FAR 381: Pharmacodynamics BINDING SITES:

? WHAT ? Orthosteric sites:

Study of the relationship of the drug concentration to pharmacological effects Active site of target to which the endogenous ligand binds
“What the drug does to the body” Orthosteric binding of ligand
Aspects of drug reactions - Influences activity of a receptor
- Receptor interactions - Facilitator results in cellular effect
- Mechanisms of action - Inhibitor prevents cellular effect
- Mechanisms of toxicity Many drugs bind to an orthosteric site
- Dose-response relationships
- Either mimics endogenous ligand or prevents it’s binding

Allosteric sites:
DRUG TARGETS:
Separate binding site that allows for conformational changes in the orthosteric site
To exert an effect, drugs must be bound to a cellular component
Allosteric modulators
Drug targets
- Binding changes orthosteric site’s shape and affinity for endogenous ligand
- Most drug targets are proteinaceous Exceptions do exist (e.g. calcium salts in o Influences efficacy of endogenous ligand
bone) o Modulator does not elicit effect on its own
- Facilitator amplifies orthosteric ligand’s effect
Binding sites allow molecules to attach to a drug target
- Inhibitor diminishes orthosteric ligand’s effect
- Generally either an orthosteric or allosteric site

PROTEIN DRUG TARGETS:

Four kinds of main regulatory proteins are drug targets

Main drug target classes

- Receptors
- Ion channels
- Enzymes
- Carrier molecules and transporters

Receptors:
Protein molecules that respond to external stimuli to produce a cellular effect through
messenger systems

Four primary types or superfamilies of receptors

- Ligand-gated ion channels (ionotropic receptors)

 - G-protein coupled receptors (GPCRs, or metabotropic receptors) Binding domains & functions:
- Kinase-linked and related receptors
- Nuclear receptors Ligand-binding sites

Ionotropic receptors: location & structure: - Extracellular sites form between specific subunit groups
- Allows for wide range of ligand specificities and affinities

Functions

- Ions are secondary messengers

- Membrane voltage changes
- Depolarisation: net positive
- Hyperpolarisation: net negative

Structure:

Resting phase:
Signal transduction:

4. Ions produce cellular effect

Binding domains & functions:

GPCRs: location & structure Ligand-binding sites

- Extracellular sites

Functions

- Variety of cellular effects depending on G-protein and effector proteins

(channels or enzymes)
- One of the most targeted drug targets in pharmacology

Resting phase:

1. Intracellular G protein-association domain associates with unattached G

protein
Signal transduction [1/3]: 1 signal transduction [3/3]:

Ligand-binding changes conformation which

increases affinity for G-protein

Kinase-linked: location & structure

Signal transduction [2/3]: Cellular kinases activated by ligand-binding

Ligand-binding sites

- Large extracellular binding site

Functions

- Variety of cellular effects depending on receptors targeted

βγ-complex remain complexed and only contributes to activity if concentrated

Resting state: Signal transduction [3/3]:

Signal transduction [1/3]:

Nuclear: location & structure

Ligand-binding results in gene modulation in nucleus

Location of receptor

- In cytosol or nucleus
- Co-factors often required for activation

Functions

- Alters genetic expression of various proteins

o Inhibition
o Induction
Signal transduction [2/3]: Resting state:
Signal transduction [1/2]: Ion channels:
Water-filled ion channel that opens upon different stimuli

Ion transfer across membrane

- Ions cannot cross unaided due to hydrophilicity

- Rate and direction of transfer controlled by electrochemical gradient of each
specific ion

Ion channels as drug targets

- Blocks transfer of molecules through channel

- Modulation of channel opening and closure

Characterisation:

Signal transduction [2/2]: Selectivity for ion species

- Size of the pore

- Nature of channel lining

Gating properties

- Stimulus that controls opening of channel

o Ligand-binding
o Voltage

Molecular architecture

- 3D structure of channel once assembled

Selectivity:

Cation channels

- Positively charged ions

- Main channels transfer Na+, Ca2+ or K+
Drug’s ability to alter activity: - Can be selective for one, or non-selective channels permeable to all
One of the most important classes of drug targets, with new information generated Anion channels
annually
- Negatively charged ions
Receptors as drug targets - Mainly permeable to Cl-
- Simulates endogenous ligand’s activity (-mimetic)
- Prevents endogenous ligand’s activity (-lytic)
- Modulation of endogenous ligand’s activity
Gating properties: Enzymes as drug targets

Various stimuli may open channels - Inhibition to reduce transmitter/substrate degradation

- Inhibition to reduce metabolite synthesis
Voltage-gated

- Opening triggered by altered cell membrane voltages

o Mostly depolarisation Diseases of high metabolite levels:
- Important for membrane excitability
Enzyme inhibition decreases product synthesis which is in excess
Ligand-gated

- Mostly extracellular ligand-binding to receptors, but some intracellular signals

exist

Enzymes:
Catalytic molecules, mostly protein, that accelerate metabolic conversion of chemical
entities

 Reduced synthesis

Enzyme inhibition decreases product synthesis which is in excess

Drug activity:

Enzymatic functions (refer to Pharmacokinetics)

- Detoxification of chemical entities for removal by excretory organs

- Activation of pro-drugs to active metabolites
- Formation of reactive, toxic metabolites
Diseases of low transmitter levels: Transfer of ions or molecules

Some diseases occur due to low concentrations of transmitters - Polar or hydrophilic molecules cannot cross membrane
- Requires facilitator to aid transport, but is saturable
- Can be energy-dependent or –independent

Transporters as drug targets

- Inhibition of transporter function

- Blocking transporter structure

Types:

 Reduced degradation

Enzyme inhibition increases substrate which is lacking

Targets of drug action:

Mimetic of endogenous ligand

- Same activity as endogenous ligand

Blockers

- Prevent transfer of molecules or binding of endogenous ligand

Allosteric modulators
Transporters & carriers:
- Influences activity of endogenous ligand either positively or negatively
Protein molecules that facilitate transfer of ions, small molecules or macromolecules
across cell membrane
THE DOSE-RESPONSE CURVE & DRUG BINDING: If large enough concentration range is used, a bottom and top plateau of tapered
effect is observed
Dose response with alcohol:

Biological effect of a drug is dependent on it’s activity, and thus will differ between
compounds

Drug response curve:

Direct relationship between concentration/dosage (x-axis logarithmic scale) and

response/effect (y-axis)

Dose-response relationship As concentration/doses increases, effect increases

Various points of information can be drawn from a dose response curve, such as
activity and toxicity

EC50

- Concentration at which 50% of effect occurs

- Absolute where 50% of maximum effect occurs
- Relative where 50% between highest and lowest plateau occurs
 THERAPEUTIC INDEX (TI):
Measure of drug’s safety as garnered from animal model

Not a perfect parameter, but

- Provides suggested safety in general population

- Narrow TI drugs are dangerous
- Broad TI drugs are considered safer

Broad/large TI drugs:

Toxicity appears at high concentrations, far above what is needed in population

Considered safe or unlikely to cause harm (e.g. penicillin)
Parameters depend on variables used

- Efficacy or Inhibitory or Lethal or Toxic (E/I/L/T)

- Concentration or Dosage (C/D)
- Other values can also be used (EC20)

Potency:

Measure of drug amount needed to elicit specific efficacy

Narrow/small TI drugs: Affinity Ability of drug to bind to receptor Higher affinity allows for greater occupation
Affinity determined by forces at play
Toxicity appears within range necessary to treat majority of population Considered
more dangerous or likely to cause harm (e.g. warfarin) Ligand-receptor forces between ligand and drug target

Activation & efficacy:

Activation Eliciting a cellular response Controlled by efficacy

DRUG-RECEPTOR INTERACTIONS:
Ligand-receptor binding:
Occupation & affinity:

Occupation Binding of drug to a receptor Controlled by affinity

Efficacy/Intrinsic activity Tendency to activate the receptor based on drug-receptor

complexes
Ligand types:
Agonists:

Ligands for receptor that result in its activation, thus possesses affinity and efficacy

1. Super agonist elicits supramaximal efficacy (>1) relative to endogenous ligand

2. Full agonist elicits maximum efficacy (1) if enough receptors occupied

3. Partial agonist elicits sub maximal efficacy (between 0 and 1) even at full
occupancy Agonists & silent antagonist:

4. Inverse agonist

Classical/silent/neutral antagonists: Co-administration of agonists:

Ligands for receptor that possess affinity, but do not activate the receptor Full & partial agonist:

(WARNING! Be careful of antagonist’s other descriptions (e.g. physiological) - Drug

capable of antagonising the effect of drug target, possibly through activation of
another cellular response)
Full and partial agonist both bind receptors and activate the system within their Constitutive activity:
efficacy spectrum
Constitutive activity (baseline activity)

- Some receptors active without bound ligand

Over-administration of agonists: o Conformational differences between active and inactive states
- States typically in equilibrium to maintain homeostasis
Full agonist overdose: o Large disequilibrium may result in disease

Inverse agonist:
Agonists (partial or full) active receptor by shifting inactive receptors to active
Full agonists activate drug targets fully and overburden system (e.g. heroin conformations
overdose)

Partial agonist as antagonist:

Partial agonist (at high doses) displaces full agonist

Inverse agonists (partial or full) reduce constitutive activity by shifting active

receptors to inactive conformation

System still activated, but sub-maximally, thus symptoms decrease Low-efficacy

partial agonist can thus be an antagonist

(e.g. buprenorphine [partial agonist] counteracts morphine [full agonist])

 ANTAGONISRIC DRUG INTERACTIONS:
Drug antagonism:
Effect of one drug is diminished or abolished by another

Selectivity & cross-reactivity:

Chemical:
Drug targets have specific conformations that allows for recognition of individual
drugs or ligands

Selectivity

- Ability to bind only specific receptors or drug targets

Cross-reactivity

- Drug acts as ligand to numerous drug targets

- Affinity and efficacy depends on the receptor type
- Produces differential effects if selectivity is lost, most often unwanted effects

As concentration increases drug loses selectivity and cross reactivity occurs (most
likely as unwanted effects) Pharmacokinetic:

Antagonist alters plasma concentration by changing any ADME pharmacokinetic

parameters

e.g. rifampicin increases oral contraceptive metabolism by inducing CYP450, thus

antagonising its contraception
Competitive:
Competition between antagonist and agonist to bind at the same binding site

Competition between molecules

- Generally structural similarity between molecules

- Target can only bind one drug molecule at a time
o Antagonist prevents binding of the agonist
o High levels of one ligand will displace the other, or natural dissociation
can occur
- May be reversible or irreversible

Beneficial use

- Prevents or reverses activity of drugs (e.g. overdoses)

Reversible competitive:

Antagonist can be displaced by another or dissociate from

Irreversible competitive:

Antagonist cannot be displaced or dissociate from receptor

 Non-competitive – allosteric antagonism:

Non-competitive: Non-competitive – secondary processes/messengers:

Drug antagonises agonist by binding to site different than active site (no
competition), reducing activation chain of events

[Increasing concentration of agonist does not displace antagonist (no competition

between them) or improve efficacy]
Physiological: Translocation of receptors

Drugs counteract one another’s effects by activating different physiological systems - Chronic agonist exposure decreases receptor expression
- Receptors internalised into cell
- As less receptors expressed on surface, less drug receptor complexes are
formed
- Gradual response

Exhaustion of receptors

- Essential intermediate substance depleted

- Drug target or second messengers cannot be synthesised

Altered drug metabolism

- Increased or decreased expression of enzymes

Physiological adaptation

- Shift of homeostatic regulators alters bodies physiological response to drug

Patient factors:
ALTERATIONS TO A PHARMACODYNAMIC PROFILE
Diminished drug effect:
Drug’s activity may decrease with continuous administering

Tachyphylaxis or desensitization

- Acute reaction, sometimes within minutes

Tolerance

- Gradual reaction, days to weeks

Refractoriness

- Resistant to treatment
- May be inherent or develop over time

Possible explanations:

Change in receptors

- Conformational change or phosphorylation of receptor prevents further activity

- Generally acute response