Handbuch der experimentellen Pharmakologie

Handbook of Experimental Pharmacology

O.Eichler Heidelberg

Heffter-Heubner New Series

Herausgegeben von / Editorial Board

A.Farah H. Herken A. D. Welch Rensselaer, NY Berlin New Brunswick, N J

Beirat/ Advisory Board

G. Acheson· E. J. Ariens . Z. M. Bacq . F. v. Briicke . P. Calabresi . S. Ebashi

E. G. Erdos . V. Erspamer . U. S. von Euler . W. Feldberg . R. Furchgott

A. Goldstein· G. B. Koelle' O. Krayer . H. Raskova . M. Rocha e Silva

F. Sakai· P. Waser . W. Wilbrandt

Vol.XX/2

Pharmacology of Fluorides

Part 2

Contributors

P. Borrevang . F. S. Caruso' P. S. Chen jr .. V. Di Stefano

A. M. G. Macdonald' R. E. Marquis' J. Matthews

E. A. Maynard' F. A. Smith, A. Wiseman

Editor

Frank A. Smith

With 59 Figures

Springer-Verlag Berlin· Heidelberg' New York 1970

ISBN-13: 978-3-642-99975-8 e-ISBN-13: 978-3-642-99973-4 DOl: 10.1007/978-3-642-99973-4

This work is subject to copyright. All rights are reserved, whether the whole or part of the material Is concerned specifically those of translation, reprinting, re·use of illustrations, broadcasting, reproduction by photocopying machine or similar means, and storage in data banks. Under § 54 of the German Copyright Law where copies are made for other than private use, a fee is payabie to the publisher, the amount of the fee to be determined by agreement with the publisher. © by Springer·Verlag Berlin· Heidelberg 1970. Library of Congress Catalog Card Number AGR 25·699. Softcover reprint of the hardcover 1st edition 1970 The use of general descriptive names, trade names, trade marks, ete. in this publication, even if the former are not especially identified, is not to be taken as a sign that such names, as understood by the Trade Marks and Mer­chandise Marks Act, may accordingly be used freely by anyone. Title No. 5731

Preface

This volume is the completion of work initially planned several years ago as a compilation of selected aspects of the biological effects of fluorides. The first portion appeared in Volume XXII (1966) of this Handbuch, under the title "Pharmacology of Fluorides, Part 1."

Inasmuch as the present volume is an integral part of the original project, justification for offering it to the scientific community remains the same as that originally set forth. This may be recapitulated as follows. The tremendous increase in the annual production of fluorides over the amounts produced thirty years ago, together with the increased diversity of their uses has correspondingly increased the hazard associated with these materials. That is to say, the possibilities of encountering their characteristic toxic effects under the conditions of their use has increased because the variety, amounts and ways of usage have increased. Their inherent toxicity, of course has not changed, but the increased hazard has led to a vast increase in the amount and scope of research reported. It is the purpose of these volumes to review selected aspects of this literature.

Rochester, N.Y., February 1970 FRANK A. SMITH

Acknowledgements

Even the casual reader of this volume will soon realize that contributing authors, as well as those contributing to its predecessor, Volume XX/I, have made good use of tables of data, figures and pertinent quotations from previously published work. The use of this material has added much to the effectiveness with which the present writers have been able to develope their reviews. In each instance the source of the material used has been acknowledged by indicating the author and citing the appropriate reference in the bibliography. Before falling back on that somewhat impersonal procedure however, the editor takes this opportunity to extend his warm thanks to each of those authors of previously published work, to journal editors, to professional societies, and to publishers who so kindly permitted us to reproduce copyrighted matter in Volumes XXII and/or XX/2 of this Handbuch. The list of journals, publishers and professional societies includes Academic Press, Inc., Acta Chemica Scandi­navica, Acta Obstetricia et Gynecologica Scandinavica, Acta Pharmacologica et Toxicologica, Air Pollution Control Association, American Chemical Society, American Association for the Advancement of Science, American Dental Association, American Journal of Botany, Ameri­can Journal of Physiology, American Journal of Public Health, American Journal of Veteri­nary Research, American Medical Association, American Physiological Society, American Society of Animal Science, American Society of Plant Physiologists, American Veterinary Medical Association, American Veterinary Publications, Inc., Analytical Chemistry, Anest­hesia, Anesthesia and Analgesia, Anesthesiology, Annals of Human Genetics, Annals of the New York Academy of Sciences, Archives of Biochemistry and Biophysics, Archives of Oral Biology, Berliner und Miinchener Tierarztliche Wochenschrift, Biochemische Zeitschrift, Biochimical et Biophysica Acta, Biochemical Journal, Biochemical Pharmacology, Boyce Thompson Institute for Plant Research, British Dental Association, British Dental Journal, British Journal of Anesthesia, British Journal of Experimental Pathology, Bulletin of the Research Council of Israel, Burgess Publishing Co., Cambridge University Press, Cancer Research, Charles C. Thomas, Publisher, Columbia University Press, Comptes rendus de l'academie des sciences, Contributions of the Boyce Thompson Institute for Plant Research, F.A. Davis Co., Deutsche Tierarztliche Wochenschrift, Elsevier Publishing Co., Endeavour, Endocrinology, Entomological Society of America, European Journal of Pharmacology, The Gerrard Press, Illinois State Geological Survey (Section of Fluorine Chemistry), Industrial and Engineering Chemistry, International Anesthetic Research Society, International Journal of Air and Water Pollution, Interscience Publishers, Inc., John Sherratt and Son, Johns Hopkins Press, Journal of Agricultural and Food Chemistry, Journal of the Air Pollution Control Association, Journal of the American Dental Association, Journal of the American Medical Association, Journal of Animal Science, Journal of Biochemistry (Tokyo), Journal of Compa­rative Pathology and Therapeutics, Journal of Dairy Science, Journal of Dental Research, Journal of Economic Entomology, Journal of Nutrition, Journal of Pathology and Bacterio­logy, Journal of Pharmacology and Experimental Therapeutics, Journal of Pharmaceutical Sciences, Journal of Physiology, H.K. Lewis, and Co., Ltd., J.B. Lippincott Co., Metabolism, Clinical and Experimental, Michigan State Medical Society, Nature, New York Academy of Sciences, Paul Parey-Verlag, Pergamon Press, Ltd., Plant Physiology, Proceedings of the American Society for Horticultural Science, Proceedings of the Royal Society (London), Proceedings of the Society for Experimental Biology and Medicine, Public Health Reports, The Royal Society, The Royal Veterinary College, M. and H. Schaper-Verlag, Science, Society of Toxicology, Springer-Verlag, Toxicology and Applied Pharmacology, University of Chicago Press, U.S. Department of Health, Education and Welfare, Williams and Wilkins Company, Wistar Institute of Anatomy and Biology, World Health Organization.

Contents

Chapter 1: Methods of Analysis for Fluorine. A.M.G. MAODONALD

I. Introduction . . . . . . . .

IT. Methods of Sample Preparation A. Organic Compounds

1. The Oxygen-Flask Method .. . . . Procedure for Oxygen-Flask Combustion

2. Fusion with Alkali Metals. . . . . • . Procedure for Alkali Metal Fusion . . . . . . .

3. Reduction with Alkali Metal in Organic Solvents . 4. Wickbold Oxyhydrogen Flame Method

B. Inorganic Samples . . . . . . C. Agricultural and Food Samples D. Miscellaneous Samples

lIT. Methods of Separation . . . . . . A. Separation of Fluorosilicic Acid by Distillation _ B. Separation by Pyrohydrolysis C. Separation by Diffusion. . . . . . . . . . . D. Separation by Ion-exchange . . . . . . . . . E. Chromatographic Separation on Paper and Thin Layers . F. Other Methods of Separation .....

IV. The Detection of Fluoride . . . _ . . . . Procedure for the 8-Hydroxyquinoline Test .

V. The Determination of Fluoride . . . . . A. Gravimetric Methods •.......•....

1. Precipitation as Lead Chlorofluoride . . . . . a) Recommended Lead Chlorofluoride Procedure

a) Reagents. . . . . . _ . . p) Procedure . . . . . . . .

2. Precipitation as Calcium Fluoride B. Titrimetric Methods . . . . . .

1. Titration with Thorium Solution a) Interferences . . . . . . . b) Direct Titration Procedure . c) Back-Titration Procedure. . .

2. Titration with Aluminum Solution . . . 3. Acid-Base Titration of Hydrofluoric Acid 4. Titration with Rare Earth Solutions . . 5. Other Titrimetric Methods. . . . . . .

a) Titration with Calcium Solutions. . . . . . . . b) Titrations based on Lead Halofluoride Formation c) Titration with lron(IIT) Solutions .. d) Titrations based on Fluorosilicic Acid.

C. Colorimetric Methods . . . . • • _ . . . 1. The Alizarin Fluorine Blue Methods . . . . . . . . . . . .

a) Recommended Lanthanum-Alizarin Fluorine Blue Procedure a) Reagents .........•............ ft) Calibration Graph . • . • • • • • • • • • • . . • . •

1

1

1 1 2 3 4 4 4 5 5 6 6

7 7 8

10 11 12 12

13 13

14 15 15 16 16 16 16 16 16 18 18 18 18 19 20 20 20 21 21 21 21 21 23 23 24

vm Contents

2. The Bleaching of Zirconium-Dye Complexes • . . . • . • . • 24 a) Recommended Zirconium-Eriochrome Cyanine R Procedure. 24

a) Reagents. . . _ . . . • • • . . . • 24 P) Calibration Graph . . • . . . . . • . 24

3. The Bleaching of Thorium-Dye Complexes. . 25 4. The Bleaching of Aluminum-Dye Complexes. 26 5. Other Colorimetric Methods . 26

a) Titanium(IV) Complexes . 26 b) Iron(III) Complexes . . . 27 c) Chloranilic Acid Methods . 27 d) Molybdenum Blue Methods 27

D. Polarographic Methods . . . 28 E. Direct Electrometric Methods 28 F. Radiometric Methods. . . . 31 G. Catalytic Methods . . . . . 31 H. Determination of Elemental Fluorine 32

VI. The Determination of Common Elements in Organic Fluorine Compounds. 32 A. Carbon, Hydrogen and Nitrogen . . . . 32 B. Other Halogens, Sulfur and Phosphorus . 33

References • . . . . . . . . . . . . . . . . . 34

Chapter 2: Effect of Inorganic Fluoride on Enzymes. A. WISEMAN. With 34 Figures 48

I. General Introduction. . . . . . . . . . . . . . . . . . . . . . . . . 48

II. Enzymes, Inhibited by Fluoride, Requiring Metal Ions, and Whose Inhibition is Enhanced by Phosphate 51 A. Enolase . . . . . . . • . . . • . . 51 B. Phosphoglucomutase . . . . . . . . 56 C. Lecithinase (from Olo8tridium welekii) . 58 D. Succinic Dehydrogenase. . . . . . . 59

III. Enzymes, Inhibited by Fluoride, Requiring Divalent Metal Ions . 66 A. Phosphatases. . . . . . 66

1. Acid Phosphatases . . 66 2. Alkaline Phosphatases 69

B. Pyrophosphatases. . . . 76 1. Acid Pyrophosphatases . 76 2. Alkaline Pyrophosphatases 76

C. Kinases . . . . 78 D. General . . . . • . • • . . 78

1. Cholinesterase . . . . . . 78 2. Glutamine Synthetase . . 80 3. Alcohol Dehydrogenase (Liver) . 81 4. a-Amylase (Ca++-activated) . . . . . . . . . . . . . • . . . . . . . 82 5. Comments on Non-Inhibition by Fluoride of some Enzymes Activated by

Divalent Metals . . . . . . . . . . . . . . . . . . . . . . . .. 83

IV. Enzymes Inhibited by Fluoride; No Requirement for Metal Ions Demonstrated 83 A. Liver Esterase . . . . . . 83 B. 5'-Adenylic Acid Deaminase 88 C. Phosphoglycerate Mutase . 88 D. Hydrazidase. . • . . . . 89

V. Enzymes, Inhibited by Fluoride, Containing Trivalent Metal Ions 90 A. Catalase. . . . . 91 B. Peroxidase. . . . 91

VI. Concluding Comments 91

References . . . . . . . . 93

Chapter 3: Changes in Cen Function Due to Inorganio Fluoride. J. MATTHEws. With 7 Figures . . • . . . . . . . . . 98

I. Introduction . . . . . . . . . . . . . 98

IT. Effect of Fluoride on Metabolic Pathways 99 A. Outline of Intermediary Metabolism. . 99 B. Glycolysis . . . . . . . . . . . . . 101 C. Pyruvate Metabolism and Oxidation in the Tricarboxylic Acid Cycle 104 D. Oxidative Phosphorylation 109 E. Respiration . . . . . . . . . . . . 112 F. Protein Synthesis ......... 113 G. The Response of Tissues to Hormones 118 H. Fat Metabolism in Adipose Tissue . . 118 I. Lipa.ses . . . . . . . . . . . . . . 121

ITl. Effects of Fluoride on Physical Activity of Cells . 122 A. Movement. . . • . . . . . . . 122

1. Motility. . . . . . . . . . . 122 2. Phagocytosis . . . . . . . . 124 3. Contractility ........ 126

B. Transport Across Cell Membranes. 131 1. Intestinal Epithelium. . . . . 131 2. Non-Polar Cells . . . . . . . 135

IV. Effects of Fluoride on the Growth and Development of Cells in Tissue Culture. 136

V. Concluding Remarks. 138

References . . . . . . . . 141

Chapter 4: Pharmacology of Sodium Fluoride. F. S. CARUSO, E. A. MAYNARD, V. DISTEFANO. With 10 Figures. 144

I. Introduction . . . . . . . . . . . . . . . . . . . • . 144

IT. Effect of Sodium Fluoride on Blood Pressure and the Heart. 145 A. Hypotensive Activity of Fluoride. . . . . . . . . . . 145 B. Pressor Effect of Fluoride . . . . . . . . . . . . . . 148 C. Stimulatory Effect of Sodium Fluoride on Isolated Heart Preparations 155

ITl. Effect of Sodium Fluoride on the Gut . . 157 A. Effect of Fluoride on the Gut, in Vivo 157 B. Effect of Fluoride on Dog Gut, in Vitro 158

IV. Effect of Sodium Fluoride on Respiration. 158

V. Summary and Conclusions 162

References. . . . . . . . . . 163

Chapter Ii: Fluoroamino Acids and Microorganisms. R. E. MARQUIS 166

I. Introduction . . . . . . . . . . . • . . . . 166

IT. Fluoroamino Acids and Cellular Microorganisms . 167 A. General Toxicity and Effects on Growing Cells 167 B. Incorporation of Fluoroamino Acids into Proteins 172 C. Effects of Fluoroamino Acids on Specific Cell Functions 177

1. Cell Division .............. 177 2. Motility . . . . • . . . . . . . . . . . . 178 3. Capsule Formation . . . . . . . . . . . . 179 4. Regulation of Enzyme Synthesis and Activity 179

x Contents

5. Transport Systems. . . . 6. Bacterial Sporogenesis .• 7. Respiration and Photosynthesis. 8. Ribosome Synthesis . • . . . . 9. Antibiotic Production . . . . . . . .

10. Maintenance of Diploid State in Fungi . 11. Virus and Bacteriocin Production ...

D. Catabolism of Fluoroamino Acids by Bacteria E. Bacterial Adaptation to Fluoroamino Acids .

1. Physiologic Adaptation. . . . . . . . . 2. Amino Acid-Secreting Mutants . . . . . 3. Mutants with Altered Transport Systems. . . . . . 4. Mutants with Altered Amino Acyl-tRNA Synthetases 5. Mutants with Altered Ribosomes

182 182 183 183 184 184 184 185 186 186 186 187 187 187

References . . . . . . . . . . . . . . . . . . . . . . . . . 188

Chapter 6: Fluorine-Containing Steroids. PHILIP S. CHEN JR., and POUL BORREVANG 193

I. Introduction . . . . . • . . . . . . . . . . . . . . . . . . . . . . . . 193

n. Fluoro Steroids with Respect to Glucogenic, Anti-In1l.ammatory and Mineralocorti-coid Effect . . . . . . . . . . 194

A. The 9a-Fluoro-Substitution in 194 1. Cortisol. . . . . . . . . 194 2 . .J1-Cortisol (Prednisolone) . 194 3 . .Js-Cortisol. . . . . . . . 195 4. Other Pregnane Derivatives 195

B. Fluorination at other Positions and in Combination with 9a-Fluoro 195 1. In Cortisone, Cortisol, Prednisone or Prednisolone 195

a) 2a-Fluoro. . . . . . . . . . . . . . . . . . . . . . . 195 b) 4-Fluoro . . . . . . . . • . . . • . . . . . . . . . . 195 c) 6a-Fluoro and 6a,9a-Difluoro . . . . . . . . . . . . . . 195 d) 15P-Fluoro and 9a,15p-Difluoro . . . . . . . . . . . . . . .. 196 e) 16a-Fluoro, 9a,16a-Difluoro, 6a,16a-Difluoro and 6a,9a,16a-Trifluoro 196 f) 16p-fluoro and 9a,16p-Difluoro . 196

2. In other Pregnane Derivatives . 196 a) 11p-Fluoro . . . . . . . 196 b) 12a-Fluoro . . . . . . . 196 c) 17a-Fluoro ... _ . . . 197 d) 21-Fluoro. . . . . . . . 197 e) Fluorine in 16a,17a-Ketal . 197

C. Other Groups in . . . . . . . . . . . . . . 197 1. 9a-Fluorocortisol and 9a-Fluoroprednisolone . 197

a) I-Hydroxy . . . . • 197 b) 2a-Methyl . . . . . 197 c) 4-Methyl and 4-Ethyl 198 d) 6a- and 6p-Chloro . 198 e) 6a-Fluoromethyl 198 f) 6a-Methoxy . 198 g) 6a-Methyl . . 198 h) 7a-Acetylthio . 198 i) 16P-Acetoxy . 198 j) 16a-Chloro . . 199 k) 16a-Ethyl . . . . 199 1) 16a-Fluoromethyl. 199 m) 16a-Hydroxy . . . 199 n) 16a,17a-Ketal 200 0) 16a-Methoxy . . . . 200 p) 16a- and 16p-Methyl. . . . . 200 q) 16-Methylene or .J15-16-Methyl 200 r) 16a, 17 a-Orthoesters (Cyclic). . 200 s) 21-Methyl . • . . . • . . . 201

2. Other 9a-Fluoropregnane Derivatives. . . . . . . . 201 a) 6a-Methyl . . . . . . . . . . . . . . . . • . 201 b) 16a-Chloro, Nitrogen and Sulfur Containing Groups 201 c) 16a-Hydroxy and Acetonide . . . . . . . . 201 d) 16a-Methyl. . . . . . . . . . . . . . . . 201 e) 17a,21-Epoxy. . . . . . . . . . . . . . . 201 f) 21-Chloro, 16a-Hydroxy and Acetonide 201 g) 21-Chloro, 16a-Hydroxy,2a-Methyl, Acetonide 202

D. With More than one Change in the 9a-Fluorocortisol or 9a-Fluoroprednisolone Molecule . . . . . . . . . . . . . . . 202

1. .16, 16a-Hydroxy . . . . . . . . . . 202 2.6a,7a-Dihydroxy . . . . . . . . . . 202 3. 6a,16a-Dimethyl. . . . . . . . . . . 202 4. 6a-Fluoro, 16a-Hydroxy and Acetonide. 202 5. 6a-Fluoro, 16a-Methyl . . . . . . . . 204 6. 6a-Fluoro, 21-Methyl. . . . . . . . . . . . . 204 7. 6-Formyl, 3-Enol Ether, 16a-Hydroxy, Acetonide 204 8. 6a-Methyl, 16a-Hydroxy and Acetonide 204 9. IIp-Acetoxy, 16a-Hydroxy. . 204

10. 16a-Methyl, 17a-Methoxy . . 204 11.17,2O:20,21-Bismethylenedioxy . . . . 204 12. 20-Ketal Derivatives. . . . . . . . . . . . . . . . 205 13. 21-Sugar Derivatives, 16a-Hydroxy . . . . . . . . . 205 14. 21-Tetrahydropyranyl Ether, 16a-Hydroxy, Acetonide . 205

E. Pyrazol Ring Attached to Ring A. . . . . . . . . . . . 205

F. Other Groups in 6a-Fluoro Compounds without 9a-Fluoro Substitution 205 1. 6a-Fluoro, 2-Methyl, 16a-Hydroxy, Acetonide, .1 1 • • 205 2. 6a-Fluoro, 16a-Hydroxy, and 6a-Fluoro, 16a-Methyl 206 3. 6a-Fluoro, 16a-Hydroxy, .1 8 and Acetonide . . 206 4. 6a-Fluoro, 3P-Acetoxy, 16a-Hydroxy Acetonide 206

G. Analytical Problems in Studying Fluorosteroids 206 1. General Aspects . . . . . . . . . . . . 206 2. Extraction. . . . . . . . . . . . . . . 207 3. Fluorosteroid Color Reaction . . . . . . 207 4. Determination in Urine as 17-Ketosteroids 207 5. Paper and Column Chromatography . . . . . 207 6. Preparation of Tritium-Labeled Triamcinolone . 208 7. Isomerization 208

H. Metabolism . . . 208 1. Absorption . . 208

a) Intestinal. . 208 b) Percutaneous 209 c) Intra-Articular 210

2. In Vitro Studies . . . . . . . 210 a) Liver Slices or Homogenates. . 210 b) Metabolism by Micro-Organisms .. .. . 212 c) Adrenal Metabolism of 9a-Fluoroandrostenedione 212

3. Plasma Protein Binding. . . . . . . . . . 212 4. In Vivo Studies . . . . . . . . . . . . . 213

a) Plasma Half-Life and Tissue Distribution. 213 b) Excretion of Metabolites . . . 214

1. Biological Effects of Fluorocorticoids 215 1. Microbiological Systems. . . . . 215

a) Growth Rate Stimulation. . . 215 b) Growth Rate Inhibition . . 216 c) Enzyme Activity - Alkaline Phosphatase 216

2. Isolated Systems in Vitro . . 216 a) Frog-Skin Sacs . . . . . 216 b) Frog Skin, Short Circuited 217 c) Perfused Rabbit Heart. . 217 d) Cat Papillary Muscle. . . 217

XII Contents

3. Effects in Animals . 217 a) Liver Enzymes • 217 b) Thymus Enzymes 218 c) Skin Enzymes. • 218 d) Skin Composition . . . . . . 218 e) Kidney. Infused Renal Artery. 218 f) Effects in Whole Animals. . . 218 g) Toxicology . . . . . . . . . 219

4. Effects in Man. . . . . . . . . .• 220 a) Extrapolation from Animal Biol1ssays to Man 220 b) Use of Corticosteroids . . . . . . . 222 c) Adrenal Suppression . . . . . . . . . . . 222 d) Side Effects of Therapeutic Doses . . . . . 223 e) Topical Effectiveness of Fluorosteroids . . . 223

III. Fluoro-Steroids with Respect to Gestagenic, Anti-Androgenic, Anti-Estrogenic, Anti-Ovulatory, and Anti-Gonadotrophic Effects. . . 224 A. The 9a-Fluoro-Substitution in . . . . . . • . . 224

1. Progesterone and ll-Oxygenated Progesterone. 224 2. 17a-Acetoxyprogesterone, ll-Oxygenated . . . 225 3. llP-Hydroxy-17a-Methyltestosterone . . . . . 225

B. Fluorination at other Positions. . . . . . . . . . . . . . . . . . .. 225 1. In Progesterone, and Its 9a-Halogenated and ll-Oxygenated Derivatives 225

a) 2a-Fluoro. . 225 b) 6a-Fluoro. . 225 c) 6,6-Difiuoro • 225 d) IIp-Fluoro . 225 e) 12a-Fluoro . 226 f) 17 a-Fluoro . 226 g) 21-Fluoro. . . 226 h) 21,21-Difiuoro. . 226 i) 21,21,21-Trifiuoro . 226

2. In 17a-Acetoxyprogesterone . . 226 a) 6a- and 6p-Fluoro, Ll1 and Ll6 226 b) 2l-Fluoro. . . . . • . . . 226

C. Other Groups in Combination with Fluorination in the Progesterone Molecule 227 1. 4-Fluoro, 6a, l6a-Dimethyl and 4,4-Difiuoro Derivatives . . . . . . . . 227 2. 6a- and 6P-Fluoro, l6a,17a-Dihydroxy and Ketals ........... 227 3. 6a-Fluoro, 16a-Methyl. . . . . . . . . . . . . . . . . . . . . . . . 227 4.6a-Fluoro, l7a-Methyl, 6-Fluoro, 17a-Ethyl, Ll6 and 6a, 2l-Difiuoro, l7a-

Methyl. . . . . . . . . . . . . . . . . . . 227 5. 6a-Trifiuoromethyl, l7a-Acetoxy . . . . . . . . . . . 228 6. 2l-Fluoro, 6-Chloro, l7a-Acetoxy,.d 6 • • • • • • • • • 228 7. 2l-Fluoro, 6a-Methyl, 17a-Acetoxy and.d6 • • • • • • • 228 8. 2l-Fluoro, 6a, l6a-Dimethyl, l7a-Hydroxy and.d6 • • • 228 9. 2l-Fluoro, 17a-Methyl, 6-Methyl and 6-Chloro, .11 1 and.d6 228

D. Fluorination in. . . . . . . . . . . . . . . . . . . 228 1. Ethisterone, 19-Norethisterone and Derivatives thereof 228

a) 5a-Fluoro, Dihydro, 19-Nor-10p-Hydroxy . . 228 b) 6a-Fluoro. . . . . . . . . . . . . . . . . . 229 c) lOp-Fluoro-19-Nor. . . . . . . . . . . . . . 229 d) l6a-Fluoro, l7a- and l7p-Chloroethinyl. . . . . 229

2.9p, 10a-Pregnanes - 6-Fluoro,.d6 and 17a-Acetoxy. 229 E. ~cellaneoUB . . . . . . . . . . . . . . . . 229 F. Biological Effects of Fluoroprogestational Steroids 229

1. Anti-Estrogenic Action . . . . . . • . . . . 230 2. Anti-Gonadotropin Activity . . . . . . . . . 230 3. Anti-Ovulatory Activity . . . . . . . . . . 230

IV. Fluoro-Steroids with Respect to Androgenic, Anabolic, Anti-Androgenic, Anti-Tumor and Anti-Gonadotropic Effect. . . . 230 A. The 9a-Fluoro Substitution in . . . . . . 230

1. llP-Hydroxy-.d'-Androstene-3,17-Dione . 230 2. l7a-Methyltestosterone, ll-Oxygenated . 231

Contents xm

B. Fluorination at other Positions in .d4-Androstene-3-one or Dihydro Compounds 231 1. 2a- and 2p-Fluoro ........ 231 2. 4-Fluoro. . . . . . . . . . . . . . . . . . . . 231 3. 6a- and 6p-Fluoro . . . . . . . . . . . . . . . . • . . . . . . . . 231 4. 6,6-Difluoro . . . . . . . . . . . • . . . . . . . . . . . . . . . . 232 5. 16a- and 16P-Fluoro and l6,l6-Difluoro ........•....... 232

C. Other Groups in .d4-Androstene-3-one or Dihydro Compounds in Combination with Fluorination. . . . . . . . . . . 232 1. 9a-Fluoro, UP-Hydroxy,7a-Acetylthio . . 232 2. 9a-Fluoro, UP-Hydroxy, 16a-Methyl . . . . . . 232 3. Up-Fluoro, 9a-Halogen . . . . . . . . . . . . 232 4. l6a-Fluoro, l6p-Methyl . . . . . . . • . . . . 232

D. Fluorination in other Androstane-3-one Derivatives . 232 1. 19-Nor-Derivatives . . . . 232

a) 4,4-Difluoro. . . . . . 232 b) 5a-Fluoro, lOP-Hydroxy 232 0) lOp-Fluoro . . . . . . 233

2. 9p,lOa . . . . . . . . . 233 6-Fluoro, ..1,.6 • • . • • • 233

E. Fluorination in Androstane Compounds without 3-Keto 233 1. In the Ring . . . . . . . . . . . . . 233

a) 3p-Fluoro. . . . . . . . . . . . . . . 233 b) 9a-Fluoro, UP-Hydroxy, 3-Methylene . . 233

2. In Side Chain . . . . . . . . . . . . . . 233 a) 2-Fluoromethyl and 2-Difluoromethyl, ..1 2 • 233 b) 2a, 3a- and 2P, 3p-Difluoromethylene . 233

F. Miscellaneous . . . . . . . . . . . . . . . 234 G. In Vitro Metabolism . . . . . . . . . . . . 234 H. Biological Effects of Fluoro Androgenio-Anabolic Steroids

1. Growth Stimulation in Man . . . . . . . . . . . . 2. Anti-Androgenic Aotion ............. .

V. Fluoro-Steroids with Respect to Estrogenio, Hypocholesterolemio, Anti-Gonado-

234 235 235

tropic and Anti-Tumor Effects . 235 A. Fluorination in. . . . . . . 235

1. Ring-A Aromatio Steroids . 235 a) 3-Fluoro . . . . . . 235 b) 4-Fluoro . . . . . . 235 0) l6a-Fluoro, l6p-Cyano 235 d) 16,16-Difluoro. . . 236

2. 19-Norethisterone . . . 236 lOp-Fluoro . . . . . . 236

3. In Side Chain . . . . . 236 17 P-(2-Fluoroethyl)Ether 236

VI. Fluoro Steroids with Respeot to Anti-Mineralocortiooid Effeot . 236

VII. Anti-Canoer Effeots of Fluoro Steroids . . . . . . . . . . . 236

VIII. Struoture-Activity Relationships and Influenoe of Fluorination 238

IX. Other Fluoro-Steroids 239

Referenoes . . . . . . . . 239

Chapter 7: Biological Properties of Selected Fluorine-Containing Organic Compounds. FRANK A. SMITH. With 8 Figures . . . . . . . . . . . . . 253

I. Introduotion . . . . . . . . . . . . . . . . . . . . . . 253 A. Preparation of Biologically Aotive Compounds of Fluorine . 254 B. Meohanisms of Aotion of Organically Bound Fluorine 255

II. The Carbon-Fluorine Bond . . . . . 256 A. Physioal Properties. . . . . . . 256 B. In Vivo Synthesis of the C-F Bond 258

XIV Contents

III. Metabolism of Organic Fluorocompounds . 260 A. In Vivo Rupture of the C-F Bond 260

1. Microbial Cleavage of the C-F Bond. 260 a) Aliphatic Compounds. . . . . . 260 b) Aromatic Compounds . . . . . 263

2. Mammalian Cleavage of the C-F Bond 264 a) Aliphatic Compounds. . . . . 264

i) Fluoroacetic acid . . . . . 264 ii) Halogenated Anesthetics 265

iii) Fluorine-Containing Sugars 267 iv) Fluoro-Oxaloacetate. . 267

b) Aromatic Compounds 268 i) Fluoronitrobenzenes 268

ii) 4-Fluorophenylalanine. . 270 iii) Acetanilides and Anilines 272 iv) 4'-Fluoro-4-dimethylaminoazobenzene . 273

3. Cleavage of the C-F Bond in Plants. . . . . 273

B. Biotransformations not Involving the C-F Bond 273 1. Bacterial Metabolism . . 273

a) Fluoronicotinic Acids 273 b) Fluorobenzoic Acids . 275 c) p-Fluorophenylalanine 277 d) 6-Fluorotryptophan . 277 e) 3-Deoxy-3-Fluoro-D-Glucose. 278

2. Mammalian Metabolism. . . . 278 a) Fluorobenzene ....... 278 b) Fluorobenzoic Acids and Amides 280 c) Fluorothiobenzoic Acids 282 d) Trifluralin . . . . . . . . . . 282 e) Fluorine-Substituted Amino Acids 284

i) m-Fluorotyrosine . . 284 ii) Fluorophenylalanines 286

iii) 5-Fluorotryptophan . 287 f) Other Compounds . . . 287

IV. Biological Effects of Organic Compounds of Fluorine. 287

A. Effects of Amino Acid Analogues on Protein Synthesis and Enzyme Induction in Mammalian Systems . . . . . . 288 1. Growth . . . . . . . . . . . . 288 2. Incorporation into Protein 288 3. Effects on Amino Acid Transport. 293 4. Effects on Activating Enzyme . . . . . . . . . 295 5. Metabolism of Fluorine-Containing Amino Acid Analogues 295

B. Effects of Organic Fluoro Compounds Upon Microorganisms 295

C. Organic Fluoro Compounds in Experimental Cancer Studies 306 1. Anticancer Agents . . . . . . . . . . . . . . . . . 306 2. Production of Experimental Cancers . . . . . . . . . 307

a) Derivatives of Dimethylaminoazobenzene. . . . . . . . . . . . . . 307 b) Fluorinated Derivatives of2-Acetylaminofluorene and Related Compounds 313 c) Fluorinated Derivatives of 10-Methyl-l,2-Benzanthracene . 316 d) Fluorotricycloquinazoline 319

D. Psychosedative Drugs. . . . . . . . . 319 1. Phenothiazines .......... 319

a) Structure - Activity Relationships 320 b) Mode of Action . . . . . . . 320

i) Erythrocyte Membrane . . 321 ii) Adrenomedullary Granules . 323

iii) ATPase . . . 323 iv) Miscellaneous. 325

c) Metabolic Fate 326 d) Toxicity 331 e) Analysis . . . 333

Contents

2. Butyrophenones . . . • . a) Structure - Activity Relationships b) Mode of Action . . . . . . . . c) Metabolic Fate •...........

i) Absorption, Distribution and Excretion ii) Biotransformations .

d) Toxicity . . . . . • .

E. Diuretic Agents . . . . . . 1. Benzothiadiazine Dioxides ....

a) Structure - Activity Relationships b) Mode of Action . . . . . . . . . c) Metabolic Fate ........... .

i) Absorption, Distribution and Excretion ii) Biotransformation. . . . . . .

d) Toxicity . . . . . . . . . . . . e) Analysis . . . . . . . . . . . .

2. Other Diuretic Agents. . . • . . . . F. Compounds Affecting the Thyroid Gland.

1. 3-Fluorotyrosine . . . . . . . . . . 2. 3-Fluoro-5-Iodotyrosine. . . . . . . 3. 3,5-DiHuorotyrosine . . . . . . . . 4. 3-Fluoro-4-Hydroxyphenylacetic Acid. 5.6-TriHuoromethyIthiouracil . . . 6. Compounds Related to Thyroxine

G. Antihypertensive Agents ..•... 1. Diuretic Drugs. . . . • . . . .

a) Benzothiadiazines . . . . . . i) Nondiuretic Benzothiadiazines

b) Phthalimidines . . . . . . 2. Catecholamine Depleting Agents

a) Guanidine Derivatives H. Adrenergic Blocking Agents . .

I. Pressor Amines. . . . . . . .

J. Monoamine Oxidase Inhibitors. 1. Derivatives of Hydrazine . . 2. Derivatives of Cyclopropylamine . 3. Indole Derivatives . . . 4. Miscellaneous . . . . .

K. Hypocholesteremic Agents . 1. Hormones. . . . . . .. .... 2. Basic Carbinols and Related Compounds 3. Miscellaneous . . .

L. Hypoglycemic Agents. 1. Sulfonylureas . . . 2. Sulfamylureas . . . 3. Triazines . . . . . . . . . 4. Thiadiazoles and Oxidiazoles . 5. Pyrazoles. •. 6. Hydantoins . . . . . . . .

M. PeniciIlins . . . . . . • . . . . . . . . . . . 1. Addition of Precursors to Fermentation Media . 2. Modification of the Penicillin Nucleus . 3. Total Synthesis . . . . . 4. Resistance to Penicillinase

N. Plant Growth Regulators

References . .

Author Index .

Subject Index •

xv

334 338 340 342 342 344 348 350 350 352 354 356 356 359 361 362 362 364 365 365 366 366 367 367 367 368 368 368 369 369 369

369

374

376 376 377 378 379

379 379 380 381

382 382 384 384 385 385 386 386 386 387 388 388 389

391

409

442

Contributors

POUL BORREVANG, Ph.D., Steroid Department, Novo Terapeutisk Laboratorium A/S, Copenhagen/Denmark

FRANK S. CARUSO, Ph.D., Bristol Laboratories, Syracuse, N.Y./USA

PmLIP S. CHEN JR., Ph.D., National Institutes of Health, Public Health Service, U.S. Department of Health, Education and Welfare, Bethesda, Md.20014/USA

VICTOR DISTEFANO, Ph. D., University of Rochester, School of Medicine and Dentistry, Department of Pharmacology and Toxicology, Rochester, N. Y. 14620jUSA

A.M.G. MACDONALD, Ph.D., University of Birmingham, Department of Chemistry, Birmingham 15/England

ROBERT E. MARQUIS, Ph.D., University of Rochester School of Medicine and Dentistry, Department of Microbiology, Rochester, N.Y. 14620/USA

J. MATTHEWS, Ph.D., CIBA Laboratories, Ltd., Horsham, Sussex/England

E.A. MAYNARD, University of Rochester, School of Medicine and Dentistry, Department of Radiation Biology and Biophysics, Rochester, N. Y. 14620/USA, Deceased

FRANK A. SMITH, Ph.D., University of Rochester School of Medicine and Dentistry, Department of Radiation Biology and Biophysics, Rochester, N.Y. 14620/USA

ALLAN WISEMAN, Ph.D., University of London, Biochemistry Unit, London SW ll/England