Assignment

Structure-Activity Relationship of a
Novel Anti-malarial Agent
Submitted for the partial fulfilment of course
Advanced Medicinal Chemistry
PHA G621

Submitted by
L Naga Rajiv
2014H147206P

Under the supervision of

Dr. Hemant R Jadhav
Associate Professor
Department of Pharmacy
BITS Pilani

Contents
1.0 Introduction ......................................................................................................................... 1
2.0 SAR studies on a novel antimalarial agent .......................................................................... 3
3.0 Conclusion ......................................................................................................................... 13
4.0 Acknowledgements ............................................................................................................ 13
5.0 References .......................................................................................................................... 14

1.0 Introduction
Discovery of new drugs initially was largely based on trial and error basis. Medicinal chemists
used to synthesize random molecules and evaluate them. Serendipity also contributed a lot to
drug discovery. But as Medicinal chemistry evolved, along with other sciences like biological
sciences, computer science, Physics etc., drug design and discovery became more and more
based on some rational. However, the time, energy and other resources which are involved in
drug design is still high. Application of Computer science in drug design greatly reduced this
effort. The probability that a molecule being designed will actually be launched in the market
was previously very low, but the in-silico methods have surely contributed in increasing this
probability.
There are several in-silico methods which have been practiced and applied till date, like 2D
QSAR, 3D QSAR, Docking, prediction of ADME properties etc. However, one of the first
applications is QSAR, where mathematical models were developed to predict the activity of a
compound of similar structure. Even before QSAR models were developed, medicinal chemists
studied the Structure Activity Relationships (SAR). As the name suggests, in SAR, the
structure of a molecule is related to its activity. That is, the influence of specific moieties in a
molecule on its activity is studied, so that this knowledge can be applied to design and
synthesize molecules of improved activity but of the same series.
SAR is based on the principle similar structures-similar effects. QSAR also has the same
principle, but it uses parameters called descriptors to describe the potency. To do an SAR study,
it just needs the structures of a series of compounds and their activity. However, QSAR study
needs some knowledge of statistics. QSAR models correlate in vivo biological activity to
physicochemical properties of compounds while SARs attempt to explain chemical moieties
or structural features that contribute to or are detrimental to the biological activity.

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SAR involves making certain changes to the lead molecule like changing the number of
methylene groups, increasing or decreasing the degree of unsaturation, introducing or removing
ring systems, halogens etc. These changes in turn effect the activity of the molecule which is
studied. For example, increasing the number of methylene groups increases the lipophilicity
which in turns increases activity. Increasing the degree of unsaturation will increase rigidity.
Substituting a methyl group with naphthyl can create steric hindrance at the site of action. Such
changes and the effect on activity are studied in SAR.
This work presents in brief the SAR studies done by a group of researchers in Germany. The
aim was this work was to optimize a lead which was found to be active against multi-drug
resistant Plasmodium falciparum strains. The work was published in 6 parts, all of which have
been summarized step-by-step in this report.

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2.0 SAR studies on a novel antimalarial agent

Malaria is one of the most threatening tropical diseases causing between 1.5 and 2.7 million
fatal cases annually. It is more prominent in children and women, particularly in Africa. This
is largely due to the emergence of resistance of Plasmodium falciparum strains. The researchers
therefore found an urgent need for new agents active against multi-drug resistant plasmodium
strains.
The researchers chose a library of 61 compounds of different structural classes from various
literature studies. They were assayed for their inhibitory activity against Plasmodium
falciparum. They found that 47 of the compounds showed activity at 100m. But only 7 of
them retained activity at 10 M. The inhibitory activity of the 61 compounds were found as
shown in Table 1.
Compunds were evaluated for their inhibitory activity against intraerythrocytic forms of
Plasmmodium falcioarum strains Dd2 and 3D7 using a semi-automated microdilution assay.
The growth of the parasite was monitored thorugh incorporation of tritium labelled
hypoxanthine. All values were estimated to be correct within 30%. This procedure was
adopted for all the assays used in this study.
Of the 61 compounds, the compound labelled 7p showed best activity and was chosen as the
lead.

Other than its activity, another reason why this compound was chosen as lead because it allows
a variety modifications. The first step after lead identification was to find the exact inhibitory
concentration of compound 7p.

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Next some dramatic modifications were done with compound 7p, as illustrated above. Each of
the three derivatives lack one of the substituents on the central benzene core. It was found that
all the three modifications resulted in marked decrease of activity, which means that all the
three substituents are necessary for activity. As all three substituents are necessary for activity,
all three are open for modifications to possible improve activity.

In the first set of modifications, the acylamino substituent at 5 position was replaced with some
substituted cinnamic acid derivatives and tested for activity. It was found that 4-phenyl and 4benzyloxy derivatives (11a and 11c) showed improved activity, while others didnt. The exact
values of inhibitory concentration of 11a and 11c were found out to be 5 times lower than that
of 7p. This improvement upon structural modifications demonstrates the potential for further
development.
At this stage, the researchers were concerned if the activity is due to cytotoxicity. So,
compounds 7p and 11a were assayed against cell lines for their potential cytotoxic effects. It
was found out that 7p was indeed cytotoxic whereas the 5-fold better compound 11a didnt
show any cytotoxicity till 300 times the concentration in which it showed activity! This made
the researchers to work on cinnamic acid derivatives.

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So considering the cytotoxic effects, the researchers chose to work on cinnamic acid derivatives
as one of the substituents on the central benzene core, although it was showed slightly reduces
activity. In the second series of modifications, they chose to study the effect of substituents in
the para position of cinnamic acid derivative. So the lead for this series is as shown below.

R:H corresponds to compound 11a. 18 substituents were tried on the para position and labelled
3a-r. The activity was assayed as described earlier. The values are as under:

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SAR: Substituting the proton in para position with electron withdrawing groups like NO2,
CF3, -Cl, -Br etc. did not improve the activity. But substituting with methyl and methoxy
groups improved the activity. So, the researchers focused on 4-alkyl and 4-alkoxy derivatives.
The 4-alkyl series showed no improvement in activity compared to 4-methyl. But the alkoxy
series showed improvement, with the peak activity shown by propoxy substitution. Further
lengthening of the alkoxy chain decreased activity.
Substitution of 3-phenylpropionyl moiety of the lead structure with 4-propoxycinnamic acid
residue resulted in better activity.

In the next set of modifications, published as part 3 of their work, the researchers replaced the
tolylacetyl residue at the 2-amino group by several acyl substituents. 11 substituents were
synthesized, labelled 6a-l, and assayed for antimalarial activity as described earlier. The initial

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compound of this series 3q is same as 6e. It has a tolyl group attached to the acetyl moiety and
an IC50 value of 340nM.

The modifications started with removal of methyl group in toluene. The activity decreased
significantly to 9000nM. Introduction of bulky groups also improve the activity. But a benzyl

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group in place of phenyl improved the activity. This shows that the methyl group is necessary
for better activity. Next, ortho substitution was tried in place of para and again the activity
decreased, suggesting that only para substitution will lead to better activity. A series
electronegative substitutions were tried. Strangely it was observed that these electronegative
groups (-Cl, -Br) had similar activity as that of CH3 substitution although it is electropositive.
In fact, -CF3 in para position improved the activity 3-fold (120nM).
The conclusion of this part was that acyl substituent does play an important role in anti-malarial
activity.
i.

A methylene attached to carbonyl carbon is necessary for activity.

ii.

Substitution of the benzyl group should be at para position only.

iii.

The substituent must of the size comparable to methyl.

iv.

CF3 substitution at para position gives best activity

In the next series of experiments, published as Part 4 by the researchers, the authors replaced
the 4-propoxy phenyl residue of the cinnamoyl moiety by biaryl structure consisting of a
terminal aryl residue and a central 2-furyl ring.

The 4-propoxy phenyl residue had an IC50 value of 340nM. The modifications done and the
found activities are shown in the following table.
The phenylfurylacrolyl derivative inhibited with an IC50 value of 415nM. This is comparable
to that of the lead (340nM). Thus, there is a scope of some substituents leading to a much better
activity profile.

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Introduction of bulky group in place of phenyl resulted in a drastic decrease of activity. This
suggests steric hinderance. However, introduction of methyl group in para position improved
the activity 3-fold (120nM). Increasing the carbon chain length to 2 and introduction of double
bond improved the activity further. However, further increase in length decreased the activity.
Alkoxy substitutions and halogens in para position also, did not show activity better than their
alkyl counterparts. Strangely, para nitro substitution showed better activity (75nM) than the
alkyl substituents, although it is polar. Repositioning of the substitution to ortho or meta
positions decreased the activity.
The conclusions of this part are:
i.

The aryl group cannot be too bulky

ii.

Substitution must be at para position only.

iii.

Non-polar moieties in para position show better activity than polar moieties, with the
exception of nitro substitution, which turned out to be the best.

In part 5 of their work, the researchers repeated their work done in part 3, but by applying the
conclusions of part 4 of their work. As found in part 3, the conclusion were:
i.

A methylene attached to carbonyl carbon is necessary for activity.

ii.

Substitution of the benzyl group should be at para position only.

iii.

The substituent must of the size comparable to methyl.

iv.

CF3 substitution at para position gives best activity

The advantage of CF3 group is that it lacks the metabolic instability that might be associated
with the methyl group.

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In the last series of experiments, the researchers repeated their work published in part 5, but
with some different substitutions. However the values were compared with the compound
which gave the best activity in part 5, i.e., the one with nitro group in para substitution.

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The trifluoromethyl and the acetyl group were identified as equipotent replacements of the
terminal nitro group in this type of anti-malarial agents. Furthermore, incorporation of a
methylsulfonyl moiety led to an inhibitor twice as active as the lead structure 1. In addition,
this inhibitor lacks the nitro group of lead structure 1 potentially associated with an
unfavourable toxicological profile.

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3.0 Conclusion
The researchers optimized the lead structure obtained by assay of 63 compunds and proposed
the following structure.

The SAR of this compound is as follows:

i.

Both the amides and benzoyl moieties are required as substituents to the central
benzene core.

ii.

At the 5th position, a CH2=CH2 is required to connect the biaryl moiety. Saturating
this bond will decrease the activity.

iii.

The substitution of terminal phenyl ring of the biaryl moiety has to be in para position
only. Ortho and meta substitutions greatly decrease the activity.

iv.

The para substitution has to be electron donating, with the exceptions of NO2 and
SO2CH3.

v.

The benzoyl substitution at position 1 is best unsubstituted.

vi.

At position 2, benzyl is required to be attached to amide bond. Phenyl in place of

benzyl will result in loss of activity.

vii.

Substitution of this phenyl ring must be in para position only.

viii.

The size of the para substitution must be of size comparable to methyl.

4.0 Acknowledgements
The author thanks Prof. Martin Schlitzer, Department of Pharmacy, University of Marburg,
Germany for providing the series of articles which they published. This work is based on the 6
publications provided by him.

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5.0 References
1) Jochen Weisner et.al., Discovery of a Novel Lead Structure of Anti-malarials,
Bioorganic and Medicinal Chemistry 9 (2001) 785-792.J
2) Jochen Weisner et.al., StructureActivity Relationships of Novel Anti-Malarial Agents.
Part 2: Cinnamic Acid Derivatives, Bioorganic & Medicinal Chemistry Letters 11
(2001) 423-424
3) Jochen Weisner et.al., StructureActivity Relationships of Novel Anti-Malarial Agents.
Part

3:

N-(4-Acylamino-3-benzoylphenyl)-4-propoxycinnamic

Acid

Amides,

Bioorganic & Medicinal Chemistry Letters 12 (2002) 543545

4) Jochen Weisner et.al., StructureActivity Relationships of Novel Anti-Malarial Agents.
Part

4:

N-(3-Benzoyl-4-tolylacetylaminophenyl)-3-(5-aryl-2-furyl)acrylic

Acid

Amides, Bioorganic & Medicinal Chemistry Letters 12 (2002) 26812683

5) Jochen Weisner et.al., StructureActivity Relationships of Novel Anti-Malarial Agents:
Part 5. N-(4-acylamino-3-benzoylphenyl)-[5-(4-nitrophenyl)-2-furyl]acrylic Acid
Amides, Bioorganic & Medicinal Chemistry Letters 13 (2003) 361363
6) Jochen Weisner et.al., StructureActivity Relationships of Novel Anti-Malarial Agents.
Part

6:

N-(3-Benzoyl-4-tolylacetylaminophenyl)-3-(5-aryl-2-furyl)acrylic

Acid

Amides with Polar Moieties. Bioorganic & Medicinal Chemistry Letters 13(2003 )
21592161

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