PHARMACODYNAMIC PRINCIPLES

Agnes Cecille G. Llamas, MD Dept. of Pharmacology UST FMS

THE THERAPEUTIC TRIANGLE

THERAPEUTICS

PHARMACODYNAMICS

PHARMACOKINETICS

EFFECTS

CONCENTRATION

DOSE

Pharmacodynamics
Study of the biochemical and physiologic processes underlying drug action Mechanism of drug action Drug-receptor interaction Efficacy Safety profile

Mechanisms of Drug Action

1.Physical Action
a. Mass of the drug e.g. bulk laxatives b. Adsorptive property e.g. charcoal, kaolin c. Osmotic activity MgSO4, mannitol d. Radioactivity I131 and other isotopes e. Radioopacity contrast media ( Barium sufate, urografin)

Mechanisms of Drug Action

2. Chemical Action a. Antacids AlOH b. Acidifying agents (NH4Cl) and alkalinizing (NaHCO3) agents react with buffers in plasma and alter pH of urine c.Oxidizing agents KMnO4, Iodine ( germicidal and inactivate ingested alkaloids) d. Chelating agents EDTA (sequester toxic metals)

PROTEINS AS TARGETS OF DRUG ACTION

Enzymes Stimulation increase in enzyme affinity for substrates, Km of the reaction decreases ; e.g. endogenous mediators and modulators Inhibition 1. Nonspecific e.g heavy metal salts, strong acids and alkalis, alcohol, formaldehyde, phenol 2. Specific a. Competitive i. Equilibrium e.g sulfonamide, physostigmine ii. Non-equilibrium e.g. organophosphates, methotrexate b. Noncompetitive - e.g. acetazolamide, aspirin, theophylline, PTU, disulfiram

PROTEINS AS TARGETS OF DRUG ACTION TRANSPORTERS Neuroamine Reuptake Inhibitors

TCA; SSRI

Loop diuretics PPIs Digitalis glycosides

Drug Receptor

A macromolecular component of a cell with which a drug interacts to produce a response

Ion channel-linked Receptors

Ion channel-linked receptors

receptor itself constitutes an ion channel activation causes channel opening (or closing) and subsequent entry (or nonentry) of ions ideal for rapid signaling between excitable cells typical ligand: neurotransmitter

Ion channel receptors NAch GABA A 5-HT3 glutamate NMDA

Structure: Protein pores in the plasma

G-PROTEIN COUPLED RECEPTORS

G-protein linked receptors

largest category of plasma membrane receptors (>500 members) seven transmembrane domains ( -helices) surface receptor indirectly regulates, via the mediation of G-proteins, the activity of another membrane-bound target protein (ion channel or enzyme)

 Basic components of G-protein linked pathway

Receptor G protein complex (heterotrimer) Target enzyme or channel

G-proteins
G proteins: bind guanine nucleotide (GTP, GDP) typically three subunits (heterotrimeric) E-subunit - binds GTP or GDP and activates G proteins F- and K-subunits; tightly bound complex switch from active/inactive state

2nd MESSENGERS AND SIGNAL TRANSDUCTION

Examples of membrane bound proteins regulated by G protein receptors

Ion channels Enzymes: Adenylate cyclase Phospholipase C phosphodiesterase

G protein associated second messengers

cAMP DAG IP3 p Ca++ Ca++ cGMP

cellular levels of second messengers may be increased or decreased by G protein activation

cAMP system

Phosphoinositol system

Arachidonic acid system Histamine

External Signal (1st messenger)

Extracellular Side

Norepinephrine

ACh

Receptor
Transducer Primary Effector

B-adrenergic receptor
Gs Adenylyl cyclase

Muscarinic Ach receptor

Gq PLC

Histamine
G? PLA2

Cytoplasmic side

Second Messenger Secondary Effector

cAMP cAMP-dependent Protein kinase

IP3 Ca2+ release

DAG PKC

Arachidonic acid
5-lipoxy- 12-lipoxyCyclogenase genase oxygenase

GPCRs

An enzyme-linked receptor is a transmembrane receptor where the binding of an extracellular ligand causes enzymatic activity on the intracellular side.

Enzyme-linked receptors
single-pass transmembrane receptors ligand-binding portion cytosolic domain: intrinsic enzymatic activity or associates directly with enzyme

 Kinase activity
transfers phosphate groups from high energy donor molecules, such as ATP to specific target molecules (substrates)

Receptors with intrinsic enzyme activity

tyrosine kinases
receptors for Insulin and growth factors IGF-1, EGF, PDGF, FGF, VEFG, NGF guanylyl cyclase serine/threonine kinase tyrosine phosphatases

Receptors that associate with enzymes (Receptor-associated kinases)

receptors have NO intrinsic enzyme activity separate cytoplasmic tyrosine kinases are associated with the receptor

Insulin receptor
Is a transmembrane receptor that is activated by insulin Consists of 2 alpha units and 2 beta units The beta subunits pass through the cellular membrane and are linked by disulfide bonds

Tyrosine kinase-associated receptor

Cytokine receptor family
e.g. interferon, IL-3, growth hormone, prolactin, erythropoietin JAK-STAT pathway:
JAK kinases phosphorylate and activate gene regulatory proteins: STATs (Signal Transducers and Activators of Transcription)

Signaling proteins that act through JAK-STAT pathway

Signaling Ligand ReceptorAssociated JAK -interferon -interferon erythropoietin Prolactin Jak1 and Jak2 Tyk2 & JAK2 Jak2 Jak1, Jak2

STATS Some Responses activated STAT1 STAT1, STAT2 STAT5 STAT5 macrophage actn inc.resistance to viral infection stim. rbc production milk production

Growth hormone Jak2 IL-3 Jak2

STAT1,S stimulates growth TAT5 STAT5 stimulates early blood cell prodn

Alberts, Molecular biology of the cell, 4th ed.

GUANYLYL CYCLASE: FUNCTION

GUANYLYL CYCLASE
NO Peptide ligands Intracellular Ca flux

GTP

cGMP

Protein kinase Ion channels phosphodiesterase

RECEPTORS SERINE/THREONINE (RSTK)

The receptors for the TGF-F superfamily of ligands have intrinsic serine/threonine kinase activity

Receptors that associate with enzymes (Receptor-associated kinases)

receptors have NO intrinsic enzyme activity separate cytoplasmic tyrosine kinases are associated with the receptor

ACTIONS OF NUCLEAR RECEPTORS

NRs modulate transcription through several mechanisms Include both activation & repression activities NR signaling is complex Can be genomic or nongenomic Ligand-dependent or ligand-independent

NUCLEAR RECEPTORS

STRUCTURE OF NUCLEAR RECEPTORS

A/B Region DNA-Binding Domain (DBD) D Region Ligand-Binding Domain (LBD) Dimer Interface Ligand-Binding Pocket (LBP) AF2 Function

Glucocorticoid Receptor Signaling

T3 MECHANISM OF ACTION

PPAR : MOA

TRANSCRIPTIONAL REGULATION

TRANSCRIPTIONAL REGULATION
DNA Recognition Transcriptional Activation Transcriptional Repression Corepressor/Coactivator Exchange Nuclear Receptor Turnover Transrepression Post-translational Modifications Nongenomic Effects

Definition of Terms
first messenger
intercellular chemical messengers or signals (e.g. hormones, neurotransmitters, paracrine/autocrine)

second messengers
nonprotein messengers that enter cytoplasm or are generated as a result of receptor activation chemical relays for signal

Signal Transduction
Receptor activation Ultimate response
q
change in plasma membrane properties cellular metabolism secretory activity cell rate of proliferation and differentiation contractile activity

Signal Transduction
 process by which a stimulus is transformed into a response TYPES OF PATHWAYS 1. Pathways initiated by intracellular receptors 2. Pathways initiated by extracellular receptors

Signal transduction
Signal transduction events start with recognition of the signal in a target tissue Receptors are proteins that bind to the signals and begin transducing the signal Depending on the type of signal, the response can be short and fast, or long and slow.

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Intracellular pathway
ligand secreted by cell A diffuses into cell B ligand binds to protein receptor in cytoplasm or nucleus receptor conformation changes ligand-receptor complex binds to DNA transcription of genes is regulated

Steps in signaling by plasma membrane receptors

1. 2. 3. Recognition of signal by receptor. Transduction of message into an intracellular signal (2nd messenger) Transmission of second messenger signal to effector(s): enzymes, ion channels, transcription factors. Modulation of the effector(s), altering activity (kinases, phosphatases). Response: summation and integration of input. Termination of response

4. 5. 6.

 Recognition Transduction Transmission

Modulation Response Termination

How do second messengers transduce signals into cellular responses?

1. activation of second-messenger dependent kinases 2. activation/closure of ion channels 3. activation of transcription factors

Role of second messengers in Signal Transduction

Specificity and Diversity
ligands activating same signaling pathway produce same effects in the same cell same signaling molecule can produce distinct responses in different cells Acetylcholine: contraction of skeletal muscle inhibits contraction of heart muscle Amplification

Signal cascade
2nd messenger initiates cascade of events amplification of signal allows first messenger to be active at low concentrations

Signal transduction
1. enzyme linked
(multiple actions)

2.

ion channel linked

(speedy)

3. G protein linked
(amplifier)

4. nuclear (gene) linked

(long lasting)

Crosstalk
Signaling pathways can interact to either upregulate or downregulate one another
For example, the insulin pathway versus the epinephrine pathway Epinephrine activates beta-adrenergic receptors in muscles, resulting in glycogenolysis Insulin stimulates the production of glycogen

These pathways work by modulating the receptors or other downstream proteins

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How is the activity in signal transduction pathways shut off?

Receptor inactivation
phosphatases GTPase phosphodiesterases

Receptor Inactivation
occurs via: decrease in first messenger concentration alteration in chemical structure of receptor (e.g. phosphorylation of receptor) endocytosis of receptor-ligand complex

Receptor Inactivation results in:

rapid breakdown or inactivation of second messengers (e.g. phosphodiesterases) GTPase breakdown of GTP to GDP in subunit pumping of calcium back into ECF or ER dephosphorylation of previously phosphorylated proteins by protein phosphatases

inactivation or reactivation of effectors (enzymes, ion channels, transcription factors)

TERMINATION OF RESPONSE

Receptor Regulation
Desensitization ( down regulation )
Phosphorylation Internalization or endocytosis Proteolysis Reduced synthesis Synthesis of inhibitor proteins

Homologous desensitization Heterologous desensitization

DESENSITIZATION BY PHOSPHORYLATION

Drug - Receptor Interaction and Pharmacodynamics (how drugs work on the body )
The action of drug on the body, including receptor interactions, doseresponse phenomena and mechanisms of therapeutic and toxic action.

Characteristics of messengerreceptor interactions

specificity

Characteristics of messengerreceptor interactions

saturation
degree to which receptors are occupied

competition
ability of different molecules very similar in structure to combine with the same receptor

Cellular response

?LigandA

Drug-receptor interaction The Binding Reaction

Drug + Free Receptor

k1 k-1

Drug-receptor Complex

(100 - DR)
Where:

DR

D = drug concentration DR= concentration of drug-receptor complex 100 - DR = free receptor concentration

Drug-receptor interaction The Binding Reaction At equilibrium: [D] x [R] x k1 = [DR] x k-1 so that: [DR] = k1 [D] [R] k-1 k-1/k1 = dissociation constant (kd)

The Binding Reaction

Dissociation constant (KD) Measure of a drugs affinity for a given receptor Defined as the concentration of drug required in solution to achieve 50% occupancy of its receptors Affinity = 1/Kd

Drug - Receptor Binding D+R DR Complex

Affinity Affinity measure of propensity of a drug to bind receptor; the attractiveness of drug and receptor Covalent bonds are stable and essentially irreversible Electrostatic bonds may be strong or weak, but are usually reversible

Drug Receptor

Affinity Refers to the strength of binding between a drug and receptor Number of occupied receptors is a function of a balance between bound and free drug

Receptor Occupancy = Kd

Terms which indicate ability of drug to bind to receptor

Potency Affinity KD or EC50

(Corresponds to Km in Michaelis-Menten analogy)

RECEPTOR OCCUPANCY THEORY

assumes that the magnitude of the response is proportional to the fraction of receptor sites occupied by the drug molecules such that e = DR Emax R total
e magnitude of response; Emax = max response; DR drug Receptor complex

EC50 and Kd

Drug Receptor Interaction DR Complex

Effect

Efficacy (or Intrinsic Activity) ability of a bound drug to change the receptor in a way that produces an effect; some drugs possess affinity but NOT efficacy

Terms which indicate ability of drug to produce a response

Efficacy Power Intrinsic Activity Emax

(Corresponds to Vmax in Michaelis-Menten analogy)

 At equilibrium: [D] x [R] x k1 = [DR] x k-1 so that: [DR] = k1 [D] [R] k-1
k-1/k1 = dissociation constant (kd)

What have we learned so far?

Ke (k1/k-1) is called the affinity constant DR is the response; D is concentration of drug when DR = 50 percent (effect is half maximal), D (or EC50) is equal to kd or the reciprocal of the affinity constant response is a measure of efficacy

The log concentration-effect relationship

POTENCY vs EFFICACY

Agonist interactions with biological receptors

Given the graded-dose response curve for 4 drugs

A. Drug B is most potent B. Drug A and C are equally potent C. Drug A is more effective than Drug C D. Drug D is most effective

LOCK and KEY AND AGONIST vs ANTAGONIST

ANTAGONISM : COMPETITIVE

Reversible Antagonist on Kd

ANTAGONISM : NONCOMPETITIVE

Irreversible Antagonist on Kd

Reversible

Irreversible

Properties of antagonist blockade of response

PARTIAL AGONISM

OCCUPANCY AND RESPONSE CURVES FOR FULL vs PARTIAL AGONIST

PARTIAL AGONIST AS ANTAGONIST

PARTIAL AGONIST ON FULL AGONIST

SPARE RECEPTORS

100% Receptor saturation = 100% Effect 100% effect at less than maximal receptor saturation Steeper curve with spare receptors both for receptor saturation effect and ligand concentration effect curves

Spare Receptors and the Effect of Irreversible Antagonism

Frequency distribution curve & quantal dose effect curve

Effectiveness, toxicity, lethality

ED50 - Median Effective Dose 50; the dose at which 50 percent of the population or sample manifests a given effect; used with quantal dose curves TD50 - Median Toxic Dose 50 - dose at which 50 percent of the population manifests a given toxic effect LD50 - Median Lethal Dose 50 - dose which kills 50 percent of the subjects

Quantification of drug safety

Therapeutic Index =

TD50 or LD50 ED50

Quantification of drug safety Therapeutic Index

TD50 or LD50 ED50

Drug A 100

sleep

death

Percent 50 Responding

0 ED50 LD50

dose

Drug B 100 sleep

death

Percent 50 Responding

0 ED50 LD50

dose

COMBINED EFFECT OF DRUGS

Summation/addition :1 + 1 = 2
E.g. an anti-histaminic agent and a benzodiazepine

Synergism : 1 + 1 = > 2
E.g. combination of sulfamethoxazole and trimethoprim

Potentiation : 0 + 1 = > 1
E.g. probenecid and penicillin

Antagonism : 1 + 1= < 2 or 1 + 0 = 0
E.g. naloxone and an opiate

DRUG ANTAGONISM
Receptor antagonism Chemical antagonism Pharmacokinetic antagonism Physiologic antagonism

PHARMA RULES ! ! !