Handbook of Experimental Pharmacology Continuation of Handbuch der experimentellen Pharmakologie

Vol. 61

Editorial Board

G. V. R. Born, London· A. Farah, Rensselaer, New York H. Herken, Berlin· A. D. Welch, Memphis, Tennessee

Advisory Board S. Ebashi . E. G. Erdos· V. Erspamer . U. S. von Euler· W S. Feldberg G. B. Koelle· O. Krayer· M. Rocha e Silva· 1. R. Vane· P. G. Waser

Chemotherapy of Viral Infections

Contributors M. 1. Bartkoski, Jr .. S. Bridges' P. E. Came' H. 1. Eggers P. H. Fischer' H. Friedman' M. Green' C. Gurgo' J. Hay B. D. Korant . J. J. McSharry· L. R. Overby· F. Pancic N.-H. Park' D. Pavan-Langston . C. 1. Pfau . L. M. Pfeffer W. H. Prusoff· J. L. Schulman· P. B. Sehgal· S. Specter B. A. Steinberg· I. Tamm . D. R. Tershak . F. H. Yin

Editors

P. E. Came and L. A. Caliguiri

Springer-Verlag Berlin Heidelberg New York 1982

Dr. PAUL E. CAME

Sterling-Winthrop Research Institute, Rensselaer, NY 12144jUSA

Dr. LAWRENCE A. CALIGUIRI

Department of Microbiology, University of Pittsburgh, School of Medicine, Pittsburgh, P A 15261 jUSA

With 133 Figures

ISBN-13: 978-3-642-68489-0 e-ISBN-13: 978-3-642-68487-6 DOl: 10.1007/978-3-642-68487-6

Library of Congress Cataloging in Publication Data. Main entry under title: Chemotherapy of viral infections. (Handbook of experimental pharmacology; vol. 61) Bibliography: p. Includes index. I. Virus diseases - Chemotherapy. 2. Antiviral agents - Testing. 1. Bartkoski, M. J. (Michael J.) II. Came, Paul E. III. Caliguiri, Lawrence. IV. Series. [DNLM: I. Viruses - Pathogenicity. 2. Antiviral agents - Pharmacodynamics. 3. Virus diseases - Drug therapy. 4. Antiviral agents - Therapeutic use. WI HA51L v. 61/QV 268.5 C517] QP905.H3 vol. 61 [RC1l4.5] 615:ls. 81-23320. ISBN-13: 978-3-642-68489-0 [616.9'25061]. AACR2

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, reproducing 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 payable to 'Verwertungsgesellschaft Wort', Munich.

© by Springer-Verlag Berlin Heidelberg 1982

Softcover reprint of the hardcover 1st edition 1982

The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use.

2122/3130-543210

Dedicated to

Bette, Paula, and Heather and Maggy, Laura, and Anne

Preface

" ... the motto for the therapeutics of the future will have to be de sedibus et causis pharmacorum. "

P. EHRLICH, 1909

Exciting events in the basic disciplines of virology, immunology, and pharmacology continue to advance the understanding of the pathogenesis and control of virus diseases. At the same time, the rational development of antiviral agents is attracting, to an increasing extent, the interest of workers in other disciplines. Improvements in technology facilitate the definition of potential target sites for antiviral intervention and unmask new viral and host genes. The outcome is a further steady development of new antiviral agents which approach the "magic bullets" first proposed by PAUL EHRLICH. Remarkable advances in protein synthetic methods that yield polypeptides which inhibit active sites of viral proteins have aided substantially in the basic and clinical study of these antiviral agents. In addition, the extremely rapid progression in recombinant DNA techniques, leading to the synthesis of large quantities of gene products, is also increasing our opportunities at a dashing pace. New information and developing technology facilitate research on the mechanism of action, toxicity, pharmacokinetics, and pharmacodynamics of new agents. The list of clinically effective antiviral agents is expanding and the number of potentially useful compounds is growing rapidly.

This book is a combined theoretical text and practical manual which, it is hoped, will be of use to all who have an interest in virus diseases, particularly scientists, physicians and graduate students. There are two major divisions of the volume: the first part deals with antiviral agents which are clinically effective and the second discusses compounds which are not, at present, widely used as chemotherapeutic agents, but are either currently under study as possible drugs or are used to elucidate the mechanism of virus replication. These major sections are preceded by a comprehensive chapter on current models of pathogenesis of virus disease produced by all the major groups of viruses. This updated coverage highlights the diversity of the important pathogens and offers insight into possible means of their control.

We have asked that the contributors of chapters consider the efforts and attempts of the chemists to cite important aspects of structure-activity relation­ships. The pharmacologic interactions including half-life, tissue distribution, and excretion rate are discussed where appropriate. The available results of clinical trials for various compounds are also discussed.

It is our goal to discuss the recent advances in historical perspective and to add a better understanding of antiviral compounds or drugs, per se. As the reader will learn, some of the chemicals which have never become drugs are the very compounds that supplied the impetus and optimism to continue the search. It is our hope that this volume will provide an awareness of previous contributions,

VIII Preface

investigations of newer agents suitable for chemotherapy of virus infections, and the excitement of the current work with the anticipation of the viral therapeutics of the future - the seat and cause of the pharmacology of antiviral agents.

We thank our colleagues who have made contributions to this work and made this volume possible, and acknowledge the valuable assistance of Ms. KATHLEEN CAVANAGH and Ms. MARY FRAZIER who provided editorial assistance in addition to excellent secretarial skills.

PAUL E. CAME LAWRENCE A. CALIGUIRI

List of Authors

Dr. M. J. BARTKOSKI, Jr., Department of Microbiology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20014/USA

Dr. S. BRIDGES, Centro di Endocrinologia e Oncologia Sperimentale.del C.N.R., Istituto di Patologia Generale, Via Sergio Pansini, 5,1-80131 Naples

Dr. P. E. CAME, Sterling-Winthrop Research Institute, Rensselaer, NY 12144/ USA

Professor Dr. H. J. EGGERS, Institut fUr Virologie der Universitiit K61n, Fiirst­Piickler-StraBe 56, D-5000 K61n 41

Dr. P. H. FISCHER, Department of Human Oncology, Wisconsin Clinical Cancer Center, University of Wisconsin, Madison, WI 53792/USA

Dr. H. FRIEDMAN, Department of Medical Microbiology, College of Medicine, University of South Florida, 12901 North 30th Street, Tampa, FL 33612/USA

Dr. M. GREEN, Institute for Molecular Virology, St. Louis University School of Medicine, 3681 Park Avenue, St. Louis, MO 6331O/USA

Dr. C. GURGO, Centro di Endocrinologia e Oncologia Sperimentale des C.N.R., Via Sergio Pansini, 5, 1-80131 Naples

Professor Dr. J. HAY, Department of Microbiology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20014/USA

Dr. B. D. KORANT, Central Research and Development Department, Experimental Station, E.!. du Pont de Nemours&Co. Inc., Wilmington, DE 19898/USA

Professor Dr. J. J. MCSHARRY, Department of Microbiology and Immunology, The Neil Hellman Medical Research Building, Albany Medical College of Union University, New Scotland Avenue, Albany, NY 12208/USA

Dr. L. R. OVERBY, Experimental Biology, Abbott Diagnostics Division, Abbott Laboratories, North Chicago, IL 60064/USA

Dr. F. PANCIC, Department of Microbiology, Sterling-Winthrop Research Institute, Columbia Turnpike, Rensselaer, NY 12144/USA

x List of Authors

Dr. N.-H. PARK, Eye Research Institute of Retina Foundation, Department of Ophthalmology, Harvard Dental School, 20 Staniford Street, Boston, MA 02ll4/USA

Professor Dr. D. PAVAN-LANGSTON, Eye Research Institute of Retina Foundation and Department of Ophthalmology, Harvard Dental School, 20 Staniford Street, Boston, MA 02ll4/USA

Dr. C. J. PFAU, Biology Department, Rensselaer Polytechnic Institute, Troy, NY 12181/USA

Dr. L. M. PFEFFER, The Rockefeller University, 1230 York Avenue, New York, NY 1002l/USA

Dr. W. H. PRUSOFF, Department of Pharmacology, Yale University, Sterling Hall, School of Medicine, 333 Cedar Street, New Haven, CT 0651O/USA

Prof. Dr. J. L. SCHULMAN, Department of Microbiology, Mt. Sinai School of Medicine, City University of New York, New York, NY 10029/USA

Dr. P. B. SEHGAL, The Rockefeller University, 1230 York Avenue, New York, NY 1002l/USA

Dr. S. SPECTER, Department of Medical Microbiology, College of Medicine, University of South Florida, 12901 North 30th Street, Tampa, FL 336l2/USA

Dr. B. A. STEINBERG, Department of Microbiology, Sterling-Winthrop Research Institute, Division of Sterlin Drug Inc., Rensselaer, NY l2l44/USA

Dr. I. TAMM, The Rockefeller University, 1239 York Avenue, New York, NY 1002l/USA

Professor Dr. D. R. TERSHAK, Microbiology and Molecular Physics, SlOl Frear Building, Department of Microbiology and Cell Biology, Biochemistry and Biophysics, The Pennsylvania State University, University Park, P A l6802/USA

Dr. F. H. YIN, Central Research and Development Department, Experimental Station, E.!. DuPont de Nemours&Co., Wilmington, DE 19898/USA

Contents

CHAPTER 1

Pathogenesis of Viral Infections. J. HAY and M. J. BARTKOSKI, JR. With 22 Figures

A. Introduction ........ . I. Defective Interfering Particles

II. Integration of Viral Genomes B. Adenoviruses ...... .

I. Structure and Replication II. Adenovirus Infections

C. Arenaviruses D. Coronaviruses . E. Viral Hepatitis

I. Hepatitis A II. Hepatitis B

F. Herpesviruses . I. Herpes Simplex Viruses I and 2

II. Herpes Simplex Virus Infections III. Cytomegalovirus IV. Varicella Zoster Virus V. Epstein-Barr Virus

G. Orthomyxoviruses .... I. Influenza A and B Viruses

II. Influenza C Virus . III. Influenza Infections

H. Human Papillomavirus J. Paramyxoviruses

I. Measles Infection II. Persistent Measles Infection

III. Mumps Infection IV. Parainfluenza Infection .

K. Picornaviruses . . . . . . . I. Structure and Replication

II. Picornavirus Infections L. Reoviruses . . . .

M. RNA Tumor Viruses . . .

I 2 3 3 4 8

9 12 14 IS 15 18 18 22 24 25 26 29 29 33 33 34 35 38 39 40 40 41 41 43

45 48

XII

N. Rhabdoviruses ..... . I. Structure and Replication

II. Rabies Virus Infection . O. Rotaviruses . . . . . . . .

I. Structure and Replication II. Rotavirus Infections . .

P. Poxviruses . . . . . . . . I. Structure and Replication

II. Poxvirus Infections Q. Togaviruses and Bunyaviruses

I. Togaviruses . II. Bunyaviruses . . . .

R. Slow Viruses . . . . . . I. Unconventional Agents

II. Conventional Agents References . . . . . . . . .

Part A

CHAPTER 2

Pyrimidine Nucleosides with Selective Antiviral Activity. P. H. FISCHER and W. H. PRUSOFF. With 1 Figure

A. Introduction ............... . B. 5-Halogenated Pyrimidine 2'-Deoxyribonuc1eosides

I. 5-Iodo-2'-Deoxyuridine 1. Synthesis ..... . 2. Antiviral Activity . . . 3. Effects on Normal Cells 4. Mechanism of Action .

II. 5-Trifluoromethyl-2'-Deoxyuridine 1. Synthesis ..... . 2. Antiviral Activity . . . 3. Effects on Normal Cells 4. Mechanism of Action

III. 5-Iodo-2'-Deoxycytidine 1. Synthesis ..... 2. Antiviral Activity . . 3. Effects on Normal Cells 4. Mechanism of Action .

C. Other 5-Substituted 2'-Deoxyuridine Derivatives I. 5-Ethyl-2'-Deoxyuridine

1. Antiviral Activity . . . 2. Effects on Normal Cells 3. Mechanism of Action .

Contents

50 51 53 54 55 57 57 58 60 61 62 65 67 67 69 70

95 98 98 98 98 98 99

100 100 100 101 101 101 101 102 102 102 103 103 103 103 103

Contents XIII

II. 5-Propyl-2'-Deoxyuridine 103 1. Antiviral Activity . . 103 2. Mechanism of Action 104

III. E-5-(2-Bromovinyl)-2'-Deoxyuridine (BVdUrd) 104 IV. 5-Methoxymethyl-2'-Deoxyuridine . . . . . . 104 V. Miscellaneous 5-Substituted 2'-Deoxyuridine Derivatives 105

D. Pyrimidine Nucleosides with an Altered Ring Structure . . . 105 E. Pyrimidine Nucleosides with an Altered Carbohydrate Moiety 105

I. Arabinofuranosylthymine . 105 1. Antiviral Activity . . . 105 2. Effects on Normal Cells 105 3. Mechanism of Action . 106

II. Arabinofuranosylcytosine . 106 III. 5-Iodo-5'-Amino-2',5'-Dideoxyuridine 107

1. Synthesis ..... 107 2. Antiviral Activity . . . . . . . . 107 3. Mechanism of Action . . . . . . 107

IV. 1-(2-Deoxy-2-Fluoro-p-D-Arabinofuranosyl)-5-Iodocytosine 108 References . . . . . . . . . . . . . . . . . . . . . . . . . 108

CHAPTER 3

Purines. N.-H. PARK and D. PAVAN-LANGSTON. With 9 Figures

A. Introduction ....... . B. 9-p-D-Arabinofuranosyladenine

I. Introduction and History II. Chemistry .....

III. Antiviral Spectrum . . . IV. Mechanism of Action .. V. Metabolism, Distribution, and Excretion

VI. Clinical and Experimental Therapeutic Aspects VII. Untoward Effects

VIII. Analogs of Ara-A ..... IX. Perspectives . . . . . . . .

C. 9-(2-Hydroxyethoxymethyl)guanine I. Introduction and History

II. Chemistry . . . . . III. Antiviral Spectrum . . . IV. Mechanism of Action .. V. Metabolism, Distribution, and Excretion

VI. Clinical and Experimental Therapeutic Aspects VII. Untoward Effects

VIII. Perspectives References . . . . . . . .

117 117 117 118 118 119 121 123 123 123 124 125 125 126 126 127 129 130 130 131

131

XIV Contents

CHAPTER 4

Amantadine and Its Derivatives. J. L. SCHULMAN. With 1 Figure

A. Introduction and History . . B. Chemical Structure .. .. C. Spectrum of Antiviral Activity D. Mechanism of Action E. Pharmacology .

I. Absorption II. Metabolism

III. Side Effects F. Animal Studies

I. Prophylaxis II. Treatment

G. Clinical Trials . I. Prophylaxis

II. Treatment H. Resistant Variants J. Conclusions and Perspectives

References . . . . . . . . . .

CHAPTER 5

The Thiosemicarbazones. C. J. PFAU. With 24 Figures

137 137 137 138 139 139 139 139 140 140 141 141 141 142 142 143 144

A. Introduction 147 B. History . . . . . . . . . . . 147 C. Chemistry ......... 148

I. Aryl Thiosemicarbazones 149 II. Quinoline, Pyridine, and Thiophene Thiosemicarbazones 151

III. Isatin-[3-Thiosemicarbazones . . . . . . 152 1. Substitution in the Aromatic Ring . . 153 2. N-Substitution in the Pyrrolidine Ring 153 3. Modification of the Pyrrolidine Ring 154 4. Modification of the TSC Side Chain 155 5. Other Compounds . . . . . . . . 155

IV. Isatin-[3-4',4'-Dialkylthiosemicarbazones 156 V. Isatin-[3-Isothiosemicarbazones . . . . 156

VI. Thiazole Thiosemicarbazones .... 157 VII. Pyrrolidine and Pyrazolone Thiosemicarbazones 158

VIII. Noncyclic Thiosemicarbazones . . 159 IX. Miscellaneous Thiosemicarbazones 160

D. Virus-Inhibitory Spectrum ...... 161 I. Aryl Thiosemicarbazones . . . . 161

II. Quinoline, Pyridine, and Thiophene Thiosemicarbazones 162 III. Isatin-[3-Thiosemicarbazones . . . . . 163 IV. Isatin-[3-4',4'-Dialkylthiosemicarbazones 167 V. Isatin-[3-Isothiosemicarbazones 168

VI. Thiazole Thiosemicarbazones .... 168

Contents XV

VII. Pyrrolidine and Pyrazolone Thiosemicarbazones 168 VIII. Noncyclic Thiosemicarbazones . . 168

IX. Miscellaneous Thiosemicarbazones 168 E. Effects on Normal Cells ....... 169

I. Aryl Thiosemicarbazones . . . . 169 II. Quinoline, Pyridine, and Thiopene Thiosemicarbazones 169

III. Isatin-fJ-Thiosemicarbazones . . . . . 169 IV. Isatin-fJ-4',4' -Dialkylthiosemicarbazones 170 V. Isatin-fJ-Isothiosemicarbazones . . . . 170

VI. Thiazole Thiosemicarbazones .... 170 VII. Pyrrolidine and Pyrazolone Thiosemicarbazones 171

VIII. Noncyclic Thiosemicarbazones . . 171 IX. Miscellaneous Thiosemicarbazones 172

F. Mechanism of Action ........ 172 I. Aryl Thiosemicarbazones . . . . 172

II. Quinoline, Pyridine, and Thiopene Thiosemicarbazones 173 III. Isatin-fJ-Thiosemicarbazones . . . . . . . 175

1. Adsorption, Penetration, and Uncoating 176 2. Transcription 177 3. Translation . . . . . . . . 178 4. Replication . . . . . . . . 179 5. Proteins and Viral Assembly 180

IV. Isatin-fJ-4',4' -Dialkylthiosemicarbazones 185 V. Isatin-fJ-Isothiosemicarbazones . . . . 186

VI. Thiazole Thiosemicarbazones .... 186 VII. Pyrrolidine and Pyrazolone Thiosemicarbazones 187 VII. Noncyclic Thiosemicarbazones . . 187 IX. Miscellaneous Thiosemicarbazones 188

G. Animal Studies . . . . . . . . . . . 188 I. Aryl Thiosemicarbazones . . . . 188

II. Quinoline, Pyridine, and Thiophene Thiosemicarbazones 189 III. Isatin-fJ-Thiosemicarbazones . . . . . 190 IV. Isatin-fJ-4',4'-Dialkylthiosemicarbazones 193 V. Isatin-fJ-Isothiosemicarbazones 193

VI. Thiazole Thiosemicarbazones 193 H. Clinical Studies . . . . . . . . . 194

I. N 1 - Methylisatin-fJ-Thiosemicarbazone 194 II. 3-Methyl-4-Bromo-5-Formylisothiazole Thiosemicarbazone 197

J. Perspectives 197 References . . . . . . . . . . . . . . . . . . . . . . . . .. 198

CHAPTER 6

Interferon and Its Inducers. P. B. SEHGAL, L. M. PFEFFER, and I. TAMM. With 5 Figures

A. Preface . . . 205 B. Introduction 205

XVI Contents

C. Production and Characterization of Human Interferons 207 I. Classification 207

II. Assay . . . . . . . . . . . 209 III. Production . . . . . . . . . 209

1. Human Leukocyte Interferon 209 2. Human Fibroblast Interferon 210 3. Human Lymphoblastoid Interferon 211 4. Alternative Sources . . . . . . . 211

IV. Characterization . . . . . . . . . . 212 1. Protein Purification and Sequencing 212 2. Cloning of cDNA Corresponding to Interferon mRNA Species 215

V. Mechanisms of Interferon Induction . . 218 1. General Comments . . . . . . . . . . . . . 218 2. Interferon Induction by Viral Inducers . . . . . 219 3. Interferon Induction by Synthetic Polynucleotides 221 4. Interferon Induction by Other Stimuli . . . . . 223 5. Genetics of Interferon Induction ....... 224 6. Characterization of Interferon mRNA Species and Their

Transcription Units . . . . . . . . VI. The Regulation of Interferon Production

1. Priming ..... . 2. Superinduction . . . . . . . . . .

D. Spectrum of Antiviral Activity . . . . . . . E. Mechanisms of Antiviral Action of Interferon

I. Interferon Binding . . . . . . II. Induction of the Antiviral State . . . .

III. Effects on the Cell Surface . . . . . . IV. Effects on Translation-Inhibitory Enzymes

1. Oligo(A) Synthetase . . . . . . 2. dsRNA-Dependent Kinase ... 3. Cap Methylation of Viral mRNA 4. The tRNA Effect . . . . . . . 5. Postscript: The Problem of Selectivity

V. Effects of Interferon in Various Virus-Host Cell Systems 1. Viral Uncoating 2. Viral Transcription 3. Viral Protein Synthesis . . 4. Viral Assembly or Release

F. Diversity ofInterferon Action: Effects on Normal and Transformed Cells I. Cell Proliferation . . . .

II. Cell Functions G. Pharmacokinetics of Interferons . . . . H. Efficacy of Interferons in Animal Models

I. Antiviral Studies II. Antitumor Studies . . . . . . . .

1. Virus-Induced Tumors . . . . . 2. Virus- and Radiation-Related "Spontaneous" Tumors 3. Transplantable Tumors ............ .

225 226 229 230 233 234 235 236 238 239 239 240 242 243 243 243 243 244 245 246 247 248 250 252 253 253 255 255 255 256

Contents

II. Toxicity of Interferon in Animals J. Efficacy of Interferons in Humans

I. Antiviral Studies ...... . II. Antitumor Studies . . . . . . .

III. Toxicity of Interferon in Humans K. Interferon Inducers ....... .

I. High Molecular Weight Substances 1. Antiviral Effects ..... . 2. Antitumor Effects . . . . . .

II. Low Molecular Weight Substances 1. Antiviral Effects 2. Antitumor Effects

L. Perspectives References . . . . . . . .

CHAPTER 7

Immunotherapy and Immunoregulation. H. FRIEDMAN and S. SPECTER

XVII

256 256 256 266 267 267 270 270 273 277 277 281 281 283

A. Introduction ........ 313 B. Immunity and Virus Infections 314 C. Antiviral Agents. . . . 316

I. Microbial Products 316 1. Protozoa . . . . 316 2. Gram-Negative Bacteria 317 3. Gram-Positive Bacteria 318 4. Mycobacteria . . . . . 320

II. Products of the Immune System 321 1. Immune Globulins 322 2. Thymic Hormones 323 3. Transfer Factor 324 4. Immune RNA 324 5. Interferon 325

III. Synthesized Immunostimulants 325 1. Levamisole 325 2. Vitamins 326 3. Tilorone . 327 4. Pyran . . 327 5. Inosiplex (lsoprinosine) 328

D. Potential of Immunostimulants as Antiviral Agents 329 References . . . . . . . . . . . . . . . . . . . 330

Part B

CHAPTER 8

Guanidine. D. R. TERSHAK, F. H. YIN, and B. D. KORANT. With 9 Figures

A. Introduction and History . . . . . . . . . . . 343 B. Synthesis and Properties of Picornavirus Proteins 343

XVIII Contents

I. "Shutoff" of Host Cell Protein Synthesis II. Synthesis of Viral Proteins

III. Mapping of the Viral Genome IV. Isolation of Viral Polypeptides

C. Replication of Viral RNA I. Viral RNA Structure

II. Viral Polymerase D. Chemistry of Guanidine E. Effects of Guanidine on Virus Replication

I. Spectrum of Inhibited Viruses . . . . II. Effects on Cells . . . . . . . . . .

III. Stage of Viral Growth Inhibited by Guanidine IV. Guanidine-Suppressive Compounds V. Site of Action of Guanidine ....

VI. Mechanism of Action of Guanidine 1. Previous Studies ...... . 2. Effects on Poliovirus Polymerase in Vitro 3. Probable Mode of Action of Guanidine

F. Guanidine as a Therapeutic Agent in Animals G. Clinical Nonvirologic Studies References . . . . . . . . . . . . . . . . .

CHAPTER 9

Benzimidazoles. H. J. EGGERS. With 22 Figures

A. Introduction and Historical Remarks B. 2-( IX-Hydroxybenzyl)-Benzimidazole

I. Virus-Inhibitory Spectrum . . II. Effects of HBB on Uninfected Cells

III. Kinetics of Antiviral Action, Effects on Viral Replication, and

344 344 347 348 348 348 351 352 352 352 353 354 356 358 359 359 361 367 368 369 370

377 378 379 381

Mechanism of Action . . . . . . . . . 383 I. Kinetic Aspects of Action . . . . . . 383 2. Effects on the Virus Replication Cycle 383 3. Mechanism of Action . . . . . . . . 388

IV. HBB Resistance and Dependence: Genetics of the System 388 V. Rescue .................... 391

VI. Superadditive Antiviral Effects of HBB and Guanidine 391 VII. Effects of HBB on Enterovirus-Infected Cells . . . . 394

C. HBB and Guanidine Chemotherapy in Animals . . . . . 395 I. Description of the Mouse System: Successful Treatment of Entero­

virus-Infected Animals . . . . . . . . . . . . . . . . . . . 395 II. Failure of Drug-Resistant Mutants to Emerge in Animals 397

III. The Importance of Drug Distribution, Metabolism, and Elimination on Therapeutic Efficacy . . . . . . . . . . . . . . . . . . 398 1. Introduction . . . . . . . . . . . . . . . . . . . . . . 398 2. Distribution and Excretion of HBB in the Mouse: Effects of

Age ......................... 400

Contents XIX

3. Metabolic Pathway of HBB in the Mouse and in Isolated Hepatocytes . . . . . . . . . . . . . . . . . . . 403

4. Distribution and Excretion of Guanidine· HCl in the Mouse . 403 5. Experiments to Define Optimum Treatment . . . . . . . . 404 6. Treatment of Mice Infected with Coxsackievirus B4 and Polio-

virus 2 ........................ 405 7. Late Treatment When First Symptoms Have Appeared:

Combined Effects of D-HBB, Guanidine, and Virus-Specific Antiserum . . . . . . . . . . . . . . . . . . . . . . . 405

D. Chemical Derivatives of HBB and Their Antiviral Activity in Cell Culture and in the Organism .. . . . . . . . . . . . . . . . . 407

I. Structure-Activity Studies . . . . . . . . . . . . . . . . . 407 II. I-Propyl-2-(oc-Hydroxylbenzyl)-Benzimidazole (l-Propyl-HBB) and

Its Hydrochloride Salt . . . . . . . . . . . . . . . .. 408 III. 2-( oc-Methyl-oc-Hydroxybenzyl)-Benzimidazole Hydrochloride

(oc-Methyl-HBB· HCl) . . . . . . . . . . . . . . . .. 409 IV. 1 ,2-bis-( 5-Methoxy-2-Benzimidazolyl)-1 ,2-Ethanediol (l,2-bis-

Benzimidazole) . . . . . . . . . . . . . . . . . . .. 410 V. 2-Amino-l-(Isopropylsulfonyl)-6-Benzimidazole Phenyl Ketone

Oxime 412 E. Conclusions 413 References . .

CHAPTER 10

Arildone: A fl-Diketone. J. J. Mc SHARRY and F. PANCIC. With 12 Figures

A. Introduction ........ . B. Chemical Structure and Synthesis

C. Antiviral Effects . . . . . . . . I. In Vitro Studies . . . . . .

1. Inhibition of Cytopathic Effects in Cell Culture 2. Plaque Reduction Tests . . . . . . . . . 3. Effect of Virus Yield . . . . . . . . . .

II. In Vivo Studies Against Herpes Simplex Virus 1. Arildone in DMSO 2. Arildone in Cream

D. Mode of Action .... I. Herpes Simplex Virus

II. Poliovirus . . .

E. Metabolism . . . . . . I. In Vitro Studies

1. Metabolism by CA TR Cells II. In Vivo Studies ..... .

1. Disposition in Laboratory Animals 2. Metabolism in Laboratory Animals

413

419 419 421 421 421 421 423 424 424 427

428 428 430

433 434 434 435 435

. 437

xx Contents

F. Toxicology . . . . ........ . 438 439 439 439

I. In Vitro Studies ........ . 1. Mutagenic Evaluation of Arildone 2. Effect of Arildone on Cellular Macromolecular Synthesis 3. Effect of Arildone and Its Metabolites on Cell Growth

II. In Vivo Studies G. Summary and Perspectives References . . . . . . . . .

CHAPTER 11

Phosphonoacetic Acid. L. R. OVERBY. With 9 Figures

· 440 · 441

442 · 443

A. Introduction and History . . . 445 B. DNA Polymerases . . . . . . . 446

I. Cellular DNA Polymerases . 446 II. Herpesvirus DNA Polymerase . 448

C. Chemistry .......... . 448 I. Structure . . . . . . . . . . 448

II. Structure-Activity Relationships . 449 1. Analogs . . . . . . . . 449 2. Phosphonoformic Acid . 452

D. Spectrum of Activity . . . . . . 454 I. Virus Replication in Tissue Culture 455

1. Productive Infections . 455 2. Transformed Cells . 456

II. Normal Cells . . . . . . 457 III. Animal Models . . . . . 461

1. Cutaneous Herpesvirus Infections . 461 2. Ocular Herpesvirus Infections 462 3. Herpes Genitalis .. . . . . . 462 4. Herpesvirus Infections of the Central Nervous System 465 5. Systemic Herpesvirus Infections 467 6. Latent Herpesvirus Infections 467 7. Varicella Infections . . . . 468 8. Lymphoproliferative Disease 468 9. Nonherpesvirus Infections . 468

10. Summary ........ 468 E. Pharmacologic and Clinical Studies 468

I. Metabolism . . . . . 469 II. Toxicology . . . . . . 469

III. Clinical Pharmacology . 469 F. Mechanism of Action . 470

I. Inhibition of Herpesvirus-Induced DNA Polymerase 470 II. Genetic Resistance . . . 471

G. Summary and Perspectives . 472 References . . . . . . . . . . 473

Contents

CHAPTER 12

Natural Products. P. E. CAME and B. A. STEINBERG. With 17 Figures

A. Introduction B. Amino Acids . . . . . . . . . . . . . .

I. Lysine . . . . . . . ....... . II. Phenylalanine and Phenylalanine Analogs

III. Methionine and Methionine Analogs . . IV. Miscellaneous Amino Acids and Analogs

C. Vitamin C D. Flavonoids . . ..... . E. Polysaccharides . . . . . .

I. Sulfated Polysaccharides II. Heparin . . . . . . .

III. Intestinal Mucopolysaccharides F. Substances from Microorganisms

I. Fungal Products 1. Aphidicolin 2. Cordycepin 3. Cytochalasin B 4. Gliotoxin, Aranotin, Sporidesmin, and Chetomin 5. Mycophenolic Acid 6. Tenuazonic Acid

II. Bacterial Products 1. Streptomyces . . 2. Other Bacteria .

G. Substances from Botanical Sources I. Alkaloids . . .

II. Tannins .... III. Lignosulfonates . IV. Other Substances

1. oc-Aminitin . . 2. Calcium Elenolate 3. Glycyrrhizic Acid

H. Substances from Marine Flora and Fauna I. Other Substances

I. Vitamin A II. Milk ..... .

III. Mosquito Tissues IV. Cobra Venom . . V. Neuraminidase Inhibitors

References .

CHAPTER 13

Rifamycins. C. GURGO, S. BRIDGES, and M. GREEN. With 2 Figures

XXI

479 479 480 481 484 486 488 489 490 490 492 492 493 493 493 494 495 495 497 497 497 497 501 502 503 504 505 505 506 506 506 507 508 508 508 509 509 510 511

A. Introduction ......... 519 B. Activity of Rifamycins on Bacteria . . . . . . . . . . . . 520

XXII Contents

I. Structural Requirements for Activity . . . . . . . . . . 520 II. Mechanism of Inhibition of RNA Polymerase . . . . . . 520

III. Effects on Psittacosis-Lymphogranuloma-Trachoma Agents 522 C. Activity of Rifamycins on Bacterial Viruses 522

I. Effect on DNA Bacteriophages . . . . . 522 II. Effect on RNA Bacteriophages . . . . . 523

D. Activity of Rifamycins on Mammalian Viruses 524 I. Growth Inhibition of Mammalian Viruses by Rifampicin 524

II. Mechanism of Vaccinia Virus Growth Inhibition by Rifampicin 525 1. Effect on DNA Synthesis of Vaccina-Infected Cells 525 2. Effect on mRNA Synthesis 526 3. Effect on Protein Synthesis . . . . . . . . 527 4. Effect on Virus Assembly ........ 527 5. Inhibition of Precursor Polypeptide Cleavage 530 6. Antivaccinia Activity of Various Rifamycin Derivatives 530 7. Effect of Rifampicin on Experimental Vaccinia Infections 531

III. Effect of Rifampicin on DNA Viruses Other Than Vaccinia 531 1. Herpesviruses. . . . . . . . . . . . . . . . . . . 531 2. Adenoviruses . . . . . . . . . . . . . . . . . . . 532

IV. Effect of Rifampicin on RNA Viruses Other Than Retroviruses 532 1. Picomaviruses . . . . . . . . . . . . . . 532 2. Vesicular Stomatitis Virus . . . . . . . . . . . . .. 533

V. Inhibition of Retrovirus Functions by Rifamycins . . . .. 533 1. Search for Selective Inhibitors of the Reverse Transcriptase 534 2. Correlation Between Lipophilicity and Antipolymerase Activity 536 3. Mechanism of Inhibition of the Reverse Transcriptase . . . . 537 4. Effects of Rifamycins on Virus Replication and Cell Trans-

formation . . . . . . . . . 538 E. Pharmacology of Rifampicin .... 543 F. Antiviral Activity of Other Ansamycins 543

I. Streptovaricins ........ 544 1. Effects on Vaccinia Virus 545 2. Effect on Retrovirus Functions 545

II. Tolypomycins . . . . . . . . . 546 III. Geldanamycin and Related Compounds 546

G. Concluding Remarks 546 References . 548

SUbject Index 557

List of Abbreviations

ABPP 2-amino-5-bromo-6-phenyl-4- BVdUrd E-5-(2-bromovinyl)-2'-pyrimidinol deoxyuridine

ADCC antibody-dependent cellular cytotoxicity CA chorioallantoic

5'-AdThd 5'-aminodeoxythymidine cAMP cyclic adenosine mono-AHF Argentinian hemorrhagic phosphate

fever CFA complete Freund's adjuvant AldUDP 5-iodo-5'-amino-2',5'- CF3dUMP 5-trifluoromethyl-2' -deoxy-

dideoxyuridine diphosphate uridine monophosphate AldUrd 5-iodo-5'-amino-2,5'- CF3dUrd 5-trifluoromethyl-2'-deoxy-

dideoxyuridine uridine AldUTP 5-iodo-5' -amino-2,5' -dideoxy- CF3dUTP 5-trifluoromethyl-2'-deoxy-

uridine triphosphate uridine triphosphate AIPP 2-amino-5-iodo-6-phenyl-4- cm cytomegalovirus inclusion

pyrimidinol disease AMP adenosine monophosphate cm Creutzfeldt-lakob disease AMV avian myeloblastosis virus CMP cytomegalovirus Ara-A arabinofuranosyladenine CPE cytopathic effect Ara-ADP arabinofuranosyladenine cpm counts per minute

diphosphate Ara-AMP arabinofuranosyladenine dATP deoxyadenine triphosphate

monophosphate dCTP deoxycytidinc triphosphate Ara-ATP arabinofuranosyladenine dCyd deoxycytidine

triphosphate DEV duck embryo vaccine Ara-C arabinofuranosylcytosine dGTP deoxyguanosine triphosphate Ara-Hx arabinofuranosylhypoxan- DHAdt 5,6-dihydro-5-azathymidine

thine DI defective interfering (particles) Ara-HxMP arabinofuranosylhypoxan- DMSO dimethylsulfoxide

thine monophosphate DNA deoxyribonucleic acid Ara-T arabinofuranosylthymine DNAP Dane particle associated RNA Ara-TMP arabinofuranosylthymine polymerase

monophosphate DNase deoxyribonuclease Ara-TTP arabinofuranosylthymine dNMP-PA deoxynucleotide mono-

triphosphate phosphate phosphonoacetate ATP adenosine triphosphate dpm disintegrations per minute

DRB 5,6-dichloro-l-J3-o-BCG bacille Calmette-Gm!rin ribofuranosylbenzimidazole BEV baboon endogenous virus dThd deoxythymidine BHF Bolivian hemorrhagic fever dTMP deoxythymidine mono-BHK baby hamster kidney (cells) phosphate BL Burkitt's lymphoma dTTP deoxythymidine triphosphate BRU-PEL Brucella abortus

preparation EA early antigen BSA bovine serum albumin EBNA Epstein-Barr nuclear antigen BTV Bluetongue virus EBV Epstein-Barr virus

XXIV List of Abbreviations

EDTA ethylenediaminetetracetic acid LCM lymphocytic chorio-EEE Eastern equine encephalo- meningitis

myelitis LHA lower hemagglutinin EIBT N1-ethylisatin-p- LPS lipopolysaccharide

thiosemicarbazone EMC encephalomyocarditis MA membrane antigen EtdUrd 5-ethyl-2'-deoxyuridine MCMV murine cytomegalovirus

MDMP 2-(4-methyl-2,6-dinitro-2'F-Ara-IC 1-(2-deoxy-2-fluoro-p-o- vanilino )-N-methylpropion-

arabinofuranosyl)-5-iodo- amide cytosine MDP muramyl dipeptide

FCS fetal calf serum MEM minimum essential medium F 3dThd trifluorothymidine MER methanol-extracted residue FMDV foot-and-mouth disease virus MHV mouse hepatitis virus FPA p-fluorophenylalanine MIBT N-methylisatin-p-thiosemi-FSV feline sarcoma virus carbazone FUDR fluorodeoxyuridine MIC minimum inhibitory

concentration GMK green monkey kidney (cells) MLV murine leukemia virus

MMdUrd 5-methoxymethyl-2'-HA hemagglutinin deoxyuridine HBB 2-«(X-hydroxybenzyl)- MMTV murine mammary tumor virus

benzimidazole m.o.i. multiplicity of infection HBcAg hepatitis B core antigen MRB 5-methyl-2-o-ribobenzimid-HBeAg hepatitis B enzyme antigen azole HBIG hepatitis B immune globulin mRNA messenger ribunucleic acid HB.Ag hepatitis B surface antigen HDCS human diploid cell strain NA neuraminidase HDL high density lipoprotein NCV noncapsid viral (protein) HFI human fibroblast interferon NDV Newcastle diseases virus HLA human leukocyte antigen NK natural killer (cells) HLI human leukocyte interferon NP nucleoprotein hnRNA heterogeneous nuclear NPC nasopharyngeal carcinoma

riboncleic acid HPLC high pressure liquid opv oral poliovirus vaccine

chromatography HPV human papillomavirus PAS periodic acid-Schiff HSV herpes simples virus PEG polyethylene glycol HVA hepatitis virus A pfu plaque forming unit HVB hepatitis virus B PML progressive multifocal

leukoencephalopathy IBT isatin-p-thiosemicarbazone PPi inorganic pyrophosphate IBV infectious bronchitis virus (phosphodiesterase) IdCyd 5-iodo-2'-deoxycytidine PrdUrd 5-propyl-2'-deoxyuridine IdUMP 5-iodo-2' -deoxyuridine PVM pneumonia virus of mice

monophosphate IdUrd 5-iodo-2' -deoxyuridine RF replicative form IgG immunoglobulin (X RI replicative intermediate IgG immunoglobulin l' RLV Rauscher leukemia virus IgM immunoglobulin J.l. RNA ribonucleic acid IMP inosine monophosphate RNP ribonuclear protein (complex) ipv inactivated poliovirus vaccine RSV respiratory synctial virus ISG immune serum globulin

SDS sodium dodecylsulfate K killer (cells) SFV Semliki Forest virus KTS kethoxal-bis-thiosemi- SGOT serum glutamate-oxalo-

carbazone acetate transaminase

List of Abbreviations XXV

SMON subacute myelo-optico- TPCK L-l-tosylamide-2-phenylethyl-neuropathy chloromethyl ketone

SRBC sheep red blood cells tRNA transfer ribonucleic acid SSPE subacute sclerosing para- TSC thiosemicarbazone

encephalitis SSV simian sarcoma virus UHA upper hemagglutinin SV simian virus UMP uridine monophosphate SVP subviral particles

VA virus-associated TCID so tissue culture median infective VCA viral capsid antigen

dose VIG vaccinia immune globulin TCT y-thiochromanone-l-thio- VPg genome-linked viral protein

semicarbazone VSV vesicular stomatitis virus TdT deoxynucleotidyl transferase VZIG varicella zoster immune TGE transmissible gastroenteric globulin

(virus of swine) VZV varicella zoster virus TK thymidine kinase TLCK N-rx-p-tosyl-L-lysinechloro- WEE Western equine encephalo-

methyl ketone myelitis TMV tobacco mosaic virus WT wild-type (virus)