Drug-Receptor Interactions

What are receptors? Traditional model was a rigid Lock and Key

Lock Receptor surface Key Drug or Ligand

Drug

Receptor

Drug-Receptor Interactions

Receptors fluid, flexible surfaces or pockets

Can change 3-D structure as ligand docks Most receptors are sites for natural ligands Small portion or surface of a macromolecule

Includes: Enzymes, components of cell membranes, intracellular protein or nucleic acid, antibodies, DNA, RNA

Ligand - Receptor docking structure changes Changes function

Drug-Receptor Interactions

Receptor - soluble molecule

Isolated and purified when overexpressed Gene inserted into microorganism

Overproduces protein Membrane bound receptor molecules much more difficult

Overexpression can result in a large number of copies Membrane is then broken into pieces (ie Ultrasonication) Affinity chromatography

Ligand covalently attached on a solid phase (Agarose) Membrane fragment with the receptor binds the solid phase Wash, wash, wash Elute receptor off the solid phase with the natural ligand

Affinity Chromatography

Drug-Receptor Interactions

Membrane bound receptors

Receptors of membranes are primarily proteins If the gene sequence is known: site directed mutagenesis Alter one or more amino acids in the receptor Has this changed the biological responses to a particular ligand?

How has altering the amino acid within the receptor altered the intramolecular forces that hold the ligand in the receptor?
Number of internal hydrogen bonds amide bonds Number of hydrophobic interactions van der Waals forces Ionic interactions

Small molecule interaction with a membrane receptor

Can result in a large change in protein conformations Structural and physical properties of membrane changes

Receptor Conformation

Drug-Receptor Interactions
Drugs can have two fates at the receptor site: 1. Irreversible covalent bonding with the receptor active site

Active-site-directed irreversible inhibition Anticancer agents such as the alkylating agents Enzyme inhibitors such as MAOI tranylcyproamine Antibacterial agents such as the beta-lactamase inhibitors

Augmentin component - Potassium Clavulanate

OH O
Beta-lactamase OH O N

H
CO2-K+

H+

Occasionally called: suicide substrates

Drug-Receptor Interactions
2. Reversible drug-receptor complex Most desirable Drug can eventually be excreted
Requires rather weak receptor / drug interactions When added together afford a stable interaction
Hydrogen bonds: 1 to 7 kcals---proteins and DNA Ionic bonding: 5 to 10 kcals Ion-dipoles bonds:1 to 7 kcals Dipole-dipole bonds: 1 to 7 kcal Van der walls: 0.5 to 1 kcal Hydrophobic bonding:1 kcal

If a molecule has each of these interactions: 9.5 to 33 kcal Covalent bonds can range from 40-140 kcal

Drug-Receptor Interactions

Agonists or stimulants
Initiate a desired response Intrinsic activity

Antagonist

Decrease/prevent the response

Response - function of number occupied receptors

Drug-Receptor Interactions
Dose response curves Reveal the affinity and effective concentration of a series of drug analogs

Drug-Receptor Theory

Occupancy Theory
Drug and receptor interact with each other Complex effects Conformational changes

Drugs structure affinity

Occupancy Theory

Rate Theory

Agonist or stimulant activity is proportional to the rate of drug-receptor combination rather than the number of occupied receptors Agonist activity is the result of a series of rapid association and dissociation of the drug and the receptor An antagonist has a high association rate but a low rate of dissociation

Other Theories
1.
1.

Induced-fit theory of enzyme-substrate interaction

Substrate or drug binding to the receptor induces 3 dimensional conformational changes in the macromolecule positioning catalytic groups in the correct position to conduct productive chemistry or altering membrane behavior (e.g. opens channels for calcium)

2.
1.

Macromolecular perturbation theory

Small molecule binding produces in a macromolecule:
1. 2. 3.

Specific conformational perturbations (Agonist) Non-specific conformational perturbations (Antagonist) An equilibrum mixture of specific and non-specific changes (partial agonist or antagonistic properties)

Reality is most likely a mixture or blend of all these theories

Agonism

Relative Potency

Relative affinity Intrinsic efficacy Neither produce maximal response in tissue

Relative Efficacy

Antagonism

Antagonism cont

Drug-Receptor Interactions
What factors influence binding? Molecular structure
Isomerism Functional groups Rigidity

Peptide bond distance = 3.61 angstroms

Drugs - spacial relationship between functional groups is typically a multiple of 3.61 Conformational changes in drugs occur to optimize this

Stereochemical features of drugs

1.

Isomerism
A.

Cis and trans isomers in double bonds

Different physical and chemical properties----distribution in a biological system are different

OH HO OH

HO trans-diethylstibesterol Estrogenic activity cis-diethylstibesterol Only 7% activity of the trans isomer

Stereochemical features of drugs

1.

Isomerism continued
B.

Conformational isomers as a result of the rotation around single bonds between two atoms

Energy barrier exists between these isomers that is sufficiently large that they can often be observed Examples:

CH3 H3 C H3 C CH3 H3 C H

CH3

Stereochemical features of drugs

1.
B.

Isomerism continued
Conformational isomers as a result of the rotation around single bonds between two atoms

Remember that this is an EQUILIBRUM process

H3C H H H H CH 3 CH3 H H CH3

Anti Lowest energy highest population

H3C H H H H3C H H CH3 HC H 3

eclipsed highest energy lowest population >>>1 in 1000

H3C H CH3 H H H H3C CH3 HH

eclipsed highest energy lowest population 1 in 1000

CH3 H3C H H H H
gauche medium energy 1 of 4 population

H H

H H

CH3 H H H H H
gauche medium energy 1 of 4 population

CH3 H

Intramolecular hydrogen bonding, dipole-dipole interactions and electrostatic forces in molecules can alter this distribution

Stereochemical features of drugs

1.

Isomerism continued
B.

Conformational isomers as a result of the rotation around single bonds between two atoms

EQUILIBRUM process - results in CONFORMATIONAL FLEXIBILITY FLEXIBILITY can lead to multiple modes of action at different receptor types Example: Acetylcholine: muscarinic and nicotinic receptors This can often lead to SIDE EFFECTS due to activity at an undesirable site of action

Stereochemical features of drugs

2.

Optical Isomerism
A.

Enantiomers - mirror image (plane of symmetry)

All physical properties are identical with the only difference is the direction each rotates plane polarized light

O H HO NH2
L-serine

O H OH HO NH2
D-serine

OH

[]D20 = +14.7 (c=10, 1 N HCl) []D20 = -14.7 (c=10, 1 N HCl)

Only the L isomer is used in protein synthesis

Stereochemical features of drugs

2.

Optical Isomerism B. Diastereomers - 2 or more chiral centers 2n = Number of diasteromers (n = # of chiral centers) Example: Ephedrine and Pseudoephedrine
CH3 H H NHCH3 OH H HO CH3 NHCH3 H
MP = 118-120 1 gram/200 mL

MP = 37-39 1 gram/20 mL

Ephedrine (Erythro)

Pseudoephedrine (Threo)

Use:

Hypotension

Decongestant

Stereochemical features of drugs

Most drugs are diastereomers Stereoisomers display different responses

Receptors - variable
blends of binding

Active transport carrier systems

Chiral, asymmetric molecules such as proteins, lipids and carbohydrates Preferential binding & transport of one diastereomer

Different lipid and water solubilities different distribution Metabolic enzymes are asymmetric
One diastereomer preferentially metabolized Important when the metabolite is the active compound

Excretion of the drug

Preferential excretion of one diastereomer over another

Isosterism in drug development

What is an isostere?? These are structural components or functional groups of a molecule whose steric, electronic and solubility characteristics are interchangeable Acyclic steric isoteres are most often exploited:
Univalent atoms and groups
CH3 NH2 OH F Cl CH3 H CH3

Cl

SH

Br

Bivalent atoms and groups

CH2 N H CO2CH2R CO2R O S

Trivalent atoms and groups

CH N

CONHR COSR

Isosterism in drug development

Cyclic steric isosteres are most often exploited:

N
Benzene Pyridine

S
Thiophene

O
Furan

N H
Pyrrole

S
Cyclopentane Tetrahydrothiophene

O
Tetrahydrofuran

N H
Pyrrolidine

Actual example: H N O O H N O S Cl OH H N O O H N O

H2 N

H 2N

Cl OH

Cefaclor - Ceclor

Loracarbef - Lorabid

Molecular modeling

In the beginning - Ball and Stick Computers revolutioned drug development

Quantum mechanics (not used much) and molecular mechanics Global minimum energy conformation Typically hydrogen bonding, ionic bonding, hydrophobic bonding will affect receptor binding

Lowest energy conformer typically NOT the most active

Solvation factors ignored generally

X-ray crystallography

Crystal match the receptor binding conformer?

Solution conformations can be determined using high-resolution nuclear magnetic resonance (NMR)

Molecular modeling

Works well in rigid molecules (steroids) Highly flexible molecules problematic Three-Dimensional Databases
Brookhaven Protein Database (1000s of proteins) Other data bases - NIH

Examples
HIV-protease and drugs available to treat AIDS
Invarase

- Saquinovir Crixivan - Indinavir Norvir - Ritonavir

Angiotensin-converting enzyme Monoamine oxidase