THE

CHOLINERGICS
 Learning Tasks
By the end of this session, students are expected to
be able to
• Describe the Nicotinic Acetylcholine receptors
• Describe the Muscarinic Acetylcholine receptor
• Discuss the Cholinergic neurochemistry
• Discuss the Cholinergic agonists their SAR and
clinically useful medicines
• Cholinergic blocking agents (antagonists) their SAR
and clinically useful medicines
 Learning tasks cont…..
• Classify the indirect acting cholinergic agonists
• Discuss the acetylcholinesterase reversible
inhibitors the diferent types and mechanism
• Discuss the acetylcholinesterase irreversible
inhibitors the different types and mechanism
• Discuss the kinetics in the mechanism of action of
indirect acting cholinergic agonist the types and
mechanism
 Buzzing

What is a receptor?
 Receptor
Receptor is a region of tissue or molecule in
the cell membrane, which responds
specifically to a particular neurotransmitter,
hormone, antigen or other substance
Cholinergic Receptors
They are characterized as:
(i)   Nicotinic and
(ii) Muscarinic
The neurotansmitter at these receptors
is the ACETYLCHOLINE
(a)    Nicotinic Acetylcholine
receptor
(i)   It is a glycoprotein in nature.
(ii)  When a neurotransmitter (Ach)
binds to it causes permeability of
the membrane to allow passage of
small cations, Ca 2+, and K+,
hence the Physiologic effect is to
elicit a temporary depolarization
of the membrane
• (iii)   And such depolarization
results in muscular
contraction at neuromuscular
junction
 (iv)     This receptors when
blocked by drugs plays an
important role in the control of
hypertension.
(b) Muscarinic acetylcholine

receptor.
(ii)  Action of Ach on muscarinic
receptors can be either stimulatory
or inhibitory
(iii)  Stimulates secretion from
salivary gland, sweat glands, and
contraction of the gut and
constriction of the airways of the
respiratory tract
• (iv) Inhibits the contraction of the
heart and relaxes smooth muscle
of blood vessels (vasodilation
effect)

• (v) A number of cholinergically

induced responces depend on K+
channel although not always in the
same way.
CHOLINERGIC NEUROCHEMISTRY
(i)   Cholinergic neurons synthesize, store and
release Ach, also they form choline acetyl
transferase (CHAT) and acetlycholinesterase
(ACHE).

(ii)  Ach is synthesized/prepared at the nerve

ending by transfer of an acetyl group from
acetyl – CoA to Choline, - the reaction is
catalyzed by choline acetyltransferase
• (iii)   Choline uptake is competitively
inhibited by several quaternary
ammonium bases e.g. Hemicholinium

and choline acetyltransferase is

inhibited in vitro by trans-N-methyl-
4-(1-naphthyl –vinyl) pyridinium
iodide.
 CH3
O N CH3
HO +

CH3 N + OH O
CH3
Hemicholinium (HC 3)
 Cholinergic Agonists
Structure for acetylcholine
O
+  
(CH3)3 N CH2 CH2 O C CH3

(i) Cholinergic Stereochemistry

The conformational properties of Ach and other
cholinergic chemicals have been studied through
various techniques
 (i)        -Roentgenographic (X-ray) crystallography
(ii)         -Nuclear magnetic resonance (NMR)
(iii)        -Molecular orbital calculations
 
 
These studies provided only
circumstantial evidence about the
conformation of the chemicals as
they act on the biological receptor
• (i)            The conformation of the
choline moiety of ACh has drawn the
most attention in attempting to relate
structure and pharmacological
activity

• (ii)          The exact conformation

adopted by ACh to give its
Cholinergic activity in Vivo is
unknown.
• (iii) So at the moment the
only alternative is to use the
more classic approaches of the
Stereochemical requirements
of drug-receptor interactions
(a) Muscarinic Receptor
• (i) A muscarinic receptor is stereo
selective in its interaction with
cholinergic agents.
 
• (ii) From this information a
hypothesis about the
conformational properties of ACh
at muscarinic receptor has been
developed
• (iii)    Muscarine (an alkaloid and
nonselective agonist of the muscurinic
acetylcholine receptor) has 8 optical
isomers or 4 geometric isomers
(enantiomeric pairs), muscarine,
epimuscarine, allomuscarine and
Epiallomuscarine.
 
• But Muscarine activity is  specific and
resides primarily in the naturally occuring
3R 5S 2S(+) muscarine;
• Structure for muscarine
The activity of muscarine
HO is highly specific and
3 4 + resides primarily in the
CH3 2 5
CH2 N (CH3)3 naturally occuring ( + )
H
1 muscarine.
O H

Muscarine

(+) (2S, 3R, 5S) – Muscarine

Note. The nicotinic receptor is not considered as highly

stereoselective as its muscarinic counterpart.
Structure – Activity Relationships of
cholinergic Agonists
 
Acetylcholine is relatively a simple molecule therefore
due to its simple Chemistry and ease of testing activity
enabled/allowed various chemical alterations
    
These alterations on the molecule are divided into 3
categories.
(i)        The Onium group
(ii)        The ester function
(iii)         The choline moiety
   
O
+
(CH3)3 N CH2 CH2 O C CH3
Buzzing O
+  
(CH3)3 N CH2 CH2 O C CH3

• Which between alfa or beta substitution will affect hydrolysis

of acetylcholine? And why?
•If the onium group is responsible for binding affinity at the
receptor. Then what type of a bond is involved and why?
Therefore:
(i)Shortening or lengthening of the chain
between the ester group and the onium moiety
reduces muscarinic activity
(ii) - Substitution on the choline moiety 
both nicotinic and muscarinic activity but
muscarine is terribly affected
()iii) -Substitution  Nicotinic activity to a
greater extent
(iv)Hydrolysis by AChE is affected more by
substitutions on the - than on the - carbon
• The Onium group is essential for the intrinsic
activity i.e.
-Contributes to the affinity of the drug to the
receptors through providing binding energy or as
a directing group.  
-The trimethylammonium groups offers the
optimal functional group for activity

• The ester group in ACh contributes to the

binding of the compound to the parasympathetic
terminal synapse probably because of its
hydrogen bond forming capacity.
It has been proposed that the muscarinic receptor contains an
anionic – binding site as shown in the hypothetical structure of
muscarinic receptor fig (a).
Site 3 o
5.7A

dt
dt

dt HO
o CH3 CH 2
+ CH 3
Site 2 3A Site 1 o
H H N
CH 3 CH 3
dt

(a) (b)

(a)- A diagram of distribution of (b) Interaction of muscarine

charges and hydrophobic with muscarinic receptor
Portions (shaded) and large
flat area for Van der Waals
interaction ( NOTE: dt= d+
– This anionic-binding site accommodates
the quaternary group of muscarine
approximately 3.0 Ă from a hydrogen-
bonding site (site 2 above) for the ring
oxygen of muscarine or the other oxygen
of the choline moiety.

– A distance of 5.7Ă separates the

quaternary nitrogen from the alcohol
group of muscarine.
– Site 3 can interact with the carbonyl
group of ACh, the ether oxygen of the
dioxane and the double bond of furan
(nucleophilc centres)

– The ether oxygen appears to be of

primary importance for high muscarinic
activity, because both choline ethyl
ether and -methylcholine ethyl ether
have high muscarinic activity.`
 +
CH3CH2 O CH2CH2 N (CH3)3

Choline Ethyl Ether

+
CH3CH2 O CH CH2 N (CH3)3
CH3
Betamethylcholine ethylether
• The classic SAR for muscarinic
agonist can be summarized as:

1.The molecule must posses a nitrogen atom

capable of bearing a positive charge,
preferably quartenary ammonium salt

2. For maximum potency, the size of the alkyl

groups substituted on the nitrogen should not
exceed the size of a methyl group
3. There should be an oxygen atom,
preferably an ester-like oxygen
capable of participating in a
hydrogen bond.

4. There should be a two-carbon unit

between the oxygen atom and the
nitrogen
 Classification of cholinergic agonists

(1) Direct Acting Cholinergic agonists - Clinically useful

(i) Acetylcholine Chloride:

CH3 O -
CL
+
CH3 N CH2CH3 O C CH3
CH3

Short duration of action has failed to be used as a cholinergic

agent. Easily hydrolysed by esterases.

-ACh Chloride is a hydroscopic powder available in admixture

 mannitol to be prepared in sterile H2O just before use.

-Acts as a short-acting miotic very useful after cataract

surgery during the placement of sutures
(ii) Methacholine Chloride
i.e. Acetyl - Methyl choline chloride

CH3 CH3 O
+ CL
CH3 N CH2 CH O C CH3
Alfa Omega
CH3

-It is an acetyl ester of -methylcholine

-Unlike the ACh, this has sufficient Stability in the body to

give sustained pararympathetic stimulation. Has almost no
nicotinic activity. Is rarely used today

-It can exist as (S) and (R) enantiomers-though the chemical

used is a racemic mixture, but its muscarinic activity resides
principally in the (S) isomer.
 Activity
Small groups

What is the source of stability in this analog?

CH3 CH3 O
+ CL
CH3 N CH2 CH O C CH3
Alfa Omega
CH3
(iii).Carbachol

CH3 O
-
+ CL
CH3 N CH2CH2 O C NH2
CH3

It is a Choline chloride carbamate

(i)Pharmacological activity similar to ACh, since is an ester of Choline,
has both muscarinic and nicotinic activity /properties

(ii)It is a miotic agents Useful in place of Pilocarpine or Neostigmine

for reducing the intraocular tension of glaucoma

(iii)Used in ocular surgery when a prolonged miosis is required when

ACh fails.

(iv)Is stable to hydrolysis than ACh due to the Carbachol Group which
 the electrophilicity of the carbonyl. Hence is stable in aqueous
solutions
• Mode of action
1- Causes release of ACh from cholinergic
nerve endings
2- Act as an inhibitor of AchE
3- It is a semi reversible inhibitor of AchE
and serves to prolong the duration of
ACh at neuromuscular function
(4) Pilocarpine HCL USP

CH2 CH3
CH3CH2 N
H H
. HCL
O O N

It is a hydrochloride of an alkaloid obtained from the dried leaflets of

Pilocarpus jaborandi or P.microphyllus in which it occurs to the extent
of about 0.5%  other alkaloid

Is hygroscopic and affected by light, may be sterilized by autoclaving.

Alkalies saponify the lactone group to give pharmacologically
inactive hydroxy acid (pilocarpic acid)

Base catalysed epimerization at the ethyl group (C-3) position occurs

to an appreciable extent and is another major pathway of degradation
Note: Both routes result in loss of pharmacological activity
Action
• It mimics the action of muscarine to stimulate
ganglia through receptor site occupation similar to
that of ACh.
• This is because the overall molecular architecture
and the interatomic distances of its functional
groups in certain conformations are similar to
those of muscarine (ie, about 4Ă from the tertiary
N-4 nitrogen to the ether oxygen or carbonyl
oxygen) and are compatible with this concept.
• Used in Rx of glacucoma.
 Chemical degradation of
pilocarpine
CH3
CH3CH2 CH3
N H2O CH3CH2
N
O
O O
N
pilocarpine O
OH OH N
pilocarpic acid
epimerization

CH3
CH3CH2 H
N
O
O N
isopilocarpine
 Assignment

• Provide the mechanism towards formation

of pilocarpic acid and the base catalized
epimerization reaction
INDIRECT – ACTING CHOLINERGIC
AGONISTS
Buzzing

Give a name of one indirect acting

cholinergic agonist you know and classify
it.
 Classification
Indirect acting cholinergic agonists are
classified into two classes:
(i) Acetylcholinesterase (ACHE) reversible
inhibitors
(ii) Acetylcholinesterase irreversible
inhibitors
 (i) Acetylcholinesterase (ACHE) reversible
inhibitors
•  Termination of ACh is primary by hydolysis by
AChE (acetylcholinesterase)
• The Products of hydrolysis are choline and acetic
acid.
• Inhibition of AChE prolong the life of the Neuro
transmitter (Ach) and produces similar
Pharmacological activity as ACh when
administered
 Anticholinesterases have been used in glaucoma
also as nerve gases and insecticides.
 O
+ +
CH3 C O CH2CH2 N (CH3 3 + H2O ACHE
HO (CH2)2 N (CH3)3+ CH3 COOH

Mechanism of hydrolysis of ACh by ACHE

The active site of AChE consists of an ammonic
and esteratic site. The ammonic site is formed by
the - carboxylate Group (COO-) of Glutamic
Acid residue.

Also present in the active site of AChE are an

acidic group HA, Pka 2, believed to be tyrosine
residue, also two imidazole groups from histidine
residue IM, and IM2 with Pka values 6,3 and 5.5
respectively; and also serine residue.
 
Steps Involved
1. Formation of the ACh + AChE reversible
complex (A)
2. Acetylation of esteratic site of the enzyme (B)
3. General base hydrolysis of acetylated
enzyme (c )
4. Free enzyme (D).
See the following diagram
 
 His C
O O
Im1 +
K +1 N(CH3)3
ACH + AChE HN N
K 1

O HA
O C CH3
Ser CH2 OH N NH
Im2
His
A
 His C
O O
I m1 +
HN N N(CH3)3

O HA
O C CH3

..
Ser CH2 OH N NH
Im2
His
B
 His C
Im1 O O

HN N
H2O
K2 H
. .o H HA
HOAC
Choline
O C CH3

O N NH
Ser CH2 IM2
His
C
 His C
Im1 O O

HN N
HA

N NH
Ser CH2 OH
IM2
His
D
 Clinically Useful medicines
( a ) P hys os tigmine

C H3
C H 3 NHC OO

N N
C H3 C H3

(i)Freely soluble in alcohol, slightly soluble in H2O

(ii)Physostigmine is an alkaloid of a dried ripe seed
of Physostigma vemenosum – white crystalline
powder.
(iii)As a free base is quite sensitive to heat, light
moisture and bases and readily undergoes
decomposition
Mechanism
It is a reversible competitive inhibitor of AChE
Use: glaucoma
 (2) Neostigmine
+
N (CH3)3

O -
Br
O CN(CH3)2

White crystalline powder soluble in water and in alcohol

Use of Physostigmine as a prototype of an indirect-acting

parasympathomimetic drug led to discovery/development
of stigmine (in which the trimethylamine group was
placed in para position.)

But better AChE inhibition is obtained if the groups are

placed Meta to each other and this gave Neostigmine a
more active and useful agent.
– The dimethyl carbamate in Neostigmine has
greater chemical stability than the methyl
Carbamate in Physostigmine towards hydrolysis

– Use and mechanism as physostigmine except

that has higher stability and greater miotic
activity

– Very useful for treating myasthenia gravis –a

condtion caused by an autoimmine mechanism
that requires an  in ACh in the neuromuscular
junction to sustain muscular activity.
4. Edrophonium Chloride (tensilon)

C2H5
+N CH A reversible Anticholineserase
CH3 3
CL
- Rrapid onset and short duration of
action then has neostigmine

OH A specific anticurane agent and actsC

in One minite to alleviale overdose of d
tubocurarine gallamine triethiodide
Pontial diagnosti agent for myasthenia gravis
• (ii) Acetylcholinesterase Irreversible
inhibitors
• Both AChE and BuChE – are inhibited
irreversibly by a group of phosphate esters
that are highly toxic (LD50 for human is 0.1
to 0.001mg/kg).  
• Actually these chemicals are nerve Poisons
and have been used in warfare and as
agriculture insecticides. 
• They permit ACh to
accumulate at nerve endings
and produce worst actions of
ACh like actions. 
• These compounds belong to a
class of organophosphorus
esters, and have a general
formula as follows:
 A
R1 = alkoxy
R1 P X R2 = alkoxyl, alkyl, tertiary amine
X = a good leaving group eg F, CN, thionate,
R2
p-nitrophenoxy

(A)– Is usually Oxygen or sulfur and

even Selenium. Other atom than O
when used, the compound needs
biological activation before it
becomes an effective cholinesterase
inhibitor.
(X)Is the leaving group when the
molecule reacts with the enzyme and
the Prototype groups include fluoride
and nitrite and P- nitrophenoxy

• (R2) – Moiety that imparts

lipophylicity to the molecule and
contributes to its absorption through
the skin
• Mechanism of Inhibition 

• Takes place in two steps namely

– Association of enzyme and Inhibitor
– The Phosphonylation step which is
completely analogous to acylation by
the substrate
• The serine residue in the esteratic site
(in the enzyme) forms a stable
phosphoryl ester with the
organophosphorus inhibitor.

• Although these are irreversible

inhibitors, it is possible to reactivate the
enzyme if action is taken soon after the
phosphorylation has occurred.
• Reactivation to the free enzyme can be
achieved by Using compounds which
can provide a nucleophilic attack on
the phosphorylated enzyme.
 
• Examples of such reactivators are
nicotinic hydroxamic acid and
Pyridine-2-aldoxime methiodide (2-
PAM).
 Phosphorylating agents such as
Sarin,
CH3
CH3 P O
CH O F
CH3

May become refractory to reactivation by cholinesterase

reactivators simply because of an aging Process which
may take place in vivo or vitro; as follows;
O O

H2O CH3
En2 P CH3 En2 P CH3 +
CH OH
CH3
O Provides OH
nucleophilic
CH attack,
CH3 CH3

aging involves partial hydrolysis of the phosphorylated

moiety that is attached to the serine residue in the esteratic
site of the enzyme.
  
• Phosphate esters used as insecticidal agents are
toxic and must be handled with extreme
caution; eg malathion though not very toxic,
will cause poisoning following ingestion of
relatively doses.

• Symptoms of toxicity are nausea, vomiting,

excessive sweating, salivation, miosis,
bradycardia; low blood pressure, and
respiratory difficulty which is the usual cause
of death.
 Malathion
CH3O S
P
CH3O S CH COOC2H5
[(dimethoxyphosphinothionyl)
CH2 COOC2H5 thio] butanedioic acid
diethylester

-Is used as an agricultural insecticide

-It is a poor inhibitor of cholinesterases
-Its effectiveness as a safe insecticide is
due to the different rates at which
humans and insects metabolize the
chemical.
Microsomal Oxidation which cause desulfuration occurs
slowly to form the phosphothioate (malaoxon) which is
10,000 times more active than malathion as a cholinesterase
inhibitor (reason for its delayed toxicity).

Insects detoxify the phosphothioate by a phosphatase –

forming dimethylphosphorothioate that is inactive as an
inhibitor(reason for poor activity)..

Human however are able to rapidly hydrolyze malathion

through a carboxyesterase enzyme yielding malathion acid
yet poorer inhibitor of AchE (reason for safety in humans).
• Other examples:
• (i) Isoflurophate,
• (ii) HETP –
Hexaethyltetraphosphate
• (iii) TEPP – Tetraethyl
pyrophosphate
• (iv)Parathion (insecticide),
• (v)Schradan (insecticide),
 An example of a reactivator

+N C=NOH
H
C H2Cl

Pralidoxime chloride (2-PAM chloride)

• used as an antidote for poisoning by parathion and related
pesticides
• effective against some phosphate that have a quaternary
nitrogen.
• an effective antagonist for some carbamates, such as
neostigmine methylsulfate and pyridostigmine bromide.

•an antidote for poisoning by an anticholinesterase and should

be used within few hours but after 36 hours is useless.
Administered IM, subcutaneous, IV
Phosphorylation and reactivation
steps
Phosphorylation of esteratic site in the enzyme and
reactivation by a reactivator such as 2-PAM involves 3
steps;
• Phosphrylation of serine residue by the inhibitor i.e.
organophosphate compound.
• Nucleophilic attack on phosphorylated residue by 2-
PAM.
• Removal of phosphorylated 2-PAM to generate free
enzyme,
 The following Fig. shows Phosphorylation
and reactivation of Cholinesterase
(a) Phosphorylation of serine residue by
isoflurophate
(b) Phosphorylated serine residue esteratic site
(c) nucleophilic attack on Phosphorylated
residue by 2-PAM;
(d) removal of phosphorylated 2-PAM to
generate free enzyme
 His C
O O His
IM1
IM1
HN N
HF HN N

H A
O H A
i C3H7O O
P i C3H7O
F P
i C3H7O
i C3H7O
.. O N NH
CH2 O H N N
IM2
Ser CH2 His
His IM2

A B
 His C
C O
O IM1 O
O
CH3 HN N
O
+N
N N CH= NO P(OiC3H2)2
H A
+N
HC
H A CH3
H O
.. N
iC3H7O NH
P O Ser CH2 O H N
iC3H7O IM2
O N NH His
CH2 IM2
His D
C
 KINETICS IN THE MECHANISMS OF
ACTION OF INDIRECT ACTING
CHOLINERGIC AGONISTS:
CHOLINESTERASE INHIBITOTS:
 
• There are 3 different chemical groupings
that are capable of inhibiting the ACHE-
these are acetyl,
• carbamyl and
• phosphoryl.
• These may react with the estaratic
sites of ACHE.
• Although the chemical reactions are
similar,
• the kinectic parameters for each of
these types of substrate are not the
same and result in the differences
between the toxicity and usefulness
of these compounds as drugs or
chemicals
 His C
O O
Im1 +
K +1 N(CH3)3
ACH + AChE HN N
K 1

O HA
O C CH3
Ser CH2 OH N NH
Im2
His
A
 His C
O O
I m1 +
HN N N(CH3)3

O HA
O C CH3

..
Ser CH2 OH N NH
Im2
His
B
 His C
Im1 O O

HN N
H2O
K2 H
. .o H HA
HOAC
Choline
O C CH3

O N NH
Ser CH2 IM2
His
C
 His C
Im1 O O

HN N
HA

N NH
Ser CH2 OH
IM2
His
D
The enzyme (ACHE) + Ach
(substrate) complex is reversible
and K+1 (association rate) and
K-1 (dissociation rate) are
respectively large ref. Fig A
above
K +1 K2 K3
E + Ach E.Ach E -A E + CH3COOH
K -1 H2O
Choline
• The enzyme substrate complex may form an
acetyl-enzyme intermediate at rate K2 ref. Fig B.
This k2 is slower than either k+1 and k-1. At this
step choline is released. This is the slowest step
hence is the rate limiting. It involves acetylation
of estaratic site of the enzyme.

• This acetyl-enzyme E-A intermediate is

hydrolysed to regenerate free enzyme. (fig C and
D). The rate of general base catalysed hydrolysis
is indicated by K3 rate constant ( ie de-acetylation
rate).
However:
•Carbamates eg in carbachol is also able to serve as
substrate for ACHE.
Therefore carbamyl esters acting as substrate for AchE form
Enzyme-Carbamyl (E-C) intermediate complex that
hydrolyse slowly and may be considered to be
semireversible inhibitors of AchE ie
•The rate of carbamylation (k2) is slower than the rate of
acetylation. The hydrolysis (K3-decarbamylation) of the
carbamyl-enzyme intermediate is 107 times slower than its
acetyl counterpart.
K +1 K K3
E + CX E .CX 2
E C
- E + C
K -1
X
 Now in this case K3 is the rate limiting step.
Note: The rate of k2 depends on the nature of the alcohol
(R-O) moiety of the ester and the carbamyl ester. Hence the order is : Esters of
carbamic acid O
( ROC NH2 )

•are better carbamylating agents of AchE than the methylcarbamyl

O
( ROC NHCH3 )

•and the dimethyl carbamyl analoques,

O
ROC N(CH2) 2

they also decabamylate from the enzyme faster

in the same order. N.B refer
physostigimine/neostigimine
• Organophosphate esters are also able to esterify the
serine residue in the active site of AchE
•Hence the organophosphorus compounds are used
as agriculcutral insecticides for the tereatment of
glaucoma and have been employed as nerve gases in
warfare. ie

K +1 K2 K3
E+ PX E.PX E- P E+ P
K -1
X
• The hydolysis rate (k3) of
phosphorylated serine is extremely
slow and hydrolysis to the free
enzyme and phosphoric acid
derivative is so limited that
inhibition is considered
irreversible
 Key Points
• Important structural features for muscarinic agonists are: the presence of
nitrogen atom capable of bearing a positive optimally a quartenary ammonium
salt
• The muscarinic receptor is highly stereospecific than the nicotinic receptor
• Carbachol provides prolonged miosis because is atable to hydrolysis

• Indirect acting cholinergic agonists are classified as reversible and irreversible

acetylcholinestrase inhibitors
• These agents are useful in the treatment of glaucoma
• Irreversible inhibitors are nerve poisons and are used in warfare and as
agricultural insecticides They belong to the organophosphorus group of
compounds
 Evaluation
• What are two cholinergic receptors?
• What is the role of the onium and ester
group of the acetylcholine
• What are the source of degradation of
pilocarpine?
 Evaluation Cont…
.
Once there was one gentleman working in a sisal farm morning to
evening. One day he did not return home till very late in the evening.
The family was worried and went on looking for him in the farm
where they found him laying down unconscious with difficulties in
breathing. While trying to find what has happened to him they saw a
spraying can containing some chemicals. They rushed him to hospital
and they met you there together with a young doctor.
Questions:
• Suggest what has happened to him
• Propose the active compound if you think any has been involved
• How does that compound work?
• Is there any room for the patient to survive? How?
• What caution will you give to the doctor attending the case
 Answer
1. Propose the type of active compound in the spray chemical→ most likely a phosphate
containing compound because he was found involved in agricultural work and the difficulties
in breathing indicates excessive muscarinic agonist acivity at the nerve endings (10 marks)
2. Describe the chemical nature and activity of these compounds → insecticides, act through
irrevesible inhibition of acetylcholine esterase enzyme (10 marks)
3. The patient can be saved because there is still room of reversing the action of this compound
by using a reactivator eg 2-PAM injection (36 hours have not elapsed as the incidence still
within same day) (10marks)
4. Explain how the 2-PAM works by the describing the 4 steps which are involved in AChE
enzyme phosphorylation by an irreversible inhibitor and reactivation reactions(10marks)
5. In your answer the structure for phosphorylated 2-PAM is necessary (10 marks)
6. Still you can caution the doctor that this is emergency and urgent and if the poison is really a
phosphate ester compound if we delay to take action another problem can arise where the 2-
PAM injection can’t work because of aging. (10 marks)
7. So briefly describe the mechanism of aging and hopefully the young doctor will inject the
patient with the 2-PAM you provide to him!(10marks)
 Reading list
Same as last session
CHOLINERGIC – BLOCKING
AGENTS
 Learning Tasks
By the end of this session, students are
expected to be able to:
• Classify the cholinergic blocking agents
• Discuss the different classes of cholinergic
antagonists and representative agents their
SAR and actions
Buzzing

• What is a cholinergic antagonist?

• And list down examples of cholinergic
blocking agents you know so far
• CHOLINERGIC – BLOCKING
AGENTS
• Cholinergic receptors are located at different places such
as at parasympathetic postganglionic nerve endings in
the smooth muscle, sympathetic and parasympathetic
ganglia and neuromuscular junction in skeletal muscle.
 
• All these receptors are exclusively activated by ACh, but
there are selective antagonists for each receptor
 
• For example Atropine is an effective, blocker at
parasympathetic postgranglionic terminal, and it acts on
both M1 and M2 receptors.
• Hexamethonium blocks transmission at the
nicotinic ACh receptor located in
autonomic ganglia.  
• D-tubocurarine blocks the effect of ACh
on skeletal muscle  the cause of
selectivity is not clearly known  
• Hence, anticholinergic action by drugs
and chemicals is apparently dependent upon
their ability to reduce the number of free
receptors that can interact with ACh.
• PARASYMPATHETIC-
POSTGANGLIONIC-BLOCKING
AGENTS
• These agents are as well known as anticholinergics,
parasympatholytics or cholinolytics
 
• The most potent anticholinergic drugs are derived from
muscarinic agonists that contain one and sometimes two
large or bulky groups,
 
• Development of these compounds has basically been based
on Atropine as a prototype that have some similarity to
ACh but contain additional substituents that enhance their
binding to the cholinergic receptor
 A

B C Chain N
A, B = bulky groups
C eg cycloalkyl,
aromatic, etc.
C = H, OH
Carboxamide.

As depicted above, the agent may contain

(i)A quaternary ammonium function or possess a tertiary
amine which becomes protonated and forms a cationic
species.
(ii)A chain which may include an ester, ether, or
hydrocarbon moiety,
(iii)The substituents A and B
which contain at least one
aromatic moiety capable of
eliciting Van der Waals
interactions to the receptor
surface and one cycloaliphatic
or other hydrocarbon moiety
for hydrophobic – bond
interactions.
(iv) C, may be hydroxyl or
carboxamide to undergo
hydrogen-bonding with the
receptor
• Cationic head  very important as a point
of attachment to the receptor
 
• The hydroxyl group = not requisite for
activity, but when suitably placed increases
activity over a similar compound without the
hydroxyl group. binding strength.

• Esteratic Group ((Ester group) =

contributes features for effective binding
 
Therapeutic indications:
There are 3 predictable and clinically
useful results from blocking the muscarinic
effect of acetylcholine (Ach)

•Mydriatic effect: - dilation of pupil and

cycloplegia

•Antispasmodic effect: lowered tone and

motility of the gastrointestinal tract and the
genitourinary tract.
•Antisecretory effect-reduced salvation
(antisialagoque), reduced persipiration
(anhidrotic), and reduced acid and gastric
secretion.

These are general effects of

parasympatholytics. Usual side effects
after oral use include dryness of the
mouth, mydriasis, and urinary retention.
• SOLANACEOUS ALKALOIDS AND
SYNTHETIC
• ANALOGUES
• (-)- Hyoscyamine, atropine( + )hyoscyamine
and scopolamine (hyoscine) are forerunners of the
class of parasympatholytic drugs – all are plant
products.
 
• Structural considerations
• All of them are esters of bicyclic aminoalcohol, 3-
hydroxytropane or of related aminoalcohols
 CH3 N

O
H
OH
SCOPINE
(6:7β-epoxy-3α-hydroxytropane or 6:7β-epoxy-3α-tropanol
CH3
N

H
O CH2OH
O C CH
Atropine
(or Hyoscyamine)

CH3
N

O H
O CH2OH
O C CH

Scopolamine of (Hyoscine)
CH3
N
1 2
CH3 N H
7 H OH
4 3
5
6
OH
(Chair) (boat)
TROPINE
(3Alfa Hydroxytopane
or 3Alfa tropanol)
• The structural formulars above show the piperidine
ring system commonly accepted i.e. the Chair
conformation-because this form has the lowest energy
requirement more stable
 
• In tropine the axially oriented hydroxyl group, trans
to the nitrogen bridge is designated as  and the
alternate Cis equatorially oriented hydroxyl group is
.
 
• And it has been pointed out that
antimuscarinic activity is associated with all
of the solanaceous alkaloids that possess
the tropine like axial orientation of the
esterified hydroxyl group.
 
• Proper enantiomorph is necessary for high
antimuscarinic activity as illustrated by the
potent (-)-hyoscyamine in comparison with
the weakly active (+) - hyoscyamine, while
the racemate, atropine has an intermediate
activity
 
• Products include; Atropine,
Atropine sulphate,
Hyoscyamine, scopolamine,
Homatropine and Ipratropium.
• Atropine
• -It is ( + )hyoscyamine ), the tropine ester of
racemic tropic acid and is optically inactive.
• Action
• It produces a mydriatic effect by paralyzing the
iris and the ciliary muscles and is used by the
oculist in iritis(inflammation of the iris) and Corneal
inflammations and lesion. Due to its effect of
drying secretion is used in rhinitis to cause relief
in colds. Treatment of arrhythmias, an antidote
for organosphosphate poisoning(inhibits muscarinic activit).
 
• Hyoscyamine
• A levorotatory form of a recemic mixture
known as atropine and used to treat disorders
of urinary tract more often than any ether
antispasmodic,
 
• N. B. Treats spasms of urinary bladder
although there is no evidence that it has any
advantages over the other balladona
preparations. Hyoscyamine preparations are
also used in therapy of peptic ulcers as
antispasmodics
• SYNTHETIC CHOLINERGIC –
BLOCKING AGENTS
• AMINOALCOHOL ESTERS 
 SAR
• A quaternary nitrogen which enhances
activity.
  Aromatic substitution lead to increase in
spasmodic activity: 2 phenyl rings are
optimal for activity more than 2 lead to
reduction in activity
Products
Clidinium Bromide
(3-hydroxyl -methylquinuclidinium bromide
benzilate),
O
C O -
Br
C
+ N
OH
CH3

•An anticholinergic agent which sometimes is marketed alone

or in combination with a minor tranquilizer chlordiazepoxide
•Used for peptic ulcer, hyperchlorhydria, ulcerative or spastic
colon
Others Dicyclomine Hydrochloride, Eucatropine
Glycopyrrolate, Mepenzolate Bromide, Methantheline,
oxyphencyclimine HCL and Propantheline Bromide (or Pro-
Banthine)
• AMINOALCOHOL ETHERS
• Agents already introduced have been used as antiparkinsonian
drugs rather than conventional anticholinergics (ie as
spasmolytics, mydriatics.
• They are closely related to the antihistamines
• Example
• Benztropine Mesylate

N CH3

C O
CH3SO3H
H

• Other agents
• Chlorphenoxamine hydrochloride, orphenadrine citrate
(C ) AMINOALCOHOLS
 These are equally efficacious in parkinsonism.
They have structural characteristics of
processing bulky groups in vicinity of
hydroxyl and cyclic amino function groups –
• Another structural feature is the (amino
propanol arrangement with three carbons
intervening between the hydroxyl and amino
functions) – all that have antiparkinson activity
are tertiary amines.
 
• SAR:
 - OH-group is necessary for
parasympathetic blocking activities.
• Quarternization destroys the
antiparkinsonian properties
• However quarternized products are used
as anticholinergic agents to produce an
antispasmodic and antisecretory
compounds e.g. tridinexethyl chloride
 • Example
• Biperidine

C CH2CH2 N

CH2 OH

Has weak visceral anticholinergic but strong nicotinolytic action,

Used in all types of Parkinson’s disease (postencephalitic, idiopathic,
arteriosclerotic) and help to eliminate akinesia, rigidity, and tremor.
Is of value in spatic disorders not related to parkinsonism e.g. multiple
sclerosis, spinal cord injury
Contraindicated in all forms of epilepsy
Others
Procyclidine HCL; Indihexethyl chloride USP, Isopropamide Iodide,
Tropicamide
(D) PAPAVERINE AND RELATED COMPOUNDS
• Papaverine exerts an antisparmodic effect on
smooth muscle – always called a non specific
antagonist.
• Papaverine interferes with the mechanism of
muscle contraction by inhibiting the enzyme
phosphodiesterase in smooth muscle cell as in the
fig below
Membrane
ATP 5 AMP
Adenylate
cyclase
Phasphodiesterase
CAMP

Smooth muscle relaxation

• Hence Papaverine is a potent Inhibitor of vascular smooth muscle , PDE and
there is a significant elevation of CAMP following the administration of it.
Hence inhibition of PDE and increased levels of CAMP is associated with
smooth muscle relaxation

R
It is an alkaloid from opium and the
N main salt is Hydrochloride. Useful in
R relieving the arterial spasm associated
CH2 with acute vascular occlusion antispa
smodic action on the vascular system
also used for bronchial spasm and
visceral spasm
R

R Papaverine
(R=OCH3)

N. B. The 4 methoxy groups are easily altered functional groups – Hence is not
well known whether these alkoxy groups are necessary for activity. But are
present in most of accepted compounds
GANGLIONIC – BLOCKING AGENTS

They are grouped as follows:

 

– (i)Depolarizing ganglionic -blocking

agents:
• Are actually ganglionic stimulants. Nicotine
in small doses gives similar action as a neuro
effector ACh, but in large doses brings about
a ganglionic block, characterized initially by
depolarization followed by typical
competitive antagonism. But such agents are
not therapeutically effective.
 
 
 – (ii)Non-depolarizing competitive
ganglionic - blocking agents.
These agents possess the
necessary affinity to attach to the
nicotinic receptor sites that are specific
for ACh – but they are unable to effect
depolarization of the cell. Examples of
therapeutically active agents are
hexamethonium, trimethophan and
tetraethylammonium salts.
 (iii)Non-depolarizing non-competitive
ganglionic - blocking agents
• Their effects are not at specific ACh
receptor site but some point further
along the chain of events –– these agents
are useful in management of peripheral
vascular diseases – a principal therapeutic
application has been treatment of
hypertension through blockade of the
sympathetic pathways.
• NEUROMUSCULAR-BLOCKING
AGENTS 
• Agents that block the transmission of ACh
at the motor end-plate are called
neuromuscular blocking agents – The
therapeutic use of these compounds is
primarily as adjuvant in surgical anesthesia
to obtain relaxation of skeletal muscle –
are also used in various orthopaedic
procedures such as alignment of fractures
and correction of dislocations.
 Products
Curare and curare alkaloids
•Tubocurarine chloride
CH3O 5 4
6 3
+
R1
7 -
N CL (1a) R1 = R2 CH3
1 2 CH3
O 8
H
OH a (1b) R1 = H; R2 = CH3
11'
12' 9
10 (1c) R1 = CH3; R2 = H
10' 11
9'
12
CH3 1' O (1b) Is the right structural
21 N OH
7'
R2 6'
( + ) tubocurarine
3' OH3
4' 5'

•An alkaloid from chondodendron tomentosum (menispermaceae)

•It is a non depolarizing blocking agent used for its paralyzing action on
voluntary muscles, the site of action being the neuromuscular junction
•Its action is inhibited or reversed by admin of AChE inhibitors such as
neostigmine or edrophonium chloride (Tensilon). 
 Key points
There are 3 groups of cholinergic blocking agents
namely
(i) Parasympatholytics e.g Hyoscyamine,
atropine( + )hyoscyamine and scopolamine
(hyoscine)] , papaverine a synthetic analog
(ii) Ganglionic – blocking agents eg
(iii) Neuromuscular-blocking agents
e.gTubocurarine chloride
 Evaluation
• What are the main actions of the cholinergic
blocking agents?
• What are the common side effects of the
cholinergic blocking agents
 Reading list
• Same as in previous session