Neglected Factors in Pharmacology and Neuroscience Research: Biopharmaceutics, Animal Characteristics, Maintenance, Testing Conditions

Techniques in the Behavioral and Neural Science

Neglected Factors in Pharmacology and Neuroscience Research

Biopharmaceutics, Animal characteristics Maintenance, Testing conditions

VOLUME 12

V. Claassen

Formerly Head of the Pharmacological Department, Solvay Duphar, Weesp, The Netherlands

ISSN  0921-0709

Volume 12 • Number suppl (C) • 1994

Table of Contents

Cover image

Title page

Previously published in Techniques in the Behavioral and Neural Sciences

Copyright page

Acknowledgements

Academic Press, Inc. - Orlando, London

Advanstar Communications, Inc. - Cleveland

Akademie Verlag - Berlin

American Association for Cancer Research - Philadelphia

American Association for Laboratory Animal Science - Cordoba

American Heart Association - Dallas

American Institute of Nutrition - Bethesda

American Pharmaceutical Association - Washington

American Physiological Society - Bethesda

American Psychological Association - Washington

Archives Internationales de Pharmacodynamie et de Thérapie - Gent

Birkhäuser Verlag AG - Basel

Blackwell Scientific Publications, Inc. - Cambridge, MA

Boxwood Press - Pacific Grove

Center for Academic Publications Japan - Tokyo

Churchill Livingstone, Inc.

Current Science - London

Editio Cantor Verlag - Aulendorf

Elsevier Science Publishers - Amsterdam, New York, Oxford, Shannon

Endocrine Society - Bethesda

European Journal of Drug Metabolism and Pharmacokinetics - Geneva

European Journal of Endocrinology -Geneva

FASEB Journal - Bethesda

Georg Thieme Verlag - Stuttgart

Gerontological Society of America - Washington

Growth Publishing Co., Inc. - Bar Harbor

Gustav Fischer Verlag - Jena

International Journal of Pharmaceutical Technology and Product Manufacture - London

Institute of Animal Technology - Cardiff

Japanese Pharmacological Society - Kyoto

John Wiley & Sons, Inc. - New York

Journal of Endocrinology Ltd. - Almondsburg

Karger AG - Basel

Macmillan Press Ltd - Basingstoke

Marcel Dekker, In. - New York

Mary Ann Liebert, In. - New York

Mosby - Year Book, Inc. - St. Louis

National Academy of Sciences

National Research Council Canada

Pharmaceutical Society of Japan - Tokyo

Pharmacology and Toxicology - Copenhagen

PJD Publications, Ltd. - Westbury

Plenum Publishing Corporation - New York

Polish Academy of Sciences - Krakow

Psychonomic Society Publications - Austin

Publishing House of Czechoslovak - Prague

Raven Press - New York

Royal Pharmaceutical Society of Great Britain - London

Royal Society of Medicine Services Ltd - London

W.B. Saunders Company - Orlando

Société d’Édition de l’Association d’Enseignement Médical des Hospitaux de Paris - Paris

Society of Pharmaceutical Sciences in Finland - Helsinki

Society for the Study of Reproduction - Champaign

Springer Verlag - Heidelberg

Swedish Pharmaceutical Press - Stockholm

Taylor and Francis, Ltd. - London

Waverly Co - Baltimore

Introduction: Reproducibility of Animal Experiments

Publisher Summary

Biopharmaceutics

Biopharmaceutics

Intravenous Drug Administration

Publisher Summary

Rate of Drug Administration

In Summary

Intramuscular Drug Administration

Publisher Summary

Intramuscular Depot and Drug Absorption

Absorption from Oily Solutions

Absorption from Suspensions

Tissue Vulnerability

In Summary

Subcutaneous Drug Administration

Publisher Summary

Subcutaneous Depot and Drug Absorption

Absorption from Aqueous Solutions

Absorption from Oily Solutions

Absorption from Aqueous Suspensions

Pellet Absorption

In Summary

Intraperitoneal Drug Administration

Publisher Summary

Errors of Intraperitoneal Injections

Drug Absorption Pathway

Absorption from Aqueous Formulations

Tolerance

In Summary

Oral Drug Administration

Publisher Summary

Intraluminal Aspects

Volume of Drug Solution

Viscosity of Drug Solution

Osmotic Pressure of Drug Solution

Cosolvents

Solubilized Systems

Lipoidal Vehicles

Solid Dosage Forms

In Summary

Animal Characteristics

Inbred strains and outbred stocks

Publisher Summary

Inbred Strains

Phenotypic Variation and Response Variability

Variations in Inbred Characteristics

Strain Differences - Some Physiological Characteristics

Strain Differences - Some Behavioural and Neurochemical Traits

Strain Differences - Pharmacokinetics

Strain Differences - Pharmacodynamics

Strain Differences - Toxicological Aspects

Random-Bred Stocks

Stock Differences

Interindividual Differences

In Summary

Male–female differences

Publisher Summary

Oestrous Cycle

Vaginal Oestrus and Sexual Behaviour

Hormone Rhythms

Neuronal Systems

Behavioural Changes

Effects of Drugs on the Oestrous Cycle

Drug-induced Behavioural Changes

Pharmacokinetics

Sex-Related Differences

Pharmacokinetics

Pharmacological Effects

Toxicological Effects

In Summary

Changes during development and aging

Publisher Summary

Growing Older

Pharmacokinetic Aspects

Pharmacodynamic Aspects

Toxicological Aspects

In Summary

Maintenance of Experimental Animals

Housing Conditions

Publisher Summary

Animal Cage and Animal Room

Social Environment

In Summary

Food composition

Publisher Summary

Diet Formulas

Natural Ingredient Diets

Purified Diets

Diet and Biological Response

Diet Monitoring

In Summary

Food and water intake

Publisher Summary

Food And Water Intake of Individually Housed Sedentary Animals

Maintenance Conditions Affecting Feeding Behaviour

Variables Modulating Drug Effects on Feeding Behaviour

In Summary

Testing Conditions

Fasting

Publisher Summary

Water Intake and Hypovolaemia

Body Weight Loss on Food Deprivation and Food Intake on Refeeding

Metabolic Effects

Hormonal and Neuronal Adaptations

Pharmacokinetics

Changes of Drug Effects

In Summary

Food Restriction

Publisher Summary

Adjustment to Restricted Feeding Schedules

Adaptation to the Restriction of Energy Intake

Metabolic Consequences of Restricted Feeding

Effect on Orcadian Rhythms

Dietary Restriction and Longevity

Effect on Pharmacokinetics

Interactions with Drug Effects

In Summary

Circadian and Other Rhythms

Publisher Summary

Daily and Orcadian Rhythms

Other Rhythms

In Summary

Anaesthesia

Publisher Summary

Induction of Anaesthesia

Effects on the Cardiovascular System

Autonomic Effects

Effect on Metabolic Processes

Effects of Anaesthesia on Pharmacokinetics

Influence of Anaesthesia on Drug Effects

In Summary

Stress

Publisher Summary

Stress Response to Preliminary Experimental Procedures

Recovery from Acute Stress Experience

Stress Response After Repeated Stress Exposure

Repeated Handling

Response Differences Between Naive and Experienced Animals

Stress Responses in Dependence on Animal Characteristics

In Summary

Import of Experimental Results

Internal Validity

Statistical Inference

External Validity

In Summary

Index

Previously published in Techniques in the Behavioral and Neural Sciences

Volume 1: Feeding and Drinking, by F. Toates and N.E. Rowland (Eds), 1987, ISBN 0-444-80895-7

Volume 2: Distribution-Free Statistics: An Application-Oriented Approach, by J. Krauth, 1988, ISBN 0-444-80934-1, Paperback ISBN 0-444-80988-0

Volume 3: Molecular Neuroanatomy, by F.W. Van Leeuwen, R.M. Buijs, C.W. Pool and O. Pach (Eds), 1989, ISBN 0-444-81014-5, Paperback ISBN 0-444-81016-1

Volume 4: Manual of Microsurgery on the Laboratory Rat, Part 1, by J.J. van Dongen, R. Remie, J.W. Rensema and G.H.J. van Wunnik (Eds), 1990, ISBN 0-444-81138-9, Paperback ISBN 0-444-81139-7

Volume 5: Digital Biosignal Processing, by R. Weitkunat (Ed.), 1991, ISBN 0-444-81140-0, Paperback ISBN 0-444-98144-7

Volume 6: Experimental Analysis of Behavior, by I.H. Iversen and K.A. Lattal (Eds), 1991, Part 1, ISBN 0-444-81251-2, Paperback ISBN 0-444-89160-9, Part 2, ISBN 0-444-89194-3, Paperback ISBN 0-444-89195-1

Volume 7: Microdialysis in the Neurosciences, by T.E. Robinson and J.B. Justice Jr. (Eds), 1991, ISBN 0-444-81194-X, Paperback ISBN 0-444-89375-X

Volume 8: Techniques for the Genetic Analysis of Brain and Behavior, by D. Goldowitz, D. Wahlsten and R.E. Wimer (Eds), 1992, ISBN 0-444-81249-0, Paperback ISBN 0-444-89682-1

Volume 9: Research Designs and Methods in Psychiatry, by M. Fava and J.F. Rosenbaum (Eds), 1992, ISBN 0-444-89595-7, Paperback ISBN 0-444-89594-9

Volume 10: Methods in Behavioral Pharmacology, by F. van Haaren (Ed.), 1993, ISBN 0-444-81444-2, Paperback ISBN 0-444-81445-0

Volume 11: Methods in Neurotransmitter and Neuropeptide Research, by S.H. Parvez (Eds), 1993, Part 1, ISBN 0-444-81369-1, Paperback ISBN 0-444-81674-7, Part 2, ISBN 0-444-81368-3, Paperback ISBN 0-444-81675-5

Copyright page

© 1994 Elsevier Science B.V. All rights reserved.

No part of this publication may be reproduced, stored in a retrieval system or transmitted in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, without the written permission of the Publisher, Elsevier Science B.V., Copyright & Permissions Department, P.O. Box 521, 1000 AM Amsterdam, The Netherlands.

No responsibility is assumed by the Publisher for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. Because of the rapid advances in the medical sciences, the Publisher recommends that independent verification of diagnoses and drug dosages should be made.

Special regulations for readers in the USA – This publication has been registered with the Copyright Clearance Center Inc. (CCC) Salem, Massachusetts. Information can be obtained from the CCC about conditions under which photocopies of parts of this publication may be made in the USA. All other copyright questions, including photocopying outside of the USA, should be referred to the Publisher.

ISBN 0-444-81871-5 (Hardback)

ISBN 0-444-81907-X (Paperback)

ISSN 0921-0709

Elsevier Science B.V.

P.O. Box 211

1000 AE Amsterdam

The Netherlands

Library of Congress Cataloging-in-Publication Data

Claassen, Volkert.

Neglected factors in pharmacology and neuroscience research / Volkert Claassen.

p. cm. -- (Techniques in the behavioral and neural sciences; v. 12)

Includes bibliographical references and Index.

ISBN 0-444-81871-5

1. Pharmacology--Research--Methodology. 2. Neurosciences--Research--Methodology. 3. Animal experimentation. I. Title. II. Series.

RM301.25.C56 1994

615′.1′072–dc20 94-16419

CIP

This book is printed on acid-free paper

Printed in The Netherlands

Acknowledgements

V. Claassen,     Zuidsingel 25, 1241 EH Kortenhoef, The Netherlands

This book was written after my retirement as pharmacologist in the pharmaceutical industry Solvay Duphar B.V.

Without the support of the Library and Scientific Information Department of Solvay Duphar I would never have embarked upon this project. Therefore my thanks are in the first place due to the management who offered me free use of these facilities, needed to realize the compilation presented herewith.

I am greatly indebted to Theo Boschman who read and critically commented upon the whole manuscript during the long period of preparation of the book. I am also grateful to An de Wachter, Wim Lammers, René Remie, Tobi Swelheim and Jaap van Harten who read carefully several chapters of the book and who eliminated through their suggestions at least a number of imperfections. I am also thankful to all other former colleagues who, from the point of view of their discipline, read and improved parts of the text.

Muriel Levenbach and Rob van Spronsen gave indispensable support during the laborious literature search. Eef van der Lee typed thoroughly from my handwritten notes the first drafts of the text; Marijke Mulder prepared very carefully the final draft. Freek Schlingmann and Ruud van Oorschot gave great support in redrawing the figures. I am much obliged to them all.

The figures in the book were redrawn on the basis of the original material. The tables are in general selected from the data presented in tables in the concerning research paper. I thank authors for their approval to use their material in this way. Alas, in some cases it was impossible, notwithstanding repeated trials, to trace the current address of the authors.

I am much obliged to the following publishers for permission to adapt copyrighted material from the indicated sources for my purposes:

Academic Press, Inc. - Orlando, London

Animal Behavior

Appetite

Behavioral and Neural Biology

Fundamental and Applied Toxicology

Hormones and Behavior

Toxicology and Applied Pharmacology

The mouse in biomedical research. Volume 1. History, genetics and wild mice. Eds. H.L. Foster, J.D. Small and J.G. Fox–1981

Advanstar Communications, Inc. - Cleveland

Neurology

Akademie Verlag - Berlin

Biomedica Biochemica Acta

American Association for Cancer Research - Philadelphia

Cancer Research

American Association for Laboratory Animal Science - Cordoba

Laboratory Animal Science

American Heart Association - Dallas

Circulation Research

American Institute of Nutrition - Bethesda

Journal of Nutrition

American Pharmaceutical Association - Washington

Journal of Pharmaceutical Sciences

American Physiological Society - Bethesda

American Journal of Physiology

American Psychological Association - Washington

Behavioral Neuroscience

Journal of Comparative Physiological Psychology

Archives Internationales de Pharmacodynamie et de Thérapie - Gent

Archives Internationales de Pharmacodynamie et de Thérapie

Birkhäuser Verlag AG - Basel

Progress in Drug Research

Blackwell Scientific Publications, Inc. - Cambridge, MA

Proceedings of the Society for Experimental Biology and Medicine

Boxwood Press - Pacific Grove

Genetic Research Strategies for Psychobiology and Psychiatry. Eds. E.S. Gershon, S. Matthysse, X.O. Breakefield and R.D. Ciaranello–1981

Center for Academic Publications Japan - Tokyo

Japanese Journal of Physiology

Churchill Livingstone, Inc.

Progress in Fibrinolysis. Volume VII. Eds. J.F. Davidson, M.B. Donati and S. Coccheri–1985

Current Science - London

Journal of Hypertension

Editio Cantor Verlag - Aulendorf

Arzneimittel-Forschung

Elsevier Science Publishers - Amsterdam, New York, Oxford, Shannon

Behavioural Brain Research

Biochimica et Biophysica Acta

Biochemical Pharmacology

Biological Psychiatry

Brain Research

Brain Research Bulletin

Comparative Biochemistry and Physiology

Developmental Brain Research

Electroencephalography and Clinical Neurophysiology

European Journal of Pharmacology

Experimental Gerontology

FEBS Letters

Food and Chemical Toxicology

Food and Cosmetics Toxicology

International Journal of Pharmacy

Life Sciences

Neurobiology of Aging

Neuropharmacology

Neuropsychopharmacology

Neuroscience and Biobehavioral Reviews

Pharmacology, Biochemistry and Behavior

Pharmacology and Therapeutics

Physiology and Behavior

Progress in Neurobiology

Thrombosis Research

Toxicology Letters

Endocrine Society - Bethesda

Endocrinology

European Journal of Drug Metabolism and Pharmacokinetics - Geneva

European Journal of Drug Metabolism and Pharmacokinetics

European Journal of Endocrinology -Geneva

Acta Endocrinologica

FASEB Journal - Bethesda

Federation Proceedings

Georg Thieme Verlag - Stuttgart

Hormone and Metabolic Research

Gerontological Society of America - Washington

Journal of Gerontology

Growth Publishing Co., Inc. - Bar Harbor

Growth

Gustav Fischer Verlag - Jena

Zeitschrift für Versuchstierkunde

International Journal of Pharmaceutical Technology and Product Manufacture - London

International Journal of Pharmaceutical Technology and Product Manufacture

Institute of Animal Technology - Cardiff

Animal Technology

Japanese Pharmacological Society - Kyoto

Japanese Journal of Pharmacology

John Wiley & Sons, Inc. - New York

Aggressive Behavior

Biopharmaceutics and Drug Disposition

Developmental Psychobiology

Teratology

Toxic Susceptibility: Male/Female Differences. Ed. E.J. Calabrese–1984

Journal of Endocrinology Ltd. - Almondsburg

Journal of Endocrinology

Karger AG - Basel

Digestion

Gerontology

Neuroendocrinology

Neuropsychobiology

Pharmacology

Macmillan Press Ltd - Basingstoke

British Journal of Pharmacology

Marcel Dekker, In. - New York

Clinical and Experimental Hypertension

Techniques of solubilization of drugs. Ed. S.H. Yalkowsky–1981

Mary Ann Liebert, In. - New York

Journal of the American College of Toxicology

Mosby - Year Book, Inc. - St. Louis

Clinical Pharmacology and Therapeutics

National Academy of Sciences

Proceedings of the National Academy of Sciences of the USA

National Research Council Canada

Canadian Journal of Physiology and Pharmacology

Pharmaceutical Society of Japan - Tokyo

Chemical and Pharmaceutical Bulletin

Pharmacology and Toxicology - Copenhagen

Acta Pharmacologica - Toxicologica

PJD Publications, Ltd. - Westbury

Research Communications in Chemical Pathology and Pharmacology

Plenum Publishing Corporation - New York

Journal of Pharmacokinetics and Biopharmaceutics

Cocaine and Other stimulants. Eds. E.H. Ellinwood and M.M. Kilbey–1977

Polish Academy of Sciences - Krakow

Polish Journal of Pharmacology and Pharmacy

Psychonomic Society Publications - Austin

Bulletin of the Psychonomic Society

Publishing House of Czechoslovak - Prague

Physiologia Bohemoslavica

Raven Press - New York

Chronobiology International

Endocrine Rhythms. Ed. D.T. Krieger–1979

Royal Pharmaceutical Society of Great Britain - London

Journal of Pharmacy and Pharmacology

Royal Society of Medicine Services Ltd - London

Laboratory Animals

W.B. Saunders Company - Orlando

Gastroenterology

Société d’Édition de l’Association d’Enseignement Médical des Hospitaux de Paris - Paris

Pathology Biologie

Society of Pharmaceutical Sciences in Finland - Helsinki

Acta Pharmaceutica Fennica

Society for the Study of Reproduction - Champaign

Biology of Reproduction

Springer Verlag - Heidelberg

Archives of Pharmacology

Diabetologica

Psychopharmacology

Swedish Pharmaceutical Press - Stockholm

Acta Pharmaceutica Secica

Taylor and Francis, Ltd. - London

Journal of Toxicology and Environmental Health

Xenobiotica

Waverly Co - Baltimore

Drug Metabolism and Disposition

The Journal of Pharmacology and Experimental Therapeutics

The substantial financial support by Solvay Duphar for the production of the book is gratefully acknowledged.

1

Introduction: Reproducibility of Animal Experiments

Publisher Summary

This chapter provides an overview of reproducibility of animal experiments. The present knowledge concerning the status and reactivity of biological systems is based on the numerous investigations by various investigators in widely differing locations. Nevertheless, integration of study results is generally accepted as valid, and its success indicates that the results of animal experiments are interchangeable among laboratories. This joint endeavor has resulted in a common view on the mechanisms which play a leading role in physiological, biochemical, and behavioral regulation processes, and on the ways in which drug effects are brought about. Often great difficulties are experienced in reproducing experiments of other investigators and sometimes even in reproducing one’s own results. Experiments are generally restricted to the determination of the state or the response pattern to internal or external stimuli of a rather small animal population. However, the phenotype of the animals, resulting from interactions of the genotype and developmental and maintenance environment, is difficult to characterize and consequently cannot be reproduced exactly from one experiment to another.

The present knowledge concerning the status and reactivity of biological systems is based on numerous investigations by various investigators in widely differing locations. Nevertheless, integration of study results is generally accepted as valid, and its success indicates that results of animal experiments are interchangeable between laboratories. This joint endeavour has resulted in a common view on the mechanisms which play a leading role in physiological, biochemical and behavioural regulation processes, and on the ways in which drug effects are brought about.

Nevertheless, it is evident that in numerous studies contradictory results are obtained. Often great difficulties are experienced in reproducing experiments of other investigators and sometimes even in reproducing one’s own results. It is clear that at least part of the difficulty is inherent in the nature of animal experimentation. Experiments are generally restricted to the determination of the state or the response pattern to internal or external stimuli of a rather small animal population. However, the phenotype of the animals, resulting from interactions of the genotype and developmental and maintenance environment, is difficult to characterize and consequently cannot be reproduced exactly from one experiment to another. The response pattern to stimuli (dramatype) may equally well vary with changes in the sometimes poorly defined proximate or immediate environment in which the response is elicited (Russell and Burch –1959). In addition, experimental stimuli, not in the least challenges with drugs, may easily vary between laboratories by using different in-house standard procedures for their application.

This uncertainty in the animal characteristics and test conditions has not necessarily hindered the development of general insight into biological regulatory mechanisms. However, conflict may occur when detailing the description of processes because not all individual test results can be generalized as being the representative response patterns of the species concerned (let alone any extrapolations to other species). In this situation it becomes increasingly important to take into account the restrictions caused by the use of a rather limited test design. The significance of the test results can be better understood by investigating the limiting conditions of the experiment.

The ideal approach to this problem would be to include genotype, developmental conditions, stimulus conditions and characteristics as variables in the experimental design. However, it is clear that such an extension of the test design would lead to unmanageable studies. At best a selection can be made of a few variables regarding animal characteristics and test conditions which than can be included in supplementary experiments to estimate the significance and possible scope of the final conclusions. Such a strategy would at least improve the chance of reproducing an experiment and thus would reduce a number of unexplained discrepancies between studies.

The choice of the variables preferentially to be investigated will differ between experiments. A selection of those variables which are most critical in a specific study could be made on the basis of descriptions in the literature of their significance in comparable experimental conditions. However, strictly relevant data of this kind are often not available. Basic information is frequently related to study areas which are not of prime interest to the investigator. Appropriate data may be abundant, but hidden in publications which normally do not come under the investigator’s attention. Selection of this information from data bases is often rather troublesome and seldom satisfactory.

These complications largely explain why the above mentioned research strategy, to validate test results in supplementary experiments covering additional variables, is difficult to put into practice. No easy solution can be given for this problem and also this book will not offer a ready made answer.

The objective of this book is limited to indicating those variables which in general may need a better control. Examples, gathered from the literature, are presented to illustrate the impact that those neglected variables may have on various characteristics. It is attempted to select a series of representative studies from a broad field of interests so that insight can be obtained about the potential effects of these parameters in experimental outcome. It is hoped that in this way an impetus will be given to the critical consideration of test design and limitations of conclusions from experimental results. In choosing examples to illustrate how variation in response patterns depends upon the parameters selected, reference has been generally made to rat experiments because most animal experiments are performed with this species. Consequently contradictory results have been reported more often with this animal than for any other species. However, there is no reason to assume that the variability in response would be less in other species.

In part, this book is written as a reaction to frustrations endured during pharmacological research of many years standing, and therefore the choice of examples from the literature is largely related to this discipline. However, as pharmacological research is to a large extent based on other life sciences, this exposé may be of interest to a much broader audience. This may certainly be the case for pharmacokineticists and toxicologists for whom drugs are the main object of study. This book may also help to improve test designs for biochemists and physiologists, not only when using drugs as tools in their experiments, but also to improve generally the control of animal characteristics and test conditions.

Reference

Russell, W.M.S., Burch, R.L.The principles of humane experimental technique. London: Methuen & Co. Ltd., 1959.

Biopharmaceutics

Outline

Biopharmaceutics

Intravenous Drug Administration

Intramuscular Drug Administration

Subcutaneous Drug Administration

Intraperitoneal Drug Administration

Oral Drug Administration

Section A

Biopharmaceutics

In many studies drugs are administered to the experimental animal. On the one hand the biological effects and/or the fate of the compounds in the organism are the subjects of these investigations. On the other hand, drugs are used as a tool to elucidate physiological processes or to simplify the response pattern to other stimuli (or other drugs). The biological effect of drugs, just like their metabolism, comes about through interaction with specific target molecules or enzymes when the compound is present in a sufficient concentration at its specific site of action. The interaction with the target molecules is determined by the molecular structure of the drug. The molecular structure, however, has at the same time a marked influence on its concentration in the biophase. Absorption, protein binding, distribution, metabolism and excretion are dependent on the molecular characteristics of the compound.

In addition, biopharmaceutical factors also play an important part in the build-up of the drug concentration in the biophase. Generally, drugs are administered as a pharmaceutical formulation, which may vary from an adjusted aqueous solution to complex graded release formulations. Such differences may significantly affect the way in which the compound becomes available for the organism.

The occurrence of a drug effect is highly dependent on the rate with which the drug is transferred from its site of administration to the circulating blood. The compound must be absorbed at a sufficient rate to result in a certain (minimal) blood concentration so that in its turn a sufficient concentration will develop in the biophase to cause the corresponding pharmacological response. So far as formulation aspects affect the transfer rate from drug depot to the circulating blood, the extent of biological responses will vary with the biopharmaceutical factors concerned. As the absorption rate has an important influence on the total blood concentration-time course, biopharmaceutical factors may also change the duration of action of a drug.

Biopharmaceutical aspects, which influence the extent of the biological response of the animal, relate to the chemical characteristics of the compound itself (salt form, complexes, ester) as well as to its physical characteristics (crystal form, particle size). The kind of solvent in which the compound is administered in combination with the additional components of the drug formulation (cosolvents, surfactants, suspending agents) are key factors for the uptake process of the drug. Not only the route of administration, but also the disposition method (site of injection, rate of injection, administration volume) may all have an influence on the rate of drug bioavailability.

The pharmacokinetics of drugs are dependent on a large series of animal characteristics which may differ significantly between studies. This applies to factors like age, sex, feeding conditions and circadian rhythms. As a consequence, the significance of biopharmaceutical factors with regard to the rate of bioavailability may differ markedly between experiments.

The extent of the bioavailability is not necessarily affected by changes of the rate of the bioavailability. This will occur, however, if the stability of the compound in the pre-absorption stage is insufficient, if the absorption is excessively retarded by precipitation or tissue binding, or if by physiological processes the duration of the absorption is restricted.

In the present section an outline is given of variations in bioavailability and biological activity as consequences of differences in the formulation of the administered drug. In the subsequent chapters these dependencies are discussed for the generally used methods of administration. Specifications of animal characteristics and test conditions are often given for the examples presented to demonstrate how variable these factors are between studies and how limited their description often is. This survey shows that bioavailability and biological activity of a compound must always be considered in connection with its formulation and route of administration. Therefore, the choice and standardization of these factors have to be considered with great care in every experiment.

2

Intravenous Drug Administration

Publisher Summary

This chapter provides an overview of the intravenous drug administration. Intravenous drug administration offers various advantages over other routes of administration. Intravenous (and intra-arterial) drug administration provides the most complete drug availability with a minimal delay. By control of the administration rate, constant plasma concentrations can be obtained at a required level. Unexpected side effects observed during the administration period can be halted by stopping the infusion (pleading for an extended infusion time). Compounds that are poorly absorbed by the gastrointestinal tract may be advantageously administered intravenously. Compounds that are unacceptably painful when administered intramuscularly or subcutaneously may present no difficulties by the intravenous route. The chapter also highlights that with intravenous drug administration strict control is needed of the pharmaceutical qualities of the drug solution. In general, it seems advisable to restrict the injection volume in rodents to a maximum of 3 ml/kg and to extend the duration of injection to minutes. When interpreting experimental results, the possibility of a disequilibrium phase during and shortly after drug administration must be taken into consideration.

Rate of drug administration

Administration volume

Tonicity of injection fluid

Drug distribution after intravenous administration

Plasma drug concentration

Complications

Intravenous drug administration offers various advantages over other routes of administration:

– intravenous (and intra-arterial) drug administration provides the most complete drug availability with a minimal delay

– by control of the administration rate, constant plasma concentrations can be obtained at a required level

– unexpected side effects observed during the administration period can be halted by stopping the infusion (pleading for an extended infusion time)

– compounds that are poorly absorbed by the gastrointestinal tract may be advantageously administered intravenously

– compounds that are unacceptably painful when administered intramuscularly or subcutaneously may present no difficulties by the intravenous route.

Nevertheless, replication of experiments may be dependent upon variables of intravenous dosing that are frequently insufficiently controlled (or reported). The rate and volume of drug administration, and the tonicity of the drug solution may affect the pharmacological responses. Artefacts may arise due to the presence of particulate matter in the injection fluid.

Moreover, when designing experiments and when interpreting experimental results, assumptions are often oversimplified regarding the mixing of drug dosage with plasma, and the equilibration of the drug between plasma and cells or tissues. A number of aspects of this transitory disequilibrium will also be discussed.

Rate of Drug Administration

The extent of the pharmacological effects after drug administration depends for most compounds on their concentration at the site of action. This local concentration-time profile is directly or more indirectly related to the course of the plasma level so that variations in the drug administration procedure can markedly affect the action profile.

Intravenous injection

One of the most important variables with intravenous drug administration is the rate of injection. However, information on the injection rate in the description of the test methods is often rather incomplete, especially in the case of a rapid injection. The injection time may then vary from a few seconds up to some minutes. As intravenously administered drug dosages do not mix instantaneously with the intravascular blood, such a variation may cause a considerable difference in local concentrations immediately following the injection phase and so give rise to differences in immediate pharmacological effects. In blood volume determinations in rats a period of 20 min (!) is allowed for complete mixing of the intravenously injected labeled serum albumin and blood (Nikodijevic et al. - 1972; Rippe et al. - 1978).

When giving a drug as a bolus injection the compound passes around as a slug for one, and possibly two, circulations (Paton-1960). Dispersion of the initial peak is influenced by the site of injection as is apparent from indicator-dilution curves (Guyton et al. - 1973). The lungs, receiving the almost undisturbed primary slug, may damp the large blood concentration to a marked extent, as a number of drugs partition readily into this tissue. In this way the lungs act as a reservoir from which drug release occurs in the subsequent period (Benet-1978).

Crawford (1966) calculated for thiopentone in the standard man the decrease of the concentration in the slug during successive circulations; experimental follow-up has not been published. Chiou (1979) compared the initial peak concentration after a rapid intravenous injection of phenylbutazone in a sheep with the theoretical zero time plasma concentration (Cp°) extrapolated from data obtained 1–2 min after dosing (data obtained by McQueen and Wardell-1971). The initial concentration was estimated to be approximately 230% of the extrapolated Cp°-values.

Occasionally the pharmacological response after a rapid intravenous injection is apparently caused by the initial peak concentration. Injection of 0.2 μg histamine in the chloralose-anaesthetized cat induces a fall in blood pressure which begins about 3 sec after the appearance of the injection slug in the carotid artery and checks 2–3 sec after the end of the slug. The blood pressure then rises steadily, in spite of the rise in concentration of histamine in the blood as recirculation occurs (Gray and Paton-1948).

The transient bradycardia before the occurrence of a sustained tachycardia after intravenous injection of atropine is suppressed when the initial peak concentration is high enough to block instantaneously the vagal postsynaptic muscarinic receptors (Crawford-1966).

The bolus injection may cause transient aspecific effects in the animal. The ‘primary fall of blood pressure’ that is liable to occur with all kinds of intravenous injections has always been an annoying complication to experimenters on animals (Sollmann–1957).

The rapid intravenous injection of methylprednisolone (30 mg/kg) in haemodynamically stable anaesthetized dogs caused immediate, transient decreases in systemic vascular resistance (–40%) and mean aortic pressure (–28%). Within 5 min after the injection these haemodynamic parameters returned to pre-injection levels. When the same dose of methylprednisolone was administered over a 5 min period, mean arterial pressure was unchanged, whereas only a minor change occurred in systemic vascular resistance (Husum et al. - 1980).

Comparable, injection rate dependent transient blood pressure falls are illustrated in table 2.1 for fluvoxamine –an antidepressant with specific serotonin uptake inhibiting properties (Claassen-1981). Whereas injection in the cat of a 3 mg/kg dose in 2 sec or 20 sec gives rise to a clear decrease in mean arterial pressure, no effect is observed when the compound is given over a period of 2 min.

Table 2.1

Effect of injection rate upon haemodynamic parameters

¹)mean values of 2 cats

Intravenous infusion

Slow intravenous infusion precludes high initial blood concentrations. Of course, under these conditions the duration of the constant-rate infusion of a fixed dose will contribute significantly to the value of the peak blood concentrationCmax and to the time-interval Teff during which blood concentration is maintained above the minimum effective concentration (Ceff).

Raymond and Morgan (1980) studied by computer simulation the relationship between infusion duration (T) and the Cmax and Teff for drugs with monoexponential elimination kinetics. In fig. 2.1 the peak blood concentration (as fraction of C0, the theoretical zero-time intercept obtained if the dose is administered as an intravenous bolus and is mixed instantaneously with the plasma) is depicted as a function of the infusion time (as ratio to drug half-life). For a given half-life there is a non-linear relationship between the peak blood concentration and the period of infusion. Infusion duration of the dose for up to one half-life results in only a 30% decrease in Cmax.

Fig. 2.1 Effect of infusion time (expressed as multiples of drug half-life) on the maximum blood concentrations of drug (as a fraction of C0− the theoretical zero-time concentration for the bolus administration) attained for single dose intravenous administration of a drug with mono-exponential elimination characteristics, (adapted from Raymond, K., and Morgan, D.J.: J. Pharmacokin. Biopharm. 8, 573–582, 1980)

The influence of the infusion time on Teff is highly dependent on the C0Ceff ratio. For smaller values of Ceff (< 0.5 C0) Teff remains relatively unchanged or even increases with longer infusion durations. A similar computer simulation study was also performed for drugs exhibiting bi-exponential elimination kinetics. The relationship between Cmax, Teff and the duration of a constant-rate intravenous infusion of a fixed dose, was more complex than for drugs with monoexponential pharmacokinetics, due to the larger number of variables. It was not possible to formulate general guidelines and the authors concluded that investigations of this relationship for bi-exponential drugs could only be done on a drug by drug basis (Morgan and Raymond-1982).

A similar computer simulation study was performed by Klockowsky and Levy (1987) to determine the relationship between the threshold dose of an infused drug and the infusion rate. A two-compartment pharmacokinetic model was used. A graph showing the relationship between infusion rate and threshold dosage is presented in fig. 2.2. From this graph it can be seen that the dose required to produce a (pharmacologically) active concentration in the peripheral compartment showed a minimum. The increase in the threshold dose of the drug with slower infusion rates is due to inactivation or excretion of the drug during the period of concentration build-up in the compartments. The increase of the threshold dose with higher infusion rates is due to a disequilibrium between the drug at the site of action and in the vascular system.

Fig. 2.2 Relationship between infusion rate into the central compartment and amount of drug (threshold dose) required to produce an effective concentration of 100 mg/l in the peripheral compartment (containing the site of action) of a two-compartment system. Pharmacokinetic parameters used are based on plasma) concentration data for phenobarbitone in dogs, (adapted from Klockowski, P.M., and Levy, G.: J. Pharm. Sci. 76, 516–520, 1987.)

Experimental data on the relationship between the rate of intravenous infusion and the threshold dose for a series of centrally acting drugs have also been provided by Levy and co-workers. In table 2.2 the total dose and the onset time of loss of righting reflex in rats is given for phenobarbitone that was infused with rates varying from 0.41 to 4.12 mg/min/rat (Danhof and Levy-1984). The total dose of phenobarbitone needed to induce the effect, ranged from 143 to 202 mg/kg and tended to increase with increasing infusion rates. Likewise, the concentration of total and free phenobarbitone in serum and the concentration of total phenobarbitone in brain at the onset of loss of righting reflex tended to increase with increasing infusion rates. On the other hand, phenobarbitone concentrations in cerebrospinal fluid (CSF) at the onset of loss of righting reflex were apparently independent of the infusion rate and averaged 108 μg/ml. This indicates that the CSF establishes a practically instantaneous equilibrium with the sites of action of phenobarbitone in the brain. The excessive phenobarbitone concentrations in plasma (and brain) are caused by the slow distribution of phenobarbitone into the relevant biophase causing a transient disequilibrium during the infusion and for some time after.

Table 2.2

Effect of phenobarbitone infusion rate on total dose required to induce and on time of onset of loss of righting reflex in rats

¹)body weight 180 g.

In a comparable study with the CNS stimulant pentylenetetrazol, constant intravenous infusion was performed at four different rates until the animals either exhibited the first myoclonic jerk or started maximal seizures (Ramzan and Levy-1985). These studies showed that infusion rate had a significant effect on infusion time and total dose needed to induce the effects. However, the concentrations of pentylenetetrazole in serum, brain and CSF at the onset of a defined pharmacological effect were independent of the infusion rate (table 2.3). Apparently, in contrast to phenobarbitone, pentylenetetrazole equilibrates very rapidly between sites of action in the brain, and plasma, CSF and the brain as a whole.

Table 2.3

Effect of infusion rate on total dose and local concentration of pentylenetetrazole at onset of first myoclonic jerk in rats

Administration Volume

The blood volume of the rat is approximately 7% and the plasma volume about 4% of the body weight (Rippe et al. - 1978). For other laboratory animals comparable figures can be found (Altman and Dittmer-1971). Intravenous drug administration causes a rapid increase of the circulating blood volume with a concomitant haemodilution. The effect of this fluid administration depends considerably on the injected volume and the rate of administration as well as the species used and the basal circulatory conditions. When administering drugs in a volume of 2 ml/kg of body weight, not unusual in rat studies, the injected volume corresponds to 3% of the blood volume. In mice larger injection volumes (e.g. 10 ml/kg) are more usual and the injection rate is generally rather rapid.

An impression of the tolerance limits for fluid loading in rodents is given by the LD50-values for aqueous salt solutions as measured with rapid intravenous injections (table 2.4) (Kampmann and Frey-1963).

Table 2.4

Lethality with rapid intravenous fluid loading (injection time 10−45 sec)

Cardiovascular effects

Cardiovascular effects upon volume loading have been studied extensively, but seldom in rodents. In these physiological studies, usually in rabbits and dogs, quite large fluid volumes are used; the period of fluid administration varies between investigators from 10 sec to 10 min and even longer. Most characteristically, there is a marked increase in cardiac output. In the cases where the basal heart rate is low this may be due to a notable increase in heart frequency (Bainbridge reflex) (Vatner and Boettcher-1978). However, under test conditions when the heart rate is high, bradycardia occurs and the increased cardiac output is then due to an increased stroke volume (Taylor et al. - 1984). The mean arterial pressure is generally not affected (Vatner and Zimpfer-1981; Taylor et al. - 1984) or moderately increased (Vatner and Boettcher-1978). In the recovery period there is no close correspondence between the normalisation of the cardiac output and the restoration of the blood volume (Prather et al. - 1969).

Only following acute expansion of the blood volume the cardiac output is increased, whereas maintenance of an elevated blood volume over a long period of time (e.g. by administering dextran) is not effective in maintaining an elevated cardiac output. Though a slow infusion of blood over periods of 30 min or more increases blood volume as much as after a rapid injection, it does not cause a significant change in cardiac output.

Correspondingly, no change in arterial pressure was observed in rats when a volume of saline equal to 5% of body weight (80% of blood volume) was infused intravenously within 4–6 min at an approximate rate of 4 ml/min (Marin-Grez et al. - 1984) (male Wistar rats, 300–400 g, pentobarbitone anaesthesia). Central venous blood pressure increased markedly reaching a maximum of about 70 mm H2O shortly after saline infusion, returning approximately to control values in about 10 min, but not quite reaching them. Urine flow and natriuresis were likewise strongly increased by this volume loading.

A decrease in mean arterial pressure (from 127 mm Hg to 119 mm Hg) was observed by Schwab et al. (1986) upon intravenous volume loading in the Inactin anaesthetized rat (isoncotic albumin equal to 25% of blood volume in 15 min). Nevertheless central venous pressure, urine flow and natriuresis were increased.

Studies in rats in which the fluid loading volume corresponds more with the generally used injection volumes are scarce. Petterson et al. (1986) reported the haemodynamic changes during 10 and 20% blood volume expansion by intra-arterial injection of whole blood over 5 min (table 2.5). Arterial blood pressure was not effected whereas the central venous pressure was markedly increased. Cardiac output was only slightly increased under a concomitant decrease of heart rate and increase of stroke volume. These haemodynamic changes of volume loading may partly be influenced by the intra-arterial injection site used. Ricksten and Thoren (1980) reported that even an intra-arterial administration of 2 ml blood/kg body weight in rats caused a 15–20% decrease of the sympathetic activity of the splanchnic nerves.

Table 2.5

Haemodynamic changes in rats during 10 and 20% blood volume expansion by intra-arterial injection of whole blood

*2-way analysis of variance followed by t-test; P < 0.05.

Lachance and Garcia (1991) measured haemodynamics after acute blood volume expansion in conscious cannulated rats after recovery from pentobarbitone anaesthesia and stabilization of their haemodynamic parameters (!). Isotonic, isoncotic volume expansion was performed i.v. with human plasma protein fraction (6 ml/kg in 1 min, corresponding to 10% of the blood volume). Heart rate was decreased in SHR (30 beats/min) but not significantly affected in WKY rats. No significant change of the mean arterial pressure occurred. Central venous pressure was transiently increased (WKY rats 1.5 mm Hg and SHR 1.8 mm Hg). Left ventricular end-diastolic pressure was likewise transiently increased; the effect was higher in SHR than in WKY rats (14 and 7 mm Hg, respectively).

Tonicity of Injection Fluid

Haemolysis

The integrity of erythrocytes and cells in general is dependent on the osmotic state of the surrounding fluid. The intravenous administration of a substantial fluid volume may significantly disturb the osmoticity of the plasma when no precautions are taken.

Body fluids, including blood, normally have an osmotic pressure corresponding to that of 0.9% NaCl solution. Erythrocytes retain their normal size and shape in such a 0.9% solution of NaCl. However, a number of clinically and/or experimentally used salts or drugs (NH4Cl, boric acid, ethanol a.o.) fail to prevent haemolysis of erythrocytes in isoosmotic concentrations (test conditions 1 vol of blood with 100 vol of solution). For substances that pass through or alter the erythrocyte membrane the isoosmotic concentration differs markedly from the isotonic concentration (Hammerlund et al. - 1961), although no haemolysis is observed when testing the effect of strongly haemolytic substances by mixing 10 vol of blood with 1 vol isoosmotic solution (NH4Cl, boric acid, ethanol, sodium carbonate and urea).

Carlini and Jurkiewicz (1966) established that the intravenous injection of distilled water in the rat provoked a transient hypotension as a consequence of bradycardia and vasodilation (rat body weight 250–350 g, injection volume 0.05–0.5 ml, duration of injection 5–10 sec). By prolonging the duration of injection to 80 sec the effect is markedly blunted. The hypotension is explained by a local haemolytic process and a concomitant release of adenylic compounds. Haemoglobin determinations in plasma indicate that about 0.08 ml of blood is haemolysed upon injection of 0.4 ml of water. Debray et al. (1967) demonstrated that such an induced haemolysis leads to an increased biliary bilirubin excretion. On the other hand, the injection of strongly hypertonic solutions also leads to hypotension. However, it is unlikely that such an aspecific effect will occur with drug solutions as even a 10% NaCl solution only causes a marginal effect in this sense (Carlini-1964).

Drug Distribution After Intravenous Administration

During and after the spreading of the drug over the total blood volume the compound will dynamically equilibrate with the perfused tissues. In general, for drugs a partition between blood and organs in combination with metabolizing and excretion processes will lead to a gradually changing distribution of the compound in the organism.

Physiological pharmacokinetic models are used for the description of the time-course of the drug concentrations in tissues and organs. These perfusion models make use of haemodynamic and tissue-plasma partition data for the prediction of drug disposition.

As an illustration, fig. 2.3 shows a simplified perfusion model simulation of the distribution of lidocaine in various tissues and its elimination, following an intravenous bolus injection in the rhesus monkey (Benowitz et al. - 1974). This simulation indicates that the lung performs an important role in early buffering by containing up to 25% of administered lidocaine in the first seconds after administration. Rapidly equilibrating tissues (RET) (including liver, brain, heart and kidney) rise to a peak level within about 1 min, whereas during further redistribution the drug accumulates in the muscle. Adipose tissue, which is only 0.5% of the body weight in the monkey, stores little lidocaine.

Fig. 2.3 Perfusion model simulation of the distribution of lidocaine in various tissues of the rhesus monkey and its elimination following an intravenous bolus injection. RET: rapidly equilibrating tissue (heart and kidney). (adapted from Benowitz, N. et al.: Clin. Pharmacol. Ther. 16, 87–98, 1974.)

Ichimura et al. (1983) developed a physiologically based pharmacokinetic model to describe the tissue distribution and elimination of pentazocine in the anaesthetized rat. The predicted time courses of the levels of pentazocine in whole blood, plasma and tissues were verified by measuring drug concentrations in arterial blood and in tissues over 6 h mainly during the elimination phase. Excellent agreement was obtained between predicted and observed concentrations after a 2 mg/kg intravenous dose (fig. 2.4). The high tissue uptake of the intravenous dose by the lung is also apparent from these data.

Fig. 2.4 Predicted and observed concentration of pentazocine in tissues and in arterial blood after a 2 mg/kg i.v. injection over 2 min into the rat. (adapted from Ichimura, F et al.: Int. J. Pharm. 15, 321–333, 1983)

How dominant the clearance function of the lungs can be is illustrated by the almost complete removal (92%) of 5-HT from the blood in one single passage through the pulmonary circulation of the dog (Thomas and Vane-1967). The primary removal by an uptake mechanism is followed by metabolisation. When monoamine oxidase is inhibited, the amine is gradually released into the circulation. The clearance function of the lungs is rather compound specific (Bakhle and Vane-1974). Like 5-HT, noradrenaline, but not adrenaline, dopamine, or histamine (!), is cleared from the circulation by uptake in the lungs followed by metabolisation. The endothelial cells of the capillaries seem to be important sites of uptake for both amines (Gilles and Pitt-1982; Junod-1985).

Uptake of drugs by the lungs especially occurs with compounds with a substantial lipophilic character and a pKa-value greater than 8. Examples of such amines include chlorphentermine, chlorpromazine, imipramine and methadone. Not seldom these retained amines are not metabolized by the lung tissue. It follows from experiments with perfused isolated rabbit lungs that a great part of these retained drugs then efflux in a multi-exponential fashion to the perfusion fluid with half-lives of tens of sec to min. In addition a part of the retained drug shows a much greater persistence (the slowly-effluxable pool); after intravenous injection of methadone in the rabbit this fraction has a half-life of about 4 h (Wilson et al. - 1976; Wilson et al. - 1979; Bend et al. - 1985).

The time-dependent distribution pattern after intravenous administration can differ markedly for various drugs. For several compounds the time to peak myocardial levels and the partition coefficient between the myocardium and plasma are summarized in table 2.6. Propranolol and verapamil show a peak myocardial concentration 1–2 min after dosing. The tissue-plasma concentration ratio reaches a value of 13.8 and 6.2 respectively. Thereafter this ratio remains at the same level as for both drugs there is a parallel decay in myocardium and in plasma concentration. Amiodarone reaches a peak myocardial concentration 10–30 min after drug administration; however, the tissue-plasma ratio shows an increase during 2 h up to a maximum value of 89 which then remains constant for the next 4 h. Bretylium shows a rather long accumulation phase of 3 to 12 h.

Table 2.6

Myocardial uptake of anti-arrhythmic drugs in the dog after intravenous drug administration

The tissue drug concentration profile is less affected by the duration of the drug infusion than plasma drug concentration levels. This more restricted dependence of the tissue drug concentration on the infusion rate is also apparent from simulation studies based on the classical two-compartment pharmacokinetic model (Uccellini et al. - 1986).

This does not exclude that a proper prolongation of the infusion time results in a longer time period during which an effective tissue concentration is maintained. When measuring the diuretic effect of furosemide in dogs an increased response (urinary output and urinary excretion of sodium) was found with increasing infusion time. The total mean 24 h urine outputs were 1102, 1464, 2190 and 3470 ml for infusion durations of 10 sec, 30 min, 2 h, and 8 h, respectively; the corresponding values for sodium excretion were 170, 175, 272 and 440 mmol, respectively (Lee et al. - 1986a) (male dogs; 7.4–17.0 kg; fasted overnight; restrained by means of a dog sling; i.v. dose 15 (one dog) or 20 mg; immediate volume replacement; all 6 dogs received the drug with the 4 rates of infusion).

Plasma Drug Concentration

Distribution phase

Plasma levels after intravenous administration of a drug will mainly be determined by the efficacy of the pulmonary first-pass elimination process.Iwamoto et al. (1987) compared the plasma concentrations after intravenous (right atrium) and intra-arterial (right pulmonary vein) administration of propranolol to rats (male Wistar rats; 7 weeks old; 210–225 g; fasted overnight; propranolol 1, 2.5, 5 or 10 mg/kg).

fig. 2.5 shows the venous concentration-time profile for both routes of administration of a dose of 1 mg/kg –similar profiles were found at other dosages. The plasma level after intra-arterial dosing was always higher than that after intravenous drug administration. The area under the curve (AUC) after intravenous dosing was always approximately half of that after intra-arterial dosing, suggesting an extensive pulmonary first-pass elimination of propranolol after intravenous administration to rats.

Fig. 2.5 Venous plasma concentration-time curve for propranolol after intra-arterial and intravenous administration at 1.0 mg/kg to 7-week-old male Wistar rats (mean±s.d.). (adapted from Iwamoto, K. et al.: J. Pharm. Pharmacol. 39, 1049–1051, 1987)

The prolonged redistribution phase after intravenous drug administration is reflected in the occurrence of significant and persistent arterial-venous (A-V) plasma concentration differences. In fig. 2.6 the plasma level profiles after intravenous bolus injection (20 sec) of 2 mg propranolol to a rabbit are shown. Immediately after dosing the drug concentration in the arterial plasma declined rapidly during the first hour, after which a slower exponential decline was seen. Venous plasma concentration started at a low level, peaked at 40 sec and declined afterwards, parallelling the terminal phase of the arterial plasma level. Initially the arterial plasma levels were higher than the venous levels; at 20 sec the ratio was 24. After 45 min the concentrations were equal, whereas in the terminal phase the venous plasma level was 1.6 fold higher than the arterial plasma level (male New Zealand rabbits; urethane narcosis; catheters in jugular vein for drug administration –and left carotid artery and vena cava for blood collection) (Lam and Chiou-1981).

Fig. 2.6 The time course of the arterial and venous plasma levels of propanolol after a 2 mg i.v. dose of propranolol. HCl in the rabbit. Inserted figure: the course in the initial 120 sec. (adapted from Lam, G., and Chiou, W.L. Res. Commun. Chem. Pathol. Pharmacol. 33, 33–48, 1981)

Comparable differences between arterial and venous plasma concentrations were seen during and following constant rate infusion of the drug (fig. 2.7). During the drug infusion period arterial and venous plasma levels rose rapidly and peaked at the end of infusion: the arterial plasma levels were always higher than the venous plasma levels. Upon stopping infusion plasma levels dropped sharply and venous plasma levels became higher than the arterial levels.

Fig. 2.7 The time course of the arterial and venous plasma levels of propranolol during and following constant infusion of propranolol. HCl (5.2 mg/h) for 60 min in a rabbit. (adapted from Lam, G., and Chiou, W.L: Res. Commun. Chem. Pathol. Pharmacol. 33, 33–48, 1981)

For a number of pharmacodynamic and pharmacokinetic considerations the use of arterial plasma concentrations is indicated, though often venous plasma levels are used (Chiou et al. - 1982; Chiou and Lam-1982).

Fluctuating plasma drug levels

The general descriptions of the drug distribution pattern after intravenous dosing predict a gradual decline of the plasma level. Fitting a curve to experimental data points has in general been done on the basis of this assumed smooth decay. This procedure seems correct in many cases but in some studies one or more peaks or undulations occur during the decline of the plasma concentration.

Secondary peaks in the plasma concentration curve were observed in the elimination phase, i.e. 0.5 to 12 h after i.v. dosing of glycyrrhizin to rats (fig. 2.8) (male Wistar rats; 240–260 g; 20–24 h fasting; glycyrrhizin 100 mg/kg i.v. right femoral vein) (Ichikawa et al. - 1986).

Fig. 2.8 The time course of the plasma level of glycyrrhizin after an i.v. dose of 100 mg/kg in the rat. (adapted from Ichikawa, T. et al.: J. Pharm. Sci. 75, 672–675, 1986)

Fluctuating and irregular patterns of blood and plasma methotrexate concentrations were apparent during intravenous infusion of the drug to unanaesthetized dogs (fig. 2.9) (Lee et al. - 1986b) (conditioned beagle-mongrel hybrid dogs; 7.4–15.2 kg; i.v. infusion during 210 min via cephalic vein, methotrexate 0.09 mg/0.14 ml kg−1min−1; blood sampling every 10 min via (another) cephalic or saphenous vein; blood sample immediately centrifuged).

Fig. 2.9 Individual variation in the course of plasma and whole blood levels during constant infusion of methotrexate (0.09 mg/kg/min) to 4 dogs. Blood was collected from 120 min to 210 min during infusion. (adapted from Lee, M.G. et al.: Biopharm. Drug Disp. 7, 487–494, 1986)

There was a marked intersubject variability in the nature and magnitude of the fluctuations or deviations from normal smooth patterns. The largest differences were found for dog E between the 170 min and 200 min samples; there was a 42.4% difference for plasma concentrations and a 27.2% difference for the blood concentrations. The authors argue that the fluctuations cannot be an artefact due to analytical problems, sampling procedures or storage effects.

Wood et al. (1979) describe such undulations in blood concentration for chlorpromazine (and one of its metabolites) following an infusion of 2.5 mg during a 30 min period into a human being. Most remarkably during the infusion a peak level (6.0 μg/l) occurs after an infusion duration of 10 min. After the end of the infusion the blood concentration rises from about 3.6 μg/l at 60 min to about 14.8 μg/l at 90 min only to drop again 30 min later! Secondary peaks have been reported by various authors for chlorpromazine and for other drugs. As Wood et al. (1979) state, in addition for various drugs an unexpected scatter of concentrations in the exponential decay phase is described which may be caused by such undulating plasma levels.

Several explanations have been suggested for this phenomenon. Smolen et al. (1975) suppose that enterohepatic recycling is responsible for secondary peaking. This would be in accordance with the absence of these concentration irregularities in fasting rabbits and humans. Ichikawa et al. (1986) found a regular bi-exponential decline of glycyrrhizin when the compound was administered to rats with a biliary fistula. The AUC was higher and the total body clearance was lower in the control rats. The cumulative biliary excretion was 80.6% of the administered dose, and intestinal absorption was confirmed by using the bile collected after intravenous dosing. The authors conclude that the secondary peaks during the elimination phase were due to enterohepatic recycling of glycyrrhizin. Wood and Leonard (1983) postulate that drug recycling occurs through bladder resorption. Misra et al. (1980) attribute fluctuations in plasma concentrations following an intravenous bolus injection to a rapid uptake of large amounts of drug in lung and heart tissue during the first 20–30 sec. Changes in blood flow would affect the release of the drug during the redistribution phase.

Lee et al. (1981a) indicate that artefact-like plasma levels can occur through a storage effect when the equilibration between blood cells and plasma levels is slow. In fig. 2.10 the plasma gentamicin concentrations are shown as a function of time elapsed between the collection and centrifugation of blood, obtained from a rabbit at 0.5 min after an intravenous bolus dose of 4 mg/kg. The difference in gentamicin plasma concentrations obtained after blood equilibration periods of 5 and 15 min resp. was 53 percent. No plasma concentration differences due to variation in incubation time were observed for blood collected 60 min after drug administration.

Fig. 2.10 Post-sampling changes in plasma concentrations of gentamicin. Blood was collected from rabbits 0.5 min after an i.v. bolus dose of 4 mg/kg gentamicin and plasma was obtained by centrifugation after various time intervals. (adapted from Lee, M.G. et al.: Biopharm. Drug Disp. 2, 89–97, 1981)

The same group of investigators have described similar storage effects for furosemide, procainamide and methotrexate (Lee et al. - 1981b; Chen et al. - 1983; Lee et al. - 1984). They suggest that the formation and subsequent reversible decomposition of a Schiff base between the primary amino group of the drug with the fatty aldehyde group on the erythrocyte membrane may explain the irregular patterns. As indicated above the storage effect cannot explain completely the undulations found in in vivo studies.

Complications

The injection of a drug solution via an intravenous needle or catheter may cause a series of aspecific, more or less serious disturbances of the vessel wall, the circulating blood and organ perfusion. The chance of occurrence of these complications is particularly high with chronic catheters or with intravenous injections which are frequently repeated. Disruption of the vessel wall brings about the risk of inflammation and of thrombus formation (thrombophlebitis). Particulate matter, present in the drug solution or formed during the drug administration, may itself cause emboli in vital organs. In addition, the presence of catheters in the blood stream may cause disturbances for example in the platelet population and disturb drug protein binding. Factors implicated in causing thrombophlebitis in man are listed in table 2.7(Turco-1975).

Table 2.7

Factors implicated in causing phlebitis associated with intravenous drug infusion in man