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A Pharmacology Primer, 6e
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ACADEMIC
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A Pharmacology Primer
Techniques for More Effective and Strategic
Drug Discovery
Sixth Edition

Terry P. Kenakin
Professor of Pharmacology
The University of North Carolina School of Medicine
Chapel Hill, NC, United States
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 Dedication

As always . for Debbie.

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Contents
Preface to sixth edition xiii
2.6.3 Differences in receptor density 35
2.6.4 Target-mediated trafficking of
stimulus 37
1. What is pharmacology? 2.7 Receptor desensitization and
1.1 About this book 1 tachyphylaxis 37
1.2 What is pharmacology? 1 2.8 The measurement of drug activity 40
1.3 The receptor concept 3 2.9 Advantages and disadvantages of
1.4 Pharmacological test systems 4 different assay formats 40
1.5 The nature of drug receptors 7 2.10 Drug concentration as an independent
1.6 From the snapshot to the movie 7 variable 41
1.7 Pharmacological intervention and the 2.10.1 Dissimulation in drug
therapeutic landscape 8 concentration 41
1.8 System-independent drug parameters: 2.10.2 Free concentration of drug 43
affinity and efficacy 11 2.11 Chapter summary and conclusions 43
1.9 What is affinity? 13 2.12 Derivations 43
1.10 The Langmuir adsorption isotherm 14 2.12.1 Series hyperbolae can be
1.11 What is efficacy? 15 modeled by a single
1.12 Doseeresponse curves 17 hyperbolic function 44
1.12.1 Potency and maximal response 18 2.12.2 Successive rectangular hyper-
1.12.2 P-scales and the representation bolic equations necessarily
of potency 18 lead to amplification 44
1.13 Chapter summary and conclusions 20 2.12.3 Saturation of any step in a
1.14 Derivations: conformational selection stimulus cascade by two
as a mechanism of efficacy 20 agonists leads to identical
References 21 maximal final responses for
the two agonists 44
2. How different tissues process drug 2.12.4 Procedure to measure free
response drug concentration in the
receptor compartment 45
2.1 The ‘eyes to see’: pharmacologic assays 23 References 45
2.2 The biochemical nature of stimuluse
response cascades 25 3. Drugereceptor theory
2.3 The mathematical approximation of
stimuluseresponse mechanisms 27 3.1 About this chapter 47
2.4 Influence of stimuluseresponse 3.2 Drugereceptor theory 47
cascades on doseeresponse curve 3.3 The use of mathematical models in
slopes 29 pharmacology 48
2.5 System effects on agonist response: 3.4 Some specific uses of models in
full and partial agonists 30 pharmacology 49
2.6 Differential cellular response to 3.5 Mass action building blocks 55
receptor stimulus 33 3.6 Classical model of receptor
2.6.1 Choice of response pathway 33 function 56
2.6.2 Augmentation or modulation of 3.7 The operational model of receptor
stimulus pathway 34 function 57

vii
viii Contents

3.8 Two-state theory 58 4.7.3 Displacement of a radioligand

3.9 The ternary complex model 59 by an allosteric antagonist 92
3.10 The extended ternary complex model 59 4.7.4 Relationship between IC50 and
3.11 Constitutive receptor activity and KI for competitive antagonists 93
inverse agonism 60 4.7.5 Maximal inhibition of binding
3.12 The cubic ternary complex model 62 by an allosteric antagonist 94
3.13 Multistate receptor models and 4.7.6 Relationship between IC50 and
probabilistic theory 63 KI for allosteric antagonists 94
3.14 Chapter summary and conclusions 65 4.7.7 Two-stage binding reactions 94
3.15 Derivations 65 4.7.8 Effect of G-Protein coupling on
3.15.1 Radioligand binding to observed agonist affinity 94
receptor dimers demonstrating 4.7.9 Effect of excess receptor in
cooperative behavior 65 binding experiments: saturation
3.15.2 Effect of variation in an HIV-1 binding curve 94
binding model 66 4.7.10 Effect of excess receptor in
3.15.3 Derivation of the operational binding experiments: displace-
model 67 ment experiments 95
3.15.4 Operational model forcing 4.7.11 Derivation of an allosteric bind-
function for variable slope 67 ing model 95
3.15.5 Derivation of two-state theory 68 References 96
3.15.6 Derivation of the extended
ternary complex model 68 5. Drug targets and drug-target
3.15.7 Dependence of constitutive molecules
activity on receptor density 69
3.15.8 Derivation of the cubic ternary 5.1 Defining biological targets 97
complex model 69 5.2 Specific types of drug targets 100
References 69 5.2.1 G-protein-coupled receptors 100
5.2.2 Ion channels 102
5.2.3 Enzymes 103
4. Pharmacological assay formats:
5.2.4 Nuclear receptors 111
binding
5.2.5 Nucleotide-based drug targets 112
4.1 The structure of this chapter 71 5.3 Small drug-like molecules 114
4.2 Binding theory and experiment 71 5.3.1 Hybrid molecules 116
4.2.1 Saturation binding 74 5.3.2 Chemical sources for potential
4.2.2 Displacement binding 76 drugs 121
4.2.3 Kinetic binding studies 79 5.4 Biologics 126
4.3 Complex binding phenomena: agonist 5.4.1 Replacement proteins 127
affinity from binding curves 80 5.4.2 Eliminating ‘undruggable’ pro-
4.4 Experimental prerequisites for correct teins through PROTACs 130
application of binding techniques 84 5.4.3 Peptides 131
4.4.1 The effect of protein concentra- 5.4.4 Antibodies 135
tion on binding curves 84 5.4.5 Immunotherapy 141
4.4.2 The importance of equilibration 5.4.6 Vaccines 141
time for equilibrium between 5.4.7 Nucleic acidebased drug species 142
two ligands 85 5.5 Summary and conclusions 147
4.5 Binding in allosteric systems 87 References 147
4.6 Chapter summary and conclusions 91 Further reading 149
4.7 Derivations 92
4.7.1 Displacement binding: 6. Agonists: the measurement of affinity
competitive interaction 92 and efficacy in functional assays
4.7.2 Displacement binding:
noncompetitive interaction 92 6.1 Functional pharmacological
experiments 151
 Contents ix

6.2 The choice of functional assays 152 7.3.5 Analyses for partial agonists 201
6.3 Recombinant functional systems 156 7.3.6 The method of Lew and Angus:
6.4 Functional experiments: dissimulation nonlinear regression analysis 203
in time 159 7.4 Noncompetitive antagonism 204
6.5 Experiments in real time versus 7.5 Agonisteantagonist hemiequilibria 208
stop-time 160 7.6 Resultant analysis 210
6.6 Quantifying agonism: the BlackeLeff 7.7 Antagonism in vivo 210
operational model of agonism 162 7.7.1 Antagonists with efficacy in
6.6.1 Affinity-dependent versus vivo 212
efficacy-dependent agonist 7.7.2 Kinetics of target coverage 214
potency 166 7.7.3 Kinetics of dissociation 216
6.6.2 Secondary and tertiary testing 7.7.4 Estimating antagonist
of agonists 168 dissociation with hemiequilibria 219
6.7 Biased signaling 169 7.8 Blockade of indirectly acting agonists 219
6.7.1 Receptor selectivity 175 7.9 Irreversible antagonism 220
6.8 Null analyses of agonism 175 7.10 Chemical antagonism 222
6.8.1 Partial agonists 175 7.11 Chapter summary and conclusions 226
6.8.2 Full agonists 179 7.12 Derivations 227
6.9 Comparing full and partial agonist 7.12.1 Derivation of the Gaddum
activities: Log(max/EC50) 182 equation for competitive
6.10 Chapter summary and conclusions 183 antagonism 227
6.11 Derivations 183 7.12.2 Derivation of the Gaddum
6.11.1 Relationship between the EC50 equation for noncompetitive
and affinity of agonists 183 antagonism 227
6.11.2 Method of Barlow, Scott, and 7.12.3 Derivation of the schild
Stephenson for affinity of equation 228
partial agonists 184 7.12.4 Functional effects of an
6.11.3 Maximal response of a partial inverse agonist with the
agonist is dependent on operational model 228
efficacy 184 7.12.5 pA2 measurement for inverse
6.11.4 System independence of full agonists 228
agonist potency ratios 184 7.12.6 Functional effects of a partial
6.11.5 Measurement of agonist agonist with the operational
affinity: method of Furchgott 184 model 229
6.11.6 Agonism as a positive 7.12.7 pA2 measurements for partial
allosteric modulation of agonists 229
receptoresignaling protein 7.12.8 Method of Stephenson for
interaction to derive partial agonist affinity
DLog(max/EC50) ratios 185 measurement 229
References 187 7.12.9 Derivation of the Method of
Gaddum for noncompetitive
7. Orthosteric drug antagonism antagonism 230
7.12.10 Relationship of pA2 and pKB
7.1 Introduction 189
for insurmountable
7.2 Kinetics of drugereceptor interaction 189
orthosteric antagonism 230
7.3 Surmountable competitive antagonism 192
7.12.11 Resultant analysis 230
7.3.1 Schild analysis 192
7.12.12 Blockade of indirectly acting
7.3.2 Patterns of DoseeResponse
agonists 231
curves that preclude schild
7.12.13 Chemical antagonism:
analysis 197
abstraction of agonist
7.3.3 Best practice for the use of
concentration 231
schild analysis 198
7.12.14 Chemical antagonism:
7.3.4 Analyses for inverse agonists in
abstraction of antagonist
constitutively active receptor
concentration 231
systems 199
References 232
x Contents

8. Allosteric modulation 9.5.1 Predicting agonism 299

9.5.2 Predicting binding 301
8.1 Introduction 233 9.5.3 Drug combinations in vivo 302
8.2 The nature of receptor allosterism 233 9.6 Summary and conclusions 303
8.3 Unique effects of allosteric modulators 235 9.7 Derivations 304
8.4 Functional study of allosteric modulators 240 9.7.1 IC50 Correction Factors:
8.4.1 Phenotypic allosteric modulation competitive antagonists 304
profiles 242 9.7.2 Relationship of pA2 and pKB for
8.4.2 Allosteric agonism 243 Insurmountable Orthosteric
8.4.3 Affinity of allosteric modulators 243 antagonism 304
8.4.4 Negative allosteric modulators 246 9.7.3 Relationship of pA2 and pKB for
8.4.5 Positive allosteric modulators 250 Insurmountable Allosteric
8.4.6 Quantifying PAM activity in vivo 254 Antagonism 305
8.4.7 NAM/PAM induced agonist bias 255 References 305
8.4.8 Optimal assays for allosteric
function 255
10. Pharmacokinetics
8.5 Functional allosteric model with
constitutive activity 256 10.1 Introduction 307
8.6 Internal checks for adherence to the 10.2 Biopharmaceutics 307
allosteric model 257 10.3 The chemistry of “drug-like”
8.7 Methods for detecting allosterism 260 character 308
8.8 Chapter summary and conclusions 262 10.4 Pharmacokinetics 313
8.9 Derivations 262 10.4.1 Drug absorption 313
8.9.1 Allosteric model of receptor 10.4.2 Route of drug
activity 262 administration 319
8.9.2 Effects of allosteric ligands on 10.4.3 General pharmacokinetics 322
response: changing efficacy 263 10.4.4 Metabolism 325
8.9.3 Schild analysis for allosteric 10.4.5 Clearance 330
antagonists 263 10.4.6 Volume of distribution and
8.9.4 Application of Log(Max/R50) half-life 332
values from R50 curves to 10.4.7 Renal clearance 338
quantify the effects of PAMs 264 10.4.8 Bioavailability 340
8.9.5 Quantifying allosterically 10.5 Nonlinear pharmacokinetics 342
mediated induced bias in 10.6 Multiple dosing 343
agonism 264 10.7 Modifying pharmacokinetics through
8.9.6 Functional allosteric model with medicinal chemistry 345
constitutive receptor activity 265 10.8 Practical pharmacokinetics 347
References 266 10.8.1 Allometric scaling 349
10.9 Placement of pharmacokinetic assays
9. The optimal design of in discovery and development 350
pharmacological experiments 10.10 The pharmacokinetics of biologics 352
10.10.1 Absorption 353
9.1 Introduction 269 10.10.2 Duration of action 354
9.2 The optimal design of pharmacological 10.10.3 Antibody PK 355
experiments 269 10.10.4 mRNA PK 355
9.2.1 Drug efficacy 270 10.11 Summary and conclusions 355
9.2.2 Affinity 283 References 356
9.2.3 Orthosteric versus allosteric
mechanisms 292 11. Safety pharmacology
9.3 Null experiments and fitting data to
models 293 11.1 Safety pharmacology 359
9.4 Interpretation of experimental data 296 11.2 Hepatotoxicity 365
9.5 Predicting therapeutic activity in all 11.2.1 Drugedrug interactions 365
systems 299 11.2.2 Direct hepatotoxicity 370
 Contents xi

11.2.3 Hepatotoxicity in context in 13.2.3 Method of Furchgott for the

vivo 372 measurement of the affinity
11.3 Cytotoxicity 372 of a full agonist 428
11.4 Mutagenicity 374 13.2.4 Schild analysis for the
11.5 hERG activity and Torsades de measurement of competitive
Pointes 376 antagonist affinity 429
11.6 Autonomic receptor profiling and 13.2.5 Method of Stephenson for
off-target effects 376 measurement of partial
11.7 General pharmacology 377 agonist affinity 431
11.8 Clinical testing and drug toxicity 379 13.2.6 Method of Gaddum for
11.9 Summary and conclusions 381 measurement of noncom-
References 381 petitive antagonist affinity 433
13.2.7 Method for estimating affin-
12. The drug-discovery process ity of insurmountable antag-
onist (dextral displacement
12.1 Some challenges for modern drug
observed) 434
discovery 383
13.2.8 Resultant analysis for
12.2 The drug-discovery process 384
measurement of affinity of
12.3 Target-based drug discovery 384
competitive antagonists
12.3.1 Target validation and the
with multiple properties 436
use of chemical tools 385
13.2.9 Measurement of the affinity
12.3.2 Recombinant systems 388
and maximal allosteric
12.4 Systems-based drug discovery 390
constant for allosteric mod-
12.5 High-throughput screening 393
ulators producing surmount-
12.5.1 Structure-based drug design
able effects 436
and virtual screening 404
13.2.10 Method for estimating
12.5.2 Phenotypic screening 405
affinity of insurmountable
12.6 The lead optimization process 409
antagonist (no dextral
12.7 Drug effectiveness 413
displacement observed):
12.7.1 Clinical testing 414
detection of allosteric effect 438
12.7.2 Determining detailed profiles
13.2.11 Measurement of pKB for
of candidate efficacy 416
competitive antagonists
12.7.3 Assays in context 417
from a pIC50 441
12.7.4 Characterization of candidate
13.2.12 Statistical assessment of
efficacies 418
selectivity 442
12.8 Summary and conclusions 419
13.2.13 Measurement of surmount-
References 420
able allosteric antagonism 447
Further reading 422
13.2.14 Measurement of
insurmountable allosteric
13. Selected pharmacological methods antagonism (second
13.1 Binding experiments 423 method) 448
13.1.1 Saturation binding 423 13.2.15 Measurement of PAM
13.1.2 Displacement binding 423 activity 450
13.2 Functional assays 426 Reference 451
13.2.1 Determination of equiactive
concentrations on Dosee
Response curves 426
13.2.2 Method of Barlow, Scott,
Appendix 1: Statistics 453
and Stephenson for
measurement of the affinity Index 483
of a partial agonist 427
This page intentionally left blank
Preface to sixth edition
Pharmacologists almost always are working in systems
they do not fully understand. This has engendered a unique
“null system” of comparisons (before and after drug) that
has sustained the field. Our view of what is actually
happening in our experiment is obtained through our assay,
and as the Nobel Laureate Sir James Black wrote “.The
prismatic qualities of the assay distort our view in obscure
ways and degrees.” (1993; Nobel Lectures: Physiology
and Medicine). What this means to the discipline is that it is
uniquely dependent upon technology unveiling what we do
not understand about physiology and as technology ad-
vances the frontier of understanding, so too does the
perception of pharmacological mechanisms and the effect
of drugs on physiology. In essence, as the acuity of the
pharmacological prism improves, so too does our under-
standing of drug mechanisms. The practical outcome of this
is that a book on pharmacology must be updated every few
years to keep up with the new understanding gained from
technologies “new eyes to see.” This volume has been
updated and has added major chapters on biologics and the
drug discovery process that reflects the changing landscape
of drug therapy as well as views of historical findings
modified by new knowledge.
Terry P. Kenakin Ph.D.
Professor of Pharmacology,
The University of North Carolina School of Medicine,
Chapel Hill, NC, United States
xiii
This page intentionally left blank
Chapter 1
What is pharmacology?
I would in particular draw the attention to physiologists to
this type of physiological analysis of organic systems which
can be done with the aid of toxic agents ..
dClaude Bernard (1813e78).
1.1 About this book
Essentially this is a book about the methods and tools used in
pharmacology to quantify drug activity. Receptor pharma-
cology is based on the comparison of experimental data and
simple mathematical models, with a resulting inference of
drug behavior to the molecular properties of drugs. From this
standpoint, a certain level of understanding of the mathe-
matics involved in the models is useful but not imperative.
This book is structured such that each chapter begins with the
basic concepts and then moves on to the techniques used to
estimate drug parameters, and, finally, for those so inclined,
the mathematical derivations of the models used. Under-
standing the derivation is not a prerequisite for understanding
the application of the methods or the resulting conclusion;
these are included for completeness and are for readers who
wish to pursue exploration of the models. In general, facility
with mathematical equations is definitely not required for
pharmacology; the derivations can be ignored without any
detriment to the use of this book.
Second, the symbols used in the models and deriva-
tions, on occasion, duplicate each other (i.e., a is an
extremely popular symbol). However, the use of these
multiple symbols has been retained, since this preserves the
context of where these models were first described and
utilized. Also, changing these to make them unique would
cause confusion if these methods were to be used beyond
the framework of this book. Therefore, care should be taken
to consider the actual nomenclature of each chapter.
Third, an effort has been made to minimize the need to
cross-reference different parts of the book (i.e., when a
particular model is described, the basics are reiterated
somewhat to minimize the need to read the relevant but
different part of the book in which the model is initially
described). While this leads to a small amount of repeated
description, it is felt that this will allow for a more unin-
terrupted flow of reading and use of the book.
A Pharmacology Primer. https://doi.org/10.1016/B978-0-323-99289-3.00015-4
Copyright © 2022 Elsevier Inc. All rights reserved.
1.2 What is pharmacology?
Pharmacology (an amalgam of the Greek pharmakos,
medicine or drug, and logos, study) is a broad discipline
describing the use of chemicals to treat and cure diseases.
The Latin term pharmacologia was used in the late 1600s,
but the term pharmacum was used as early as the 4th
century to denote the term drug or medicine. In the Greek
translations “Pharmakeia” refers to Sorcery/Witchcraft
which no doubt was evident when particular herbal treat-
ments were effective. There are subdisciplines within
pharmacology representing specialty areas. Pharmacoki-
netics deals with the disposition of drugs in the human
body. To be useful, drugs must be absorbed and transported
to their site of therapeutic action. Drugs will be ineffective
in therapy if they do not reach the organs(s) to exert their
activity; this will be discussed specifically in Chapter 9,
Pharmacokinetics, of this book. Pharmaceutics is the study
of the chemical formulation of drugs to optimize absorption
and distribution within the body. Pharmacognosy is the
study of plant natural products and their use in the treat-
ment of disease. A very important discipline in the drug-
discovery process is medicinal chemistry, the study of the
production of molecules for therapeutic use. This couples
synthetic organic chemistry with an understanding of how
biological information can be quantified and used to guide
the synthetic chemistry to enhance therapeutic activity.
Pharmacodynamics is the study of the interaction of the
drug molecule with the biological target (referred to
generically as the “receptor,” vide infra). This discipline
lays the foundation of pharmacology since all therapeutic
application of drugs has a common root in pharmacody-
namics (i.e., as a prerequisite to exerting an effect, all drug
molecules must bind to and interact with receptors).
The history of pharmacology is tied to the history of
drug discoverydsee Chapter 9, The Optimal Design of
Pharmacological Experiments. As put by the Canadian
physician Sir William Osler (1849e919; the “father of
modern medicine”), “. the desire to take medicine is
perhaps the greatest feature which distinguishes man from
animals ..” Pharmacology as a separate science is
approximately 120e140 years old. The relationship be-
tween chemical structure and biological activity began to be
studied systematically in the 1860s [1]. It began when
1
2 A Pharmacology Primer
physiologists, using chemicals to probe physiological sys-
tems, became more interested in the chemical probes than
the systems they were probing. By the early 1800s, phys-
iologists were performing physiological studies with
chemicals that became pharmacological studies more aimed
at the definition of the biological activity of chemicals. The
first formalized chair of pharmacology, indicating a formal
university department, was founded in Estonia by Rudolf
Bucchiem in 1847. In North America, the first chair was
founded by John Jacob Abel at Johns Hopkins University
in 1890. A differentiation of physiology and pharmacology
was given by the pharmacologist Sir William Paton [2]:
If physiology is concerned with the function, anatomy with
the structure, and biochemistry with the chemistry of the
living body, then pharmacology is concerned with the
changes in function, structure, and chemical properties of
the body brought about by chemical substances
dW.D.M. Paton (1986).
Many works about pharmacology essentially deal in
therapeutics associated with different organ systems in the
body. Thus, in many pharmacology texts, chapters are
entitled drugs in the cardiovascular system, the effect of
drugs on the gastrointestinal (GI) system, the central ner-
vous system (CNS), and so on. However, the underlying
principles for all of these is the same, namely, the phar-
macodynamic interaction between the drug and the bio-
logical recognition system for that drug. Therefore, a
prerequisite to all of pharmacology is an understanding of
the basic concepts of doseeresponse and how living cells
process pharmacological information. This generally is
given the term pharmacodynamics or receptor pharma-
cology, where receptor is a term referring to any biological
recognition unit for drugs (membrane receptors, enzymes,
DNA, and so on). With such knowledge in hand, readers
will be able to apply these principles to any branch of
therapeutics effectively. This book treats doseeresponse data
generically and demonstrates methods by which drug ac-
tivity can be quantified across all biological systems irre-
spective of the nature of the biological target.
A great strength of pharmacology as a discipline is that
it contains the tools and methods to convert “descriptive
data,” i.e., data that serve to characterize the activity of a
given drug in a particular system, to “predictive data.” This
latter information can be used to predict that drug’s activity
in all organ systems, including the therapeutic one. This
defines the drug-discovery process which is the testing of
new potential drug molecules in surrogate systems (where a
potentially toxic chemical can do no lasting harm) before
progression to the next step, namely, testing in human
therapeutic systems. The models and tools contained in
pharmacology to convert drug behaviors in particular
organs to molecular properties (see Chapter 2: How
Different Tissues Process Drug Response) are the main
subject of this book, and the step-by-step design of phar-
macologic experiments to do this are described in detail in
Chapter 8, The Optimal Design of Pharmacological Ex-
periments (after the meaning of the particular parameters
and terms is described in previous chapters).
The human genome is now widely available for drug-
discovery research. Far from being a simple blueprint of
how drugs should be targeted, it has shown biologists that
receptor genotypes (i.e., properties of proteins resulting
from genetic transcription to their amino acid sequence) are
secondary to receptor phenotypes (how the protein interacts
with the myriad of cellular components and how cells tailor
the makeup and functions of these proteins to their indi-
vidual needs). Since the arrival of the human genome, re-
ceptor pharmacology as a science is more relevant than ever
in drug discovery. Current drug therapy is based on less
than 500 molecular targets, yet estimates utilizing the
number of genes involved in multifactorial diseases suggest
that the number of potential drug targets ranges from 2000
to 5000 [3]. Thus, current therapy is using only 5%e10%
of the potential trove of targets available in the human
genome.
A meaningful dialog between chemists and pharma-
cologists is the single most important element of the drug-
discovery process. The necessary link between medicinal
chemistry and pharmacology has been elucidated by
Paton [2]:
For pharmacology there results a particularly close rela-
tionship with chemistry, and the work may lead quite
naturally, with no special stress on practicality, to thera-
peutic application, or (in the case of adverse reactions) to
toxicology.
dW.D.M. Paton (1986).
Chemists and biologists reside in different worlds from
the standpoint of the type of data they deal with. Chemistry
is an exact science with physical scales that are not subject
to system variance. Thus, the scales of measurement are
transferable. Biology deals with the vagaries of complex
systems that are not completely understood. Within this
scenario, scales of measurement are much less constant and
much more subject to system conditions. Given this, a gap
can exist between chemists and biologists in terms of un-
derstanding and also in terms of the best method to progress
forward. In the worst circumstance, it is a gap of credibility
emanating from a failure of the biologist to make the
chemist understand the limits of the data. Usually, however,
credibility is not the issue, and the gap exists due to a lack
of common experience. This book was written in an
attempt to limit or, hopefully, eliminate this gap.
 What is pharmacology? Chapter | 1 3

1.3 The receptor concept
One of the most important concepts emerging from early
pharmacological studies is the concept of the receptor.
Pharmacologists knew that minute amounts of certain
chemicals had profound effects on physiological systems.
They also knew that very small changes in the chemical
composition of these substances could lead to huge dif-
ferences in activity. This led to the notion that something
on or in the cell must specifically read the chemical infor-
mation contained in these substances and translate it into a
physiological effect. This something was conceptually
referred to as the “receptor” for that substance. Pioneers
such as Paul Ehrlich (1854e915, Fig. 1.1A) proposed the
existence of “chemoreceptors” (actually he proposed a
collection of amboreceptors, triceptors, and polyceptors) on
cells for dyes. He also postulated that the chemoreceptors
on parasites, cancer cells, and microorganisms were
different from healthy host and thus could be exploited
therapeutically. The physiologist turned pharmacologist
John Newport Langley (1852e926, Fig. 1.1B), during his
studies with the drugs jaborandi (which contains the alka-
loid pilocarpine) and atropine, introduced the concept that
receptors were switches that received and generated signals
and that these switches could be activated or blocked by
specific molecules. The originator of quantitative receptor
theory, the Edinburgh pharmacologist Alfred Joseph Clark
(1885e941, Fig. 1.1C), was the first to suggest that the
data, compiled from his studies of the interactions of
acetylcholine and atropine, resulted from the unimolecular
interaction of the drug and a substance on the cell surface.
He articulated these ideas in the classic work The Mode of
Action of Drugs on Cells [4], later revised as the Handbook
of Experimental Pharmacology [5]. As put by Clark
It appears to the writer that the most important fact shown
by a study of drug antagonisms is that it is impossible to
explain the remarkable effects observed except by assuming
that drugs unite with receptors of a highly specific pattern
.. No other explanation will, however, explain a tithe of
the facts observed.
dA.J. Clark (1937).
Clark’s next step formed the basis of receptor theory by
applying chemical laws to systems of “infinitely greater
complexity” [4]. It is interesting to note the scientific at-
mosphere in which Clark published these ideas. The
dominant ideas between 1895 and 1930 were based on
theories such as the law of phasic variation essentially
stating that “certain phenomena occur frequently.” Ho-
meopathic theories like the ArndteSchulz law and
WebereFechner law were based on loose ideas around
surface tension of the cell membrane, but there was little
physicochemical basis for these ideas [6]. In this vein,
prominent pharmacologists of the day, such as Walter
Straub (1874e944), suggested that a general theory of
chemical binding between drugs and cells utilizing re-
ceptors was “. going too far . and . not admissible”
[6]. The impact of Clark’s thinking against these concepts
cannot be overemphasized to modern pharmacology.

FIGURE 1.1 Pioneers of pharmacology. (A) Paul Ehrlich (1854e915). Born in Silesia, Ehrlich graduated from Leipzig University to go on to a
distinguished career as head of institutes in Berlin and Frankfurt. His studies with dyes and bacteria formed the basis of early ideas regarding recognition
of biological substances by chemicals. (B) John Newport Langley (1852e926). Though he began reading mathematics and history in Cambridge in 1871,
Langley soon took to physiology. He succeeded the great physiologist M. Foster to the chair of physiology in Cambridge in 1903 and branched out into
pharmacological studies of the autonomic nervous system. These pursuits led to germinal theories of receptors. (C) Alfred J. Clark (1885e941). Beginning
as a demonstrator in pharmacology in King’s College (London), Clark went on to become Professor of pharmacology at University College London. From
there he took the chair of pharmacology in Edinburgh. Known as the originator of modern receptor theory, Clark applied chemical laws to biological
phenomena. His books on receptor theory formed the basis of modern pharmacology.
4 A Pharmacology Primer
It is possible to underestimate the enormous signifi-
cance of the receptor concept in pharmacology until it is
realized how relatively chaotic the study of drug effect
was before it was introduced. Specifically, consider the
myriad of physiological and pharmacological effects of
the hormone epinephrine in the body. As shown in
Fig. 1.2, a host of responses are obtained from the CNS,
cardiovascular system, smooth muscle, and other organs.
It is impossible to see a thread which relates these very
different responses until it is realized that all of these are
mediated by the activation of a single protein receptor,
namely, in this case, the b-adrenoceptor. When this is
understood, a much better idea can be gained as to how to
manipulate these heterogeneous responses for therapeutic
benefit; the receptor concept introduced order into physi-
ology and pharmacology.
Drug receptors can exist in many forms, including cell
surface proteins, enzymes, ion channels, membrane trans-
porters, DNA, and cytosolic proteins (see Fig. 1.3). There
are examples of important drugs for all of these. This book
deals with general concepts which can be applied to a range
of receptor types, but most of the principles are illustrated
with the most tractable receptor class known in the human
genome, namely, seven transmembrane (7TM) receptors
(7TMRs). These receptors are named for their characteristic
structure that consists of a single protein chain that tra-
verses the cell membrane seven times to produce extra-
cellular and intracellular loops. These receptors activate
G-proteins to elicit response, thus they are also
commonly referred to as G-protein-coupled receptors
(GPCRs); this should now be considered a limiting moniker
as these proteins signal to a wide variety of signaling
molecules in the cell and are not confined to G-protein
FIGURE 1.2 A sampling of the heterogeneous physiological and pharmacological response to the hormone epinephrine. The concept of receptors links
these diverse effects to a single control point, namely, the b-adrenoceptor.
effects. There are between 800 and 1000 [7] of these in
the genome [the genome sequence predicts 650 GPCR
genes, of which approximately 190 (on the order of 1% of
the genome of superior organisms) are categorized as
known 7TMRs [8] activated by some 70 ligands]. In the
United States, in 2000, nearly half of all prescription drugs
were targeted toward 7TM receptors [3]. These receptors,
comprising between 1% and 5% of the total cell protein,
control a myriad of physiological activities. They are
tractable for drug discovery because they are on the cell
surface, and therefore drugs do not need to penetrate the
cell to produce effect. In the study of biological targets such
as 7TMRs and other receptors, a “system” must be
employed that accepts chemical input and returns biological
output. It is worth discussing such receptor systems in
general terms before their specific uses are considered.
1.4 Pharmacological test systems
Molecular biology has transformed pharmacology and the
drug-discovery process. As little as 20 years ago, screening
for new drug entities was carried out in surrogate animal
tissues. This necessitated a rather large extrapolation to
span the differences in genotype and phenotype. The belief
that the gap could be bridged came from the notion that the
chemicals recognized by these receptors in both humans
and animals were the same (vide infra). Receptors are
unique proteins with characteristic amino acid sequences.
While polymorphisms (spontaneous alterations in amino
acid sequence, vide infra) of receptors exist in the same
species, in general the amino acid sequence of a natural
ligand-binding domain for a given receptor type largely
may be conserved. There are obvious pitfalls of using

FIGURE 1.3 Schematic diagram of potential drug targets. Molecules can affect the function of numerous cellular components both in the cytosol and on
the membrane surface. There are many families of receptors that traverse the cellular membrane and allow chemicals to communicate with the interior of
the cell.
surrogate species receptors for predicting human drug ac-
tivity, and it never can be known for certain whether
agreement for estimates of activity for a given set of drugs
ensures accurate prediction for all drugs. The agreement is
very much drug and receptor dependent. For example, the
human and mouse a2-adrenoceptors are 89% homologous
and thus considered very similar from the standpoint of
amino acid sequence. Furthermore, the affinities of the a2-
adrenoceptor antagonists atipamezole and yohimbine are
nearly indistinguishable (atipamezole human a2-C10
Ki ¼ 2.9  0.4 nM, mouse a2-4H Ki ¼ 1.6  0.2 nM;
yohimbine human a2-C10 Ki ¼ 3.4  0.1 nM, mouse a2-
4H Ki ¼ 3.8  0.8 nM). However, there is a 20.9-fold
difference for the antagonist prazosin (human a2-C10
Ki ¼ 2034  350 nM, mouse a2-4H Ki ¼ 97.3  0.7 nM)
[9]. Such data highlight a general theme in pharmacological
research, namely, that a hypothesis, such as one proposing
that two receptors which are identical with respect to their
sensitivity to drugs are the same, cannot be proven, only
disproven. While a considerable number of drugs could be
tested on the two receptors (thus supporting the hypothesis
that their sensitivity to all drugs is the same), this hypoth-
esis is immediately disproven by the first drug that shows
differential potency on the two receptors. The fact that a
series of drugs tested show identical potencies may mean
only that the wrong sample of drugs has been chosen to
unveil the difference. Thus, no general statements can be
made that any one surrogate system is completely predic-
tive of activity on the target human receptor. This will al-
ways be a drug-specific phenomenon.
The link between animal and human receptors is the fact
that both proteins recognize the endogenous transmitter
(e.g., acetylcholine, norepinephrine), and therefore the hope
What is pharmacology? Chapter | 1 5
is that this link will carry over into other drugs that
recognize the animal receptor. This imperfect system
formed the basis of drug discovery until human cDNA for
human receptors could be used to make cells express hu-
man receptors. These engineered (recombinant) systems are
now used as surrogate human-receptor systems, and the
leap of faith from animal receptor sequences to human-
receptor sequences is not required (i.e., the problem of
differences in genotype has been overcome). However,
cellular signaling is an extremely complex process and cells
tailor their receipt of chemical signals in numerous ways.
Therefore, the way a given receptor gene behaves in a
particular cell can differ in response to the surroundings in
which that receptor finds itself. These differences in
phenotype (i.e., properties of a receptor produced by
interaction with its environment) can result in differences in
both the quantity and quality of a signal produced by a
concentration of a given drug in different cells. Therefore,
there is still a certain, although somewhat lesser, leap of
faith taken in predicting therapeutic effects in human tis-
sues under pathological control from surrogate recombinant
or even surrogate natural human-receptor systems. For this
reason, it is a primary requisite of pharmacology to derive
system-independent estimates of drug activity that can be
used to predict therapeutic effect in other systems.
A schematic diagram of the various systems used in
drug discovery, in order of how appropriate they are to
therapeutic drug treatment, is shown in Fig. 1.4. As dis-
cussed previously, early functional experiments in animal
tissue have now largely given way to testing in recombinant
cell systems engineered with human-receptor material. This
huge technological step greatly improved the predictability
of drug activity in humans, but it should be noted that there
6 A Pharmacology Primer
FIGURE 1.4 A history of the drug-discovery process. Originally, the only biological material available for drug research was animal tissue. With the
advent of molecular biological techniques to clone and express human receptors in cells, recombinant systems supplanted animal-isolated tissue work. It
should be noted that these recombinant systems still fall short of yielding drug response in the target human tissue under the influence of pathological
processes.
still are many factors that intervene between the genetically
engineered drug-testing system and the pathology of human
disease.
A frequently used strategy in drug discovery is to ex-
press human receptors (through transfection with human
cDNA) in convenient surrogate host cells (referred to as
“target-based” drug discovery; see Chapter 10: Safety
Pharmacology for further discussion). These host cells are
chosen mainly for their technical properties (i.e., robust-
ness, growth rate, stability) and not with any knowledge of
verisimilitude to the therapeutically targeted human cell
type. There are various factors relevant to the choice of
surrogate host cell, such as a very low-background activity
(i.e., a cell cannot be used that already contains a related
animal receptor for fear of cross-reactivity to molecules
targeted for the human receptor). Human receptors are often
expressed in animal surrogate cells. The main idea here is
that the cell is a receptacle for the receptor, allowing it to
produce physiological responses, and that activity can be
monitored in pharmacological experiments. In this sense,
human receptors expressed in animal cells are still a theo-
retical step distanced from the human receptor in a human
cell type. However, even if a human surrogate is used (and
there are such cells available), there is no definitive evi-
dence that a surrogate human cell is any more predictive of
a natural receptor activity than an animal cell when
compared to the complex receptor behavior in its natural
host cell type expressed under pathological conditions.
Receptor phenotype dominates in the end organ, and the
exact differences between the genotypic behavior of the
receptor (resulting from the genetic makeup of the receptor)
and the phenotypic behavior of the receptor (due to the
interaction of the genetic product with the rest of the cell)
may be cell specific. Therefore, there is still a possible gap
between the surrogate systems used in the drug-discovery
process and the therapeutic application. Moreover, most
drug-discovery systems utilize receptors as switching
mechanisms and quantify whether drugs turn on or turn off
the switch. The pathological processes that we strive to
modify may be more subtle. As put by pharmacologist Sir
James Black [10]:
. angiogenesis, apoptosis, inflammation, commitment of
marrow stem cells, and immune responses. The cellular
reactions subsumed in these processes are switch like in
their behavior . biochemically we are learning that in all
these processes many chemical regulators seem to be
involved. From the literature on synergistic interactions, a
control model can be built in which no single agent is
effective. If a number of chemical messengers each bring
information from a different source and each deliver only a
subthreshold stimulus but together mutually potentiate each
other, then the desired information-rich switching can be
achieved with minimum risk of miscuing.
dJ.W. Black (1986).
Such complex end points are difficult to predict from
any one of the component processes leading to yet another
leap of faith in the drug-discovery process. For these rea-
sons, an emerging strategy for drug discovery is the use of
natural cellular systems. This approach is discussed in some
detail in Chapter 11, The Drug Discovery Process.
Even when an active drug molecule is found and ac-
tivity is verified in the therapeutic arena, there are factors
that can lead to gaps in its therapeutic profile. When drugs
are exposed to huge populations, genetic variations in this
population can lead to discovery of alleles that code for
mutations of the target (isogenes), and these can lead to
variation in drug response. Such polymorphisms can lead to

resistant populations (i.e., resistance of some asthmatics to
the b-adrenoceptor bronchodilators [11]). In the absence of
genetic knowledge, these therapeutic failures for a drug
could not easily be averted since they in essence occurred
because of the presence of new biological targets not
originally considered in the drug-discovery process. How-
ever, as new epidemiological information becomes avail-
able, these polymorphisms can now be incorporated into
the drug-discovery process.
There are two theoretical and practical scales that can
be used to make system-independent measures of drug
activity on biological systems. The first is a measure of the
attraction of a drug for a biological target, namely, its
affinity for a receptor. Drugs must interact with receptors
to produce an effect, and the affinity is a chemical term
used to quantify the strength of that interaction. The sec-
ond is much less straightforward and is used to quantify
the degree of effect imparted to the biological system after
the drug binds to the receptor. This is termed efficacy. This
property was named by Stephenson [12] within classical
receptor theory as a proportionality factor for the tissue
response produced by a drug. There is no absolute scale
for efficacy, but rather it is dealt with in relative terms
(i.e., the ratio of the efficacy of two different drugs on a
particular biological system can be estimated and, under
ideal circumstances, will transcend the system and be
applicable to other systems as well). It is the foremost task
of pharmacology to use the translations of drug effect
obtained from cells to provide system-independent esti-
mates of affinity and efficacy. Before specific discussion
of affinity and efficacy, it is worth considering the mo-
lecular nature of biological targets.
1.5 The nature of drug receptors
While some biological targets such as DNA are not protein
in nature, most receptors are. It is useful to consider the
properties of receptor proteins to provide a context for the
interaction of small molecule drugs with them. An impor-
tant property of receptors is that they have a 3D structure.
Proteins are usually composed of one or more peptide
chains; the composition of these chains makes up the pri-
mary and secondary structure of the protein. Proteins also
are described in terms of a tertiary structure, which defines
their shape in 3D space, and a quaternary structure, which
defines the molecular interactions between the various
components of the protein chains (Fig. 1.5). It is this 3D
structure which allows the protein to function as a recog-
nition site and effector for drugs and other components of
the cell; in essence, the ability of the protein to function as a
messenger, shuttling information from the outside world to
the cytosol of the cell. For 7TMRs, the 3D nature of the
receptor forms binding domains for other proteins such as
What is pharmacology? Chapter | 1 7
G-proteins (these are activated by the receptor and then go
on to activate enzymes and ion channels within the cell; see
Chapter 2: How Different Tissues Process Drug Response)
and endogenous chemicals such as neurotransmitters, hor-
mones, and autacoids that carry physiological messages.
This important class of drug target is named for a charac-
teristic structure consisting of 7TM domains looping into
the extracellular and intracellular spacedsee Fig. 1.6.
These molecules are the main transfer points of information
from the outside to the inside of the cell, and such transfers
occur through changes in the conformation of the receptor
protein (vide infra). For other receptors, such as ion chan-
nels and single transmembrane enzyme receptors, the
conformational change per se leads to a response, either
through an opening of a channel to allow the flow of ionic
current or the initiation of enzymatic activity. Therapeutic
advantage can be taken by designing small molecules to
utilize these binding domains or other 3D binding domains
on the receptor protein in order to modify physiological and
pathological processes.
1.6 From the snapshot to the movie
Drugs interact with living physiology and the outcome of
the interaction is controlled by a combination of the
intrinsic properties of the drug and the sensitivity of the
system to intervention. This being the case, drugs can have
different profiles of activity in different tissues depending
on the tissue sensitivity and setpoint of physiology.
Through the mechanics of mathematical models of drug
activity and the system-independent scales of drug activity
(i.e., affinity, efficacy), pharmacological procedures are
uniquely able to convert a single observation of drug ac-
tivity in a test system (the ‘snapshot’) to a prediction of the
complete realm of activities for that same drug in a range of
tissues of varying setpoints of physiology (the ‘movie’).
This is an essential property of pharmacology in drug dis-
covery as all initial evaluations of new drug activity are
made in isolated systems and assessments of what the new
molecule will do in other systems must be made. In
essence, the cellular host system completely controls what
the experimenter observes regarding the events taking place
at the drug receptor. Drug activity is thus revealed through
a “cellular veil” that can, in many cases, obscure or sub-
stantially modify drugereceptor activity (Fig. 1.7). Minute
signals, initiated either at the cell surface or within the
cytoplasm of the cell, are interpreted, transformed, ampli-
fied, and otherwise altered by the cell to tailor that signal to
its own particular needs. The application of pharmacolog-
ical principles and modeling enable ‘snapshots’ of drug
activity obtained is experiments to guide the progress of
molecules toward drug candidate statusdsee Chapter 3 for
further details.
8 A Pharmacology Primer
FIGURE 1.5 Increasing levels of protein structure. A protein has a given amino acid sequence to make peptide chains. These adopt a 3D structure
according to the free energy of the system. Receptor function can change with changes in tertiary or quaternary structure.
1.7 Pharmacological intervention and
the therapeutic landscape
It is useful to consider the therapeutic landscape with
respect to the aims of pharmacology. As stated by Sir
William Ossler (1849e919) “. the prime distinction be-
tween man and other creatures is man’s yearning to take
medicine.” The notion that drugs can be used to cure dis-
ease is as old as history. One of the first written records of
actual “prescriptions” can be found in the Ebers Papyrus
(c.1550 BCE): “. for night blindness in the eyes . liver
of ox, roasted and crushed out . really excellent!“dsee
Fig. 1.8. Now it is known that liver is an excellent source of
vitamin A, a prime treatment for night blindness, but that
chemical detail was not known to the ancient Egyptians.
Disease can be considered under two broad categories:
those caused by invaders such as pathogens and those
caused by intrinsic breakdown of normal physiological
function. The first generally is approached through the
invader (i.e., the pathogen is destroyed, neutralized, or
removed from the body). The one exception of where the
host is treated when an invader is present is the treatment of
HIV-1 infection leading to AIDS. In this case, while there
are treatments to neutralize the pathogen, such as anti-
retrovirals to block viral replication, a major new approach
is the blockade of the interaction of the virus with the
protein that mediates viral entry into healthy cells, the
chemokine receptor CCR5. In this case, CCR5 antagonists
are used to prevent HIV fusion and subsequent infection.
The second approach to disease requires an understanding
of the pathological process and repair of the damage to
return to normal function.
The therapeutic landscape onto which drug discovery
and pharmacology in general combat disease can gener-
ally be described in terms of the major organ systems of
the body and how they may go awry. A healthy cardio-
vascular system consists of a heart able to pump deoxy-
genated blood through the lungs and to pump oxygenated
blood throughout a circulatory system that does not
unduly resist blood flow. Since the heart requires a high

FIGURE 1.6 Depiction of the structure of seven transmembrane domain
receptors, one of the most if not the most important therapeutic targets
available in the human genome. Chemicals access the receptor through the
extracellular space by binding to the extracellular domains of the protein.
This causes a conformational change in the protein that alters the inter-
action of signaling proteins in the cell cytosol. This latter process results in
the initiation of cellular signaling.
degree of oxygen itself to function, myocardial ischemia
can be devastating to its function. Similarly, an inability to
maintain rhythm (arrhythmia) or loss in strength with
concomitant inability to empty (congestive heart failure)
can be fatal. The latter disease is exacerbated by elevated
arterial resistance (hypertension). A wide range of drugs
are used to treat the cardiovascular system, including
coronary vasodilators (nitrates), diuretics, renine
angiotensin inhibitors, vasodilators, cardiac glycosides,
calcium antagonists, beta and alpha blockers, antiar-
rhythmics, and drugs for dyslipidemia. The lungs must
extract oxygen from the air, deliver it to the blood, and
release carbon dioxide from the blood into exhaled air.
Asthma, chronic obstructive pulmonary disease (COPD),
and emphysema are serious disorders of the lungs and
airways. Bronchodilators (beta agonists), antiin-
flammatory drugs, inhaled glucocorticoids, anticholiner-
gics, and theophylline analogs are used for treatment of
these diseases. The CNS controls all conscious thought
and many unconscious body functions. Numerous dis-
eases of the brain can occur, including depression, anxi-
ety, epilepsy, mania, degeneration, obsessive disorders,
and schizophrenia. Brain functions such as those
What is pharmacology? Chapter | 1 9
controlling sedation and pain also may require treatment.
A wide range of drugs is used for CNS disorders,
including serotonin partial agonists and uptake inhibitors,
dopamine agonists, benzodiazepines, barbiturates, opi-
oids, tricyclics, neuroleptics, and hydantoins. The GI tract
receives and processes food to extract nutrients and
removes waste from the body. Diseases such as stomach
ulcers, colitis, diarrhea, nausea, and irritable bowel syn-
drome can affect this system. Histamine antagonists,
proton pump blockers, opioid agonists, antacids, and se-
rotonin uptake blockers are used to treat diseases of the GI
tract.
The inflammatory system is designed to recognize self
from nonself, and to destroy nonself to protect the body. In
diseases of the inflammatory system, the self-recognition
can break down, leading to conditions in which the body
destroys healthy tissue in a misguided attempt at protection.
This can lead to rheumatoid arthritis, allergies, pain, COPD,
asthma, fever, gout, graft rejection, and problems with
chemotherapy. Nonsteroidal antiinflammatory drugs,
aspirin and salicylates, leukotriene antagonists, and hista-
mine receptor antagonists are used to treat inflammatory
disorders. The endocrine system produces and secretes
hormones crucial to the body for growth and function.
Diseases of this class of organs can lead to growth and
pituitary defectsddiabetes; abnormality in thyroid, pitui-
tary, adrenal cortex, and androgen function; osteoporosis;
and alterations in estrogeneprogesterone balance. The
general approach to treatment is through replacement or
augmentation of secretion. Drugs used are replacement
hormones, insulin, sulfonylureas, adrenocortical steroids,
and oxytocin. In addition to the major organ and physio-
logical systems, diseases involving neurotransmission and
neuromuscular function, ophthalmology, hemopoiesis and
hematology, dermatology, immunosuppression, and drug
addiction and abuse are amenable to pharmacological
intervention.
Cancer is a serious malfunction of normal cell growth.
In the years from 1950 to 1970, the major approach to
treating this disease was to target DNA and DNA pre-
cursors according to the hypothesis that rapidly dividing
cells (cancer cells) are more susceptible to DNA toxicity
than normal cells. Since that time, a wide range of new
therapies based on manipulation of the immune system,
induction of differentiation, inhibition of angiogenesis, and
increased killer T-lymphocytes to decrease cell prolifera-
tion has greatly augmented the armamentarium against
neoplastic disease. Previously, lethal malignancies such as
testicular cancer, some lymphomas, and leukemia are now
curable.
Three general treatments of disease are surgery, genetic
engineering (still an emerging discipline), and pharmaco-
logical intervention. While early medicine was subject to
the theories of Hippocrates (460e357 BCE), who saw
10 A Pharmacology Primer

FIGURE 1.7 The cellular veil. Drugs act on biological receptors in cells to change cellular activity. The initial receptor stimulus usually alters a
complicated system of interconnected metabolic biochemical reactions, and the outcome of the drug effect is modified by the extent of these in-
terconnections, the basal state of the cell, and the threshold sensitivity of the various processes involved. This can lead to a variety of apparently different
effects for the same drug in different cells. Receptor pharmacology strives to identify the basic mechanism initiating these complex events.

FIGURE 1.8 The Ebers Papyrus is a 110-page scroll (20 m long) thought to have been written in 1550 BCE but containing information dating from
3400 BCE. It is a record of Egyptian medicine and contains numerous “prescriptions” some of which, though empirical, are valid therapeutic approaches
to diseases.

health and disease as a balance of four humors (i.e., black
and yellow bile, phlegm, and blood), by the 16th century
pharmacological concepts were being formulated. These
could be stated concisely as the following [13]:
l Every disease has a cause for which there is a specific
remedy.
l Each remedy has a unique essence that can be obtained
from nature by extraction (“doctrine of signatures”).
l The administration of the remedy is subject to a dosee
response relationship.
The basis for believing that pharmacological interven-
tion can be a major approach to the treatment of disease is
the fact that the body generally functions in response to
chemicals. Table 1.1 shows partial lists of hormones and
neurotransmitters in the body. Many more endogenous
chemicals are involved in normal physiological function.
 What is pharmacology? Chapter | 1 11

TABLE 1.1 Some endogenous chemicals controlling normal physiological function.

Neurotransmitters
Acetylcholine 2-Arachidonylglycerol Anandamide
ATP Corticotropin-releasing hormone Dopamine
Epinephrine Aspartate Gamma-aminobutyric acid
Galanin Glutamate Glycine
Histamine Norepinephrine Serotonin
Hormones
Thyroid-stimulating hormone Follicle-stimulating hormone Luteinizing hormone
Prolactin Adrenocorticotropin Antidiuretic hormone
Thyrotropin-releasing hormone Oxytocin Gonadotropin-releasing hormone
Growth-hormone-releasing hormone Corticotropin-releasing hormone Somatostatin
Melatonin Thyroxin Calcitonin
Parathyroid hormone Glucocorticoid(s) Mineralocorticoid(s)
Estrogen(s) Progesterone Chorionic gonadotropin
Androgens Insulin Glucagon
Amylin Erythropoietin Calcitriol
Calciferol Atrial-natriuretic peptide Gastrin
Secretin Cholecystokinin Neuropeptide Y
Insulin-like growth factor Angiotensinogen Ghrelin

Leptin

ATP, adenosine triphosphate.

The fact that so many physiological processes are
controlled by chemicals provides the opportunity for
chemical intervention. Thus, physiological signals medi-
ated by chemicals can be initiated, negated, augmented, or
modulated. The nature of this modification can take the
form of changes in the type, strength, duration, or location
of signal.
1.8 System-independent drug
parameters: affinity and efficacy
The process of drug discovery relies on the testing of mol-
ecules in systems to yield estimates of biological activity in
an iterative process of changing the structure of the molecule
until optimal activity is achieved. It will be seen in this book
that there are numerous systems available to do this, and that
each system may interpret the activity of molecules in
different ways. Some of these interpretations can appear to
be in conflict with each other, leading to apparent capricious
patterns. For this reason, the way forward in the drug
development process is to use only system-independent in-
formation. Ideally, scales of biological activity should be
used that transcend the actual biological system in which the
drug is tested. This is essential to avoid confusion and also
because it is quite rare to have access to the exact human
system under the control of the appropriate pathology
available for in vitro testing. Therefore, the drug-discovery
process necessarily relies on the testing of molecules in
surrogate systems and the extrapolation of the observed ac-
tivity to all systems. The only means to do this is to obtain
system-independent measures of drug activity, namely, af-
finity and efficacy.
If a molecule in solution associates closely with a re-
ceptor protein, it has affinity for that protein. The area where
it is bound is the binding domain or locus. If the same
molecule interferes with the binding of a physiologically
active molecule such as a hormone or a neurotransmitter
(i.e., if the binding of the molecule precludes activity of the
physiologically active hormone or neurotransmitter), the
molecule is referred to as an antagonist. Therefore, a phar-
macologically active molecule that blocks physiological ef-
fect is an antagonist. Similarly, if a molecule binds to a
receptor and produces its own effect, it is termed an agonist.
It also is assumed to have the property of efficacy. Efficacy is
12 A Pharmacology Primer

detected by observation of pharmacological response.
Therefore, agonists have both affinity and efficacy.
Classically, agonist response is described in two stages,
the first being the initial signal imparted to the immediate
biological target, namely, the receptor. This first stage is
composed of the formation, either through interaction with
an agonist or spontaneously, of an active state receptor
conformation. This initial signal is termed the stimulus
(Fig. 1.9). This stimulus is perceived by the cell and pro-
cessed in various ways through successions of biochemical
reactions to the end point, namely, the response. The sum
total of the subsequent reactions is referred to as the
stimuluseresponse mechanism or cascade (see Fig. 1.10).
Efficacy is a molecule-related property (i.e., different
molecules have different capabilities to induce a physio-
logical response). The actual term for the molecular aspect
of response-inducing capacity of a molecule is intrinsic
efficacy (see Chapter 3: DrugeReceptor Theory for how
this term evolved). Thus, every molecule has a unique
value for its intrinsic efficacy (in cases of antagonists this
could be zero). The different abilities of molecules to
induce response are illustrated in Fig. 1.10. This figure
shows doseeresponse curves for four 5-HT (hydroxytryp-
tamine) (serotonin) agonists in rat jugular vein. It can be
seen that if response is plotted as a function of the percent
receptor occupancy, different receptor occupancies for
the different agonists lead to different levels of response.
For example, while 0.6 g force can be generated by
5-HT by occupying 30% of the receptors, the agonist 5-
cyanotryptamine requires twice the receptor occupancy to
generate the same response (i.e., the capability of 5-
cyanotryptamine to induce response is half that of 5-HT
[14]). These agonists are then said to possess different
magnitudes of intrinsic efficacy.

FIGURE 1.9 Schematic diagram of response production by an agonist. An initial stimulus is produced at the receptor as a result of agonistereceptor
interaction. This stimulus is processed by the stimuluseresponse apparatus of the cell into observable cellular response.

FIGURE 1.10 Differences between agonists producing contraction of rat jugular vein through activation of 5-HT receptors. (A) Doseeresponse curves
to 5-HT receptor agonists, 5-HT (filled circles), 5-cyanotryptamine (filled squares), N,N-dimethyltryptamine (open circles), and N-benzyl-5-
methoxytryptamine (filled triangles). Abscissae: logarithms of molar concentrations of agonist. (B) Occupancy response curves for curves shown in
panel A. Abscissae: percent receptor occupancy by the agonist as calculated by mass action and the equilibrium dissociation constant of the agoniste
receptor complex. Ordinates: force of contraction in g. Data drawn from P. Leff, G.R. Martin, J.M. Morse, Differences in agonist dissociation constant
estimates for 5-HT at 5-HT2-receptors: a problem of acute desensitization? Br. J. Pharmacol. 89 (1986) 493e499.

It is important to consider affinity and efficacy as
separately manipulatable properties. Thus, there are chem-
ical features of agonists that pertain especially to affinity
and other features that pertain to efficacy. Fig. 1.11 shows a
series of key chemical compounds made en route to the
histamine H2 receptor antagonist cimetidine (used for
healing gastric ulcers). The starting point for this discovery
program was the knowledge that histamine, a naturally
occurring autacoid, activates histamine H2 receptors in the
stomach to cause acid secretion. This constant acid secre-
tion is what prevents the healing of lesions and ulcers. The
task was then to design a molecule that would antagonize
the histamine receptors mediating acid secretion and pre-
vent histamine H2 receptor activation to allow the ulcers to
heal. This task was approached with the knowledge that
molecules, theoretically, could be made that retained or
even enhanced affinity but decreased the efficacy of hista-
mine (i.e., these were separate properties). As can be seen
in Fig. 1.11, molecules were consecutively synthesized
with reduced values of efficacy and enhanced affinity until
the target histamine H2 antagonist cimetidine was made.
This was a clear demonstration of the power of medicinal
chemistry to separately manipulate affinity and efficacy for
which, in part, the Nobel Prize in Medicine was awarded in
1988.
1.9 What is affinity?
The affinity of a drug for a receptor defines the strength of
interaction between the two species. The forces control-
ling the affinity of a drug for the receptor are thermody-
namic (enthalpy as changes in heat and entropy as changes
FIGURE 1.11 Key compounds synthesized to eliminate the efficacy (burgundy red) and enhance the affinity (green) of histamine for histamine H2
receptors to make cimetidine, one of the first histamine H2 antagonists of use in the treatment of peptic ulcers. Quotation from J.W. Black, A personal view
of pharmacology, Ann. Rev. Pharmacol. Toxicol. 36 (1996) 1e33.
What is pharmacology? Chapter | 1 13
in the state of disorder). The chemical forces between the
components of the drug and the receptor vary in impor-
tance in relation to the distance of the drug from the re-
ceptor’s binding surface. Thus, the strength of
electrostatic forces (attraction due to positive and negative
charges and/or complex interactions between polar
groups) varies as a function of the reciprocal of the dis-
tance between the drug and the receptor. Hydrogen
bonding (the sharing of a hydrogen atom between an
acidic and basic group) varies in strength as a function of
the fourth power of the reciprocal of the distance. Also
involved are van der Waals’ forces (weak attraction be-
tween polar and nonpolar molecules) and hydrophobic
bonds (interaction of nonpolar surfaces to avoid interac-
tion with water). The combination of all of these forces
causes the drug to reside in a certain position within the
protein-binding pocket. This is a position of minimal free
energy. It is important to note that drugs do not statically
reside in one uniform position. As thermal energy varies
in the system, drugs approach and dissociate from the
protein surface. This is an important concept in pharma-
cology as it sets the stage for competition between two
drugs for a single binding domain on the receptor protein.
The probability that a given molecule will be at the point
of minimal free energy within the protein-binding pocket
thus depends on the concentration of the drug available to
fuel the binding process and also the strength of the in-
teractions for the complementary regions in the binding
pocket (affinity). Affinity can be thought of as a force of
attraction and can be quantified with a very simple tool,
first used to study the adsorption of molecules onto a
surface, namely, the Langmuir adsorption isotherm.
14 A Pharmacology Primer
1.10 The Langmuir adsorption isotherm
Defined by the chemist Irving Langmuir (1881e957,
Fig. 1.12), the model for affinity is referred to as the
Langmuir adsorption isotherm. Langmuir, a chemist at
General Electric, was interested in the adsorption of mol-
ecules onto metal surfaces for the improvement of lighting
filaments. He reasoned that molecules had a characteristic
rate of diffusion toward a surface (referred to as conden-
sation and denoted a in his nomenclature) and also a
characteristic rate of dissociation (referred to as evapora-
tion and denoted as V1; see Fig. 1.12). He assumed that the
amount of surface that already has a molecule bound is not
available to bind another molecule. The surface area bound
by molecule is denoted q1, expressed as a fraction of
the total area. The amount of free area open for the binding
of molecule, expressed as a fraction of the total area, is
denoted as 1  q1. The rate of adsorption toward the sur-
face therefore is controlled by the concentration of drug in
the medium (denoted m in Langmuir’s nomenclature)
multiplied by the rate of condensation on the surface and
the amount of free area available for binding:
Rate of diffusion toward surface ¼ amð1  q1 Þ. (1.1)
The rate of evaporation is given by the intrinsic rate of
dissociation of bound molecules from the surface multi-
plied by the amount already bound:
Rate of evaporation ¼ V1 q1 .
Once equilibrium has been reached, the rate of
adsorption equals the rate of evaporation. Equating (1.1)
and (1.2) and rearranging yields
FIGURE 1.12 The Langmuir adsorption isotherm representing the binding of a molecule to a surface. Photo shows Irving Langmuir (1881e957), a
chemist interested in the adsorption of molecules to metal filaments for the production of light. Langmuir devised the simple equation still in use today for
quantifying the binding of molecules to surfaces. The equilibrium is described by condensation and evaporation to yield the fraction of surface bound (q1)
by a concentration m.
am
q1 ¼ . (1.3)
am þ V1
This is the Langmuir adsorption isotherm in its original
form. In pharmacological nomenclature, it is rewritten ac-
cording to the convention
½AR ½A
r¼ ¼ ; (1.4)
½Rt  ½A þ KA
where [AR] is the amount of complex formed between the
ligand and the receptor, and [Rt] is the total number of re-
ceptor sites. The ratio r refers to the fraction of maximal
binding by a molar concentration of drug [A] with an equi-
librium dissociation constant of KA. This latter term is the
ratio of the rate of offset (in Langmuir’s terms V1 and
referred to as k2 in receptor pharmacology) divided by
the rate of onset (in Langmuir’s terms a denoted k1 in re-
ceptor pharmacology).
It is amazing to note that complex processes such as
drugs binding to protein, activation of cells, and observa-
tion of syncytial cellular response should apparently so
closely follow a model based on these simple concepts.
This was not lost on A.J. Clark in his treatise on druge
receptor theory The Mode of Action of Drugs on Cells [4]:
It is an interesting and significant fact that the author in
1926 found that the quantitative relations between the con-
centration of acetylcholine and its action on muscle cells, an
(1.2) action the nature of which is wholly unknown, could be most
accurately expressed by the formulae devised by Langmuir
to express the adsorption of gases on metal filaments.
dA.J. Clark (1937).

The term KA is a concentration, and it quantifies af-
finity. Specifically, it is the concentration that binds to 50%
of the total receptor population [see Eq. (1.4) when [A] ¼
KA]. Therefore, the smaller is the KA, the higher is the
affinity. Affinity is the reciprocal of KA. For example, if
KA ¼ 108 M, then 108 M binds to 50% of the receptors.
If KA ¼ 104 M, a 10,000-fold higher concentration of the
drug is needed to bind to 50% of the receptors (i.e., it is of
lower affinity).
It is instructive to discuss affinity in terms of the
adsorption isotherm in the context of measuring the amount
of receptor bound for given concentrations of drug. Assume
that values of fractional receptor occupancy can be visu-
alized for various drug concentrations. The kinetics of such
binding is shown in Fig. 1.13. It can be seen that initially
the binding is rapid, in accordance with the fact that there
are many unbound sites for the drug to choose. As the sites
become occupied, there is a temporal reduction in binding
until a maximal value for that concentration is attained.
Fig. 1.13 also shows that the binding of higher concentra-
tions of drug is correspondingly increased. In keeping with
the fact that this is first-order binding kinetics (where the
rate is dependent on a rate constant multiplied by the
concentration of reactant), the time to equilibrium is shorter
for higher concentrations than for lower concentrations.
The various values for receptor occupancy at different
concentrations constitute a concentration binding curve
(shown in Fig. 1.14A). There are two areas in this curve of
particular interest to pharmacologists. The first is the
maximal asymptote for binding. This defines the maximal
number of receptive binding sites in the preparation. The
binding isotherm [Eq. (1.4)] defines the ordinate axis as the
fraction of the maximal binding. Thus, by definition, the
maximal value is unity. However, in experimental studies,
real values of capacity are used since the maximum is not
FIGURE 1.13 Time course for increasing concentrations of a ligand with a KA of 2 nM. Initially, the binding is rapid but slows as the sites become
occupied. The maximal binding increases with increasing concentrations as does the rate of binding.
What is pharmacology? Chapter | 1 15
known. When the complete curve is defined, the maximal
value of binding can be used to define fractional binding at
various concentrations and thus define the concentration at
which half-maximal binding (binding to 50% of the re-
ceptor population) occurs. This is the equilibrium dissoci-
ation constant of the drugereceptor complex (KA), the
important measure of drug affinity. This comes from the
other important region of the curve, namely, the midpoint.
It can be seen from Fig. 1.14A that graphical estimation of
both the maximal asymptote and the midpoint is difficult to
perform with the graph in the form shown. A much easier
format to present binding, or any concentrationeresponse
data, is a semilogarithmic form of the isotherm. This allows
better estimation of the maximal asymptote and places the
midpoint in a linear portion of the graph where intra-
polation can be done (see Fig. 1.14B). Doseeresponse
curves for binding are not often visualized, as they require a
means to detect bound (over unbound) drug. However, for
drugs that produce a pharmacological response (i.e., ago-
nists), a signal proportional to bound drug can be observed.
The true definition of a doseeresponse curve is the
observed in vivo effect of a drug given as a dose to a whole
animal or human. However, it has entered into the common
pharmacological jargon as a general depiction of drug and
effect. Thus, a doseeresponse curve for binding is actually
a binding concentration curve, and an in vitro effect of an
agonist in a receptor system is a concentrationeresponse
curve.
1.11 What is efficacy?
The property that gives a molecule the ability to change a
receptor, such that it produces a cellular response, is termed
efficacy. Early concepts of receptors likened them to locks
and keys. As stated by Paul Ehrlich,
16 A Pharmacology Primer
FIGURE 1.14 Doseeresponse relationship for ligand binding according to the Langmuir adsorption isotherm. (A) Fraction of maximal binding as a
function of concentration of agonist. (B) Semilogarithmic form of curve shown in panel A.
Substances can only be anchored at any particular part of
the organism if they fit into the molecule of the recipient
complex like a piece of mosaic finds its place in a pattern.
This historically useful but inaccurate view of receptor
function has in some ways hindered development models of
efficacy. Specifically, the lock-and-key model implies a
static system with no moving parts. However, one feature
of proteins is their malleability. While they have structure,
they do not have a single structure but rather many potential
shapes referred to as conformations. A protein stays in a
particular conformation because it is energetically favorable
to do so (i.e., there is minimal free energy for that
conformation). If thermal energy enters the system, the
protein may adopt another shape in response. Stated by
Linderstrom-Lang and Schellman [15]:
. a protein cannot be said to have “a” secondary struc-
ture but exists mainly as a group of structures not too
different from one another in free energy .. In fact, the
molecule must be conceived as trying every possible
structure ..
dLindstrom and Schellman (1959).
Not only are a number of conformations for a given
protein possible, but the protein samples these various
conformations constantly. It is a dynamic and not a static
entity. Receptor proteins can spontaneously change
conformation in response to variations in the energy of the
system. An important concept here is that small mole-
cules, by interacting with the receptor protein, can bias the
conformations that are sampled. It is in this way that drugs
can produce active effects on receptor proteins (i.e.,
demonstrate efficacy). A thermodynamic mechanism by
which this can occur is through what is known as
conformational selection [16]. A simple illustration can be
made by reducing the possible conformations of a given
receptor protein to just two. These will be referred to as
the “active” (denoted [Ra]) and “inactive” (denoted [Ri])
conformations.
Thermodynamically it would be expected that a ligand
may not have identical affinity for both receptor confor-
mations. This was an assumption in early formulations of
conformational selection. For example, differential affinity
for protein conformations was proposed for oxygen binding
to hemoglobin [17] and for choline derivatives and nico-
tinic receptors [18]. Furthermore, assume that these con-
formations exist in an equilibrium defined by an allosteric
constant L (defined as [Ra]/[Ri]) and that a ligand [A] has
affinity for both conformations defined by equilibrium as-
sociation constants Ka and aKa, respectively, for the inac-
tive and active states.
It can be shown that the ratio of the active species Ra in
the presence of a saturating concentration (rN) of the
ligand versus in the absence of the ligand (r0) is given by
the following (see Section 1.14):
rN að1 þ LÞ
¼ . (1.5)
r0 ð1 þ aLÞ
It can be seen that if the factor a is unity (i.e., the af-
finity of the ligand for Ra and Ri is equal [Ka ¼ aKa]), then
there will be no change in the amount of Ra when the ligand
is present. However, if a is not unity (i.e., if the affinity of
the ligand differs for the two species), then the ratio
necessarily will change when the ligand is present. There-
fore, its differential affinity for the two protein species will
alter their relative amounts. If the affinity of the ligand is
higher for Ra, then the ratio will be >1 and the ligand will
enrich the Ra species. If the affinity for the ligand for Ra is
less than for Ri, then the ligand (by its presence in the
system) will reduce the amount of Ra. For example, if the
affinity of the ligand is 30-fold greater for the Ra state, then
in a system where 16.7% of the receptors are spontaneously
in the Ra state, the saturation of the receptors with this
agonist will increase the amount of Ra by a factor of 5.14
(16.7%e85%).
This concept is demonstrated schematically in Fig. 1.15.
It can be seen that the initial bias in a system of proteins
containing two conformations (square and spherical) lies
 What is pharmacology? Chapter | 1 17

FIGURE 1.15 Conformational selection as a thermodynamic process to bias mixtures of protein conformations. (A) The two forms of the protein are
depicted as circular and square shapes. The system initially is predominantly square. Gaussian curves to the right show the relative frequency of
occurrence of the two conformations. (B) As a ligand (blue dots) enters the system and prefers the circular conformations, these are selectively removed
from the equilibrium between the two protein states. The distributions show the enrichment of the circular conformation at the expense of the square one.
(C) A new equilibrium is attained in the presence of the ligand favoring the circular conformation because of the selective pressure of affinity between the
ligand and this conformation. The distribution reflects the presence of the ligand and the enrichment of the circular conformation.

far toward the square conformation. When a ligand (filled
circles) enters the system and selectively binds to the cir-
cular conformations, this binding process removes the cir-
cles driving the backward reaction from circles back to
squares. In the absence of this backward pressure, more
square conformations flow into the circular state to fill the
gap. Overall, there is an enrichment of the circular con-
formations when unbound and ligand-bound circular con-
formations are totaled.
This also can be described in terms of the Gibbs free
energy of the receptoreligand system. Receptor confor-
mations are adopted as a result of attainment of minimal
free energy. Therefore, if the free energy of the collection
of receptors changes, so too will the conformational
makeup of the system. The free energy of a system
composed of two conformations ai and ao is given by the
following [19]:
X P
DGi ¼ DG0i  RT
X Doseeresponse curves depict the response to an agonist
 lnð1 þ Ka;i ½AÞ=lnð1 þ Ka;0 ½AÞ;
where Ka,i and Ka,0 are the respective affinities of the ligand
for states i and o. It can be seen that unless Ka,i ¼ Ka,0, the
logarithmic term will not equal zero and the free energy of
P P  along the concentration axis, slope, and maximal asymptote
the system will change DGi s DG0i . Thus, if a
ligand has differential affinity for either state, then the
free energy of the system will change in the presence of
the ligand. Under these circumstances, a different confor-
mational bias will be formed by the differential affinity of
the ligand. From these models comes the concept that bind-
ing is not a passive process, whereby a ligand simply ad-
heres to a protein without changing it. The act of binding
can itself bias the behavior of the protein. This is the ther-
modynamic basis of efficacy.
1.12 Doseeresponse curves
The concept of “doseeresponse” in pharmacology has been
known and discussed for some time. A prescription written
in 1562 for hyoscyamus and opium for sleep clearly states,
“If you want him to sleep less, give him less” [13]. It was
recognized by one of the earliest physicians, Paracelsus
(1493e1541), that it is only the dose that makes something
beneficial or harmful: “All things are poison, and nothing is
without poison. The dose [sic] alone makes a thing not
poison.”
(1.6) in a cellular or subcellular system as a function of the
agonist concentration. Specifically, they plot response as a
function of the logarithm of the concentration. They can be
defined completely by three parameters, namely, location
(Fig. 1.16). At first glance, the shapes of doseeresponse
curves appear to closely mimic the line predicted by the
Langmuir adsorption isotherm, and it is tempting to assume
18 A Pharmacology Primer
FIGURE 1.16 Doseeresponse curves. Any doseeresponse curve can be
defined by the threshold (where response begins along the concentration
axis), the slope (the rise in response with changes in concentration), and
the maximal asymptote (the maximal response).
that doseeresponse curves reflect the first-order binding
and activation of receptors on the cell surface. However, in
most cases, this resemblance is happenstance, and dosee
response curves reflect a far more complex amalgam of
binding, activation, and recruitment of cellular elements of
response. In the end, these may yield a sigmoidal curve, but
in reality they are far removed from the initial binding of
drug and receptor. For example, in a cell culture with a
collection of cells with varying thresholds for depolariza-
tion, the single-cell response to an agonist may be complete
depolarization (in an all-or-none fashion). Taken as a
complete collection, the depolarization profile of the culture
where the cells all have differing thresholds for depolari-
zation would have a Gaussian distribution of depolarization
thresholdsdsome cells being more sensitive than others
(Fig. 1.17A). The relationship of depolarization of the
complete culture to the concentration of a depolarizing
agonist is the area under the Gaussian curve. This yields a
sigmoidal doseeresponse curve (Fig. 1.17B) that resembles
the Langmuirian binding curve for drugereceptor binding.
The slope of the latter curve reflects the molecularity of the
drugereceptor interaction (i.e., one ligand binding to one
receptor yields a slope of unity for the curve). In the case of
the sequential depolarization of a collection of cells, it can
be seen that a narrower range of depolarization thresholds
yields a steeper doseeresponse curve, indicating that the
actual numerical value of the slope for a doseeresponse
curve cannot be equated to the molecularity of the binding
between agonist and receptor. In general, shapes of
doseeresponse curves are completely controlled by cellular
factors and cannot be used to discern drugereceptor
mechanisms. These must be determined indirectly by null
methods.
1.12.1 Potency and maximal response
There are certain features of agonist doseeresponse curves
that are generally true for all agonists. The first is that the
magnitude of the maximal asymptote is totally dependent
on the efficacy of the agonist and the efficiency of the
biological system to convert receptor stimulus into tissue
response (Fig. 1.18A). This can be an extremely useful
observation in the drug-discovery process when attempting
to affect the efficacy of a molecule. Changes in chemical
structure that affect only the affinity of the agonist will have
no effect on the maximal asymptote of the doseeresponse
curve for that agonist. Therefore, if chemists wish to opti-
mize or minimize efficacy in a molecule, they can track the
maximal response to do so. Second, the location, along
the concentration axis of doseeresponse curves, quantifies
the potency of the agonist (Fig. 1.18B). The potency is the
molar concentration required to produce a given response.
Potencies vary with the type of cellular system used to
make the measurement and the level of response at which
the measurement is made. A common measurement used to
quantify potency is the EC50, namely, the molar concen-
tration of an agonist required to produce 50% of the
maximal response to the agonist. Thus, an EC50 value of
1 mM indicates that 50% of the maximal response to the
agonist is produced by a concentration of 1 mM of the
agonist (Fig. 1.19). If the agonist produces a maximal
response of 80% of the system maximal response, then
40% of the system maximal response will be produced by
1 mM of this agonist (Fig. 1.19). Similarly, an EC25 will be
produced by a lower concentration of this same agonist; in
this case, the EC25 is 0.5 mM.
1.12.2 P-scales and the representation of
potency
Agonist potency is an extremely important parameter in
drugereceptor pharmacology. Invariably it is determined
from log-doseeresponse curves. It should be noted that
since these curves are generated from semilogarithmic
plots, the location parameter of these curves is log nor-
mally distributed. This means that the logarithms of the
sensitivities (EC50) and not the EC50 values themselves
are normally distributed (Fig. 1.20A). Since all statistical
parametric tests must be done on data that come from
normal distributions, all statistics (including comparisons
of potency and estimates of errors of potency) must come
from logarithmically expressed potency data. When log
normally distributed EC50 data (Fig. 1.20B) are con-
verted to EC50 data, the resulting distribution is seriously
skewed (Fig. 1.20C). It can be seen that error limits on
the mean of such a distribution are not equal [i.e., one
standard error of the mean unit (see Chapter 12: Statistics
and Experimental Design) either side of the mean gives

FIGURE 1.19 Doseeresponse curves. Doseeresponse curve to an
agonist that produces 80% of the system maximal response. The EC50
(concentration producing 40% response) is 1 mM, the EC25 (20%) is
0.5 mM, and the EC80 (64%) is 5 mM.
different values on the skewed distribution (Fig. 1.20C)].
This is not true of the symmetrical normal distribution
(Fig. 1.20B).
One representation of numbers such as potency estimates
is with the P-scale. The P-scale is the negative logarithm of
number. For example, the pH is the negative logarithm of a
What is pharmacology? Chapter | 1 19
FIGURE 1.17 Factors affecting the
slope of doseeresponse curves. (A)
Gaussian distributions of the thresholds
for depolarization of cells to an agonist
in a cell culture. Solid line shows a
narrow range of threshold, and the
lighter line a wider range. (B) Area
under the curve of the Gaussian distri-
butions shown in panel A. These would
represent the relative depolarization of
the entire cell culture as a function of
the concentration of agonist. The more
narrow range of threshold values cor-
responds to the doseeresponse curve of
steeper slope.
FIGURE 1.18 Major attributes of
agonist doseeresponse curves.
Maximal responses solely reflect
efficacy (left), while the potency
(location along the concentration
axis) reflects a complex function of
both efficacy and affinity (right).
hydrogen ion concentration (105 M ¼ pH ¼ 5). It is essential
to express doseeresponse parameters as P-values (log of
the value, as in the pEC50) since these are log normal.
However, it sometimes is useful on an intuitive level to
express potency as a concentration (i.e., the antilog value).
One way this can be done and still preserve the error esti-
mate is to make the calculation as P-values and then convert
to concentration as the last step. For example, Table 1.2
shows five pEC50 values, giving a mean pEC50 of 8.46 and a
standard error of 0.21. It can be seen that the calculation of
the mean as a converted concentration (EC50 value) leads to
an apparently reasonable mean value of 3.8 nM, with a
standard error of 1.81 nM. However, the 95% confidence
limits (range of values that will include the true value) of the
concentration value is meaningless, in that one of them (the
lower limit) is a negative number. The true value of the EC50
lies within the 95% confidence limits given by the mean -
þ 2.57  the standard error, which leads to the values 8.4
and 0.85 nM. However, when pEC50 values are used for
the calculations, this does not occur. Specifically, the mean
of 8.46 yields a mean EC50 of 3.47 nM. The 95% confidence
limits on the pEC50 are 7.8e9.0. Conversion of these limits
to EC50 values yields 95% confidence limits of 1e11.8 nM.
Thus, the true potency lies between the values of 1 and
11.8 nM 95% of the time.
20 A Pharmacology Primer

FIGURE 1.20 Log normal distributions of sensitivity of a pharmacological preparation to an agonist. (A) Doseeresponse curve showing the distribution
of the EC50 values along the log concentration axis. This distribution is normal only on a log scale. (B) Log normal distribution of pEC50 values (log
EC50 values). (C) Skewed distribution of EC50 values converted from the pEC50 values shown in panel B.

l System-independent measures of drug activity coupled

TABLE 1.2 Expressing mean agonist potencies with with pharmacological models of drug mechanisms can
error. combine to convert a single ‘snapshot’ of activity in
pEC50a EC50 (nM)b one system into the complete ‘film’ of what the drug
will do in vivo.
8.5 3.16
l Affinity is the strength of binding of a drug to a receptor.
8.7 2 It is quantified by an equilibrium dissociation constant.
8.3 5.01 l Affinity can be depicted and quantified with the Lang-
8.2 6.31 muir adsorption isotherm.
l Efficacy is measured in relative terms (having no abso-
8.6 2.51
lute scale) and quantifies the ability of a molecule to
Mean ¼ 8.46 Mean ¼ 3.8 produce a change in the receptor (most often leading
SE ¼ 0.21 SE ¼ 1.81 to a physiological response).
a
l Doseeresponse curves quantify drug activity. The
Replicate values of 1/N log EC50’s.
b
Replicate EC50 values in nM. maximal asymptote is totally dependent on efficacy,
while potency is due to an amalgam of affinity and
efficacy.
l Measures of potency are log normally distributed. Only
1.13 Chapter summary and conclusions P-scale values (i.e., pEC50) should be used for statistical
tests.
l Some ideas on the origins and relevance of pharma-
cology and the concept of biological “receptors” are
discussed. 1.14 Derivations: conformational
l Currently, there are drugs for only a fraction of the selection as a mechanism of
druggable targets present in the human genome.
l While recombinant systems have greatly improved the
efficacy
drug-discovery process, pathological phenotypes still Consider a system containing two receptor conformations
are a step away from these drug-testing systems. Ri and Ra that coexist in the system according to an allo-
l Because of the fact that drugs are tested in experimental, steric constant denoted L.
not therapeutic, systems, system-independent measures Assume that ligand A binds to Ri with an equilibrium
of drug activity (namely, affinity and efficacy) must association constant Ka, and Ra by an equilibrium associ-
be measured in drug discovery. ation constant aKa. The factor a denotes the differential

affinity of the agonist for Ra (i.e., a ¼ 10 denotes a 10-fold
greater affinity of the ligand for the Ra state). The effect of
a on the ability of the ligand to alter the equilibrium be-
tween Ri and Ra can be calculated by examining the amount
of Ra species (both as Ra and ARa) present in the system in
the absence of ligand and in the presence of ligand. The
equilibrium expression for ([Ra]þ[ARa])/[Rtot], where
[Rtot] is the total receptor concentration given by the con-
servation equation [Rtot] ¼ [Ri]þ[ARi]þ[Ra]þ[ARa], is
Lð1 þ a½A=KA Þ
r¼ ; (1.7)
½A=KA ð1 þ aLÞ þ 1 þ L
where L is the allosteric constant, [A] is the concentration
of ligand, KA is the equilibrium dissociation constant of
the agonistereceptor complex (KA ¼ 1/Ka), and a is the
differential affinity of the ligand for the Ra state. It can
be seen that in the absence of agonist ([A] ¼ 0), r0 ¼ L/
(1 þ L), and in the presence of a maximal concentration
of ligand (saturating the receptors; [A]/N),
rN¼(a(1 þ L))/(1 þ aL). The effect of the ligand on
changing the proportion of the Ra state is given by the ratio
r/r0. This ratio is given by
rN að1 þ LÞ
¼ . (1.8)
r0 ð1 þ aLÞ
Eq. (1.8) indicates that if the ligand has an equal affinity
for both the Ri and Ra states (a ¼ 1), then rN/r0 will equal
unity, and no change in the proportion of Ra will result
from maximal ligand binding. However, if a > 1, then the
presence of the conformationally selective ligand will cause
the ratio rN/r0 to be > 1, and the Ra state will be enriched
by presence of the ligand.
References
[1] A.-H. Maehle, C.-R. Prull, R.F. Halliwell, The emergence of the
drug-receptor theory, Nat. Rev. Drug Discov. 1 (2002) 1637e1642.
[2] W.D.M. Paton, On becoming a pharmacologist, Annu. Rev. Phar-
macol. Toxicol. 26 (1986) 1e22.
What is pharmacology? Chapter | 1 21
[3] J. Drews, Drug discovery: a historical perspective, Science 287
(2000) 1960e1964.
[4] A.J. Clark, The Mode of Action of Drugs on Cells, Edward Arnold,
London, 1933.
[5] A.J. Clark, A. Heffter, General Pharmacology Handbuch der Exper-
imentellen Pharmakologie, Springer, Berlin, 1937, pp. 165e176, 4.
[6] B. Holmstedt, G. Liljestrand, Readings in Pharmacology, Raven
Press, New York, NY, 1981.
[7] A. Marchese, S.R. George, L.F. Kolakowski, K.R. Lynch,
B.F. O’Dowd, Novel GPCRs and their endogenous ligands:
expanding the boundaries of physiology and pharmacology, Trends
Pharmacol. Sci. 20 (1999) 370e375.
[8] J.C. Venter, M.D. Adams, E.W. Myers, P.W. Li, R.J. Mural,
G.G. Sutton, The sequence of the human genome, Science 291
(2001) 1304e1351.
[9] R. Link, D. Daunt, G. Barsh, A. Chruscinski, B. Kobilka, Cloning of
two mouse genes encoding a2-adrenergic receptor subtypes and
identification of a single amino acid in the mouse a2-C10 homolog
responsible for an interspecies variation in antagonist binding, Mol.
Pharmacol. 42 (1992) 16e17.
[10] J.W. Black, A personal view of pharmacology, Annu. Rev. Phar-
macol. Toxicol. 36 (1996) 1e33.
[11] R. Buscher, V. Hermann, P.A. Insel, Human adrenoceptor poly-
morphisms: evolving recognition of clinical importance, Trends
Pharmacol. Sci. 20 (1999) 94e99.
[12] R.P. Stephenson, A modification of receptor theory, Br. J. Pharma-
col. 11 (1956) 379e393.
[13] S. Norton, Origins of pharmacology, Mol. Interv. 5 (2005) 144e149.
[14] P. Leff, G.R. Martin, J.M. Morse, Differences in agonist dissociation
constant estimates for 5-HT at 5-HT2-receptors: a problem of acute
desensitization? Br. J. Pharmacol. 89 (1986) 493e499.
[15] A. Linderstrom-Lang, P. Schellman, Protein conformation, Enzymes
1 (1959) 443e471.
[16] A.S.V. Burgen, Conformational changes and drug action, Fed. Proc.
40 (1966) 2723e2728.
[17] J.J. Wyman, D.W. Allen, The problem of the haem interaction in
haemoglobin and the basis for the Bohr effect, J. Polym. Sci. 7
(1951) 499e518.
[18] J. Del Castillo, B. Katz, Interaction at end-plate receptors between
different choline derivatives, Proc. Roy. Soc. Lond. B. 146 (1957)
369e381.
[19] E. Freire, Can allosteric regulation be predicted from structure? Proc.
Natl. Acad. Sci. U.S.A. 97 (2000) 11680e11682.
This page intentionally left blank
Chapter 2
How different tissues process drug
response
[Nature] can refuse to speak but she cannot give a wrong
answer.
d Dr. Charles Brenton Hugins (1966).
We have to remember that what we observe is not nature in
itself, but nature exposed to our method of questioning .
dWerner Heisenberg (1901e76).
2.1 The ‘eyes to see’: pharmacologic
assays
If a drug possesses the molecular property of efficacy, then
it produces a change in the receptor that may be detected by
the cell. However, this can occur only if the stimulus is of
sufficient strength and the cell has the amplification ma-
chinery necessary to convert the stimulus into an observ-
able response. In keeping with the mandatory partnership of
the sensitivity of the cell system and the intrinsic power of
the agonist to produce a response, the cellular assay be-
comes a key component that controls the amount of in-
formation that can be gained from the experiment, in
essence, the assay becomes the ‘eyes to see’ the change
imparted to the cell by the drug. Cellular assays can be
natural (and thus the sensitivity and components are set by
Nature) or recombinant whereby the experimenter can
manipulate the levels of response components and thus the
sensitivity of the system. Fig. 2.1 shows some of the factors
that play into the design of a pharmacologic assay whether
as applied to binding studies (Chapter 4) or functional
studies (Chapter 6). When building recombinant systems,
the first option is the type of cell to be used. Different cells
have different components and some of these may be
critical to the response of a given agonist. A case in point is
the response to the hormone amylin which interacts with a
receptor formed by a dimer of the calcitonin receptor and
the membrane protein RAMP3 (Receptor Activity modi-
fying Protein 3). Thus, if a cell is not used that contains
RAMP3, then transfection of calcitonin receptors will not
constitute receptors for amylin and the assay will yield
erroneous results (for further details see Fig. 5.3). Similarly,
the relative stoichiometry of signaling components in a cell
A Pharmacology Primer. https://doi.org/10.1016/B978-0-323-99289-3.00013-0
Copyright © 2022 Elsevier Inc. All rights reserved.
may affect observed bias of agonist response (for further
details see Chapter 6). For complex binding curves
whereby the binding complex is an amalgam of the receptor
with other components (i.e., G-protein), differences in the
complimentary protein can lead to differences in binding
profiles. For instance, overexpression of receptor to the
point where the G-protein components in a cell are insuf-
ficient to produce adequate levels of ternary complex, then
complex binding curves will be produced (for further de-
tails see Fig. 4.19). If cell response is measured, then
various cells reflect changes in function in different ways
thus ‘business rules’ for the definition of what will be
considered drug response must be determined and adhered
to throughout the experiment. A major determinant of the
sensitivity of cells for agonists acting on receptors is the
level of receptor density expressed in the cell, i.e., a high
receptor density produces a sensitive tissue whereas a low
density an insensitive tissue. The impact of functional assay
composition will be considered repeatedly in this book as it
is of paramount importance for the discernment of drug
activity in in vitro systems.
Fig. 2.2A shows a functional doseeresponse curve for
human calcitonin in human embryonic kidney (HEK) cells
transfected with cDNA for human calcitonin receptor type
2 [1]. The response being measured here is the hydrogen
ion release by the cells, a sensitive measure of cellular
metabolism. Also shown (dotted line) is a curve for calci-
tonin binding to the receptors (as measured with radio-
ligand binding). A striking feature of these curves is that the
curve for function is shifted considerably to the left of the
binding curve. Calculation of the receptor occupancy
required for 50% maximal tissue response indicates that
less than 50% occupancy, namely, more on the order of
3%e4%, is needed. In fact, a regression of tissue response
upon the receptor occupancy is hyperbolic in nature
(Fig. 2.2B), showing a skewed relationship between re-
ceptor occupancy and cellular response. This skewed
relationship indicates that the stimulation of the receptor
initiated by binding is amplified by the cell in the process of
response production.
The ability of a given agonist to produce a maximal
system response can be quantified as a receptor reserve.
23
24 A Pharmacology Primer

FIGURE 2.1 Main options available for the design of recombinant assay systems are the type of cell to be used and the level of receptors available to
response to agonists.

FIGURE 2.2 Binding and doseeresponse curves for human calcitonin on human calcitonin receptors type 2. (A) Doseeresponse curves for micro-
physiometry responses to human calcitonin in HEK cells (open circles) and binding in membranes from HEK cells (displacement of [125I]-human
calcitonin). (B) Regression of microphysiometry responses to human calcitonin (ordinates) upon human calcitonin fractional receptor occupancy
(abscissae). Dotted line shows a direct correlation between receptor occupancy and cellular response. HEK, human embryonic kidney. (A) Data from W.-J.
Chen, S. Armour, J. Way, G.C. Chen, C. Watson, P.E. Irving, Expression cloning and receptor pharmacology of human calcitonin receptors from MCF-7
cells and their relationship to amylin receptors, Mol. Pharmacol. 52 (1997) 1164e1175.

The reserve refers to the percentage of receptors not
required for production of maximal response (sometimes
referred to as spare receptors). For example, a receptor
reserve of 80% for an agonist means that the system
maximal response is produced by activation of 20% of the
receptor population by that agonist. Receptor reserves can
be quite striking. Fig. 2.3 shows guinea pig ileal smooth
muscle contractions to the agonist histamine before and
after irreversible inactivation of a large fraction of the re-
ceptors with the protein alkylating agent phenoxybenz-
amine [2]. The fact that the depressed maximum
doseeresponse curve is observed so far to the right of the
control doseeresponse curve indicates a receptor reserve of
98% [i.e., only 2% of the receptors must be activated by
histamine to produce the tissue’s maximal response
(Fig. 2.3B)]. In teleological terms, this may be useful, since
it allows neurotransmitters to produce rapid activation of
organs with minimal receptor occupancy leading to optimal
and rapid control of function.
Receptor reserve is a property of the tissue (i.e., the
strength of amplification of receptor stimulus inherent to the
cells) and it is a property of the agonist (i.e., how much
stimulus is imparted to the system by a given agonist receptor
occupancy). This latter factor is quantified as the efficacy of
the agonist. A high-efficacy agonist need occupy a smaller
fraction of the receptor population than a lower efficacy
agonist to produce a comparable stimulus. Therefore, it is
incorrect to ascribe a given tissue or cellular response system
with a characteristic receptor reserve. The actual value of the
receptor reserve will be unique to each agonist in that system.
For example, Fig. 2.4 shows the different amplification hy-
perbolae of Chinese hamster ovary (CHO) cells transfected
with b-adrenoceptors in producing cyclic adenosine mono-
phosphate (AMP) responses to three different b-adrenoceptor
 How different tissues process drug response Chapter | 2 25

FIGURE 2.3 Guinea pig ileal responses to histamine. (A) Contraction of guinea pig ileal longitudinal smooth muscle (ordinates as a percentage of
maximum) to histamine (abscissae, logarithmic scale). Responses obtained before ( filled circles) and after treatment with the irreversible histamine
receptor antagonist phenoxybenzamine (50 mM for 3 minutes; open circles). (B) Occupancyeresponse curve for data shown in (A). Ordinates are per-
centage of maximal response. Abscissae are calculated receptor occupancy values from an estimated affinity of 20 mM for histamine. Note that maximal
response is essentially observed after only 2% receptor occupancy by the agonist (i.e., a 98% receptor reserve for this agonist in this system). Data
redrawn from T.P. Kenakin, D.A. Cook, Blockade of histamine-induced contractions of intestinal smooth muscle by irreversibly acting agents, Can. J.
Physiol. Pharmacol. 54 (1976) 386e392.

mechanisms in the cell. Each has its own function and

FIGURE 2.4 Occupancyeresponse curves for b-adrenoceptor agonists in
transfected CHO cells. Occupancy (abscissae) calculated from binding af-
finity measured by displacement of [125I]-iodocyanopindolol. Response
measured as increases in cyclic AMP. Drawn from S. Wilson, J.K. Cham-
bers, J.E. Park, A. Ladurner, D.W. Cronk, C.G. Chapman, Agonist potency
at the cloned human beta-3 adrenoceptor depends on receptor expression
level and nature of assay, J. Pharmacol. Exp. Ther. 279 (1996) 214e221.
agonists [3]. It can be seen that isoproterenol requires many
times less receptors to produce 50% response than do both
the agonists BRL 37344 and CGP 12177. This underscores
the idea that the magnitude of receptor reserves is very much
dependent on the efficacy of the agonist (i.e., one agonist’s
spare receptor is another agonist’s essential one).
2.2 The biochemical nature of
stimuluseresponse cascades
Cellular amplification of receptor signals occurs through a
succession of saturable biochemical reactions. Different
receptors are coupled to different stimuluseresponse
operates on its own timescale. For example, receptor
tyrosine kinases (activated by growth factors) phosphory-
late target proteins on tyrosine residues to activate protein
phosphorylation cascades such as mitogen-activated protein
(MAP) kinase pathways. This process, on a timescale on
the order of seconds to days, leads to protein synthesis from
gene transcription with resulting cell differentiation and/or
cell proliferation. Nuclear receptors, activated by steroids,
operate on a timescale of minutes to days and mediate gene
transcription and protein synthesis. This leads to homeo-
static, metabolic, and immunosuppression effects. Ligand-
gated ion channels, activated by neurotransmitters, operate
on the order of milliseconds to increase the permeability of
plasma membranes to ions. This leads to increases in
cytosolic Ca2þ, depolarization, or hyperpolarization of
cells. This in turn results in muscle contraction, release of
neurotransmitters, or inhibition of these processes.
G-protein-coupled receptors (GPCRs) react with a wide
variety of molecules, from small ones such as acetylcholine
to some as large as the protein SDF-1a. Operating on a
timescale of minutes to hours, these receptors mediate a
plethora of cellular processes. A common reaction in the
activation cascade for GPCRs is the binding of the activated
receptor to a trimeric complex of proteins called G-proteins
(Fig. 2.5). These proteinsdcomposed of three subunits
named a, b, and gdact as molecular switches for a number
of other effectors in the cell. The binding of activated re-
ceptors to the G-protein initiates the dissociation of GDP
from the a-subunit of the G-protein complex, the binding of
guanosine monophosphate (GTP), and the dissociation of the
complex into a- and bg-subunits. The separated subunits of
the G-protein can activate effectors in the cell such as ade-
nylate cyclase and ion channels. Amplification can occur at
these early stages if one receptor activates more than one
26 A Pharmacology Primer

FIGURE 2.5 Activation of trimeric G-proteins by activated receptors. An agonist produces a receptor active state that goes on to interact with the G-
protein. A conformational change in the G-protein causes bound GDP to exchange with GTP. This triggers dissociation of the G-protein complex into a-
and bg-subunits. These go on to interact with effectors such as adenylate cyclase and calcium channels. The intrinsic GTPase activity of the a-subunit
hydrolyzes bound GTP back to GDP, and the inactivated a-subunit reassociates with the bg-subunits to repeat the cycle.

FIGURE 2.6 Production of cyclic AMP from ATP by the enzyme adenylate cyclase. Cyclic AMP is a ubiquitous second messenger in cells activating
numerous cellular pathways. The adenylate cyclase is activated by the a-subunit of Gs-protein and inhibited by the a-subunit of Gi-protein. Cyclic AMP is
degraded by phosphodiesterases in the cell.

G-protein. The a-subunit also is a GTPase, which hydrolyzes
the bound GTP to produce its own deactivation. This ter-
minates the action of the a-subunit on the effector. It can be
seen that the length of time for which the a-subunit is active
can control the amount of stimulus given to the effector, and
that this also can be a means of amplification (i.e., one a-
subunit could activate many effectors). The a- and bg-sub-
units then reassociate to complete the regulatory cycle
(Fig. 2.5). Such receptor-mediated reactions generate cellular
molecules called second messengers. These molecules go on
to activate or inhibit other components of the cellular ma-
chinery to change cellular metabolism and state of activation.
For example, the second messenger (cyclic AMP) is generated
by the enzyme adenylate cyclase from ATP. This second
messenger furnishes fuel, through protein kinases, for the
phosphorylation of serine and threonine residues on a number
of proteins such as other protein kinases, receptors, metabolic
enzymes, ion channels, and transcription factors (see Fig. 2.6).
Activation of other G-proteins leads to the activation of
phospholipase C. These enzymes catalyze the hydrolysis of
 How different tissues process drug response Chapter | 2
FIGURE 2.7 Production of second messengers IP3 and DAG through activation of the enzyme phospholipase C. This enzyme is activated by the a-
subunit of Gq-protein and also by bg-subunits of Gi-protein. IP3 stimulates the release of Ca2 from intracellular stores, while DAG is a potent activator of
protein kinase C. DAG, diacylglycerol; IP3, inositol 1,4,5-triphosphate.
phosphatidylinositol 4,5-bisphosphate to 1,2-diacylglycerol
(DAG) and inositol 1,4,5-triphosphate (see Fig. 2.7). This
latter second messenger interacts with receptors on intracel-
lular calcium stores, resulting in the release of calcium into the
cytosol. This calcium binds to calcium sensor proteins such as
calmodulin or troponin C, which then go on to regulate the
activity of proteins such as protein kinases, phosphatases,
phosphodiesterase, nitric oxide synthase, ion channels, and
adenylate cyclase. The second messenger DAG diffuses in the
plane of the membrane to activate protein kinase C isoforms,
which phosphorylate protein kinases, transcription factors, ion
channels, and receptors. DAG also functions as a source of
arachidonic acid, which goes on to be the source of eicosanoid
mediators such as prostanoids and leukotrienes. In general, all
these processes can lead to a case where a relatively small
amount of receptor stimulation can result in a large
biochemical signal. An example of a complete stimuluse
response cascade for the b-adrenoceptor production of blood
glucose is shown in Fig. 2.8 [4].
There are numerous second messenger systems such as
those utilizing cyclic AMP and cyclic guanosine mono-
phosphate (GMP), calcium and calmodulin, phosphoinositides,
and DAG with accompanying modulatory mechanisms. Each
receptor is coupled to these in a variety of ways in different cell
types. Therefore, it can be seen that it is impractical to attempt
to quantitatively define each stimuluseresponse mechanism
for each receptor system. Fortunately, this is not an important
prerequisite in the pharmacological process of classifying ag-
onists, since these complex mechanisms can be approximated
by simple mathematical functions.
2.3 The mathematical approximation of
stimuluseresponse mechanisms
Each of the processes shown in Fig. 2.8 can be described by
a MichaeliseMenten type of biochemical reaction, a stan-
dard generalized mathematical equation describing the
27
interaction of a substrate with an enzyme. Michaelis and
Menten realized in 1913 that the kinetics of enzyme re-
actions differed from those of conventional chemical re-
actions. They visualized the reaction of substrate and an
enzyme yielding enzyme plus product as a form of this
equation: reaction velocity ¼ (maximal velocity of the
reaction  substrate concentration)/(concentration of sub-
strate þ a fitting constant Km). The constant Km (referred to
as the MichaeliseMenten constant) characterizes the
tightness of the binding of the reaction between substrate
and enzyme, essentially a quantification of the coupling
efficiency of the reaction. Km is the concentration at which
the reaction is half the maximal value or, in terms of ki-
netics, the concentration at which the reaction runs at half
its maximal rate. This model forms the basis of enzymatic
biochemical reactions and can be used as a mathematical
approximation of such functions.
As with the Langmuir adsorption isotherm, which in
shape closely resembles MichaeliseMenten type
biochemical kinetics, the two notable features of such re-
actions are the location parameter of the curve along the
concentration axis (the value of Km or the magnitude of the
coupling efficiency factor) and the maximal rate of the re-
action (Vmax). In generic terms, MichaeliseMenten re-
actions can be written in the form
½substract$Vmax ½input$MAX
Velocity ¼ ¼ (2.1)
½substract þ Km ½input þ b
where b is a generic coupling efficiency factor. It can be
seen that the velocity of the reaction is inversely propor-
tional to the magnitude of b (i.e., the lower the value of
b, the more efficiently is the reaction coupled). If it is
assumed that the stimuluseresponse cascade of any given
cell is a series succession of such reactions, there are two
general features of the resultant that can be predicted math-
ematically. The first is that the resultant of the total series of
reactions will itself be of the form of the same hyperbolic
28 A Pharmacology Primer

FIGURE 2.8 Stimuluseresponse cascade for the production of blood glucose by activation of b-adrenoceptors. Redrawn from N.D. Goldberg, G.
Weissman, R. Claiborne, Cyclic nucleotides and cell function, in: G. Weissman, R. Claiborne (Eds.), Cell Membranes Biochemistry, Cell Biology, and
Pathology, H. P. Publishing, New York, NY, 1975, pp. 185e202.

shape (see Section 2.12.1). The second is that the location

parameter along the input axis (magnitude of the coupling
efficiency parameter) will reflect a general amplification
of any single reaction within the cascade (i.e., the magni-
tude of the coupling parameter for the complete series
will be lower than the coupling parameter of any single re-
action; see Fig. 2.9). The magnitude of btotal for the series
sum of two reactions (characterized by b1 and b2) is given
by (see Section 2.12.2):
b1 b2
btotal ¼ . (2.2)
1 þ b2
It can be seen from Eq. (2.2) that for positive nonzero FIGURE 2.9 Amplification of stimulus through successive rectangular
values of b2, btotal < b1. Therefore, the location parameter hyperbolae. The output from the first function (b ¼ 0.3) becomes the input
of the rectangular hyperbola of the composite set of re- of a second function with the same coupling efficiency (b ¼ 0.3) to yield a
more efficiently coupled overall function (b ¼ 0.069). Arrows indicate the
actions in series is shifted to the left (increased potency) of potency for input to yield 50% maximal output for the first function and
that for the first reaction in the sequence (i.e., there is the series functions.
amplification inherent in the series of reactions).
The fact that the total stimuluseresponse chain can be the relationship between the strength of signal imparted
approximated by a single rectangular hyperbola furnishes the to the receptor between two agonists is accurately reflected
basis of using an end-organ response to quantify an agonist by the end-organ response (Fig. 2.10). This is the primary
effect in a nonsystem-dependent manner. An important reason that pharmacologists can circumvent the effects of the
feature of such a relationship is that it is monotonic (i.e., cellular veil and discern system-independent receptor events
there is only one value of y for each value of x). Therefore, from translated cellular events.
 How different tissues process drug response Chapter | 2 29

FIGURE 2.10 The monotonic nature of stimuluseresponse mechanisms. (A) Receptor stimulus generated by two agonists designated 1 and 2 as a
function of agonist concentration. (B) Rectangular hyperbola characterizing the transformation of receptor stimulus (abscissae) into cellular response
(ordinates) for the tissue. (C) The resulting relationship between tissue responses to the agonists as a function of agonist concentration. The general rank
order of activity (2 > 1) is preserved in the response as a reflection of the monotonic nature of the stimuluseresponse hyperbola.

FIGURE 2.11 Agonist-stimulated phosphorylation process with unsaturable dephosphorylation. Panel A shows dephosphorylation (solid ascending
line) as a linear unsaturable process and phosphorylation (descending dotted lines) for a range of agonist concentrations. Rates decrease as the substrate is
depleted. Where these curves intersect denotes a steady-state response (open circles). Panel B: Steady-state responses (ordinates) as a function of agonist
concentration. A sigmoidal curve of slope ¼ 1 describes steady-state responses. Redrawn from J.J. Tyson, K.C. Chen, B. Novak, Sniffers, buzzers, toggles
and blinkers: dynamics of regulatory and signaling pathways in the cell, Curr. Opin. Cell Biol. 15 (2003) 221e231.

2.4 Influence of stimuluseresponse
cascades on doseeresponse curve
slopes
For standard mass action kinetics whereby a single molecule
binds to a single receptor, the resulting binding curves have
Hill coefficient slopes of unity (providing cooperativity is
not present in the binding reactions). However, for agonists
with such simple Langmuirian binding kinetics (slope ¼ 1),
the cellular response curves for that agonist in functional
systems often will have slopes different from unity; this is
not due to cooperativity of binding but rather through signal
processing by the stimuluseresponse cascades in the cell
cytosol [5]. For example, a simple cytosolic biochemical
reaction such as the phosphorylation and dephosphorylation
of an enzyme can change the slope of concentrationeresponse
curve of agonists affecting the reaction. Fig. 2.11 shows
a concentrationeresponse curve for an agonist promoting the
phosphorylation of an enzyme. Fig. 2.11A shows an
ascending solid line depicting the rate of enzyme dephos-
phorylation and multiple descending dotted lines depicting
rates of phosphorylation for different concentrations of
agonist. The open circles represent steady states where the rate
of phosphorylation equals the rate of dephosphorylation; these
are the observed response points for the agonist and are
shown, as a function of agonist concentration, in Fig. 2.11B.
30 A Pharmacology Primer

FIGURE 2.12 Agonist-stimulated phosphorylation process with saturable dephosphorylation process described by MichaeliseMenten kinetics. Curve
descriptions as for Fig. 2.11. Panel B shows that the relationship between steady-state responses and agonist concentration is described by a sigmoid
function of slope ¼ 3. Redrawn from J.J. Tyson, K.C. Chen, B. Novak, Sniffers, buzzers, toggles and blinkers: dynamics of regulatory and signaling
pathways in the cell, Curr. Opin. Cell Biol. 15 (2003) 221e231.

This figure shows a case where the dephosphorylation is

nonsaturable; the slope of the resulting concentratione
response curve has a slope of unity consistent with mass ac-
tion binding kinetics. However, if the dephosphorylation
process is saturable and described by MichaeliseMenten ki-
netics (as might be expected in a cellular biochemical
reaction), then a different pattern emerges. Specifically, the
resulting concentrationeresponse curve (obtained from
the steady-state intersections of the phosphorylation and
dephosphorylation rates) has a slope of 3, considerably steeper
than that mediating agonist binding to the receptor. Thus, it
can be seen that the processing of receptor stimulus by the cell
can control the slopes of concentrationeresponse curves
making inferences about cooperativity fruitless in functional
systems. Depending on the nature of the feedback loops found FIGURE 2.13 Receptor occupancy curves for activation of human
in cellular stimuluseresponse cascades, a wide variety of calcitonin type 2 receptors by the agonist human calcitonin. Ordinates:
response outcomes can result from varying concentratione response as a fraction of the maximal response to human calcitonin.
Abscissae: fractional receptor occupancy by human calcitonin. Curves
response curve slope, to transient and phasic activity [5] shown for receptors transfected into three cell types: HEK cells, CHO
(Fig. 2.12). cells, and Xenopus laevis melanophores. It can be seen that the different
cell types lead to differing amplification factors for the conversion from
agonist receptor occupancy to tissue response. CHO, Chinese hamster
2.5 System effects on agonist response: ovary; HEK, human embryonic kidney.
full and partial agonists
For any given receptor type, different cellular hosts should for amplification of receptor stimuli. This is illustrated by
have characteristic efficiencies of coupling, and these the strikingly different magnitudes of the receptor reserves
should characterize all agonists for that same receptor for calcitonin and histamine receptors shown in Figs. 2.2
irrespective of the magnitude of the efficacy of the agonists. and 2.3. Fig. 2.13 shows the response produced by human
Different cellular backgrounds have different capabilities calcitonin activation of the human calcitonin receptor type
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imprudent. Avec le revenu...
—Cela suffira, cela suffira, Aleck! Que tu es bonne et généreuse! Nous
aurons un beau revenu et si nous pouvons le dépenser...
—Pas tout, mon ami, pas tout, mais seulement une partie, une
raisonnable, une petite partie. Mais tout le capital, chaque centime du
capital doit être employé en de bons placements. Tu trouves bien cela
raisonnable, n’est-ce pas?
—Mais... oui... oui, bien sûr. Mais il nous faudra attendre si longtemps—
au moins six mois—avant de toucher les premiers intérêts.
—Oui, même plus, peut-être.
—Attendre plus, Aleck? Pourquoi? Est-ce qu’on ne paie pas les intérêts
par semestres?
—A certains placements, oui, mais je ne placerai pas de cette façon-là.
—De quelle façon alors?
—A de gros intérêts.
—Gros? Voilà qui est bien. Continue, Aleck, explique-toi.
—Charbon; les nouvelles mines... Je compte y mettre dix mille.
—Par saint Georges! Voilà qui sonne bien, Aleck! Les actions vaudraient
combien alors? Et quand?
—Au bout d’un an environ; elles rapportent dix pour cent, et vaudront
trente mille. Je suis toute renseignée. L’annonce est dans le journal de
Cincinnati.
—Juste ciel! Trente mille pour dix mille en une année! Mettons-y tout le
capital et nous aurons quatre-vingt-dix mille dollars au bout d’un an. Je vais
écrire et souscrire tout de suite. Demain il pourrait être trop tard.
Il courait déjà vers sa table à écrire, mais Aleck l’arrêta et le remit sur sa
chaise en lui disant:
—Ne perds pas ainsi la tête; nous ne pouvons souscrire avant d’avoir
l’argent, ne le sais-tu pas?
Sally se calma un peu, mais à la surface seulement...
—Mais, Aleck, nous l’aurons, tu sais, et bientôt. Ses peines sont
probablement terminées à l’heure qu’il est. Je parierais gros qu’en ce
moment on creuse sa tombe.
Aleck dit en frissonnant:
—Comment peux-tu agir ainsi, Sally? Ne parle pas de la sorte; c’est tout
à fait scandaleux.
—Ah! bien, mets qu’on prépare son auréole, si tu veux. Je me soucie fort
peu de tout l’attirail; je parlais tout simplement. Ne peux-tu pas laisser
parler les gens?
—Mais comment peux-tu trouver un plaisir quelconque à parler d’une
façon aussi légère? Aimerais-tu qu’on dise cela de toi avant même que tu
sois glacé dans la tombe?
—Il n’y a pas de danger, de quelque temps, je pense, surtout si ma
dernière action avait été de faire cadeau de ma fortune à des gens dans le
seul but de leur faire du mal. Mais ne nous inquiétons pas de Tilbury, Aleck,
et parlons de choses plus terre à terre... Il me semble vraiment que cette
mine est le meilleur placement pour toute la somme... Quelle est ton
objection?
—Tous les œufs dans le même panier, voilà l’objection.
—Très bien, si tu juges ainsi. Et pour les autres vingt mille? Que
comptes-tu en faire!
—Rien ne presse. Je vais regarder autour de moi avant d’en rien faire du
tout.
—Très bien, si tu es décidée, soupira Sally qui resta quelque temps
plongé dans ses méditations.
 —Il y aura donc, dit-il enfin, vingt mille de revenu sur les dix mille
engagés dès l’année prochaine. Nous pourrons dépenser cela, n’est-ce pas?
Aleck fit un signe négatif:
—Non, non, mon chéri, dit-elle. Cela ne se vendra pas bien jusqu’à ce
que nous ayons touché le dividende semestriel. Nous pourrons en dépenser
une petite partie...
—Une pacotille, pas davantage? Et toute une année à attendre... Peste!
Je...
—Sois patient, je t’en prie! Sais-tu que cela pourrait même être versé au
bout de trois mois? C’est tout à fait dans le domaine des choses possibles.
—Oh! bonheur. Oh! merci.
Sally bondit pour aller embrasser sa femme avec reconnaissance.
—Nous toucherions trois mille alors! Pense, trois mille! Et combien
pourrons-nous en dépenser, Aleck? Sois généreuse, va, tu seras une si
gentille petite femme!
Aleck se trouva flattée, si flattée qu’elle céda et abandonna mille dollars,
ce qui, selon elle, était une folie. Sally l’embrassa une bonne douzaine de
fois et n’arrivait pas à témoigner toute sa gratitude et toute sa joie. Cette
nouvelle marque d’affection fit franchir à Aleck les limites de la prudence
et avant qu’elle ait pu se reprendre, elle avait fait une autre concession à son
heureux époux en lui accordant deux mille dollars sur les cinquante ou
soixante mille qu’elle comptait retirer au bout d’une année des vingt mille
restant de l’héritage. Les yeux remplis de larmes de joie, Sally s’écria:
—Oh! je veux te serrer bien fort!
Et il le fit. Il reprit ensuite son bloc-notes et se prit à marquer, pour les
premiers achats, les objets qu’il désirait acheter tout d’abord: cheval,
voiture, traîneau, couverture de fourrure, souliers vernis, chien, chapeau de
soie, dentier...
—Dis-moi, Aleck?
—Eh bien?
—Tu fais des comptes, n’est-ce pas? Voilà qui va bien. As-tu trouvé le
placement des vingt mille maintenant?
—Non. Ça ne presse pas. Il me faut chercher et réfléchir encore.
—Mais, je te vois faire des chiffres... Que combines-tu alors?
 —Mais il faut bien que je trouve l’emploi des trente mille qui viennent
des mines, n’est-ce pas?
—Bonté divine! Quelle tête! Je n’y aurais jamais songé. Comment t’en
tires-tu? Où en es-tu arrivée?
—Pas très loin encore. Deux années ou trois. J’ai placé l’argent deux
fois, une fois dans les huiles, une fois dans les blés.
—Mais, Aleck, c’est splendide. Et à combien arrives-tu maintenant?
—Je crois... Eh bien, pour être sûre de ne pas me tromper... à environ
cent vingt-quatre mille net... mais ce sera probablement plus.
—Grand Dieu, comme c’est beau! La chance a tourné de notre côté
après tant de labeur! Aleck?
—Eh bien?
—Je vais compter au moins trois cents dollars pour les œuvres des
missions en pays païens. Nous n’avons pas le droit de ne penser qu’à nous-
mêmes.
—Tu ne pouvais faire une plus noble chose. Voilà bien ta nature, mon
généreux ami!
A cette louange, Sally tressaillit de joie, mais étant juste et honnête, il se
dit que le mérite en revenait plus à Aleck, qu’à lui-même, puisque sans elle
il n’aurait jamais eu tout cet argent.
Ils montèrent se coucher et, dans leur délirant bonheur, ils laissèrent la
bougie allumée au salon. Ils n’y pensèrent que lorsqu’ils furent déshabillés.
Sally alors fut d’avis de la laisser brûler; ils pouvaient s’accorder cela
maintenant, même s’il y avait eu mille bougies allumées. Tel ne fut pas
l’avis d’Aleck: elle descendit pour l’éteindre et ce fut même une excellente
chose, car, en route, elle trouva tout à coup une combinaison qui lui
permettait de convertir immédiatement ses cent quatre-vingt mille dollars
en un demi-million.

III
Le petit journal auquel Aleck s’était abonnée paraissait le jeudi. Il devait
faire douze cents kilomètres avant d’arriver du village de Tilbury et on ne
pouvait le recevoir que le samedi. La lettre du Tilbury était partie un
vendredi, plus d’un jour trop tard pour que le bienfaiteur ait pu mourir et
que la nouvelle paraisse dans le numéro de cette semaine-là, mais c’était
largement à temps pour que cela ait la possibilité de paraître la semaine
suivante. Donc, les Foster furent obligés d’attendre presque une semaine
entière avant de savoir s’il leur était arrivé quelque chose de satisfaisant ou
non. Ce fut une bien longue semaine et l’attente fut très douloureuse. Ils
auraient à peine pu la supporter s’ils n’avaient eu de nombreuses pauses de
diversion. Nous avons vu qu’ils savaient en trouver. La femme empilait des
fortunes sans discontinuer. Le mari les dépensait ou du moins en dépensait
la partie dont sa femme voulait bien lui permettre de disposer.
Enfin arriva le samedi et en même temps le Sagamore hebdomadaire.
Mme Eversby se trouvait là. C’était la femme du pasteur presbytérien et elle
«travaillait» les Foster pour obtenir d’eux des contributions aux œuvres
charitables. A l’arrivée du journal, la conversation s’arrêta net—du côté des
Foster. Mme Eversby s’aperçut bientôt que ses hôtes n’écoutaient plus un
mot de ce qu’elle disait; elle se leva aussitôt et s’en alla étonnée et indignée
au plus haut point. Dès qu’elle fut hors de la maison, Aleck déchira avec
impatience le bande du journal et elle parcourut hâtivement des yeux la
colonne des décès. Déception! Tilbury n’était nulle part mentionné!
Aleck était chrétienne depuis son berceau. Le devoir et la force de
l’habitude lui imposèrent donc une fausse attitude. Elle se raidit et dit avec
une pieuse joie forcée:
—Soyons humblement reconnaissants qu’il ait été épargné et...
—Au diable sa sale carcasse! Je voudrais...
—Sally, quelle honte!
—Je m’en moque! repartit l’homme furieux. Tu penses la même chose et
si tu n’étais pas si immoralement pieuse, tu serais honnête et tu dirais la
même chose.
Sur un ton de dignité blessée, Aleck répondit:
—Je ne sais pas comment tu peux dire des choses si injustes et si
blessantes. La piété immorale n’existe pas!
Sally eut un remords. Il voulut le cacher en essayant confusément de se
disculper et pour cela il chercha à changer la forme de ses paroles. Il avait
le tort de croire qu’il pourrait apaiser son experte moitié en abandonnant la
forme pour retenir le fond.
 —Je ne voulais pas dire quelque chose de si affreux que cela, Aleck. Je
ne voulais pas dire de la piété immorale... Je voulais dire seulement...
seulement... eh bien, la piété conventionnelle, tu sais... la piété d’église; le...
la... mais tu sais ce que je veux dire, Aleck; le... quand on donne un article
plaqué pour du vrai métal, tu sais, sans penser à mal mais par habitude
invétérée, manière apprise, coutume pétrifiée, loyauté envers... envers...
peste! Je ne trouve pas les bons mots, mais, encore une fois, tu sais ce que
j’ai voulu dire, Aleck, et je n’y vois pas de mal en somme. Je vais
t’expliquer; tu vois, c’est comme si une personne...
—Tu en as tout à fait assez dit, dit Aleck froidement, laisse tomber le
sujet.
—Moi, je veux bien, répondit Sally avec ferveur en s’épongeant le front.
Et une grande reconnaissance qu’il ne savait exprimer se lisait en ses yeux.
Vaincu selon toute apparence, il était désormais convenablement maté et
soumis, aussi lut-il son pardon dans les yeux d’Aleck.
Le grand sujet, leur intérêt suprême revint immédiatement sur le tapis.
Rien ne pouvait le retenir bien longtemps dans l’ombre. Le couple examina
ensemble le problème que posait l’absence de l’avis de décès de Tilbury. Ils
l’examinèrent dans tous les sens, avec plus ou moins d’espoir, mais ils
furent obligés d’en revenir par où ils avaient commencé, c’est-à-dire de
reconnaître que la seule explication raisonnable de l’absence de cet avis
était décidément que Tilbury n’était pas mort. Il y avait là quelque chose
d’un peu triste, même d’un peu injuste peut-être, mais la vérité était là et il
fallait l’accepter. Ils furent d’accord là-dessus.
A Sally, la chose parut être une dispensation insondable, plus insondable
que de raison, pensait-il. C’était bien là une des choses les plus inutilement
insondables qu’il pût se remettre dans l’esprit. Il exprima cette idée avec
quelque sentiment, mais s’il espérait par là «faire marcher» Aleck, il n’y
réussit point. Si elle avait une opinion, elle la garda pour elle.
Les Foster n’avaient plus qu’à attendre le journal de la semaine suivante.
Tilbury avait évidemment mis longtemps à mourir. Telle fut leur pensée
commune. Le sujet fut donc abandonné et ils se remirent à travailler d’aussi
bon cœur que possible.
Maintenant il faut dire que, sans le savoir, ils avaient tout ce temps fait
tort à Tilbury. Tilbury avait tenu sa promesse, il l’avait tenue à la lettre, il
était mort comme il l’avait annoncé. Il y avait alors quatre jours qu’il était
mort et il avait eu le temps de s’y habituer; il était entièrement mort,
parfaitement mort, aussi mort que tous les habitants du cimetière... et son
décès s’était produit largement à temps pour être mentionné dans le
Sagamore de cette semaine-là. Il n’avait été oublié que par pur accident.
Ce fut un accident qui n’arrive jamais aux journaux des grandes villes,
mais qui est fort commun dans les petites feuilles de village. Au moment de
mettre sous presse, les confiseurs de l’endroit envoyèrent au journal une
garde boîte de bonbons et immédiatement la petite nécrologie du triste
Tilbury fut remplacée par les remerciements enthousiastes du directeur.
Mais, dans leur précipitation, les typographes desserrèrent la colonne, et
les caractères composant la notice nécrologique se trouvèrent dispersés.
Sans cela, elle aurait paru dans un autre numéro, car les textes composés
sont immortels dans les imprimeries de ces petits journaux, mais une
composition détruite n’est plus jamais reprise et ainsi, quels que pussent
être les événements futurs, jamais, dans le cours des siècles, le Sagamore ne
devait annoncer le décès de Tilbury!

IV
Cinq semaines s’écoulèrent lentement. Le Sagamore arrivait
régulièrement le samedi, mais jamais il ne mentionna Tilbury Foster. La
patience de Sally fut alors à bout et il s’écria avec colère:
—Maudites soient ses entrailles! Il est immortel!
Aleck le reprit sévèrement et ajouta d’une voix solennelle et glacée:
—Que dirais-tu si tu étais retiré brusquement de la vie après avoir laissé
échapper une telle pensée?
Sally répondit sans avoir suffisamment réfléchi:
—Mais je m’estimerais encore heureux qu’elle se soit échappée.
Il lança cela par orgueil, voulant répondre quelque chose et ne trouvant
rien de mieux sur le moment. Puis il s’esquiva, craignant d’être écrasé dans
une discussion avec sa femme.
Six mois passèrent. Le Sagamore gardait toujours un silence obstiné sur
le sort de Tilbury. En attendant, Sally avait plusieurs fois tenté de «jeter une
sonde», c’est-à-dire de suggérer qu’il faudrait savoir. Aleck s’était montrée
tout à fait indifférente à ces suggestions. Sally résolut alors de réunir toutes
ses forces et d’attaquer de front. Il proposa donc de se déguiser et d’aller au
village de Tilbury pour y découvrir subrepticement quelles pouvaient être
leurs espérances. Aleck s’opposa à ce projet dangereux avec beaucoup
d’énergie et de décision. Elle dit à son mari:
—A quoi peux-tu bien penser? Vraiment tu ne me laisses pas respirer. Il
faut te surveiller tout le temps comme un petit enfant pour l’empêcher de
marcher dans les flammes. Tu resteras exactement où tu es.
—Mais voyons, Aleck! Je pourrais très bien le faire sans que personne le
sache, j’en suis certain...
—Sally Foster, ne sais-tu pas que pour cela il te faudrait poser des
questions sur notre parent?
—Bien sûr. Mais, et puis après? Personne ne se douterait de qui je suis.
—Ah! entendez-le. Un jour, il faudra que tu prouves aux exécuteurs que
tu ne t’es jamais informé. Et alors?
Il avait oublié ce petit détail. Il ne répondit rien; il n’y avait rien à
répondre.
Aleck ajouta:
—Et maintenant, sors cette idée de ta tête et n’y pense plus. Tilbury t’a
préparé ce piège. Ne vois-tu pas que c’est un piège? Il est sur ses gardes et
il compte bien que tu te laisseras prendre. Eh bien! il sera déçu, du moins
tant que je tiendrai le gouvernail, Sally!
—Eh bien?
—Tant que tu vivras, serait-ce cent ans, ne fais jamais d’enquête!
Promets.
—Je promets, je promets, dit le brave homme avec un soupir et à contre-
cœur.
Aleck se radoucit alors et dit:
—Ne sois pas si impatient. Nous prospérons. Nous pouvons attendre,
rien ne presse. Notre petit revenu certain augmente tout le temps. Quant à
l’avenir, je n’ai pas encore fait fausse route, cela s’empile par mille et dix
mille. Il n’y a pas une autre famille dans le district qui ait de si belles
espérances. Déjà nous commençons à rouler dans l’abondance. Tu sais cela,
n’est-ce pas?
—Oui, Aleck, c’est certainement vrai.
 —Alors sois reconnaissant de tout ce que le bon Dieu a fait pour nous et
cesse de te tourmenter. Tu ne t’imagines pas, n’est-ce pas, que nous aurions
pu arriver à ces résultats prodigieux sans son secours et son aide?
—N... non, dit-il en hésitant, je suppose que non. Puis, il ajouta avec
beaucoup de sentiment et d’admiration: et cependant, je crois du fond du
cœur que tu n’as besoin d’aucune aide dans l’élaboration de tes
combinaisons financières.
—Oh! tais-toi. Je sais bien que tu ne penses pas à mal et que tu ne
cherches pas à être irrévérencieux, pauvre homme, mais tu ne sembles pas
pouvoir ouvrir la bouche sans laisser sortir des choses qui font trembler. Tu
me tiens dans un perpétuel émoi. Je crains maintenant pour toi plus que
pour nous tous. Autrefois je n’avais pas peur du tonnerre, mais maintenant,
quand je l’entends, je...
Sa voix sombra, elle commença à pleurer et ne put achever sa phrase.
Cela alla au cœur de Sally; il la prit dans ses bras, la caressa et la consola. Il
lui promit une meilleure conduite et, tout en se réprimandant lui-même, il
implora son pardon en se frappant la poitrine. Il était de bonne foi et peiné
de ce qu’il avait fait; il était prêt à n’importe quel sacrifice pour y remédier.
Par conséquent, il y réfléchit longuement et profondément dans la
solitude et il se décida à faire ce qui lui paraîtrait le mieux. Il était facile de
promettre de changer de conduite, il avait tant de fois promis déjà... Mais
cela ferait-il quelque bien et surtout un bien permanent? Non, ce ne serait
que provisoire, il connaissait sa faiblesse et l’avouait avec chagrin. Il ne
pourrait pas tenir sa promesse, il fallait trouver quelque chose de mieux et
de plus sûr: au prix de ruses fort habiles, il économisa longtemps sou par
sou et lorsqu’il eut assez d’argent, il mit un paratonnerre sur la maison.
Quels miracles l’habitude ne peut-elle faire accomplir! Et comme les
habitudes sont vite et facilement prises! aussi bien les habitudes
insignifiantes que celles qui nous transforment complètement. Si, par
accident, nous nous réveillons à deux heures du matin deux nuits de suite,
nous avons raison de nous inquiéter, car un accident semblable peut créer
une habitude: l’usage du whisky pendant un mois peut... mais inutile
d’insister: nous connaissons, tous, ces faits ordinaires de la vie.
L’habitude de bâtir des châteaux au pays des songes, l’habitude de rêver
en plein jour, comme elle grandit vite! Quelle jouissance elle devient!
Comme nous volons à des enchantements nouveaux et délicieux à chaque
moment de loisir, comme nous aimons nos chimères, comme nous savons
endormir nos âmes et nous enivrer de nos propres fantaisies trompeuses!
Oh! oui, et combien notre vie irréelle s’emmêle et se fusionne vite et
facilement avec notre vie matérielle de telle façon que nous ne pouvons
plus les distinguer l’une de l’autre!
Aleck s’abonna bientôt à un journal quotidien de Chicago et à
l’Indicateur des Finances. Douée d’un singulier flair financier, elle les
étudia aussi consciencieusement, toute la semaine, qu’elle étudiait sa Bible
le dimanche. Sally était perdu d’admiration en remarquant avec quelle
sûreté se développaient le génie et le jugement de sa femme en tout ce qui
concernait le soin de leurs capitaux tant matériels que spirituels. Il était
aussi fier de la voir exploiter audacieusement les affaires de ce monde que
de bénéficier de la conscience avec laquelle elle savait se prémunir en vue
de l’avenir éternel. Il remarqua qu’elle n’avait jamais cessé de tenir la
balance égale entre ses affaires terrestres et ses affaires religieuses. Dans les
deux cas, comme elle le lui expliqua un jour, il s’agissait de capitaux; mais
en ce qui concerne les capitaux terrestres, elle ne s’en préoccupait, et ne les
plaçait que pour les déplacer en vue de la spéculation, tandis que, dans le
second cas, elle plaçait ses capitaux spirituels une fois pour toutes et dans
une affaire de tout repos. Ainsi, elle ne perdait pas la tête et elle savait se
garantir un bon avenir—dans tous les cas et de toutes les façons.
Il ne fallut que quelques mois pour former l’imagination de Sally et
d’Aleck. Chaque jour l’ébullition de leur cerveau se faisait plus intense. En
conséquence, Aleck gagnait de l’argent imaginaire beaucoup plus vite
qu’elle n’avait rêvé de pouvoir le faire au début, et l’habileté de Sally à en
dépenser le surplus grandit en proportion. Aleck s’était tout d’abord accordé
douze mois pour spéculer sur les charbons, tout en reconnaissant que ce
délai pourrait peut-être se réduire à neuf mois. Mais c’était là un petit
travail, un travail d’enfant, dû à des facultés financières qui n’avaient
encore rien appris, rien expérimenté... qui ne connaissaient pas tous les
perfectionnements possibles: les perfectionnements arrivèrent bientôt; alors
les neuf mois s’évanouirent et le placement imaginaire des dix mille dollars
revint triomphant avec trois cents pour un de profit derrière lui en moins de
trois mois!
Ce fut une grande journée pour les Foster. Ils en restèrent muets de joie...
muets aussi pour une autre raison: après avoir beaucoup surveillé le marché,
Aleck avait fait dernièrement avec crainte et tremblement son premier essai.
Elle risqua les derniers vingt mille de l’héritage. En esprit, elle vit grimper
la cote, point par point—avec la possibilité constante d’une chute
imprévue... A la fin, son anxiété devint trop grande; elle était encore novice
dans l’art de l’achat à découvert et non endurcie encore... Elle avait donc,
par une dépêche imaginaire, donné l’ordre de vendre. Elle dit que quarante
mille dollars de bénéfice étaient suffisants. La vente fut effectuée le jour
même où ils apprirent l’heureuse issue de l’affaire des charbons.
Donc, ce soir-là, ils restèrent ébahis et bien heureux, tâchant de
s’habituer à leur bonheur et de se faire à l’idée qu’ils valaient actuellement
cent mille dollars en bon argent solide et imaginaire.
Ce fut la dernière fois qu’Aleck se laissa épouvanter par la spéculation
ou plutôt son anxiété ne parvint plus comme cette fois-ci à troubler son
sommeil et à pâlir sa joue.
Ce fut vraiment une soirée mémorable. Petit à petit, l’idée qu’ils étaient
riches prit profondément racine dans leurs âmes et ils se mirent à chercher
des placements. Si nous avions pu voir avec les yeux de ces rêveurs, nous
aurions vu leur petit cottage en bois, si gentil et si propret, faire place à une
belle bâtisse où briques à deux étages et entourée d’une grille en fer; nous
aurions vu un triple lustre accroché au plafond; nous aurions vu l’humble
devant de foyer devenir un Brussels resplendissant à 10 francs le mètre;
nous aurions vu la cheminée plébéienne remplacée par un phare orgueilleux
aux portes de mica. Nous aurions vu bien d’autres choses encore, par
exemple, le cheval, la voiture, le traîneau, le chapeau haut-de-forme... et
tout le reste.
A partir de ce moment-là et bien que leurs filles et les voisins n’aient
toujours continué à voir que le petit cottage, ce petit cottage était devenu
une maison à deux étages pour Aleck et Sally et pas une nuit ne se passa
sans qu’Aleck ne se fît un grand souci des imaginaires notes de gaz et ne
reçût pour toute consolation que l’insouciante réponse de Sally:
—Eh bien, quoi! nous pourrons toujours payer ça!
Avant d’aller se coucher ce premier soir de leur richesse, le couple
décida qu’il fallait se réjouir de quelque façon... Ils donneraient un grand
dîner... Oui, c’était une bonne idée. Mais quelle raison donner aux enfants et
aux voisins? Ils ne pouvaient songer à dévoiler le fait qu’ils étaient riches.
Sally y aurait consenti, il le désirait même, mais Aleck ne perdit pas la tête
et s’y opposa catégoriquement. Elle dit que l’argent était aussi réel que s’il
était dans leur coffre-fort, mais qu’il fallait attendre qu’il y fût en réalité.
Elle établit sa ligne de conduite sur cette base et elle demeura inébranlable.
Pour elle, le grand secret devait être gardé vis-à-vis des enfants et du monde
entier.
Ils furent donc très perplexes. Il fallait se réjouir, ils y tenaient à tout
prix, mais puisqu’ils devaient garder le secret, quel prétexte donner? Aucun
anniversaire n’était proche. Sally, poussé à bout, s’impatientait. Mais
brusquement il eut ce qu’il lui parut être une magnifique inspiration. Tout
leur ennui s’évanouit en une seconde; ils pourraient célébrer la découverte
de l’Amérique. C’était une idée splendide!
Aleck fut extrêmement fière de Sally. Elle dit qu’elle n’y aurait jamais
songé, mais Sally, bien que gonflé de joie et d’orgueil, essaya de n’en
laisser rien voir et dit que ce n’était vraiment rien, que n’importe qui aurait
pu y penser. A quoi l’heureuse Aleck répondit avec un élan de fierté:
—Oh, certainement, n’importe qui! oh, n’importe qui! Hosanna Dilknis
par exemple ou peut-être Adelbert Pistache... Oh, oui! Eh bien, je voudrais
bien les y voir, voilà tout! Bonté divine, s’ils arrivaient à penser à la
découverte d’une île de quarante kilomètres carrés, c’est, je parie bien, tout
ce qu’ils pourraient faire; mais pour ce qui est d’un continent tout entier,
voyons, Sally Foster, tu sais parfaitement qu’ils se feraient une entorse au
cerveau, et même alors, ils n’y arriveraient pas!
... La chère âme! Elle savait que son mari était intelligent et si, par
affection, elle jugeait cette intelligence au-dessus de sa valeur, sûrement elle
était pardonnable pour une erreur si douce et aimable.

V
La fête projetée se passa à merveille. Les amis de Foster, jeunes et vieux,
étaient tous présents. Parmi les jeunes se trouvaient Flossie et Grâce
Pistache et leur frère Adelbert qui était un jeune ferblantier plein d’avenir. Il
y avait aussi Hosanna Dilkis jeune, un plâtrier ayant à peine fini son
apprentissage. Depuis bien des mois, Adelbert et Hosanna s’étaient montrés
très assidus auprès de Gwendolen et Clytemnestra Foster, et les parents des
deux jeunes filles en avaient témoigné leur entière satisfaction. Mais ils se
rendirent compte alors que ce sentiment n’existait plus. Ils sentaient que
leur nouvelle condition financière avait élevé une barrière sociale entre
leurs filles et les jeunes ouvriers. Leurs filles pouvaient désormais regarder
plus haut, c’était même leur devoir. Elles ne devaient pas viser à épouser
moins qu’un homme d’affaires ou un avocat... Mais papa et maman s’en
chargeraient et il n’y aurait pas de mésalliance.
Néanmoins, ces projets et ces pensées demeuraient cachés et ne firent
pas ombrage aux réjouissances de la fête. Tout ce que les invités purent lire
sur la physionomie de leurs hôtes, ce fut un contentement calme et hautain.
Leur maintien grave et digne força l’admiration et l’étonnement de tous les
assistants. Tous s’en aperçurent, tous en parlèrent, mais personne n’en
devina le secret. C’était un profond mystère. Trois personnes différentes
firent sans malice la même remarque sur cette apparente prospérité: «On
dirait qu’ils ont fait un héritage!»
Et c’était bien vrai.
La plupart des mères auraient pris en main la question matrimoniale à
l’ancienne façon. Elles auraient fait un long discours solennel et sans tact—
un discours fait pour manquer son but en causant des larmes et des révoltes
secrètes; et les mêmes mères auraient gâté davantage leur jeu en défendant
aux jeunes ouvriers de continuer leurs assiduités auprès de leurs filles. Mais
la mère des petites Foster était d’une nature différente, elle était pratique.
Elle ne parla de la chose ni aux jeunes gens, ni à personne, sauf à Sally. Il
l’écouta et comprit, il comprit et il admira. Il dit:
—Je saisis ton idée. Au lieu de mépriser les échantillons offerts et de
gâter le marché en blessant des sentiments légitimes, tu cherches
simplement d’autres échantillons pour le même prix et tu laisseras faire le
reste à la nature. C’est la sagesse, Aleck, la sagesse solide comme un roc.
Où est ton élu? As-tu déjà jeté l’hameçon?
Elle n’avait encore rien fait. Il fallait qu’ils examinent ensemble l’état de
ce que Sally appelait le «marché». Pour commencer, ils discutèrent le jeune
Brodish, avocat de talent, et Fullon, un jeune dentiste. Il aurait fallu que
Sally les invitât à déjeuner.
—Mais, pas tout de suite, rien ne presse, dit Aleck. Il faut les surveiller
et attendre, une grande lenteur ne nuirait jamais dans une affaire si
importante.
Il arriva que cette façon de voir fut sage aussi, car, avant trois semaines,
Aleck fit une affaire qui porta ses imaginaires cent mille à quatre cent mille
de la même espèce. Pour la première fois, les Foster eurent du champagne à
dîner... Pas du vrai, mais une qualité assez ressemblante pour contenter leur
imagination montée. Ce fut Sally qui le proposa et Aleck consentit par
faiblesse. Au fond, ils étaient tous les deux troublés et honteux, car M.
Foster faisait partie de la Société de tempérance et aux enterrements il en
portait les insignes: un large baudrier qui faisait peur aux chiens. Sa femme
appartenait à la Ligue des W. C. T. U. et adhérait à tout ce que cette ligue
implique de vertu sans tache et de rigide sainteté. Mais voilà! La vanité des
richesses commençait à se loger dans leurs cœurs et à y faire son travail
accoutumé! Ils agissaient, pour prouver une fois de plus la triste vérité
démontrée déjà bien des fois dans le monde: savoir que la pauvreté surpasse
tous les bons principes pour défendre l’homme contre les vices et la
dégradation. Ils valaient désormais plus de quatre cent mille dollars. Ils
reprirent la question matrimoniale; il ne fut question, ni du dentiste, ni de
l’avocat, il n’y avait plus lieu, ils étaient forcément disqualifiés. Ils
causèrent du fils du banquier et du fils du docteur, mais, comme dans le cas
précédent, ils décidèrent d’attendre et de réfléchir, d’aller lentement et
sûrement.
La chance vint de nouveau de leur côté. Aleck, toujours en éveil, vit une
occasion, très risquée à la vérité, mais elle eut l’audace de faire le pas... Un
temps de doute, de frissons, de terrible malaise survint, car l’insuccès aurait
équivalu à la ruine complète... Puis vint le résultat et Aleck, à demi morte
de joie, pouvait à peine maîtriser sa voix lorsqu’elle dit à Sally:
—L’incertitude est passée, Sally, et nous valons un million!
Sally pleura de reconnaissance et dit:
—Oh! Electra, joyau entre toutes les femmes, chérie de mon cœur, nous
sommes enfin libres, nous nageons dans la richesse, nous n’aurons plus
besoin de compter. Ce serait le cas où jamais d’acheter une bouteille de
vraie «veuve Cliquot».
Et il sortit de l’armoire une bouteille de bière brune qu’il offrit en
sacrifiée, en disant:
—Au diable la dépense!
Elle le réprimanda doucement, les yeux tout humides de joie.
Ils reléguèrent le fils du banquier et le fils du docteur et s’assirent pour
considérer les titres du fils du gouverneur et du fils d’un sénateur.
 VI
Il serait oiseux de rapporter par le menu toutes les bonnes affaires
fictives qui enrichirent si rapidement les Foster. Leurs progrès furent
rapides, foudroyants, merveilleux. N’importe quelle combinaison trouvée
par Aleck devenait une mine d’or. Les millions s’entassèrent sur les
millions et le Pactole aux flots merveilleux qui les submergeait augmentait
sans cesse de volume en roulant à pleins bords. Les Foster eurent
successivement cinq millions, dix millions, puis vingt, trente millions... Où
devait donc s’arrêter leur fortune?
Deux années passèrent pour eux dans ce délire; c’est à peine si, dans leur
rêve, ils sentaient le temps passer. Ils possédaient alors trois cents millions
de dollars. Ils faisaient partie de toutes les grandes banques de l’État et, bien
qu’ils n’en eussent aucun souci, ils augmentaient sans cesse leurs capitaux...
C’était une fois cinq millions, une autre fois dix, qui venaient tomber dans
leur caisse... par la force des choses. Les combinaisons, les coups de bourse
leur étaient toujours profitables. Leurs trois cents millions se trouvèrent
doublés, puis une seconde fois, puis une troisième.
Ils eurent plus de deux milliards!
Toutes ces affaires n’allaient pas sans une certaine quantité d’écritures.
A ce moment-là, la comptabilité parut s’embrouiller. Les Foster virent et
comprirent qu’il serait désastreux de laisser cet état s’aggraver davantage,
ils savaient bien qu’une chose bien commencée doit être continuée avec
soin dans toutes ses parties, mais pour bien tenir leurs comptes en règle,
c’était une affaire de plusieurs heures de travail par jour. Et où trouver ces
heures-là? Sally vendait des épingles et du sucre tout le long du jour et,
pendant ce temps, Aleck faisait la cuisine, lavait la vaisselle, faisait les lits...
et cela chaque jour de l’année! Bien plus, elle ne réclamait pas l’aide de ses
filles, car ses filles avaient à garder des mains blanches pour jouer leur rôle
futur dans la haute société. Les Foster savaient, chacun d’eux savait bien
qu’il n’y avait qu’une façon de trouver les douze ou quinze heures par
semaine qu’il fallait employer aux comptes, mais tous deux avaient honte
d’y penser et tous deux attendaient que l’autre en parlât le premier.
Enfin, Sally dit:
—Il faudra y arriver. C’en est trop pour moi. Fais comme si j’avais dit la
chose, n’importe si j’ai prononcé le mot ou non.
 Aleck rougit, mais fut reconnaissante. Sans plus discourir ils tombèrent
dans le péché: ils violèrent le jour du repos[B], ils l’employèrent à leurs
calculs; il n’y avait que ce moyen pour ne pas se perdre dans leurs registres.
Et ce premier péché en appela d’autres. Il n’y a que le premier pas qui
coûte. De grandes richesses constituent des tentations terribles et elles
mènent fréquemment à la perdition les gens qui n’y sont pas habitués. Donc
les Foster se plongèrent dans le péché et profanèrent le saint jour. Ils se
mirent au travail avec acharnement et compulsèrent leur portefeuille. Quelle
magnifique liste de valeurs! Les Chemins de Fer, les Transatlantiques, les
Pétroles, les Câbles inter-océaniens, les De Beers, les Cuivres, l’Acier!
Deux milliards! Et tout excellemment placé en des affaires sûres, en des
entreprises connues, prospères, donnant de gros intérêts et des dividendes
superbes. Leur revenu était de cent vingt millions par an! Aleck, à ce
résultat, poussa un long soupir de joie et dit:
—Est-ce assez?
—Oui, Aleck.
—Que faut-il faire maintenant?
—Tout arrêter.
—Se retirer des affaires?
—C’est cela.
—Je suis de cet avis. Le gros travail est achevé. Il nous faut prendre un
long repos et jouir de notre argent.
—Parfait!... Aleck?
—Quoi donc, chéri?
—Combien pouvons-nous dépenser de notre revenu?
—Tout.
A ce mot, son mari se sentit débarrassé d’un énorme fardeau, il ne
répondit pas un mot, il était trop heureux pour le manifester en paroles.
Dès lors, ils continuèrent à violer le jour de repos chaque semaine. Ce
n’est que le premier pas qui coûte, hélas, répétons-le! Chaque dimanche ils
passaient la journée tout entière à élaborer les plans des différentes choses
qu’ils feraient pour dépenser leur revenu. Et ils ne s’arrêtaient dans ces
délicieuses occupations que fort avant dans la nuit. A chaque séance, Aleck
mettait des millions à la disposition des grandes œuvres philanthropiques et
religieuses, et Sally employait autant de millions à des affaires auxquelles il
donna des noms dans le commencement, mais dans le commencement
seulement, car, par la suite, il les rangea toutes sous la catégorie commode
et complète de «dépenses diverses». Ces occupations furent la cause de
sérieuses dépenses en pétrole. Pendant quelque temps, Aleck en fut un peu
ennuyée, puis, après quelques semaines, elle cessa d’en prendre souci, car
elle n’en eut plus le prétexte; elle fut seulement peinée, affligée, honteuse,
mais elle ne dit rien. C’était si peu de chose! En réalité, Sally prenait ce
pétrole au magasin, il se faisait voleur. Il en est toujours ainsi. De grandes
richesses, aux personnes qui ont été pauvres, sont de terribles tentations.
Avant que les Foster ne devinssent riches, on pouvait leur confier un litre de
pétrole, mais maintenant... jetons un voile sur ces petites faiblesses de leurs
consciences. Du reste, du pétrole aux pommes il n’y a pas loin. Sally
rapporta quelques pommes. Puis un morceau de savon, puis une livre de
sucre. Ah! comme il est facile d’aller du mal au pire, lorsqu’une fois on est
parti du mauvais pied!
Entre temps, d’autres événements avaient marqué le chemin où les
Foster couraient avec leur fortune. Leur irréelle maison de briques avait fait
place à un magnifique château de pierres de taille couvert d’ardoises. Peu
après, ce château lui-même devint trop petit, d’autres demeures s’élevèrent,
royales et étonnantes, toujours plus hautes, plus grandes, plus belles et
chacune à son tour fut abandonnée pour un palais plus vaste encore et d’une
architecture plus merveilleuse. En ces derniers jours, nos rêveurs venaient
de faire construire le château idéal. Il s’élevait, dans une région lointaine,
sur une colline boisée au-dessus d’une vallée sinueuse où une rivière
déroulait ses méandres gracieux... tout le pays lui servait de parc... C’était
bien le palais des amants du rêve.
Toujours animée par de nombreux hôtes de distinction, cette splendide
demeure se trouvait à l’orient, vers Newport Rhode Island, en pleine Terre-
Sainte de l’aristocratie américaine. D’habitude, ils passaient une grande
partie de leur dimanche dans cette magnifique demeure seigneuriale et le
reste du temps ils allaient en Europe ou faisaient quelque délicieuse
croisière dans leur yacht étincelant. Six jours de vie basse et matérielle,
étroite et mesquine, et le septième jour tout entier passé en pleine cité des
songes... ainsi s’était ordonnée leur existence.
Et dans leur vie de la semaine, durant les six jours de travail vulgaire, ils
demeurèrent diligents, soigneux, pratiques, économes. Ils continuèrent à
faire partie de leur petite église presbytérienne et à travailler pour elle et
pour le succès des dogmes austères qu’elle enseignait. Mais, dans leur vie
de rêve, ils n’obéissaient plus qu’à leurs fantaisies, quelles qu’elles fussent,
et même si elles étaient changeantes. En cette question d’église, les
fantaisies d’Aleck ne furent pas très désordonnées ni bien fréquentes, mais
celles de Sally firent compensation. Aleck, dans son existence imaginaire,
s’était ralliée tout de suite à l’église épiscopale dont les attaches officielles
l’attiraient. Mais peu après, elle entra dans la haute église à cause du grand
luxe de cérémonies, et enfin elle se fit catholique pour la même raison.
Les libéralités des Foster commencèrent dès le début de leur prospérité,
mais à mesure que croissait leur fortune, elles devinrent extraordinaires,
énormes. Aleck bâtissait une ou deux Universités par dimanche, un hôpital
ou deux aussi, quelques églises, très souvent une cathédrale. Une fois, Sally
s’écria gaiement, mais sans avoir bien réfléchi:
—Il faut qu’il fasse froid pour qu’un jour se passe sans que ma femme
n’envoie un bateau chargé de missionnaires pour décider ces excellents
Chinois à troquer leur confucianisme contre le christianisme!
Cette phrase peu courtoise et ironique blessa les sentiments profonds
d’Aleck et elle se retira en pleurant. Cela le fit rentrer en lui-même et lui
alla au cœur; dans sa triste honte, il aurait donné des mondes pour que cette
malencontreuse phrase ne lui fût pas venue à la bouche. Elle n’avait
formulé aucun blâme et cela le mettait encore plus mal à l’aise. Une telle
situation pousse fatalement au retour sur soi-même... Ne l’avait-il pas
attristée déjà d’autres fois? Ce silence généreux qu’elle lui avait seul opposé
l’angoissa plus que tout, et, reportant ses pensées sur lui-même, il revit
défiler devant lui, en procession lamentable, les tableaux de sa vie, depuis
l’origine de leur fortune. Et comme les souvenirs revenaient en foule, il
sentit que ses joues s’empourpraient de honte et qu’il s’était plus avili qu’il
n’aurait cru. L’existence de sa femme, qu’elle était belle, toute semée de
généreuses actions, toute tendue vers les choses idéales; et la sienne, qu’elle
était frivole, égoïste, vide, ignoble... et toute orientée vers les plus bas et
vils soucis? Jamais il n’avait fait un pas vers le mieux mais quelle descente
vertigineuse vers le pire!
Il compara ses propres actes avec ceux de sa femme et il s’indigna de
s’être moqué d’elle, lui. Ah! c’était bien à lui de reprocher quelque chose à
sa généreuse Aleck! Que pouvait-il dire? Qu’avait-il fait jusqu’à présent?
Voici: lorsqu’elle bâtissait sa première église, lui, il fondait avec d’autres
multimillionnaires blasés un Poker club où il perdait des centaines de mille
dollars, et il était fier de la célébrité que cela lui amenait. Lorsqu’elle
bâtissait sa première université, que faisait-il? Il menait une vie de
dissipation et de plaisirs secrets dont le scandale avait été grand.
Lorsqu’elle fondait un asile pour les infirmes, que faisait-il?—Hélas!
Lorsqu’elle établissait les statuts de cette noble Société pour la purification
des sexes, que faisait-il? Oui, vraiment, que faisait-il? Et lorsqu’elle se fit
accompagner de toutes les W. C. T. U. pour briser les flacons de la
pernicieuse liqueur, lui, que faisait-il? Ivre trois fois par jour! Enfin, au
moment où cette femme au grand cœur, qui avait bâti plus de cent
cathédrales, était reçue avec grand honneur par le pape et recevait la Rose
d’or qu’elle avait si bien gagnée... lui, lui, où était-il? Il faisait sauter la
banque à Monte-Carlo.
Il s’arrêta. Il n’eut pas la force d’aller plus loin. Il ne pouvait pas
supporter tous ces affreux souvenirs. Il se leva soudain. Une grave
résolution s’affirmait en lui: tous ces secrets devaient être révélés, toutes ces
fautes devaient être confessées. Il ne mènerait plus une vie à part. Et il allait
d’abord droit à elle pour lui dire tout.
C’est ce qu’il fit. Il lui raconta tout. Il pleura sur son sein, il sanglota,
gémit et implora son pardon. Ce fut pour elle une bien dure minute et le
choc qu’elle éprouva fut d’une violence inouïe, elle blêmit et chancela, mais
elle se reprit... Après tout, n’était-il son mari, son bien, son tout, le cœur de
son cœur, l’amour de ses yeux, le sien, à elle depuis toujours et en toutes
choses? Elle lui pardonna. Elle sentit cependant qu’il ne serait plus jamais
pour elle tout ce qu’il avait été jusqu’alors; elle savait qu’il pouvait se
repentir, mais non se réformer. Mais encore, tout avili et déchu qu’il fût,
n’était-il pas son tout, son unique, son idole, son amour? Elle lui dit qu’elle
n’était que son esclave et lui ouvrit tout grands ses bras.

VII
Quelque temps après ces événements, les époux Foster étaient
mollement étendus sur des fauteuils à l’avant de leur yacht de rêve qui
croisait dans les mers du Sud. Ils gardaient le silence, chacun d’eux ayant
fort à faire avec ses propres pensées. Ces longs silences étaient devenus de
plus en plus fréquents entre eux, et leur vieille et familière camaraderie qui
leur faisait se partager toutes leurs idées s’était insensiblement évanouie.
Les terribles révélations de Sally avaient fait leur œuvre; Aleck avait fait de
terribles efforts pour en chasser le souvenir, mais elle n’avait pas réussi à
s’en débarrasser et la honte et l’amertume qu’elle avait ressenties à ces
tristes aveux demeuraient dans son âme empoisonnant peu à peu sa noble
vie de rêve. Elle s’apercevait maintenant (le dimanche) que son mari
devenait un être arrogant et fourbe. Elle ne voulait pas le voir, aussi tâchait-
elle de ne pas lever les yeux sur lui le dimanche.
Mais elle-même ne méritait-elle aucun blâme? Hélas! elle savait bien ce
qui en était. Elle avait un secret pour son mari, elle n’agissait pas
loyalement envers lui et cela lui causait une grande angoisse. Elle n’avait
pas osé lui en parler! Elle avait été tentée par une occasion exceptionnelle
qui se présentait de mettre la main sur l’ensemble des chemins de fer et des
mines de tout le pays et elle s’était remise à la spéculation et elle avait
risqué leur fortune entière. Maintenant, elle tremblait tout le jour chaque
dimanche de laisser percer son inquiétude. Elle avait trahi la confiance de
son mari et dans son misérable remords elle se sentait remplie de pitié pour
lui, elle le voyait là devant elle tout heureux, satisfait, confiant en elle!
Jamais il n’avait eu le moindre soupçon et elle se désespérait en elle-même
à penser qu’une calamité formidable où sombrerait toute leur fortune ne
tenait maintenant qu’à un fil...
—Dis donc, Aleck!
Cette interruption soudaine de son rêve causée par l’appel de son mari
lui fut un soulagement. Elle lui en fut reconnaissante et quelque chose de
son ancienne tendresse perça dans le ton de sa douce réponse:
—Eh bien, chéri!
—Sais-tu, Aleck, je pense que nous nous sommes trompés, c’est-à-dire
que tu t’es trompée. Je parle de cette affaire de mariages.
Il s’assit, se cala bien dans un grand fauteuil et continua vivement:
—Considère bien tout. Voilà plus de cinq ans que cela traîne. Tu as
toujours agi de la même façon depuis le commencement: à mesure que nous
devenions plus riches, tu as élevé tes prétentions. Toutes les fois que je
m’imagine avoir bientôt à préparer les noces, tu aperçois un meilleur parti
pour tes filles et je n’ai plus qu’à rengainer mes préparatifs avec mon
désappointement. Je crois que tu es trop difficile. Quelque jour nous nous
en repentirons. D’abord, nous avons écarté le dentiste et l’avocat... Cela,