Patrick

An Introduction to Medicinal Chemistry 3/e

Chapter 19

CHOLINERGICS, ANTICHOLINERGICS
& ANTICHOLINESTERASES
Part 1: Cholinergics & anticholinesterases

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Contents

Part 1: Cholinergics & anticholinesterases

1. Nerve Transmission (3 slides)

2. Neurotransmitter
3. Transmission process (10 slides)
4. Cholinergic receptors (2 slides)
4.1. Nicotinic receptor (2 slides)
4.2. Muscarinic receptor - G Protein coupled receptor (2 slides)
5. Cholinergic agonists
5.1. Acetylcholine as an agonist
5.2. Nicotine and muscarine as cholinergic agonists
5.3. Requirements for cholinergic agonists
6. SAR for acetlcholine (6 slides)
7. Binding site (muscarinic) (3 slides)
8. Active conformation of acetylcholine (2 slides)
9. Instability of acetylcholine
10. Design of cholinergic agonists (7 slides)
11. Uses of cholinergic agonists (2 slides)

[46 slides]
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CHOLINERGIC
NERVOUS
SYSTEM

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1. Nerve Transmission
Peripheral nervous system

CNS

Brain

Peripheral nerves

Muscle

Heart

Gastro-
intestinal
tract
(GIT)

Spinal cord

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http://trc.ucdavis.edu/biosci10v/bis10v/media/ch25/nervous_divisio
ns.swf

http://trc.ucdavis.edu/biosci10v/bis10v/media/ch25/ne
rvous_divisions.swf

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1. Nerve Transmission
Peripheral nervous system
Skeletal
muscle
CNS
(Somatic)
Ach
(N)
CNS
(Autonomic) Synapse
Ach (N) NA
Sympathetic

Adrenaline
Ach Adrenal
(N) medulla AUTONOMIC
Parasympathetic Synapse

Ach
Ach (M)
(N)
Smooth muscle
Cardiac muscle

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http://entochem.tamu.edu/neurobiology/index.html

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 1. Nerve Transmission
Synapses

100-500A
Receptors

Nerve impulse New signal

Nerve Nerve

Vesicles containing Release of Receptor binding

neurotransmitters neurotransmitters and new signal

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2. Neurotransmitter
Acetylcholine (Ach)

O
+
C NMe 3
H 3C O

Acetyl Choline

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3. Transmission process
Signal in nerve 1

Nerve 2
Nerve 1
...
Signal ...
...

. Acetylcholine
Acetylcholinesterase enzyme

Vesicle
Cholinergic receptor ©1
3. Transmission process
Vesicles fuse with membrane and release Ach

Nerve 2
Nerve 1

Signal

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3. Transmission process

Nerve 2
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3. Transmission process

• Receptor binds Ach 2o Message

• Induced fit triggers 2o message
• Triggers firing of nerve 2
• Ach undergoes no reaction

Nerve 2
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 3. Transmission process

• Ach departs receptor

• Receptor reverts to resting state
• Ach binds to acetylcholinesterase

Nerve 2
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 3. Transmission process

Ach hydrolysed
by acetylcholinesterase

O O

C C HO
H 3C O
NMe3
H 3C OH + NMe3 Nerve 2
Acetylcholine Acetic acid Choline ©1
3. Transmission process
Choline binds to carrier protein
Choline

Nerve 2
Nerve 1

Carrier protein for choline

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3. Transmission process
Choline transported into nerve

Nerve 2
Nerve 1

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3. Transmission process
Ach resynthesised

Nerve 2
Nerve 1

E 1 = Choline acetyltransferase
O O

C E1 C
NMe3
H 3C SCoA + HO CH2 CH2 NMe3 H 3C O
Choline Acetylcholine

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3. Transmission process
Ach repackaged in vesicles

Nerve 2
Nerve 1

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4. Cholinergic receptors
Receptor types
• Not all cholinergic receptors are identical
• Two types of cholinergic receptor - nicotinic and muscarinic
• Named after natural products showing receptor selectivity
HO

NMe

Me CH2NMe3
O
N

Nicotine L-(+)-Muscarine

Activates cholinergic Activates cholinergic

receptors at nerve synapses receptors on smooth
and on skeletal muscle muscle and cardiac muscle
Acetylcholine is natural messenger for both receptor types
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Peripheral nervous system
Skeletal
Muscle
CNS
(Somatic) SOMATIC
Ach
(N)
CNS
(Autonomic) Synapse
Ach (N) NA
Sympathetic

Adrenaline
Ach Adrenal
(N) medulla AUTONOMIC
Parasympathetic Synapse

Ach
Ach (M)
(N)
Smooth Muscle
Cardiac Muscle

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 4.1 Nicotinic receptor

Control of Cationic Ion Channel:

Binding
Receptor site Messenger

Induced
Cell fit Cell
membrane membrane
‘Gating’
(ion channel
opens)
Five glycoprotein subunits
traversing cell membrane

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 4.1 Nicotinic receptor

The binding sites

Binding
sites

Ion channel
 
 

Cell  
membrane   


Two ligand binding sites

x subunits mainly on -subunits

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 4.2 Muscarinic receptor - G Protein coupled receptor
Activation of a signal protein
• Receptor binds messenger leading to an induced fit
• Opens a binding site for a signal protein (G-protein)
messenger
induced
fit

closed open
G-protein
bound

G-protein
split ©1
 4.2 Muscarinic receptor - G Protein coupled receptor
Activation of membrane bound enzyme
• G-Protein is split and subunit activates a membrane bound
enzyme
• Subunit binds to an allosteric binding site on enzyme
• Induced fit results in opening of an active site
• Intracellular reaction is catalysed

Enzyme Enzyme

active site
active site (open)
(closed)
subunit
Intracellular
reaction
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 5. Cholinergic agonists
5.1 Acetylcholine as an agonist
Advantages
• Natural messenger
• Easily synthesised

O HO NMe3 AcO NMe3

+ NMe 3
Ac2O

Disadvantages
• Easily hydrolysed in stomach (acid catalysed hydrolysis)
• Easily hydrolysed in blood (esterases)
• No selectivity between receptor types
• No selectivity between different target organs
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 5. Cholinergic agonists
5.2 Nicotine and muscarine as cholinergic agonists
Advantages
• More stable than Ach
• Selective for main cholinergic receptor types
• Selective for different organs

Disadvantages
• Activate receptors for other chemical messengers
• Side effects

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 5. Cholinergic agonists
5.3 Requirements for cholinergic agonists

• Stability to stomach acids and esterases

• Selectivity for cholinergic receptors
• Selectivity between muscarinic and nicotinic receptors
• Knowledge of binding site
• SAR for acetylcholine

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 6. SAR for acetlcholine
Ethylene
bridge

NMe3
Me O

Acetoxy 4 o Nitrogen

Quaternary nitrogen is essential

O O

CMe3 NMe2
H3C O H3C O

Bad for activity

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6. SAR for acetylcholine
Ethylene
bridge

NMe3
Me O

Acetoxy 4 o Nitrogen

• Distance from quaternary nitrogen to ester is important

• Ethylene bridge must be retained

O O

H 3C O H 3C O NMe3
NMe3

Bad for activity ©1

6. SAR for acetylcholine
Ethylene
bridge

NMe3
Me O

Acetoxy 4 o Nitrogen

Ester is important

NMe3 NMe3
H 3C O H 3C

Bad for activity

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6. SAR for acetylcholine
Ethylene
bridge

NMe3
Me O

Acetoxy 4 o Nitrogen

Minimum of two methyl groups on quaternary nitrogen

O Et
O Et
N Me
N Et H 3C O
H 3C O Me
Et

Bad for activity Active

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6. SAR for acetylcholine
Ethylene
bridge

NMe3
Me O

Acetoxy 4 o Nitrogen

Methyl group of acetoxy group cannot be extended

O

H 3C
NMe3
O

Bad for activity

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6. SAR for acetylcholine
Ethylene
bridge

NMe3
Me O

Acetoxy 4 o Nitrogen

Conclusions:
• Tight fit between Ach and binding site
• Methyl groups fit into small hydrophobic pockets
• Ester interacting by H-bonding
• Quaternary nitrogen interacting by ionic bonding

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7. Binding site (muscarinic)

hydrophobic
pocket Trp-307
Asp311
CH3 CH3
CO2

N CH3
O O hydrophobic
pockets
CH3
Trp-616
Trp-613
H
H
O N hydrophobic
pocket

Asn-617

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7. Binding site (muscarinic)

vdw Trp-307
Asp311
CH3 CH3
CO2
Ionic bond
N CH3 vdw
O O
H-bonds CH3 vdw
Trp-616
Trp-613
H
H
O N

Asn-617

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7. Binding site (muscarinic)

• Possible induced dipole dipole interaction between quaternary

nitrogen and hydrophobic aromatic rings in binding site
• N+ induces dipole in aromatic rings

+ +
R
NMe3
- -

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8. Active conformation of acetylcholine

O H H Me
Me

C N
Me O Me

H H

• Several freely rotatable single bonds

• Large number of possible conformations
• Active conformation does not necessarily equal the most
stable conformation

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 8. Active conformation of acetylcholine
Rigid Analogues of acetylcholine
HO
O O H
NMe3
Me CH2NMe3 Me CH2NMe3 Me
O O O
MUSCARINE H

• Rotatable bonds ‘locked’ within ring

• Restricts number of possible conformations
• Defines separation of ester and N
4.4A 5.9A
O O
N N

Muscarinic Nicotinic
receptor receptor
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9. Instability of acetylcholine

Me 3N

 O

 C

H 3C O

• Neighbouring group participation

• Increases electrophilicity of carbonyl group
• Increases sensitivity to nucleophiles

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 10. Design of cholinergic agonists

Requirements

• Correct size

• Correct pharmacophore - ester and quaternary nitrogen

• Increased stability to acid and esterases

• Increased selectivity

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 10. Design of cholinergic agonists
Use of steric shields

Rationale

• Shields protect ester from nucleophiles and enzymes

• Shield size is important

• Must be large enough to hinder hydrolysis

• Must be small enough to fit binding site

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 10. Design of cholinergic agonists
hinders binding to esterases
O Me and provides a shield to
nucleophilic attack
Methacholine NMe3
Me O *

Properties asymmetric centre

• Three times more stable than acetylcholine
• Increasing the shield size increases stability but decreases
activity
• Selective for muscarinic receptors over nicotinic receptors
• S-enantiomer is more active than the R-enantiomer
• Stereochemistry matches muscarine
• Not used clinically
HO
O Me O H

Me CH2NMe3 Me CH2NMe3
Me CH2NMe3 O O
O H Me
(S) (R)
MUSCARINE
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 10. Design of cholinergic agonists
Use of electronic factors

• Replace ester with urethane

• Stabilises the carbonyl group


O O O

H 2N
C
H 2N
C
= 
H 2N
C

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10. Design of cholinergic agonists

C NMe3
H2N O Carbachol

Properties
• Resistant to hydrolysis
• Long lasting
• NH2 and CH3 are equal sizes. Both fit the hydrophobic pocket
• NH2 = bio-isostere
• Muscarinic activity = nicotinic activity
• Used topically for glaucoma

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10. Design of cholinergic agonists

Steric + Electronic factors

O Me
C Bethanechol
NMe3
H2N O *

Properties
• Very stable
• Orally active
• Selective for the muscarinic receptor
• Used to stimulate GI tract and urinary bladder after surgery

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10. Design of cholinergic agonists

Nicotinic selective agonist

C
* NMe3
Me O * asymmetric centre
Me

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11. Uses of cholinergic agonists

Nicotinic selective agonists

Treatment of myasthenia gravis
- lack of acetylcholine at skeletal muscle causing weakness

Muscarinic selective agonists

• Treatment of glaucoma
• Switching on GIT and urinary tract after surgery
• Treatment of certain heart defects. Decreases heart muscle
activity and decreases heart rate

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Peripheral nervous system
Skeletal
Muscle
CNS
(Somatic) SOMATIC
Ach
(N)
CNS
(Autonomic) Synapse
Ach (N) NA
Sympathetic

Adrenaline
Ach Adrenal
(N) medulla AUTONOMIC
Parasympathetic Synapse

Ach
Ach (M)
(N)
Smooth Muscle
Cardiac Muscle

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