PVA hydrolyzes the phenoxyacetamide bond of MelPO too slowly to generate a toxic level of melphalan.
Similar content being viewed by others
References
Bagshawe KD, Springer CJ, Searle F, Antoniw P, Sharma SK, Melton RG, Sherwood RFA (1988) Cytotoxic agent can be generated selectively at cancer sites. Br J Cancer 58: 700–703
Baker WL (1984) A sensitive procedure for screening microorganisms for the presence of penicillin amidase. Aust J Biol Sci 37: 257–265
Baurain R, Masqeulier M, Deprez-De Campaneere D, Trouet A (1980) Amino acid and dipeptide derivatives of daunorubicin. 2. Cellular pharmacology and antitumor activity of L1210 leukemic cells in vitro and in vivo. J Med Chem 23: 1171–1174
Blakey DC, Wawrzynczak EJ, Wallace PM, Thorpe PE (1988) Antibody toxin conjugates: a perspective. Monoclonal antibody therapy. Prog Allergy 45: 50–90
Clark EA, Shu G, Ledbetter JA (1985) Role of the bp35 cell surface polypeptide in human B-cell activation. Proc Natl Acad Sci USA 82: 1766–1770
Di Marco A (1982) Anthracycline antibiotics. In: Cancer Medicine, 2nd edn. Lea and Febiger, Philadelphia, pp 872–906
Edwards DC (1983) Targeting potential of antibody conjugates. Pharmacol Ther 23: 147–177
Ghose T, Blair AH (1987) The design of cytotoxic-agent-antibody conjugates. CRC Crit Rev Therap Drug Carrier Syst 3: 263–359
Hellström KE, Hellström I, Brown JP (1982) Human tumor-associated antigens identified by monoclonal antibodies. In: Springer Seminars in Immunopathology series, Mechanisms of host resistance in cancer, vol. 5. Springer-Verlag, New York, pp 127–146
Hellström I, Horn D, Linsley P, Brown JP, Brankovan V, Hellström KE (1986) Monoclonal mouse antibodies raised against human lung carcinoma. Cancer Res 46: 3917–3923
Hellström KE, Hellström I, Goodman GE (1987) Antibodies for Drug Delivery. In: Controlled drug delivery, fundamentals and applications, 2nd edn. Dekker, New York, pp 623–653
Jain RK (1989) Delivery of novel therapeutic agents in tumors: physiological barriers and strategies. J Natl Cancer Inst 81: 570–576
Lambert JM, Senter PD, Yau-Young A, Blättler WA, Goldmacher VS (1985) Purified immunotoxins that are reactive with human lymphoid cells. J Biol Chem 260: 12035–12041
Levin Y, Sela BA (1979) Studies on amino acid and peptide derivatives of daunorubicin. FEBS Lett 98: 119–122
Lowe DA, Romancik G, Elander RP (1986) Enzymatic hydrolysis of penicillin-V to 6-aminopenicillanic acid byFusarium oxysporum. Biotechnol Lett 8: 151–156
Pastan I, Willingham MC, Fitzgerald DJP (1986) Immunotoxins. Cell 47: 641–648
Senter PD, Saulnier MG, Schreiber GJ, Hirschberg DL, Brown JP, Hellström I, Hellström KE (1988) Anti-tumor effect of antibody-alkaline phosphatase conjugates in combination with etoposide phosphate. Proc Natl Acad Sci USA 85: 4842–4846
Senter PD, Schreiber GJ, Hirschberg DL, Ashe SA, Hellström KE, Hellström I (1990) Enhancement of the in vitro and in vivo antitumor activities of phosphorylated mitomycin C and etoposide derivatives by monoclonal antibody-alkaline phosphatase conjugates. Cancer Res 49: 5789–5792
Smyth MJ, Pietersz GA, McKenzie IFC (1987) Selective enhancement of antitumor activity ofN-acetyl MEL upon conjugation with monoclonal antibodies. Cancer Res 47: 62–69
Takahashi T, Yamaguchi T, Kitamura K, Suzuyama H, Honda M, Yokota t, Kotanagi H, Takahashi M, Hashimoto Y (1988) Clinical application of monoclonal antibody-drug conjugates for immunotargeting chemotherapy of colorectal carcinoma. Cancer 61: 881–888
Thorpe PE (1985) Antibody carriers of cytotoxic agents in cancer therapy: a review. In: Monoclonal antibodies '84: biological and clinical applications. Editrice Kurtis, Milan, pp 475–506
Upeslacis J, Hinman L (1988) Chemical modifications of antibodies for cancer chemotherapy. Annu Rep Med Chem 23: 151–169
Vallera DA, Ash RC, Zanjani ED, LeBien TW, Beverly PCL, Neville DM, Youle RJ (1983) Anti-T-cell reagents for human bone marrow transplantation: ricin linked to three monoclonal antibodies. Science 222: 512–515
Vitetta ES, Fulton RJ, May RD, Till M, Uhr JW (1987) Redesigning nature's poisons to create anti-tumor reagents. Science 238: 1098–1104
Wheeler GP (1982) Alkylating Agents. In: Cancer Medicine, 2nd ed., Lea and Febiger, Philadelphia, pp 824–843
Yang HM, Reisfeld RA (1988) Doxorubicin conjugated with a monoclonal antibody directed to a human melanoma-associated proteoglycan suppresses the growth of established tumor xenografts in nude mice. Proc Natl Acad Sci USA 85: 1189–1193
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kerr, D.E., Senter, P.D., Burnett, W.V. et al. Antibody-penicillin-V-amidase conjugates kill antigen-positive tumor cells when combined with doxorubicin phenoxyacetamide. Cancer Immunol Immunother 31, 202–206 (1990). https://doi.org/10.1007/BF01789169
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01789169