CIMETIDINE

a rational approach to drug design

 TRADITIONAL DRUG DESIGN

Lead generation:
Natural ligand / Screening

Biological Testing

Drug Design Cycle

If promising
Synthesis of New Compounds

Pre-Clinical Studies
 ULCER AND THERAPY
• Ulcers : erosions of the
mucous membranes of the
stomach or duodenum.

• The presence of gastric acid

aggravates the problem and
delays recovery.

• In the early 1960s: neutralize gastric acid in the stomach by

administering bases such as sodium bicarbonate or calcium
carbonate.
 ULCER : GASTRIC JUICE
Acetylcholine
oesophagus
Histamine Gastrin

M
H Cck2
cAMP Gastric juices release is
+ promoted by AcH,
+ +
Parietal
Cells Histamine, and Gastrin.

Stomach H+ Cl -

Stomach
HCl
Antrum
Pyloric Receptors
Sphincter
Ion channel
Duodenum
Proton pump

Release of gastric acid should, therefore, be inhibited by antagonists blocking either the
cholinergic receptor or the receptor for gastrin or histamine receptor
 HISTAMINE
Experimentally histamine stimulated gastric
acid release in vitro

antihistamine agent might be

effective in treating ulcers
 HISTAMINE

NH3 NH3
 
HN N
Nš NH
Imidazole ring

• Two possible tautomers

• pKa for the α-NH2 group = 9.80 (99.6% ionize at
plasma pH)
• pKa for the imidazole ring = 5.74 (mostly not ionised
at plasma pH)
 ANTI-HISTAMINE
Commonly used to treat symptoms such as inflammation & itching
MeO NMe2 NMe2
O
N

Mepyramine Diphenhydramine
Benadryl
• No effect on gastric acid release
• Failed to inhibit all the then-known actions of histamine
• SK&F propose two types of histamine receptor (H1 and H2)
• H1 - responsible for classical actions of histamine
• H2 - proposed as the receptor on the parietal cells
• Claim that H2 receptors are unaffected by classical antihistamines
• Implies classical antihistamines are H1 specific
 LEAD : HISTAMINE
• No known H2 antagonist at the time - no lead
compound  use histamine itself as the lead
compound
• Aim : vary structure of histamine (an agonist) into
become an antagonist
• Need to know SAR requirements for H2 agonists
• Analogues tested by their ability to promote
gastric acid release
 SAR OF HISTAMINE
• Side chain had to have a positively charged
NH3 nitrogen
HN • Flexible chain between the above cation and a
N
heteroaromatic ring

NH3 NH3
HN HN
N N

H1 Receptor H2 Receptor

Two nitrogen atoms are required for H1 agonist activity

All three nitrogen atoms are required for H2 agonist activity
 AGONIST TO ANTAGONIST
• Add extra functional groups to find extra binding
interactions with the binding site
• Extra binding interactions may result in a different
mode of binding resulting in a different induced fit for
the receptor
• Different induced fit may fail to activate the receptor
• As result, analogue binds but fails to activate the
receptor
• Analogue likely to bind more strongly than an agonist
AGONIST TO ANTAGONIST

HN
Histamine

N NH3 HN

N NH3

Receptor (Inactive) Induced Fit - Receptor 'Switched on'

Extra
HN Functionality
N NH2
HN

N NH2

Receptor (Inactive)

Different induced fit

 AGONIST TO ANTAGONIST
Examples - extra hydrophobic groups

NHR1R2
N NHR1R2
N HN
R3 N

• No antagonist activity observed with extra hydrophobic groups

• Try adding extra hydrophilic groups instead
• Aim is to search for extra polar binding regions
 Nα-Guanylhistamine
H Guanidine moiety
N NH2
geometry of bonding might be
HN
N H2N altered to produce an antagonist.

• Exibit Partial agonist

• Prevents histamine from fully receptor activation
• Nα-guanylhistamine is binding to the proposed H2 receptor,
resulting in weak activation
• Whilst present, Nα -guanylhistamine blocks histamine from
binding
 Nα-Guanylhistamine
H
NH3 N NH2
HN
HN N H2N
N
The guanidine group is basic and ionised
But has TAUTOMER!

H H H
N NH2 N NH2 N NH2
HN HN HN
N H2N N H2N N H2N

H
N NH2
HN
N H2N

The positive charge is more diffuse and can be further away from the imidazole ring
 BINDING REGION
Antagonist
binding region

H
N NH2 Imidazole ring
HN binding region

N H2N Agonist
binding region

Polar Binding Region

• One of the polar binding regions is accessed by agonists and the

other by antagonists
• The antagonist polar region is further from the imidazole binding
region
 BINDING REGION
Antagonist
binding region

H
N NH2 Imidazole ring
binding region
HN
N H2N
Agonist
binding region

Antagonist Antagonist
binding region binding region

NH3
Imidazole ring HN HN
binding region
N N NH3
Agonist
binding region

No interaction as an antagonist Strong interaction as an agonist

 BINDING REGION
Antagonist
binding region

H
N NH2 Imidazole ring
binding region
HN
N H2N
Agonist
binding region

Antagonist
H2N
binding region
NH2
HN NH
Imidazole ring HN
binding region
N N NH
Agonist NH2
binding region
H2N

Binding as an antagonist Binding as an agonist

Receptor not activated Receptor activated
 CHELATION STRATEGY
Find an analogue which would bind to the antagonist region only.

H
H Proposal : The guanidine moiety
N N Strong interaction
HN
H interacts with a carboxylate ion in the
N N O
H H antagonist binding region by means of
O
Receptor two H-bonds and an ionic interaction
X=NH,S

H
S NH2 N NH2
HN HN
N NH2 N B X
A
X= SMe, Me
Both are partial agonists, but A has greater antagonist properties
 CHAIN EXTENSION
Aim : push the polar guanidine group further out and to increase the
interaction with the antagonist binding region
3C Bridge NH2 NH2
3C Bridge

N NH2 S NH2
HN H HN
N N
Guanidine Isothiourea
Partial agonist Partial agonist
Antagonist activity increases Antagonist activity decreases!

NH2 NH2

N NH S NH
HN HN
N H H N H
Strong interaction Weak interaction
O O O O
Different form of
Receptor hydrogen bonding Receptor
taking place
 CHAIN EXTENSION
Aim : push the polar guanidine group further out and to increase the
interaction with the antagonist binding region

NH2 Antagonist
binding region
HN
NH2
HN HN
N N
Imidazole ring NH2
binding region HN
Agonist
binding region NH2

Good binding as an antagonist Binding as an agonist

 CHAIN EXTENSION
Aim : push the polar guanidine group further out and to increase the
interaction with the antagonist binding region
X

N NH
Partial agonists with good antagonist
HN activity (X= Me or SMe)
N H H

N NH
HN
N H H
Strong interaction

O O

Receptor
X=NH2, SMe, Me
 POLAR BINDING REGION
is it really necessary for the chelating group to be charged?

Replace the ionic guanidine group with a neutral H-

bonding group
Thiourea
HN
H
N NH2
N
S

No agonist activity, very weak antagonist

 POLAR BINDING REGION
Similarities - Planarity, geometry, size, polarity, H-bonding ability
Differences - Thiourea is neutral while guanidine is basic and ionised

Thiourea Guanidine
H H
N NH2 N NH2

S e-withdrawing NH2
Neutral Basic

HN
HN S
H Chain extension
N NH2
N
N N NHMe
S H

burimamide
 CHAIN EXTENSION

Conclusions
• Chain extension leads to a pure antagonist with good
activity
• Chain extension allows a better overlap of the thiourea
group with the antagonist binding region
• Establishes the existence of H2 receptors
 DEVELOPMENT
burimamide was not suitable for clinical trials because its
activity was still too low for oral administration
HN S
Chain extension
N N NHMe
H

H H
H 
 N N
N N -H
š
I š N R N R II
N R N R
H H
H
III
H
 DEVELOPMENT
burimamide was not suitable for clinical trials because its
activity was still too low for oral administration
e-withdrawing e-donating
HN HN HN
CH2CH2NH3 H CH2CH2CH2CH2NHCSNHMe
N N N

Histamine Imidazole Burimamide

pKa = 5.74 pKa = 6.80 pKa = 7.25
Ionisation = 3% Ionisation = 40%

Decreasing the basicity and ionisation of the imidazole ring in

burimamide closer to that of histamine may increase the binding
interactions to the imidazole binding region
 DEVELOPMENT
Electron withdrawing
HN S pKa = 6.25
S Increase in antagonist activity
N N NHMe
H
Non-ionised imidazole is favoured
Thiaburimamide

Tautomer of Histamin
H  Inductive effect
N
• Favoured tautomer for histamine is I š R
N
• Side chain is electron withdrawing
• Inductive effect decreases with distance
• Np is less basic than N
• N is more likely to be protonated
 DEVELOPMENT
Metiamide Electron donating
Me
HN S
S
N N NHMe
H
Electron withdrawing

• 10 fold increase in antagonist activity w.r.t burimamide

• Increase in pKa to 6.80
• Increase in ionisation to 20%
• Unacceptable side effects - kidney damage
 DEVELOPMENT
• The side effects of metiamide may be due to the thiourea
group
• The thiourea group is not a natural functional group
• Replacing thiourea with a natural functional group may
remove the side effects

S O NH

N NHMe N NHMe N NHMe

H H H

Thiourea Urea Guanidine

Toxic side effects Drop in activity Drop in activity but
no agonist activity!
 DEVELOPMENT

H2N
NH2
NH
S
HN HN

N N S
NH
Agonist
binding region NH2
H2N
Binding as an antagonist No binding as an agonist
 DEVELOPMENT
• Retain the guanidine group
• Guanidine is a natural group present in the amino acid
arginine
• Increase activity by making the guanidine group neutral
• Add a strong electron withdrawing group to decrease basicity
(e.g. NO2 or CN)
Cimetidine
Me
H
N NHMe
S
HN
N N
CN
Electron withdrawing
cyanide group
 DEVELOPMENT
The cyanoguanidine moiety H
N NHMe
• Acts as a bio-isostere for
the thiourea group N
CN
H
N NHMe
• Both groups are polar but essentially neutral
• Both groups have low partition coefficients S

• The cyanoguanidine group is weakly acidic and weakly

basic - amphoteric
• The cyanogaunidine group is not ionised at pH 7.4
 DESOLVATION
H H
O

H
R G R G
R G O
Desolvation Binding
H
Energy penalty Energy released

O
H H

• A guanidine unit is highly polar and highly solvated

• The ease of desolvation may affect strength of binding and

activity

• A urea group is more hydrophilic than a cyanoguanidine group

• May explain lower activity of the urea analogue
 DESOLVATION
log (Act)
NCN

NNO2
HN
. .
NHMe
S
6.0
HN NHMe . HN NHMe

.
S
NNO2
HN
NCN .. NH2
HN NH2

5.0
HN NH2
.
N
O

O
. O HN N
H
N HN NHMe
Me
HN . HN X
.
N
H O
4.0 S
HN NH2 N N NHMe
H
Aminal system (Z)

-1.8 -1.4 -1.0 -0.6

log P of HZ
Log (activity) = 2.0 log P + 7.4
 DESOLVATION
O
Me Me
HN CHNO2 HN N

S S
N NHMe N N N
N H
H H

Greater activity than expected Lower activity than expected

Hydrophilic group should based on the hydrophobicity of
lower activity the group present
DIPOLE MOMENT
Approach and orientation Strong hydrogen bonding
O 2N

H
N NMe
R H O 2N

H
N NMe
R H

Receptor Receptor
surface surface

Dipole
H-Bonding regions Moments

Approach and orientation Weak hydrogen bonding

O

N
N
H O
H N
R N
N
H
H N
R

Receptor Receptor
surface surface
 CIMETIDINE
• The starting point is the guanyl-histamine that possesses weak antagonistic
properties againt the gastric secretion induced by histamine.
• The lengthening of the side chain of this compound increased clearly the H2-
antagonistic activity, but a residual agonist effect remained.
• In replacing the strongly basic guanidino function by a neutral thiourea, burimamide
is obtained. Although very active, this compound is rejected for its low oral
bioavailability.
• The addition of a methyl group in position 4 of the imidazolic ring, followed by the
introduction of an electron-withdrawing sulfur atom in the side chain, lead finally to a
compound that is both very active and less ionized, properties which improved its
absorption by the oral route.
• The derivative thus obtained, metiamide is excellent and, moreover, 10 times more
potent than burimamide. However, metiamide, because of its thiourea grouping, is
tainted with side effect (agranulocytosis, nephrotoxicity), that would limit its clinical
use.
• The replacement of the thiourea by an isosteric grouping having the same pKa (N-
cyanoguanidine) lead finally to cimetidine.
 RANITIDINE
4 3
Me2N CHNO2
5
2 S
O N NHMe
H

• Contains a nitroketeneaminal group

• Different heterocyclic ring
• Took over from cimetidine as the most widely sold
prescription drug in the world