Carbon-Fluorine Compounds Chemistry, Biochemistry & Biological Activities A Ciba Foundation Symposium 1972 Elsevier - Excerpta Medica North-Holland Associated Scientific Publishers . Amsterdam - London . New York Carbon-Fluorine Compounds Chemistry, Biochemistry & Biological Activities The Ciba Foundation for the promotion of international cooperation in medical and chemical research is a scientific and educational charity established by CIBA Limited - now CIBA-GEIGYLimited - of Basle. The Foundation operates independently in London under English trust law. Ciba Foundation Symposia are published in collaboration with Associated Scientific Publishers (Elsevier Scientific Publishing Company, Excerpta Medica, North-Holland Publishing Company) in Amsterdam. Associated Scientific Publishers, P.O. Box 3489, Amsterdam Carbon-Fluorine Compounds Chemistry, Biochemistry & Biological Activities A Ciba Foundation Symposium 1972 Elsevier - Excerpta Medica North-Holland Associated Scientific Publishers . Amsterdam - London . New York 0 Copyright 1972 Ciba Foundation All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying and recording, or by any information storage and retrieval system, without permission in writing from the publishers. ISBN Excerpta Media 90 219 4002 7 ISBN American Elsevier 0-444-10373-2 Library of Congress Catalog Card Number 72-76005 Published in 1972 by Associated Scientific Publishers, P.O. Box 3489, Amsterdam, and 52 Vanderbilt Avenue, New York, N.Y. 10017. Suggested series entry for library catalogues: Ciba Foundation Symposia. Printed in The Netherlands by Mouton & Co., The Hague Contents SIR RUDOLPH PETERS Introduction 1 B. c. SAUNDERS Chemical characteristics of the carbon-fluorine bond 9 Discussion 27 A. G. SHARPE The physical properties of the carbon-fluorine bond 33 Discussion 49 SIR RUDOLPH PETERS Some metabolic aspects of fluoroacetate especially related to fluorocitrate 55 Discussion 70 E. M. GAL Effects of fluoro compounds on metabolic control in brain mitochondria 77 Discussion 90 J. E. G. BARNETT Fluorine as a substituent of oxygen in biological systems: examples in mammalian membrane transport and glycosidase action 95 Discussion 1 10 General discussion I 117 c. HEIDELBERGER The nucleotides of fluorinated pyrimidines and their biological activities 125 Discussion 137 L. FOWDEN Fluoroamino acids and protein synthesis 141 Discussion 155 vi Contents General discussion I1 161 P. w. KENT Synthesis and reactivity of fluorocarbohydrates 169 Discussion 209 N. F. TAYLOR The metabolism and enzymology of fluorocarbohydrates and related compounds 21 5 Discussion 235 R. A. DWEK Nuclear magnetic resonance studies of macromolecules with fluorine nuclei as probes 239 Discussion 271 A. WETTSTEIN Chemistry of fluorosteroids and their hormonal properties 281 Discussion 298 P. BUFFA, V. GUARRIERA-BOBYLEVA and I. PASQUALI-RONCHETTI Biochemical effects of fluoroacetate poisoning in rat liver 303 Discussion 327 General discussion 111 331 P. GOLDMAN The use of microorganisms in the study of fluorinated com- pounds 335 Discussion 349 J. L. SHUPE Clinical and pathological effects of fluoride toxicity in animals 357 Discussion 383 SIR RUDOLPH PETERS Chairman’s concluding remarks 389 Index of Contributors 391 Subject Index 392 Contributors Sy'mposium on Carbon-Fluorine Compounds: Chemistry, Biochemistry and Biological Activities held at the Ciba Foundation, London, 13th-15th September 1971 Chairman: SIR RUDOLPH PETERS Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 IQW w. D. ARMSTRONG Department of Biochemistry, University of Minnesota, 227 Millard Hall, Minneapolis, Minnesota 55455, USA J. E. G. BARNETT Department of Physiology and Biochemistry, The University of Southampton, Highfield, Southampton SO9 5NH E. D. BERGMANN Department of Organic Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel P. BUFFA Istituto di Patologia Generale, Universith degli Studi di Modena, Via Campi 287, 41 100 Modena, Italy R. B. CAIN Biological Laboratory, The University of Kent at Canterbury, Beverley Farm, Canterbury, Kent R. A. DWEK Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU A. B. FOSTER Department of Chemistry, Chester Beatty Research Institute, Royal Cancer Hospital, Fulham Road, London SW3 L. FOWDEN Department of Botany and Microbiology, University College London, Gower Street, London WC1 E. M. GAL Division of Neurobiochemistry, Department of Psychiatry, The University of Iowa, 500 Newton Road, Iowa City, Iowa 52240, USA P. GOLDMAN Department of Health, Education and Welfare, Public Health Service, Building 10, Room 8D17, National Institutes of Health, Bethesda, Maryland 20014, USA L. D. HALL Department of Biochemistry, The University of Sheffield, Sheffield S 10 2TN ... Vlll Contributors D. B. HARPER Department of Biological Sciences, University of Kent at Canterbury, Canterbury, Kent c. HEIDELBERGER McArdle Laboratory for Cancer Research, Medical Center, University of Wisconsin, Madison, Wisconsin 53706, USA P. w. KENT Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU E. KUN Department of Pharmacology, University of California School of Medicine, San Francisco, California 94122, USA G. w. MILLER Huxley College of Environmental Studies, West Washington State College, Bellingham, Washington 98225, USA B. c. SAUNDERS Department of Organic and Inorganic Chemistry, University of Cambridge, Cambridge CB2 1QW A. G. SHARPE Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB5 8BL M. SHORTHOUSE Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW J. L. SHUPE Department of Veterinary Science, Utah State University, Logan, Utah 84321, USA D. R. TAVES Department of Pharmacology and Toxicology, The University of Rochester School of Medicine and Dentistry, 260 Crittenden Boulevard, Rochester, New York 14620, USA N. F. TAYLOR Biochemistry Group, School of Biological Sciences, The University of Bath, Claverton Down, Bath BA2 7AY P. F. v. WARD Biochemistry Department, Agricultural Research Council Institute of Animal Physiology, Babraham, Cambridge A. WETTSTEIN CIBA-GEIGY Limited, CH-4002 Basle, Switzerland Editors: KATHERINE ELLIOTT and JOAN BIRCH (The editors gratefully acknowledge the invaluable assistance of Dr P. W. Kent in the preparation of this book) Introduction SIR RUDOLPH PETERS Department of Biochemistry, University of Cambridge Some two years ago interest in the carbon-fluorine bond seemed to be increasing and Dr P. W. Kent and I thought that a meeting with an interdisciplinary approach to these problems might bring the present studies to a focus and provide a spring-board for fresh attacks on these problems. Interest in the monofluorocarbon bond goes back to the curiosity of an organic chemist in Belgium in the last century. It was later picked up in Germany FIG.1. Dichapetalum cymosum (gifblaar). (Photograph kindly presented by Dr Dyer.) 2 Sir Rudolph Peters FIG.2. Seed of Dichapetalum toxicarium. The kernels contain fluoro-oleic acid and some other acid (from Peters et al. 1960). as a lead to possible insecticides; and compounds with the carbon-fluorine bond became known in the United Kingdom early in the 1940’s when it was imperative to look at the toxicity of these compounds, to be forewarned in case they were used against the allies in World War 11. The main work was done in Cambridge by Dr Saunders and his group, and we shall hear from him about this (pp. 9-27). Until 1943 fluoroacetate was just an interesting chemical com- pound, possibly toxic, with a very stable carbon-fluorine bond, resistant even to hot concentrated sulphuric acid. But then the interest was suddenly widened by the discovery by J. S. C. Marais (1943) that fluoroacetate was the toxic principle of a well-known South African plant, Dichapetalum cymosum or gifblaar (Fig. 1). This plant grows in the Transvaal region and causes considerable losses of cattle, especially in the spring, when they eat the fresh shoots coming up among the rocks. These plants are spread by rhizomes and are very difficult to eradicate. The toxicity varies from a maximum in the South African spring to a minimum in their autumn. Many other varieties of Dichapetalum are now known in Africa-most of them less toxic. An interesting one is D. toxicarium from W. Introduction 3 FIG.3. Acacia georginae (gidyea) trees growing in the vicinity of Alice Springs. (Photograph kindly presented by Mr L. R. Murray.) Africa; the plant contains traces of fluoroacetate but the main toxicity is in the seeds (Fig. 2) and is due to fluoro-oleic acid (18 carbon atoms) and small amounts of shorter-chained fluoro acids with even numbers of carbon atoms. In these compounds the fluoride ion is in the o-position. Nearly 70 years ago, Renner (1904) described how the ‘local’ doctors used the seeds of this plant to produce loss of motor activity and sensation, and presumably death. Up to 1961 this was the extent of our plant knowledge, though it led to much interesting research and to the extensive use of fluoroacetate (called Compound 1080) to poison rodents and rabbits, especially in Australia and New Zealand. It was also found to be a good systemic insecticide for aphids in concentrations which did not kill the plants. In 1961, while in Australia, I was invited to visit Drs J. H. Whittem and J. D. McConnell and Mr L. R. Murray at Alice