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Studies of Radioactive Cisplatin ( Pt) for Tumour Imaging and Therapy

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Studies of Radioactive Cisplatin ( Pt) for Tumour Imaging and Therapy SE0000130 Studies of radioactive cisplatin ( Pt) for tumour imaging and therapy Johan Areberg Department of Radiation Physics, Malmo Lund University LUND UNIVERSITY Malmo 2000 31-14 Studies of radioactive cisplatin (191Pt) for tumour imaging and therapy Johan Areberg LUND UNIVERSITY Malmo 2000 Department of Radiation Physics Lund University Malmo University Hospital SE-205 02 Malmo SWEDEN Doctoral Dissertation Lund University Institute of Radiology and Physiology Department of Radiation Physics, Malmo Malmo University Hospital SE-205 02 Malmo, SWEDEN Copyright© 2000 Johan Areberg (pp 1-60) ISBN 91-628-3989-6 Printed in Sweden by Elanders Skogs Grafiska, Malmo 2000 To Cecilia and Adam ABSTRACT A radioactive variant of the cytostatic agent «x-dichlorodiammineplatinvim(ir), cisplatin, was synthesised from 191PtCU. The 191Pt-cisplatin was found to be a sterile product of high radionuclide, radiochemical and chemical purity. The pharmacokinetics of platinum in tumour tissue and organs at risk of fourteen patients undergoing treatment with cisplatin were studied by exchanging a small fraction of the prescribed amount of cisplatin with 191Pt-cisplatin. The uptake and retention of platinum were investigated by gamma camera measurements up to ten days after infusion of 191Pt-cisplatin. Highest concentration of platinum was found in the liver, on average 5.7 ± 0.5 ug/g normalised to a given amount of 180 mg cisplatin. Corresponding value for the kidneys was 1.9 + 0.3 ug/g. Uptake of platinum in tumours was visualised in five patients with an average maximum concentration of 4.9 ±1.0 ug/g normalised to a given amount of 180 mg cisplatin. The data from the pharmacokinetic study was used together with data from the literature to estimate the absorbed dose and effective dose to patients receiving radioactive cisplatin. The effective doses were calculated to be 0.10 ± 0.02 mSv/MBq, 191 0.17 ± 0.04 mSv/MBq and 0.23± 0.05 mSv/MBq for Pt-, i53mpt.; ^d i95mpt.ciSpiatin respectively. The combined effect of the radio- and chemotoxicity from 191Pt-cisplatin was investigated both in vitro and in vivo. A cervical cancer cell line was incubated with cisplatin or 191Pt-cisplatin with various concentrations and specific activities. It was shown that the surviving fraction was smaller for cells treated with 191Pt-cisplatin than for cells treated with the same concentration of non-radioactive cisplatin. The surviving fraction decreased with increasing specific activity. Isobologram technique showed that the radio- and chemotoxicity interacted in a supra-additive (synergistic) manner. In an in vivo model, nude mice with xenografted tumours were given physiological saline (controls), cisplatin or 191Pt-cisplatin. The growth was significantly more retarded for the tumours on the mice given 191Pt-cisplatin compared with the tumours on the other mice. No significant change in general toxic effect, manifested as weight change or mortality, could be detected between the groups of mice given cisplatin and 191Pt- cisplatin respectively. 191Pt-cisplatin was shown to be a radiopharmaceutical suitable for both tumour imaging and therapy. There is potential for an increased use of radioactive cisplatin in the future. LIST OF PAPERS This thesis is based on the following papers, which will be referred to in the text by their Roman numerals: I Gamma camera imaging of platinum in tumours and tissues of patients after administration of *™Pt-cisplatin. Areberg J, Bjorkman S, Einarsson L, Frankenberg B, Lundqvist H, Mattsson S, Norrgren K, Scheike O, Wallin R Ada Oncol 38:221-228,1999 II Absorbed doses to patients from l^Pt-, 193mpt_ an<j 195mpt.ciSpjatm Areberg J, Norrgren K, Mattsson S ApplRadiat hot 5l(5):581-586,1999 III In vitro toxicity of 191Pt-labelled cisplatin to a human cervical carcinoma cell line (ME-180). Areberg J, Johnsson A, Wennerberg J Accepted for publication in Int] Radiat Oncol Biol Pbys TV The antitumour effect of radioactive cisplatin (191Pt) on nude mice. Areberg J, Wennerberg J, Johnsson A, Norrgren K, Mattsson S Submitted to IntJRadzat Oncol Biol Phys 191 V Radiation dosimetry of Pt-, i»3mpt_ atl(j wsmpt.cigpiatin fa tumour- bearing mice. Areberg J, Hindorf C, Liu X, Norrgren K, Ljungberg M, Mattsson S, Strand S-E Internal report, MA-RADFYS 2000:01 Preliminary reports were given at: • The Annual Meeting of the Swedish Medical Society, Stockholm, Sweden, 29 November - 1 December, 1995 • Radioactive Isotopes in Clinical Medicine and Research, Badgastein, Austria, 9 — 12 January, 1996 • First Easter School in Radiopharmaceutics, Liverpool, United Kingdom, 24 - 29 March, 1996 • Second International Conference on Isotopes (2ICI), Sydney, Australia, 12-16 October, 1997 • The Annual Meeting of the Swedish Medical Society, Goteborg, Sweden, 24 - 26 November, 1998 ABBREVIATIONS AAS atomic absorption spectrometry Adv adsorptive voltammetry CT computed tomography DNA deoxyribonucleic acid EC electron capture Gy gray HPGe high-purity germanium HPLC high-performance liquid chromatography IC50 50% inhibition concentration ICP-AES inductively coupled plasma atomic emission spectrometry ICP-MS inductively coupled plasma mass spectrometry ICRP International Commission on Radiological Protection IT isomeric transition MIRD Medical Internal Radiation Dose Committee (USA) MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylformazan bromide PET positron emission tomography SD standard deviation SEM standard error of the mean SGD specific growth delay Sv sievert TD time for a tumour to double its volume TLC thin-layer chromatography XRF X-ray fluorescence CONTENTS INTRODUCTION 11 AIMS OF THE STUDY 12 BACKGROUND 13 An overview of the use of radioactive cytostatic agents 13 Cisplatin in clinical use 13 Chemistry of cisplatin and formation of DNA-adducts 14 Toxicity of cisplatin 16 Other platinum analogues for chemotherapy 16 Methods for measuring platinum in biological material 16 Pharmacokinetics of platinum after cisplatin treatment 17 Platinum isotopes and their production 18 Gamma camera imaging of radioactive platinum and other radionuclides 20 Combination of cisplatin and radiation for therapeutic purpose 21 Calculation of absorbed and effective doses 24 MATERIAL AND METHODS 26 Production of 19IPt 26 Synthesis of 191Pt-cisplatin 26 Pharmacokinetic study of platinum in patients 29 In vitro toxicity of 19IPt-cisplatin 29 In vivo toxicity of 191Pt-cisplatin 30 Absorbed dose and effective dose calculations 30 RESULTS AND DISCUSSION 33 Synthesis of 191Pt-cisplatin 33 Gamma camera imaging of 191Pt 34 Pharmacokinetics of platinum after treatment with cisplatin 34 In vitro toxicity of 191Pt-cisplatin 37 General toxicity and effect on tumours of 191Pt-cisplatin in vivo 38 Absorbed dose and effective dose 41 General comments 42 FUTURE ASPECTS 44 Chemoembolization 44 Intratumoural administration 45 Electrochemotherapy 45 CONCLUSIONS 46 ACKNOWLEDGEMENTS 47 REFERENCES 49 NEXT PAGE(S) left BLANK INTRODUCTION It has been estimated that in Sweden about 40% of the population will develop cancer at some time in their life. Despite recent developments and improvements in the treatment, 23% of all deaths in Sweden are related to cancer (Swedish National Board of Health and Welfare, 1998). The dominating treatment modalities for cancer are surgery, radiotherapy, chemotherapy and hormone therapy. Hyperthermia and photodynamic therapy give just a marginal contribution and new methods like immunotherapy and genetherapy are still at a stage of research and very limited clinical trials. In chemotherapy, cytostatic agents that inhibit the proliferation of tumour cells are used. The optimal cytostatic agent acts on the tumour cell, but is inactive in normal cells. Unfortunately no such drug exists, why side-effects of chemotherapy are common. It is therefore of great importance to develop methods that can give information about how effective a treatment with a cytostatic agent will be and/or how the organs at risk will react. The chemotherapy regimens given today are very general and the sensitivity of the tumour cells to the drug is normally not taken into consideration in the planning of the treatment. A more individualised treatment planning like that in external radiotherapy would be one way to improve the therapeutic gain. The different treatment modalities are often combined to optimise the outcome of the treatment. The combination of external radiotherapy and chemotherapy has markedly improved the therapeutic results for those tumours which are both chemosensrtive and radiosensitive. By using cytostatic agents labelled with radioactive compounds, chemo- and radiotherapy can be combined, but such agents have so far not become an alternative for therapy. The benefit of using radioactive cytostatic agents could be twofold: 1. The radiation emitted from the decay of the radionuclide can be detected, e.g. by a gamma camera, whereby the uptake and distribution of the drug in the patient can be monitored after the administration. 2. The effect of the radiation, i.e. the radiotoxicity, interacts with the chemotoxic effect of the drug and optimally an enhanced effect of the combined treatment is reached. If the enhancement is more pronounced for tumour cells than for normal cells, there is potential for a therapeutic gain. 11 AIMS OF THE STUDY The main purpose of this thesis was to explore the usefulness of radioactive cisplatin for imaging, for pharmacokinetic studies and for tumour therapy. The specific aims were to • develop a method to synthesise sterile radioactive cisplatin, 191Pt-cisplatin,
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