Drug Receptors & Pharmacodynamics

Nancy Bruton-Maree, CRNA, MS Raleigh School of Nurse Anesthesia UNCG

Pharmacodynamics
The component of a cell or organism that interacts with a drug & initiates the chain of events leading to the drugs observed effect is a receptor Receptors have become the central focus of investigation of drug effects & their mechanisms of action (Pharmacodynamics)

Receptor Concept
Receptors largely determine the quantitative relations between dose or concentration of drug & pharmacologic effects Receptors are responsible for selectivity of drug action Receptors mediate the actions of pharmacologic agonists & antagonists

Common Definitions
Orphan receptors their ligands have not been identified Regulatory proteins Proteins which medicate the actions of endogenous chemical signals
Classes of proteins
Enzymes one of the types of receptors that has been identified which may be inhibited by binding a drug Transport proteins Structural proteins

Relation Between Drug Concentration & Response

This can be described with mathematical precision This is an idealized relationship that underlies the more complex relations between dose & effect that occur when a drug is given to a patient.

Concentration-Effect Curve & Receptor Binding of Agonists

Responses to low doses of a drug usually increase in direct proportion to dose As doses increase, the response increment diminishes Doses may be reached at which no further increase in response can be achieved

Equation Used to Develop Curve

E = Emax X C C + EC50 E = effect observed at concentration C EMAX = the maximal response that can be produced by the drug EC50 = the concetration of drug that produces 50% of maximal effect

Relationship Between Drug Concentration & Drug Effect or Receptor Bound Drug

The Drug Concentration at Which Effect or Receptor Occupancy is Half-Maximal Are Denoted by EC50 & Kd

Hyperbolic Relationship
Resembles the mass action law, which describes association between 2 molecules of a given affinity This resemblance suggests that drug agonists act by binding to a distinct class of biologic molecules with a characteristic affinity for the drug receptor Drug bound to receptors (B) relates to the concentration of free (unbound) drug (C) as depicted in the previous slide

Previous Curve
Equation
B = Bmax X C C + Kd Bmax = indicates the total concentration of receptor sites Kd = the concentration of free drug at which half-maximal binding is observed. This constant characterizes the receptors affinity for binding the drug in a reciprocal fashion If the Kd is low, the binding affinity is high and vice versa The EC50 & Kd may be identical, but need not be

Other Facts About Curves

Dose-response data are often presented as a plot of the drug effect (ordinate) against the logarithm of the dose or concentration (abscissa) This makes the curve sigmoid in shape with a linear midportion This expands the scale of the concentration axis at low concentrations (where the effect is changing rapidly) & compresses it at high concentrations (where the effect is changing slowly), but has no special biologic or pharmacologic significance.

Receptor-Effector Coupling & Spare Receptors

Coupling the transduction process that links drug occupancy of receptors & pharmacologic response The relative efficiency of occupancy-response coupling is partially determined by the initial conformational change in the receptor The effect of full agonists can be considered more efficiently coupled to receptor occupancy than can the effects of partial agonists

Receptor-Effector Coupling & Spare Receptors

Receptor occupancy into cellular response coupling efficiency is also determined by the biochemical events that transduced Sometimes the biologic effect of the drug is linearly related to the number of receptors bound This is often true for the drug-regulated ion channels in which the ion current produced by the drug is directly proportional to the number of receptors bound

Receptor-Effector Coupling & Spare Receptors

Factors that contribute to non-linear occupancyresponse coupling
spare receptors receptors are spare for a given pharmacologic response if it is possible to elicit a maximal biologic response at a concentration of agonist that does not result in occupancy of the full complement of available receptors

Receptor-Effector Coupling & Spare Receptors

What does Temporal mean?
Maximal response can be elicited by activation of relatively few receptors because the response initiated by an individual ligand-receptor binding event persists longer than the binding event itself Spare in number If the concentration or amount of cellular components other than the receptors limits the coupling of receptor occupancy to response, then a maximal response can occur without occupancy of all receptors; thus the sensitivity of a cell or tissue to a particular concentration of agonist depends not only on the affinity of the receptor for binding the agonist but also on the degree of spareness (the total number of receptors present compared with the number actually needed to elicit a maximal response.

Receptor-Effector Coupling

Concept of Spare Receptor Coupling

B Bmax = C C + Kd The Dd of the agonist-receptor interaction determines what fraction (B/Bmax) of total receptors will be occupied at a given free concentration (C) of agonist regardless of the receptor concentration

Competitive & Irreversible Antagonists

Receptor antagonists bind to receptors but do not activate them The primary action of antagonists is to prevent agonists from activating receptors Inverse agonist also an antagonist but can reduce receptor activity below basal levels observed in the absence of bound ligand

Competitive & Irreversible Antagonists

Competitive Antagonist In the presence of a fixed concentration of agonist, increasing concentrations of a reversible antagonist progressively inhibit the agonist response High antagonist concentration prevent response completely. However, sufficiently high concentrations of agonist can surmount the effect of a given concentration of the antagonist. Because the antagonism is competitive, the presence of antagonist increases the agonist concentration required for a given degree of response and so the agonist concentration-effect curve is shifted to the right.

Competitive Antagonism

Competitive v. Irreversible Antagonism

The concentration of the agonist required to produce a given effect in the presence of a fixed concentration of competitive antagonist is greater than the concentration of the agonist required to produce the same effect when the antagonist is not present. The ratio of the two agonists concentration is related to the dissociation constant of the antagonist by the Schild equation

Noncompetitive Antagonist

Schild Equation
C C = 1 + [I] Ki
C is the concentration of an agonist required to produce a given effect in the presence of a fixed concentration [I] of competitive antagonist [I] is the fixed concentration of the antagonist C the agonist concentration required to produce the same effect in the absence of the antagonist Ki is the dissociation constant of the antagonist

Importance of Schild Equation

The degree of inhibition produced by a competitive antagonist depends on the concentration of antagonist Clinical response to a competitive antagonist depends on the concentration of agonist that is competing for binding to receptors

Irreversible Antagonism
This type of antagonism may be irreversible or nearly irreversible It involves forming covalent bonds or binding so tightly to receptors that the receptor is unavailable to the agonist

Possible Reversal of Irreversible Antagonism

Advantages & Disadvantages of Irreversible Antagonism

Once the irreversible antagonist has occupied the receptor, it need not be present in unbound form to inhibit agonist responses Therefore, the duration of action of such an irreversible antagonist is relatively independent of its own rate of elimination & more dependent on the rate of turnover of receptor molecules

Noncompetitive Antagonists
Noncompetitive antagonists can function in different ways:
By binding to a site on the receptor protein separate from the agonist binding site which prevents receptor activation without blocking agonist binding Although these drugs can act noncompetitively, their action is reversible if they do not bind covalently Allosteric modulators function without inactivating the receptor & alter receptor function without inactivating the receptor

Partial Agonists
There are two classes of agonists based on the maximal pharmacologic response that occurs when all receptors are occupied. The two classes are full agonist & partial agonist

Receptor-Effector Coupling

Partial Agonist v. Full Agonist

Partial Agonists
Facts
Failure of a partial agonist to produce a maximal response is not due to decreased affinity for binding receptors Partial agonists competitively inhibit the responses produced by full agonists Many drugs used as antagonists are actually weak partial agonists

Partial Agonist

Other Mechanisms of Drug Antagonism

Chemical antagonist one drug acts as a chemical antagonist of the other simply by ionic binding that makes the other drug unavailable for interactions with the proteins involved in the outcome (i.e., blood clotting) Physiologic antagonist between endogenous regulatory pathways mediated by different receptors. The use of physiologic antagonists produces effects that are less specific & less easy to control than are the effects of a receptor-specific antagonism

Signaling Mechanisms & Drug Action

Questions to answer:
Why do some drugs produce effects that persist for minutes, hours, or even days after the drug is no longer present in the body? Why do responses to other drugs rapidly diminish with prolonged or repeated administration? How do cellular mechanisms for amplifying external chemical signals explain the phenomenon of spare receptors? Why do chemically similar drugs often exhibit extraordinary selectivity in their actions? Do these mechanism provide targets for enveloping new drugs?

Five Basic Mechanisms of Transmembrane Signaling

Lipid-soluble ligand that crosses the membrane & acts as an intracellular receptor Transmembrane receptor protein whose IC enzymatic activity is allosterically regulated by a ligand that binds to a site on the proteins EC domain Transmembrane receptor that binds & stimulates a protein tyrosine kinase Ligand-gated transmembrane ion channel that can be induced to open or close by the binding of ligand Transmembrane receptor protein that stimulates a GTP-binding signal transducer protein (G protein)which in turn modulates production of an IC second messenger.

Five Basic Mechanism

IC Receptors for Lipid-Soluble Agents

Classes include:
Steroids Thyroid hormone

The receptors for these stimulate transcription of genes by binding to response elements on DNA near the gene whose expression is to be regulated

IC Receptors

Two Therapeutically Important Consequences

All of these hormones produces their effect after a characteristic lag period of 30 minutes to several hours
The time required to synthesize new proteins

The effects of these agents can persist for hours or days after the agonist concentration has been reduced to zero
The persistence is due to slow turnover of most enzymes & proteins, which can remain active in cells for hours or days after synthesis.

Ligand-Regulated Transmembrane Enzymes Including Receptor Tyrosine Kinases

Signalling is by trophic hormones Examples are insulin, epidermal growth factor (EGF), platelet-deriving growth factor (PDGF), atrial natriuretic peptide (ANP), & transforming growth factor- (TGF-) Receptors are polypeptides consisting of an EC hormonebinding domain & a cytoplasmic enzyme domain May be a protein tyrosine kinase, a serine kinase, or a guanylyl cyclase The two domains are connected by a hydrophobic segment of the polypeptide that crosses the lipid bilayer of the plasma membrane

Ligand-Regulated Transmembrane Enzymes

What Limits Intensity & Duration of Action

Down-regulation occurs when tyrosine kinases are involved Ligand-binding induces accelerated endocytosis of receptors from the cell surface, followed by the degradation of receptors from the cell surface. When this process occurs at a rate faster than de novo synthesis of receptors, the total number of cell-surface receptors is reduced.

Cytokine Receptors
These receptors respond to a heterogeneous group of peptide ligands The protein tyrosine kinase activity is not intrinsic to the receptor molecule A separate protein tyrosine kinase, from the januskinase (JAK) family, binds non-covalently to the receptor Cytokine receptors dimerize after they bind the activating ligand, allowing the bound JAKs to become activated & to phosphorylate tyrosine residues on the receptor

Cytokine Receptors
Phosphorylated tyrosine residues on the receptors cytoplasmic surface then set in motion a complex signaling dance by binding another set of proteins, called STATs The bound STATs are themselves phosphorylated by the JAKs, 2 STAT molecules dimerize the STAT/STAT dimer dissociates form the receptor & travels to the nucleus, where it regulates transcription of specific genes.

Cytokine Receptors

Ligand- and Voltage-Gated Channels

Natural ligands are
Acetylcholine Serotonin GABA Glutamate

Each of their receptors transmits its signal across the plasma membrane by increasing transmembrane conductance of the relevant ion & thereby altering the electrical potential across the membrane

Ligand- & Voltage-Gated Receptors

The time elapsed between the binding of the agonist to a ligand-gated channel & the cellular response can often be measured in milliseconds Ligand-gated ion channels can be regulated by multiple mechanisms, including phosphorylation and endocytosis Voltage-gated channels do not bind neurotransmitters directly but are controlled by membrane potential Such channels are also important drug targets

Ligand- & Voltage-Gated Channels

G Proteins & Second Messengers

Second messengers are cyclic adenosine-3,5 monophophate (cAMP), calcium ion, & phophoinositides G proteins usually use a transmembrane signaling system with 3 separate components
EC ligand Receptor Activated G protein The concentration of second messenger changes

G Protein & Second Messenger

For cAMP, the effector enzyme is adenylyl cyclase Examples of G protein receptors
adrenoceptors Glucagon receptors Thyrotropin receptors Dopamine receptors Serotonin receptors

G Protein & Second Messenger

Duration of activation of adenylyl cyclase depends on the longevity of GTP binding to the G protein Receptors coupled to G proteins are GPCRs They make up a family of seven-transmembrane (7TM) or serpentine receptors

G Proteins & Second Messengers

G Proteins & Second Messenger

GPCRs exist as homodimers & heterodimers

Receptor Regulation - Desensitization

Receptor Regulation Down Regulation

Well Established Second Messengers

cAMP Calcium & Phophoinositides cGMP

cAMP Second Messenger Pathway

Calcium & Phosphoinositides

Some of the hormones, neurotransmitters, & growth factor that trigger this pathway bind to receptors liked to G proteins; others bind to receptor tyrosine kinases Crucial step is stimulation of a membrane enzyme, phospholipase C which splits a minor phospholipid component of the membrane, phophatidylinosital4,5-bisphophate, into 2 second messengers, diacylglycerol and inositol-1,4,5-trisphosphate

Ca2+ -Phosphoinositide Signaling Pathway

Cyclic Guanosine Monophosphate

cGMP establishes signals in only a few cells Cell-surface receptors stimulate membrane-bound guanylyl cyclase to produce cGMP cGMP acts by stimulating a cGMP-dependent protein kinase Termination is by enzymatic degradation of the cyclic nucleotide & by phosphorylation of kinase substrates

cGMP

Interplay Among Signaling Mechanisms

Calcium-phosphoinositide & cAMP signaling pathways oppose one another in some cells but compliment each other in other cells

Phosphorylation
Supplies amplification & flexible regulation Amplification the attachment of a phosphoryl group to a serine, threonine, or tyrosine residue powerfully amplifies the initial regulatory signal by recording a molecular memory that the pathway has been activated Dephosphorylation erases the memory

Phosphorylation
Flexible regulation differing substrate specificities of the multiple protein kinases regulated by second messengers provide branch points in signaling pathways that may be independently regulated

Receptor Classes & Drug Development

Structure-activity relationship the agonist or antagonist that occupies the same receptor as the agonist must fit the receptor like a lock & key If two agonists exhibit identical relative potencies in producing 2 distinct effects, it is likely that the 2 effects are mediated by similar or identical receptor molecules The same neurotransmitter can act on different receptor types

Dose & Response In Patients

Graded Dose-Response Relationships to choose among drugs & to determine appropriate doses of a drug, one must know
Pharmacologic potency Maximal efficacy In relation to the desired therapeutic effect wanted

Graded Dose-Response Curves

Potency
Refers to the concentration (EC50 ) or dose (ED50 ) of a drug required to produce 50% of that drugs maximal effect Potency determines the administered dose of drug Potency depends in part on
Affinity of receptors for binding the drug The Efficiency with which drug-receptor interaction is coupled to response

Potency
Potency of a drug should be stated in dosage units Relative potency, the ratio of equi-effective doses, may be used in comparing 2 drugs

Efficacy
A drugs clinical effectiveness depends not on its potency but on it maximal efficacy & its ability to reach the relevant receptors This can depend on
Its route of administration Absorption Distribution Clearance from blood Clearance from its site of action

Maximal Efficacy
Efficacy is shown on the response axis The drugs propensity to cause toxic effects may limit the ability to utilize its maximal efficiacy Efficacy may be determined by
The drugs mode of interactions with the receptors By characteristics of the receptor-effector system involved

Graded Dose-Response Curves

Shape of Dose-Response Curves

Curves A, B, & C in the preceding slide approximate the shape of a simple Michaelis-Menton relation. Not all clinical responses do this Extremely steep dose-response curves like curve D may have important clinical consequences if the upper portion of the curve represents an undesirable extent of response Steep dose-response curves can result from cooperative interactions of several different actions of a drug

Quantal Dose-Effect Curves

These curves determine the dose of drug required to produce a specified magnitude of effect in a large number of patients or experimental animals They plot the cumulative frequency distribution of responders v. the log dose A specific quantal effect may be chosen on the basis of clinical relevance, for preservation of safety of experimental subjects, or it may be an inherently quantal event

Quantal Dose-Effect Curves

The frequency distribution of such responses are plotted against the log of the dose, which produces a gaussian normal curve variation When these responses are summated, the resulting cumulative frequency distribution constitutes a quantal dose-effect curve or the proportion or percentage of individuals who exhibit the effect plotted as a function of log dose

Quantal Dose-Effect Curve

 Both curve types give information about potency & selectivity of drugs Graded dose-response curves indicate maximal efficacy of a drug Quantal dose-effect curves indicate potential variability of responses among individuals

Variation In Drug Responsiveness

Idiosyncratic drug responses Idiosyncratic responses Immunologic mechanisms (hypersensitivity) Quantitative variations Hyporeactive Hyperreactive Tolerance Tachyphylaxis Need to consider: Propensity of a drug to produce tolerance or tachyphylaxis Effects of sex, age, body size, disease states, genetic factors, simultaneous administration of other drugs

Four Mechanisms That Contribute To Variation In Drug Response

Alteration in concentration of drug that reaches receptor Variation in concentration of an endogenous receptor ligand Alterations in number or function of receptors Changes in components of response distal to the receptor

Clinical Selectivity: Beneficial v. Toxic Effects of Drugs

Beneficial & toxic effects mediated by the same receptor-effector mechanism Beneficial & toxic effects mediated by identical receptors but in different tissues or by different effector pathways Beneficial & toxic effects mediated by different types of receptors