Summary
Combination chemotherapy is widely employed in clinical oncology; however, there is no generally accepted model to evaluate individual tumour susceptibility to a given drug combination protocol. We therefore investigated the drug interaction of ifosfamide (4-hydroxyperoxy-ifosfamide) and ACNU in a recently developed in vitro model of paired sequential combination chemotherapy in primary and metastatic malignant brain tumours. A long-term standard [6,3-3H]-thymidine-incorporation assay, employing a liquid scintillation counting protocol, was selected to assess the drug sensitivity of human tumours. In vitro drug exposures were derived from correlating in vivo-(systemic and CNS) and in vitro-pharmacokinetic drug parameters. In combination experiments tumour cells were treated sequentially by two drugs in both sequences: drug exposures were calculated for 2 h with a 1-h drug-free interval in between. “Cut-off” concentrations (maximum in vitro exposure doses) were calculated as 1.74μM (for primary CNS tumours: 0.58μM) for ifosfamide and 5.4μM (for primary CNS tumours: 1.33μM) for ACNU. Dose/response relations were derived from isotope incorporation rates after cells had grown for approximately five population doubling times. Combination isoboles were plotted after drug doses had been transformed into “equieffective doses”, enabling comparison of drug combination effects. In all three glioblastomas (with CNS exposure dose) an additive or supra-additive effect could be observed in either sequence (in one tumour a biphasic additive isobole was found for both sequences). Out of three bronchial carcinomas (small-cell type, brain metastases) in two nonidentical sequences a supra-additive effect was observed in two tumours, with antagonistic effects in the third tumour. In all three malignant melanomas and in one renal carcinoma antagonistic effects were observed, whereas in a second renal carcinoma supra-additive effects were demonstrated for both sequences. We conclude that drug combination chemotherapy effects at the cellular level may be extremely heterogeneous.
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Abbreviations
- ACNU:
-
3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-1-(2-chloroethyl)-1-nitrosourea
References
Ali-Osman F, Maurer HR (1983) In vitro cytostatic drug testing in the human stem cell assay: a modified method for the determination of the sensitivity index. Turn Diagn Ther 4:1–5
Ali-Osman F, Giblin J, Dougherty DV, Rosenblum ML (1987) Application of in vivo and in vitro pharmacokinetics for physiologically relevant drug exposure in a human tumor clonogenic cell assay. Cancer Res 47:3718–3724
Arndt CAS, Balis FM, McCully CL, Colvin OM, Poplack DG (1988) Cerebrospinal fluid penetration of active metabolites of cyclophosphamide and ifosfamide in rhesus monkeys. Cancer Res 48:2113–2115
Bogdahn U (1983) Chemosensitivity of malignant human brain tumors: preliminary results. J Neurooncol 1:149–166
Bogdahn U, Zapf J, Weber H, Dünisch G, Löbering HG, Martin R, Mertens HG (1987a) Vidarabin-monophosphate, BCNU, VM 26 — an in vitro comparative study of active agents in the treatment of malignant human brain tumours. Br J Cancer 55:153–158
Bogdahn U, Zapf J, Weber H, Dünisch G (1987b) An in vitro comparative study of BCNU, VM-26, 5-FU and Ara-A5′P in treatment of primary and secondary CNS-neoplasms; potential clinical relevance and clinical correlations. In: Chatel M, Pecker J (eds) Brain oncology. Nijhoff, Dordrecht, pp 355–361
Bogdahn U, Lutz M, Zapf J, Weber H (1988) Individualisierte Hirntumortherapie — Kritische Aspekte. In: Bamberg M, Sack H (eds) Therapie primärer Hirntumoren. Zuckschwerdt, München, pp 27–31
Brade W (ed) (1987) Ifosfamide in tumor therapy. Karger, Basel
Chen G, Ensor CR (1953) The combined anticonvulsant activity and toxicity of dilantin and N-methyl-5-phenylsuccinimide. J Lab Clin Med 41:78–83
Chou TC, Talalay P (1981) Generalized equations for the analysis of inhibitions of Michaelis-Menten and higher-order kinetic systems with two or more mutually exclusive and nonexclusive inhibitors. Eur J Biochem 115:207–216
Chou TC, Talalay P (1983) Analysis of combined drug effects: a new look at a very old problem. Trends Pharmacol Sci 4:450–454
Deen DF, Williams ME (1979) Isobologram analysis of X-ray — BCNU interactions in vitro. Radiat Res 79:483–491
Elion GB, Singer S, Hitchings GH (1954) Antagonists of nucleic acid derivatives. VIII. Synergism in combinations of biochemically related antimetabolites. J Biol Chem 208:477–488
Finney DJ (1971) Probit analysis, 3rd edn. Cambridge University Press, Cambridge, pp 146–153
Hamburger AW, Salmon SE (1977) Primary bioassay of human tumor stem cells. Science 197:461–463
Hildebrand J (1986) Radiotherapy and chemotherapy of malignant brain gliomas. Drugs Exp Clin Res 12:167–175
Hill AV (1910) The possible effects of the aggregation of molecules of hemoglobin on its dissociation curve. J Physiol (Lond) 40:47
Hori T, Muraoka K, Saito Y, Sasahara K, Ianagaki H, Inoue Y, Adachi S, Anno Y (1987) Influence of modes of ACNU administration on tissue and blood drug concentration in malignant brain tumours. J Neurosurg 66:372–378
Jellinger K (ed) (1987) Therapy of malignant brain tumors. Springer, Berlin Heidelberg New York, pp 286–293; 358–369
Kern DH, Bertelsen CA, Mann BD et al. (1983) Clinical application of the clonogenic assay. Ann Clin Lab Sci 13:10–15
Kern DH, Morgan CR, Hildebrand-Zanki SU (1988) In vitro pharmacodynamics of 1-β-d-arabinofuranosylcytosine: synergy of antitumor activity withcis-diamminedichloroplatinum (II). Cancer Res 48:117–121
Kimmel DW, Shapiro JR, Shapiro WR (1987) In vitro drug sensitivity testing in human gliomas. J Neurosurg 66:161–171
Kitahara M, Katakura R, Mori T, Suzuki J, Sasaki T (1986) Combined effect of ACNU and 5-FU on rat glioma cells in spheroids and monolayer cultures. Int J Cancer 38:215–222
Kitahara M, Katakura R, Suzuki J, Sasaki T (1987) Experimental combination chemotherapy of ACNU and 5-FU against cultured glioma model (spheroid) and subcutaneous rat glioma. Int J Cancer 40:557–563
Kohno N, Ohnuma T, Biller HF, Holland JF (1988) Effects of cisplatin plus fluorouracil vs cisplatin plus cytarabine on head and neck squamous multicellular tumor spheroids. Arch Otolaryngol Head Neck Surg 114:157–161
Loewe S, Muischnek H (1926) Über Kombinationswirkungen: I. Mitteilung: Hilfsmittel der Fragestellung. Naunyn-Schmiedebergs Arch Exp Pathol Pharmakol 114:313–326
Moon TE (1980) Quantitative and statistical analysis of the association between in vitro and in vivo studies. In: Salmon SE (ed) Cloning of human tumour stem cells. Liss, New York, pp 209–221
Nakamura K, Asami M, Kawada K, Sasahara K (1977) Quantitative determination of ACNU, a new water-soluble anti tumor nitrosourea, in biological fluids and tissues of patients by HPLC. Annu Rep Sankyo Res Lab 29:66–74
Nowell PC (1977) The clonal evolution of tumour cell populations. Science 194:23–28
Rosenblum ML, Gerosa MA, Wilson CB, Barger GR, Pertuiset BF, Tribolet N de, Doughterty DV (1983) Stem cell studies of human malignant brain tumors: part 1. Development of the stem cell assay and its potential. J Neurosurg 58:170–176
Salmon SE, Hamburger AW, Soehnlen B, Durie BG, Alberts DS, Moon TE (1978) Quantitation of differential sensitivity of human tumor stem cells to anticancer drugs. N Engl J Med 298:1321–1327
Shapiro WR (1986) Therapy of adult malignant brain tumors: what have the clinical trials taught us? Semin Oncol 13:38–45
Steel GG, Peckham MJ (1979) Exploitable mechanisms in combined radiotherapy — chemotherapy: the concept of additivity. Int J Radiat Oncol Biol Phys 5:85–91
Thomas DGT, Darling JL, Paul EA (1985) Assay of anticancer drugs in tissue culture: relationship of relapse free interval and in vitro chemosensitivity in patients with malignant glioma. Br J Cancer 51:525–532
Valeriote F, Lin H (1975) Synergistic interaction of anticancer agents: a cellular perspective. Cancer Chemother Rep 59:895–900
Von Hoff DD, Clark GM, Stogdill BJ, Sarosdy MF, Mattox DE, Page DP, Cruz AB, Sandbach JF (1983) Prospective clinical trial of a human tumor cloning system. Cancer Res 43:1926–1931
Walker MD, Green SB, Byar DP et al. (1980) Randomized comparisons of radiotherapy and nitrosureas for the treatment of malignant glioma after surgery. N Engl J Med 303:1323–1329
Webb JL (1963) Enzymes and metabolic inhibitors, vol 1. Academic Press, New York, pp 66–79; 487–512
Wong JTF (1975) Kinetics of enzyme mechanisms. Academic Press, London, pp 28–33; 134–137; 151–154
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Bogdahn, U., Drenkard, D., Lutz, M. et al. In vitro studies on drug interaction of ifosfamide and ACNU in primary and metastatic human brain tumours. J Cancer Res Clin Oncol 117 (Suppl 4), S157–S163 (1991). https://doi.org/10.1007/BF01613223
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DOI: https://doi.org/10.1007/BF01613223