Safety Pharmacology in Pharmaceutical Development
Approval and Post Marketing Surveillance, Second Edition -
2nd Edition
Visit the link below to download the full version of this book:
https://medipdf.com/product/safety-pharmacology-in-pharmaceutical-development-ap
proval-and-post-marketing-surveillance-second-edition-2nd-edition/
Click Download Now
Second Edition
Safety
Pharmacology
in Pharmaceutical
Development
Approval and Post Marketing
Surveillance
Shayne C. Gad
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
CRC Press
Taylor & Francis Group
6000 Broken Sound Parkway NW, Suite 300
Boca Raton, FL 33487-2742
© 2012 by Taylor & Francis Group, LLC
CRC Press is an imprint of Taylor & Francis Group, an Informa business
No claim to original U.S. Government works
Version Date: 20120320
International Standard Book Number-13: 978-1-4398-4568-4 (eBook - PDF)
This book contains information obtained from authentic and highly regarded sources. Reasonable efforts have been
made to publish reliable data and information, but the author and publisher cannot assume responsibility for the
validity of all materials or the consequences of their use. The authors and publishers have attempted to trace the
copyright holders of all material reproduced in this publication and apologize to copyright holders if permission to
publish in this form has not been obtained. If any copyright material has not been acknowledged please write and let
us know so we may rectify in any future reprint.
Except as permitted under U.S. Copyright Law, no part of this book may be reprinted, reproduced, transmitted,
or utilized in any form by any electronic, mechanical, or other means, now known or hereafter invented, includ-
ing photocopying, microfilming, and recording, or in any information storage or retrieval system, without written
permission from the publishers.
For permission to photocopy or use material electronically from this work, please access www.copyright.com
(http://www.copyright.com/) or contact the Copyright Clearance Center, Inc. (CCC), 222 Rosewood Drive, Danvers,
MA 01923, 978-750-8400. CCC is a not-for-profit organization that provides licenses and registration for a variety
of users. For organizations that have been granted a photocopy license by the CCC, a separate system of payment
has been arranged.
Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for
identification and explanation without intent to infringe.
Visit the Taylor & Francis Web site at
http://www.taylorandfrancis.com
and the CRC Press Web site at
http://www.crcpress.com
Dedication
To Novie Beth—
for bringing so much
light and joy
to this life
Contents
Preface..................................................................................................................xi
About the Author..............................................................................................xv
Chapter 1 Safety pharmacology: Background, history, issues,
and concerns................................................................................. 1
1.1 General versus safety pharmacology.................................................... 2
1.2 History........................................................................................................ 4
1.3 Reasons for poor predictive performance............................................. 7
1.4 Why tiers?.................................................................................................. 8
1.5 Study designs and principles.................................................................. 9
1.5.1 Selection of methodology and species.................................... 9
1.6 Issues........................................................................................................ 12
1.7 Integral versus separate......................................................................... 13
1.8 Summary................................................................................................. 15
References.......................................................................................................... 15
Chapter 2 Regulatory requirements: ICH, FDA, EMA, and Japan...... 19
2.1 Regulatory requirements....................................................................... 19
References.......................................................................................................... 23
Chapter 3 Principles of screening and study design............................. 25
3.1 Introduction............................................................................................. 25
3.2 Characteristics of screens...................................................................... 27
3.3 Uses of screens........................................................................................ 30
3.4 Types of screens...................................................................................... 32
3.5 Criterion: Development and use.......................................................... 32
3.6 Analysis of screening data.................................................................... 35
3.6.1 Univariate data.......................................................................... 35
3.6.1.1 Control charts........................................................... 35
3.6.1.2 Central tendency plots............................................ 37
3.6.2 Multivariate data...................................................................... 38
3.6.2.1 Analog plot............................................................... 39
3.7 Study design............................................................................................ 41
vii
viii Contents
3.7.1 Animal model........................................................................... 41
3.7.2 Group size.................................................................................. 42
3.7.3 Statistical design....................................................................... 42
3.7.3.1 Dose levels and test concentrations...................... 42
References.......................................................................................................... 42
Chapter 4 Cardiovascular system.............................................................. 45
4.1 Introduction............................................................................................. 45
4.2 History...................................................................................................... 45
4.2.1 Special case (and concern)—QT prolongation..................... 45
4.2.1.1 Regulatory developments....................................... 47
4.2.2 Patch-clamp studies using recombinant cells
expressing hERG channels...................................................... 57
4.2.3 hERG protein expression system............................................ 57
4.2.3.1 Relevance of hERG to QT prolongation................ 58
4.2.3.2 Expression and recording systems........................ 58
4.2.4 Cardiovascular function testing............................................. 59
4.2.5 Conscious rodent, dog, and primate telemetry studies...... 64
4.2.5.1 Six-lead ECG measurement in the
conscious dog........................................................... 64
4.2.6 Systems for recording cardiac action potentials.................. 65
4.2.6.1 Cloned human potassium channels..................... 65
4.2.6.2 Cardiac action potential in vitro—Purkinje
fibers.......................................................................... 65
4.2.6.3 Monophasic action potential in anesthetized
guinea pigs................................................................ 65
4.2.6.4 ECG by telemetry in conscious guinea pigs........ 65
4.2.6.5 Hemodynamics and ECG in anesthetized
or conscious dogs or primates............................... 66
4.3 Summary................................................................................................. 66
References.......................................................................................................... 66
Chapter 5 Central nervous system............................................................ 69
5.1 Core battery CNS procedures............................................................... 72
5.1.1 General behavioral observation............................................. 72
5.1.2 Functional observational battery........................................... 73
5.2 Rat............................................................................................................. 74
5.2.1 Observational assessments..................................................... 74
5.2.2 Locomotor activity.................................................................... 75
5.2.3 Motor coordination.................................................................. 75
5.2.4 Pain sensitivity.......................................................................... 76
5.2.5 Convulsive threshold............................................................... 76
5.3 Dog............................................................................................................ 77
5.3.1 Neurobehavioral screen........................................................... 77
Contents ix
5.4 Sleep induction and interaction with hypnotics................................ 81
5.5 Higher cognitive function..................................................................... 81
5.5.1 Passive avoidance..................................................................... 81
5.5.2 Morris maze.............................................................................. 82
5.5.2.1 Materials.................................................................... 82
5.5.2.2 Set up apparatus and begin acquisition testing.... 83
5.5.2.3 Continue trials.......................................................... 83
5.6 Isolated tissue assays............................................................................. 84
5.6.1 Electrophysiology methods..................................................... 84
5.7 CNS function: Electroencephalography.............................................. 86
5.8 Neurochemical and biochemical assays............................................. 87
References.......................................................................................................... 88
Chapter 6 Respiratory system.................................................................... 91
6.1 Plethysmography.................................................................................... 96
6.2 Design of respiratory function safety studies.................................... 99
6.2.1 General considerations............................................................ 99
6.2.2 Study design.............................................................................. 99
6.2.3 Capnography........................................................................... 100
6.3 Study design considerations............................................................... 101
6.3.1 Dose selection.......................................................................... 101
6.3.2 Species selection..................................................................... 101
6.4 Summary............................................................................................... 101
References........................................................................................................ 102
Chapter 7 Renal function.......................................................................... 105
7.1 Major functions of the kidney............................................................ 106
7.2 Acute renal failure................................................................................ 108
7.3 Functional reserve of the kidney........................................................ 109
7.4 Clearance................................................................................................ 109
7.5 Free water clearance and renal concentrating ability......................114
7.5.1 Renal blood flow......................................................................116
7.5.2 Fractional excretion of sodium..............................................116
7.6 Clinical chemistry measures...............................................................116
7.7 Animal models.......................................................................................117
7.7.1 Rat..............................................................................................117
7.7.2 Dog.............................................................................................118
7.8 Examples of species differences in drug sensitivity........................118
References.........................................................................................................119
Chapter 8 The gastrointestinal system................................................... 121
8.1 Drug-induced alterations of GI transit or motility.......................... 122
8.2 Gastrointestinal function.................................................................... 122
8.3 Assessment of intestinal transit......................................................... 124
x Contents
8.4 Determination of intestinal absorption............................................. 126
8.4.1 Methods of administering test substance........................... 126
8.4.2 Methods for quantitating degree of absorption................. 127
8.4.2.1 Appearance in systemic fluids............................. 127
8.5 Gastric emptying rate and gastric pH changes: A new model...... 130
8.6 Effects of drugs on gut immune system (jejunum, ileum, colon).... 131
8.7 Candidate drugs to evaluate for effects on gut immune system.... 131
8.8 Conclusions........................................................................................... 132
References........................................................................................................ 132
Chapter 9 The immune system................................................................ 135
9.1 Introduction to the immune system and adverse modulation
activities of drugs................................................................................. 135
9.1.1 Passive cutaneous anaphylaxis:
Test for potential antigenicity of compound...................... 137
9.1.1.1 Test method for pulmonary sensitization.......... 140
9.1.2 Center for Drug Evaluation and Research guidance
for investigational new drugs................................................141
9.2 Overview of the immune system....................................................... 142
9.3 Immunotoxic effects............................................................................. 148
9.3.1 Immunosuppression.............................................................. 149
9.3.2 Immunosuppressive drugs................................................... 151
9.3.2.1 Antimetabolites...................................................... 151
9.3.2.2 Glucocorticosteroids.............................................. 154
9.3.2.3 Cyclosporine........................................................... 155
9.3.2.4 Nitrogen mustards................................................. 155
9.3.2.5 Estrogens................................................................. 156
9.3.2.6 Heavy metals.......................................................... 156
9.3.2.7 Antibiotics............................................................... 157
9.3.3 Immunostimulation............................................................... 157
9.3.3.1 Hypersensitivity.................................................... 159
9.3.3.2 Photosensitization.................................................. 164
9.3.3.3 Autoimmunity........................................................ 165
References........................................................................................................ 169
Appendix A: Acronyms................................................................................. 175
Appendix B: Laboratories conducting safety pharmacology testing...... 179
Preface
Safety pharmacology is the evaluation and study of the pharmacological
effects of a potential drug that are unrelated to the desired therapeutic
effect and, therefore, may present a hazard—particularly in individuals
who already have one or more compromised or limited organ system
functions, as is generally the case for those receiving pharmacotherapy.
Unlike other nonclinical evaluations of a drug’s safety, these safety phar-
macology evaluations are usually conducted at doses close to the intended
clinical dose. Continued International Conference on Harmonisation of
Technical Requirements for Registration of Pharmaceuticals for Human
Use (ICH) guidelines, followed by versions by national regulatory author-
ities including those of the U.S. Food and Drug Administration (FDA),
have made such evaluations mandatory before a potential drug is intro-
duced into humans—while also failing to provide clear guidance as to
how the requirements are to be met.
General/safety pharmacology has been an emerging discipline
within the pharmaceutical industry over the last 30 years in which unan-
ticipated effects of new drug candidates on major organ function (i.e.,
secondary pharmacological effects) are critically assessed in a variety of
animal models. A survey was conducted to obtain customer input on the
role and strategies of this emerging discipline. Overlooked in importance
by all but a few (Zbinden, 1966, 1984) for many years, the Japanese clearly
became the leaders in developing and requiring such information, while
the United States was (and remains) in a position behind Japan and the
European Union (EU) in establishing formal requirements and in imple-
menting industrial programs.
Most companies have traditionally conducted evaluations of cardio-
vascular and central nervous system (CNS) functions, while few have
conducted respiratory, gastrointestinal, and renal function evaluations;
a few conduct a ligand-binding/activity panel as part of their pharma-
cological profiling. Resources needed to complete a company’s standard
safety pharmacology program are approximately one to four full-time
people per compound. One third of companies use a maximum tolerated
dose (MTD) for safety pharmacology studies; two thirds use multiples
xi
xii Preface
of pharmacological or therapeutic doses. Until 2002, only half conducted
safety pharmacology studies with Good Laboratory Practices (GLPs),
using the 1992 Japanese guidelines as a guide or outline. Company clini-
cians were most often cited as the primary customers for whom safety
pharmacology studies were done, followed by research and development
scientists and then regulatory authorities. These results suggest that most
companies primarily conducted safety pharmacology for its contribution
to risk assessment and critical care management.
It is important that the tests employed detect bidirectional drug effects
and be validated in both directions with appropriate reference (control)
substances. This requirement is less appropriate for multiparameter pro-
cedures. Blind testing could be an advantage. Ethical considerations are
important, but the ultimate ethical criterion is the assessment of risk for
humans. Safety pharmacology studies should not be overly inclusive but
should be performed to the most exacting standards, including GLP com-
pliance. This is, of course, backward; such human tolerance is properly an
extension (and expression) of the nonclinical safety pharmacology.
The other point of view has been that properly executed, repeated-dose
preclinical safety studies meeting the current design theoretically could
fill these needs, recognizing that undesired pharmacological activities of
novel drugs or biologicals may limit development of a therapeutic agent
prior to the characterization of any toxicological effects. In rodent species,
general pharmacology assays have traditionally been used to screen new
agents for pharmacological effects on the central and peripheral nervous
systems, the autonomic nervous system and smooth muscles, the respira-
tory and cardiovascular systems, the digestive system, and the physiolog-
ical mechanisms of water and electrolyte balance. In large animal species,
such as dogs and nonhuman primates, smaller numbers of animals per
study limit their use for screening assays, but these species may play an
important role in more detailed mechanistic studies. For drugs and bio-
logicals that must be tested in nonhuman primates because of species-
specific action of the test agent, functional pharmacology data are often
collected during acute or subacute toxicity studies. This requires careful
experimental design to minimize any impact that pharmacological effects
or instrumentation may have on the assessment of toxicity. In addition,
with many new therapies targeted at immunological diseases, the phar-
macological effect of therapeutics on the immune system presents new
challenges for pharmacology profiling and has now been addressed by
its own ICH guidance (ICH S8). The applications of pharmacology assay
by organ system in both rodent and large animal species are discussed,
as well as practical issues in assessing pharmacological end points in the
context of toxicity studies (Martin et al. 1997; Matsuzawa et al. 1997).
In Europe, the numbers of registered drugs and drug expenditure are
increasing rapidly. Within the EU, regulations no longer require that new
Preface xiii
drugs have to be better than old ones. At the same time, pharmacoepide-
miology studies in Europe and the United States continue to show that
adverse drug reactions now may account for up to 10% of the admissions
of patients to hospitals at a cost of hundreds of millions of U.S. dollars
annually (Sjouist, 2000). Compared with 30 years ago, this represents a
considerable increase. The many shortcomings of clinical trials and their
relevance to health care provide a partial explanation. Adverse drug reac-
tions are often poorly studied and documented in these trials and sel-
dom included in health economical analyses of the value of new drugs.
Pharmacovigilance is product oriented rather than utilization oriented
and quite invisible in clinical medicine. This is regrettable because up to
50% of adverse drug reactions (ADRs) are dose dependent and thus pre-
ventable. Hopefully, the rapid progress in molecular and clinical phar-
macogenetics will provide new tools to enable clinicians to choose and
dose drugs according to the needs of individual patients. A good starting
point for those not well versed in pharmacology and the range of poten-
tial mechanisms of action and interaction can be found in Goodman and
Gilman’s The Pharmacological Basis of Therapeutics (Brunton, Lazo, and
Parker, eds., 11th ed., McGraw-Hill, 2006).
In this essential and rapidly changing field, it is hoped that this first
volume on the subject will answer many questions and add clarity to
existing requirements.
References
Martin, L.I., Horvath, C.J., and Wyand, M.S., Safety pharmacology screening:
Practical problems in drug development, Int. J. Toxicol., 1997, 16:41–65.
Matsuzawa, T., et al., Current status of conducting function tests in repeated dose
toxicity studies in Japan, J. Toxicol. Sci., 1997, 22:374–382.
Sjouist, F., Drug-related hospital admissions, Ann. Pharmacol., 2000, 34:832–839.
Zbinden, G., Neglect of function and obsession with structure in toxicity testing.
In: Proc. 9th Int. Cong. Pharmacol., vol. 1, New York: Macmillan, 1984, pp.
43–49.
Zbinden, G., The significance of pharmacologic screening tests in the preclinical
safety evaluation of new drugs, J. New Drugs, 1966, 6:1–7.
About the Author
Shayne C. Gad, B.S. (Whittier College, California, chemistry and biol-
ogy, 1970) and Ph.D. in pharmacology/toxicology (University of Texas
at Austin, 1977), Diplomate of the American Board of Toxicologists,
Academy of Toxicological Sciences, is the principal of Gad Consulting
Services, a 19-year-old consulting firm with six employees and more
than 500 clients (including 120 pharmaceutical companies in the United
States and 50 overseas). Prior to this, he served in director-level and above
positions at Searle, Synergen, and Becton Dickinson. He has authored
or edited more than 44 published books and more than 360 chapters,
articles, and abstracts in the fields of toxicology, statistics, pharmacol-
ogy, drug and medical device development, and safety assessment. He
has more than 34 years of broad-based experience in toxicology, drug
and device development, statistics, and risk assessment. He has specific
expertise in neurotoxicology, in vitro methods, cardiovascular toxicol-
ogy, inhalation toxicology, immunotoxicology, and genotoxicology. He is
a past president of the American College of Toxicology, the Roundtable
of Toxicology Consultants, and three of Society of Toxicology’s specialty
sections and is a recipient of the American College of Toxicology Lifetime
Contribution Award. He has had direct involvement in the preparation
of investigational device exemptions (INDs) (95 successfully to date), new
drug applications (NDAs), product license applications (PLAs), abbre-
viated new drug applications (ANDAs), 510(k), investigational device
exemptions (IDEs), comparative toxicogenomics database (CTD), product
marketing applications (PMAs) and clinical data bases for Phase 1 and
2 studies. He has consulted for the FDA, U.S. Environmental Protection
Agency (EPA), and National Institutes of Health (NIH) and has trained
reviewers and been an expert witness for the FDA. He has also conducted
the triennial toxicology salary survey as a service to the profession for the
past 22 years.
xv
chapter one
Safety pharmacology
Background, history,
issues, and concerns
Although our current approach to nonclinical evaluation of the safety of
drugs generally serves us well in identifying potential adverse effects of
drugs before they go into man, almost all drugs in clinical use have side
effects, serious and not so serious, that are only identified after they are
in clinical use.
The adverse drug reactions which the standard tox-
icological test procedures do not aspire to recognize
include most of the functional side-effects. Clinical
experience indicates, however, that these are much
more frequent than the toxic reactions due to mor-
phological and biochemical lesions [1].
Safety pharmacology is the evaluation and study of the pharmacologic
effects of a potential drug (or excipient) that are unrelated to the desired
therapeutic effect and therefore may present a hazard—particularly
in individuals who already have one or more compromised or limited
organ system functions. Unlike other nonclinical evaluations of the safety
of a drug, these evaluations are usually conducted at doses close to the
intended clinical dose.
Such pharmacologically based adverse effects have not figured in
Phase I (first in man) volunteer deaths for some years. While such volun-
teer deaths are very rare (only two have been reported in the United States
since 1990), severe adverse events are not.
In the United States, from 1954 until 1980, 7000 healthy volunteers
participated in Phase I studies with only one death (in 1980, not due to
adherence to study participation) [2]. Additionally, in the United States
there was one other reported volunteer death in 2001 [3].
In a typical 12-month period, surveyed in Great Britain, there were no
deaths in 8163 healthy volunteers, with the three reported severe adverse
events being a severe dermatitis, an anaphylactic shock, and perforation of
an ulcer [4]. And in a Phase I center in France, among 1015 healthy young
