Merged

“DESIGN, SYNTHESIS AND MOLECULAR
DOCKING OF SOME QUINOLINE DERIVATIVES

AS NLRP3 INHIBITORS ”
Final year B. Pharm Project submitted to

SAVITRIBAI PHULE PUNE UNIVERSITY, PUNE
By
Mr. Dinesh V. Patil and Mr. Shubham M. Patil
B. Pharm
Under the Guidance of

Dr. Vitthal V. Chopade
M. Pharm, PhD
Associate Professor
DEPARTMENT OF PHARMACEUTICAL
CHEMISTRY
Progressive Education Society’s Modern College of
Pharmacy, Nigdi, Pune (2021-2022)
(Accredited by NAAC and NBA)
SAVITRIBAI PHULE PUNE UNIVERSITY

DECLARATION
We are Mr. Dinesh V. Patil and Mr. Shubham M. Patil
declared that the thesis entitled “DESIGN, SYNTHESIS
AND MOLECULAR DOCKING OF SOME
QUINOLINE DERIVATIVES AS NLRP3
INHIBITORS ” submitted as per curriculum of Bachelor
of Pharmacy is the record of work carried out by us under
the guidance of Dr. Vithhal V.Chopade and has not
formed the basis for the award of any degree or diploma
or other titles in this or any other university.
Date: Mr. Shubham M. Patil,

Place: Nigdi, Pune Mr. Dinesh V. Patil
PES Modern College of Pharmacy
Nigdi, Pune-44
Dr. Vithhal V.Chopade
Associate Professor
Department of Chemistry
PES Modern College of Pharmacy
Nigdi, Pune-44
Certificate by the Guide
This is to certify that the thesis entitled, “Design,

Synthesis and Molecular Docking of some quinoline
derivatives as NLRP3 Inhibitors” submitted as per
curriculum of Bachelor of Pharmacy is the record of
research work carried out by Mr. Dinesh V. Patil and Mr.
Shubham M. Patil under my guidance and supervision and
that has not formed the basis for the award of any degree
or diploma or other titles in this or any other university.
Date: Dr. Vithhal V.Chopade
Place: Nigdi, PUNE M. Pharm, PhD.

PES’s Modern College of Pharmacy
Nigdi, Pune-44
Dr. P. D. Chaudhari
Principal
P.E. S Modern College of Pharmacy
Nigdi, Pune – 44
Ex-Dean, Faculty of Pharmaceutical Sciences,
Savitribai Phule Pune University, Pune.
Endorsement by the Principal
This is to certify that the partial research work for the

dissertation entitled “Design, Synthesis and Molecular
Docking of some quinoline derivatives as NLRP3
Inhibitors” is a record of bonafide research work carried
out by Mr. Dinesh V. Patil and Mr. Shubham M. Patil
under the guidance of Dr. Vithhal V.Chopade at P.E.S
Modern College of Pharmacy
Nigdi, Pune.
Date: Dr. P. D. Chaudhari

Place: Nigdi, PUNE Principal
ACKNOWLEDGEMENT
It is a moment of gratification and pride to look back with

a sense of contentment at the long travelled path, to be
able to recapture some of the fine moments, there are too
many people who helped lead both of us here and
supported us.
First of all, we take this opportunity to express our
heartfelt & deep sense of gratitude with sincerity to our
esteemed Research Guide Dr. V. V. Chopde, assistant
professor, department of pharmaceutical chemistry for
their constant help, support, encouragement and valuable
suggestions throughout project work. we are very much
thankful for their unflinching support, affection. which
helped us a lot in exploring our abilities. he supported us
at each and every step of the research work. I must say
thanks for his patience and courage to help us out
throughout the project and providing good environment
for the research work. And made it possible for us to carry
out the present work which otherwise, we would have
been very difficult. It was our luck to get an opportunity
to work under his guidance.
We express our sincere thanks to Dr. Praveen D.
Chaudhari, Principal, P.E. Society's Modern College of
Pharmacy, Nigdi, Pune, for making available every
facility needed for research work.
Our next work of gratitude is for Dr. SHAILAJA b.
JADHAV HOD of Department of Pharmaceutical
Chemistry for his valuable guidance throughout the
graduation journey helped me out to complete my
research work.
We would like to thank Mr. S. Y. Choudhari M. Pharm,
Assistant Professor Department of pharmaceutical
chemistry all teaching and non - teaching staff of P.E.
Society's Modern College of Pharmacy, Nigdi, Pune for
their help to complete our project work successfully.
We would like to express my profound feelings of
gratitude and affection to our parents for their everlasting
blessing, love, patience and support they gave us. This
dissertation is simply impossible without them.
We are grateful to our friends and all M.Pharm
chemistry students who helped me directly or indirectly
in the successful completion of project .
Nothing can resist a determined will and without intense
desire to succeed, we will be always thankful to the GOD
for giving us the strength, will and wisdom to carry out
the project successfully.
Mr. Dinesh V. Patil,

Mr. Shubham M. Patil
Index
Sr. Name Page No.

No.
1 Introduction 1-7
2 Literature survey 8-9
3 Rationale 10-13
4 Experimental Work 14-29
5 Result and Discussion 30-41
6 Conclusion 42
7 Future Prospect 43
8 Reference 42-45
List of Figures
Sr. Title Page.

No. No.
1 Schematic representation on docking 7
2 tetra quinoline derivative 10
3 Structure of MCC950 11
4 Structure of Tranilast
5 PDB ID-7ALV 17
6 3D structure of binding interaction of ligand 31
(MCC950)
7 2D structure of binding interaction of ligand 31
(MCC950)
8 3D structure of binding interaction of 32
ligand1 (acyl derivative)
ligand2 (Propionyl derivative)
ligand3 (Butyryl derivative)
List of Tables
Sr. No. Title Page.

No.
1 Target Protein 7ALV 16
2 Pharmacokinetic properties 35
(ADME) as shown in following
3 An anti-inflammatory study is 37
shown in the following
INTRODUCTION
1.INTRODUCTION
1.1 Inflammation: -
Inflammation can be detected when a wound swells up,
turns red, and hurts. Inflammation is the body's immune
system's reaction to an irritant in broad terms. The irritant
could be a bacterium, but it could also be a foreign item
in your finger, such as a splinter.
This suggests that an inflammation does not begin

solely when a wound has been infected with bacteria, is
gushing pus, or is not healing properly. It begins when the
body tries to defend itself against the toxic irritation.
Inflammation can be caused by a variety of factors. The
following are the most typical:
Pathogens (germs) like bacteria, viruses or fung
• External injuries like scrapes or damage through
foreign objects (for example a thorn in your
finger)
• Effects of chemicals or radiation
• General responses in the body: -
• If the inflammation is severe, the body may react
in a variety of ways. The following signs and
symptoms may be present:
• Blood changes, such as an increase in the number
of immune system cells.
Page | 1
Design, Synthesis and Molecular Docking of some quinoline
derivatives as NLRP3 Inhibitors
INTRODUCTION
1.2 Inflammasomes: -
Inflammasomes are receptors/sensors
in the innate immune system that regulate caspase-1
activation and inflammation in response to infectious
microorganisms and compounds produced from host
proteins. It's been linked to a variety of inflammatory
conditions. Recent advances have considerably improved
our understanding of the molecular pathways that activate
various inflammasomes. Furthermore, mounting evidence
in animal models, backed up by human data, clearly
suggests that the inflammasome is involved in the
initiation or advancement of diseases with major public
health implications, such as metabolic disorders and
neurodegenerative diseases. Finally, there have been
recent studies that indicate to prospective therapies that
target inflammasome activity in inflammatory illnesses.
These three areas of inflammasome research will be the
focus of this review.
1.3 NLRP3 Inflammasomes: -

The NLRP3 inflammasome,
which consists of NLRP3 (nucleotide-binding domain
leucine-rich repeat (NLR) and pyrin domain containing
receptor 3), ASC (apoptosis-associated speck-like protein
containing a caspase recruitment domain), and
Page | 2
INTRODUCTION
procaspase-1, is by far the best studied and characterised

inflammasome.
The NLRP3 inflammasome is activated by a wide range
of stimuli. When the NLRP3 protein is activated, it
recruits the adapter ASC protein, which then recruits the
procaspase-1, causing it to be cleaved and activated,
causing inflammatory cytokine maturation and secretion,
as well as pyroptosis. However, abnormal activation of
the NLRP3 inflammasome has been linked to a variety of
disorders, including diabetes, atherosclerosis, metabolic
syndrome, cardiovascular, and neurodegenerative
diseases, sparking a huge interest in finding NLRP3
inflammasome inhibitors.
Various inhibitors of the NLRP3 inflammasome

pathway have been discovered recently, and they have
been validated in vitro studies and in vivo trials in animal
models of NLRP3-related illnesses. Some of these
inhibitors target the NLRP3 protein directly, while others
target additional inflammasome components and
products. Directly targeting the NLRP3 protein may be a
superior option since it avoids off-target
immunosuppressive effects and so limits tissue death. The
many pharmacological inhibitors of the NLRP3
inflammasome will be discussed in this study, as well as
their mechanisms of action
Page | 3
INTRODUCTION
1.4 Quinoline-
Properties of Quinoline
Chemical formula C9H7N
Molar mass 12.16g/mol
Appearance Colorless oily liquid
Density 1.093g/ml
Melting point -15 degree Celsius
Boiling point 237 degree Celsius
Solubility in water Slightly soluble
Acidity (pKa) 4.85
Quinoline and its fused heterocyclic derivatives, which

have been evaluated for a variety of pharmacological
activities, are an important class of chemicals for the
creation of novel drugs. As a result, a number of scientists
have synthesised these molecules as target structures and
Page | 4
INTRODUCTION
tested their biological effects. The current study examines

the current state of knowledge on quinolines and their
biological activity, including anticancer,
antimycobacterial, antibacterial, anticonvulsant, anti-
inflammatory, and cardiovascular effect.
Recent studies found that 2-substituted 3 aryl quinoline
derivative, Pyrazole [4,3-C] quinolines derivative
tetrahydroquinolines, 8-hydroxy quinoline derivative
have some anti-inflammatory activity.
1.5 Drug design:-
The development of pharmacophore-based and molecular

docking and scoring techniques is known as drug design
(CADD). Docking is a technique for predicting the
binding orientation of small molecule therapeutic
candidates to their protein targets, as well as the small
molecule's affinity and activity. As a result, docking is
critical in rational drug design. Given the biological and
pharmacological importance of molecular docking,
significant attempts have been made to improve docking
prediction algorithms. The results are examined using a
statistical scoring procedure that calculates the interaction
energy and turns it into numerical values known as the
docking score. Different visualisation tools, like as Pymol
and Rasmol, can be used to see the 3D posture of the
bound ligand. This could aid in determining the optimum
Page | 5
INTRODUCTION
ligand match. Predicting the manner of protein ligand

interaction can help in protein annotation by assuming the
active site of the protein molecule. Furthermore,
molecular docking has a significant role in drug design
and discovery.
1.6 Molecular Docking:-
Molecular docking is one of the most widely used virtual

screening approaches, particularly when the target
protein's 3D structure is available. This technique may
predict the ligand–protein binding affinity as well as the
structure of the protein–ligand complex, which is helpful
information for lead optimization.
Protein-ligand Docking:
The purpose of protein-ligand docking is to find the
optimal binding between a small molecule (ligand) and a
protein. It is generally applied to the drug discovery and
development process with the aim of finding a potential
drug candidate. First, a target protein is identified. This
protein is usually linked to a disease and is known to bind
small molecules. Second, a ‘library’ of possible ligands is
assembled. Ligands are small molecules that bind to a
protein and may interfere with protein function. Each of
the compounds in the library is then ‘docked’ into the
protein to find the optimal binding position and energy.
Page | 6
INTRODUCTION
Fig 1.1: Schematic representation on docking
Page | 7
LITERATURE SURVEY
2.LITERATURE SURVEY:-
Author Title Year Structure
name
Zhen Dai et Development of 2021
al. novel
tetrahydroquinoline
inhibitors of
NLRP3
inflammasome for
potential treatment
of DSS-Induced
mouse colitis
Mohd Discovery of 2019

quinazoline-4(3H)-
abdulla et
ones as NLRP3
al.
Inflammasome
inhibitors:
computational
design, metal free
synthesis& in vitro
biological
evaluation
Page | 8
LITERATURE SURVEY
Xiaoli Chen et al. The 2017

antimalarial
chloroquine
suppresses
LPS-induced
NLRP3
inflammasome
activation &
confers
protection
against murine
endotoxic
shock
Page | 9
RATIONALE
3. RATIONALE:-
In present scenario quinoline nucleus has
emerged out as a potential pharmacophore for design &
development of novel derivatives. Literature studies
revealed that compounds bearing this heterocyclic
bioactive core have diverse pharmacological activities.
Zhen Dai et al. discovered that series of
tetrahydroquinoline inhibited the NLRP3 inflammasome.
The structure of drug is given below it contain the
quinoline moiety.
Fig 3.1: tetra quinoline derivative
Rebecca C. Coli et al discovered that The NLRP3 could

be involved in the pathogenesis of several inflammatory
and autoimmune diseases such as type 2 diabetes mellitus,
multiple sclerosis (MS), atherosclerosis, Alzheimer’s
disease (AD), cryopyrin-associated periodic syndrome
(CAPS), and infectious diseases. Therefore, the inhibition
Page | 10
RATIONALE
of NLRP3 can be a useful treatment option for

inflammatory diseases. In this regard, MCC950, as a
small molecule, is capable of inhibiting NLRP3 and,
following inhibition of NLRP3, production of interleukin-
1β (IL-1β) and IL-18 as pro-inflammatory cytokines
reduced. The structure contain the amide moiety.
(C=ONH)
Fig3.2: Structure of MCC950

Ali saeedi-boroujeni at al found that potential anti-
inflammatory & NLRP3 inflammasome inhibitor drug
inflammatory cytokinin signaling caused by SAR-CoV -2
can be mitigated following the administration of tranilast
& other therapeutic agents by targeting
NLRP3inflammasome, resulting in improvement of
patient. It contain the amide moiety, (C=ONH ) moiety
Page | 11
RATIONALE
Fig 3.3: Structure of Tranilast
David harrison et al. discovered that ester moiety act as a

highly permeable delivery vehicle & offer potential in
treatment of NLRP3 driven diseases, particularly when
tissue penetration is required.
We have designed the drug by combining the amide,

quinoline & ester moiety
Page | 12
RATIONALE
+ +
Page | 13
EXPERIMENTAL WORK
4. EXPERIMENTAL WORK
4.1 Design o of Molecule :-

1) Molecular docking study for anti-
inflammatory activity:
Docking is a method that predicts the preferred
orientation of one molecule to a second when
bound to each other to form a stable complex.
Docking studies were performed on the hits with
the help of AutoDock software version AutoDock
1.5.6.
The various steps involved in docking are as follows :

➢ Preparing the ligand:
The ligand structure opens in the " Marvin view
application ". Cleaned this structure as 2D and
3D. Saved the file in Mol2 format i.e. ligand.
mol2 AutoDock accepts the ligand structure in
either Mol2 format or PDB format. Using
AutoDock Tools the Mol2 format of the file was
converted to a PDBQT file, which was the actual
input to the docking procedure. Ligand flexibility
was achieved by choosing a root atom. This root
atom acts as the center of rotation during
coordinate transformation in the docking
simulation.
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EXPERIMENTAL WORK
➢ Preparing the Protein:

Before preparation of the protein, the most
appropriate protein site or the binding site was
selected. Two approaches are commonly taken
one is to select a single well-suited representative
protein structure to dock and the other is to use an
ensemble of representative structures, into which
each of the candidate ligands is docked, here the
second approach was used for our study. Once
the site was selected then the protein preparation
was started. There was one crystal structure of all
the target proteins that are downloaded from the
PDB ( www.pdb.org ). The PDB code 7ALV was
used for the crystal structures of NLRP3.
The crystal structures were selected
based on the crystal resolution. Resolution is the
measure of the quality of data that was collected
on the crystal containing the protein. When all the
proteins in a crystal are aligned identically, they
result in the formation of a very perfect crystal
and all of the proteins will scatter the X - rays the
next same way. On the other hand, the diffraction
pattern does not contain much information when
the proteins in the crystal are slightly different due
to local flexibility or motion. Hence, the
resolution is considered as a level of detail present
in the diffraction pattern and the level of detail
Page | 15
EXPERIMENTAL WORK
that would be seen in an electron density map

calculation.
The crystal structures with the
selected PDB codes were examined to check for
the presence of mutations and the complete side
chains. The PDB codes for the current molecular
docking study, the structures were observed for
the correct three-dimensional conformations. The
structures were also filtered based on the presence
of ligands, crystalline waters, and cofactors.
Table4.1.1: Target Protein 7ALV

7ALV
PDB ID 7ALV
Name
Crystal Structure of NLRP3 NACHT domain in
complex with a potent inhibitor
Method
X-RAY Diffraction
Resolution
2.83
R-Value Free
0.265
R-Value Work
0.213
Page | 16
EXPERIMENTAL WORK
R-Value Observed
0.215
Fig 4.1.1: (PDB ID-7ALV) Protein structure of

7ALV was imported from the protein data bank
from the web.
The protein i.e. 3ALV opened in
Discovery Studio ( Figure). After that, by pressing
CTRL + H, the side panel showed a component of
the PDB structure . From the side, panel removed
water molecules, hetero atoms, and ligand groups.
This was done by clicking on " water ", " ligand
group", and pressing the delete button
respectively. The final structure file PDB with the
same 7ALV_edited.pdb.
Page | 17
EXPERIMENTAL WORK
In the next step, 7ALV_edited.pdb

opened in AutoDock Ca Tools. The Polar
hydrogen atom is added to the molecule. This file
saves as 7ALV_H.pdb.
➢ Preparation of grid: The receptor grid was

generated for the active site amino acids of the
protein. The active site of a protein was embedded
in a 3D grid. The grid preparation for protein. The
protein file 7ALV_H.pdb open in AutoDock. The
various atoms were added. After adding the
atoms, entered the XYZ coordinates obtained
from the PDB file in the boxes marked X, Y, and
Z centers respectively. If more than one co -
ordinates were found took the mean of the
coordinates. The 3D grid is shown in the center of
the protein structure with X, Y, and Z dimensions
of 20.89, - 35.1625, and 17.136 respectively.
AutoGrid pre - calculates the grid maps for each
atom type present in the docked ligand. In the "
number of points in x, y, and z dimensions ", slide
to 60, 60, and 60 respectively. The grid file saves
as output 7ALV.gpf.
➢ Preparation of docking: After preparing the

protein and ligand files, performing molecular
docking is the next step. The docking procedure
using AutoDock requires the following files as
input:
Page | 18
EXPERIMENTAL WORK
1) A grid map file, that is generated by

AutoGrid ,
2) A PDBQT file for the ligand
3) A docking parameter file that specifies the
files and parameters for the docking
calculation. ADT is used to generate the
docking parameter file.
4) The final docked coordinates were written
by AutoDock in the docking log file
(DLG). The docking protocol was set as
follows.
a) Redocking as a Validation Method of Docking: -
Redocking was performed as a validation method for
protein ligand complexes . The 3D structures of the crystal
ligands were obtained by removing the crystal ligand from
protein crystallographic complexes . The crystal ligands
were then redocked with all the proteins using AutoDock.
b) Evaluating the Results of Docking :-
At the end of a docking simulation , the clustering
information and the internal energies were written by
AutoDock into a DLG file . AutoDock performed the
cluster analysis of the different docked conformations .
The number of runs was set based on the structure of
ligand and protein . Initially AutoDock was run using the
default parameters for docking . After analyzing the
results , if the docked complexes did not yield binding
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EXPERIMENTAL WORK
poses with lowest binding affinities and active site amino

acid interactions , the parameters were modified as
follows . Some protein - ligand complexes yielded lowest
binding energy conformations within 10 runs while some
complexes too
k 50-100 runs. For example , active compound when
docked with NLRP3 produce the conformations in ten
runs . Likewise , it yields the lowest binding energy
conformations until 10 docking runs . Sometimes the
docking runs the complex started generating positive
binding energies . Since positive binding energies are the
measure of false results the docking was stopped at 10
runs . Thus , the sufficient sampling required for
molecular docking was provided based upon the trial
docking experiments conducted . The sufficient sampling
depends on the ligand structure and the number of torsions
in the ligand structure . The minimum energy found in
each run was reported as a histogram from the DLG file ,
which also includes a table of RMSD values within each
cluster . The results were analyzed using ADT and Visual
Molecular Dynamics ( VMD ) . The similar structure
ligands resulted in less difference in their binding
affinities .
c) Analyzing AutoDock Results :
The DLG file is opened and observed with the help of
ADT . The interactions between the docked conformation
of the ligand and the receptor were studied during this step
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EXPERIMENTAL WORK
. By default , ADT showed the ligand as a ball and stick

surrounded by a molecular surface . The colored surface
distinguishes the regions that were in contact with the
receptor from gray colored regions that were not in
contact . Portions of the receptor that were in contact with
the ligand were shown with the ball and stick. Hydrogen
bonds were shown as a string of small spheres.
d) Checked the conformation Info window to see if
ligand was interacting with the receptor and the
amino acids involved in the interaction . If the
amino acids are same as that of the LigPlot /
Active Site , the RUN is successful . If there is no
interaction or the interacting amino acid is
different selected the second RUN with minimum
bending energy Continued this step until the
appropriate RUN was obtained.
During this step , the conformations were
studied to check binding affinity interactions of
the ligand with proteins were identified and
grouped into the result data.
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EXPERIMENTAL WORK
Table4.1.2. : Softwares and websites required:
S Software/
r. Logo server Links
N
o.
1. PASSonline http://www.way2drug.com/passo
nline/
2. PubChem https://pubchem.ncbi.nlm.nih.gov
/
3. RCSB https://rcsb.org/
PDB
4. Swiss http://www.swissadme.ch/
ADME
5. Marvin https://chemaxon.com/products/
Sketch marvin
6. https://chemistrydocs.com/chemd
Chemdraw raw-pro-8-0/
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EXPERIMENTAL WORK
7. Biovia https://discover.3ds.com/discover
Discovery y-studio-visualizer-download
Studio
8. AutoDock https://autodock.scripps.edu/
Page | 23
EXPERIMENTAL WORK
4.2 General scheme of synthesis:-
Step 1: Synthesis of 4-methyl acetanilide
Step 2: Synthesis of 2-chloro 3-formyl 6-methyl quinoline
Page | 24
EXPERIMENTAL WORK
Step3: Synthesis-chloroquinoline-3-carbaldehyde-6-
methyl phenyl hydrazone
Step 4: a) Synthesis of acyl derivative
Page | 25
EXPERIMENTAL WORK
Step 4: b) Synthesis of propionyl derivative
Page | 26
EXPERIMENTAL WORK
Step 4: c) Synthesis of butyryl derivative
4.3 Procedure of synthesis:-
Step 1: acetic anhydride (10 mL,0.1 mol) & glacial acetic

acid (10 ml, 0.175 mol) were added to substituted anilines
(0.1 mol), heated gently under reflux for 1 hr, & poured
into ice water (200ml), where crystals of acetylated
product were formed. The crystals were filtered &
recrystallized with acetic acid: water (1:2), producing
acetanilide in yields of between 94-100% Heating hr- 5-
6hr
Step 2: Place dimethyl formamide (DMF)(3 eq) in a flask
equipped with a drying tube cooled to 0C temperature,
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EXPERIMENTAL WORK
then POCl3(Phosphorous oxychloride) (7 eq) was added

drop wise with stirring to it. To this solution add
acetanilide (1 mmol). After few minutes the reaction
mixture was refluxed for 6-8 h. After completion of
requiring time reaction, the mixture was cooled and
poured in ice-Coldwater and stirred about half an hour
then filtered to offer powder of compound.
(Heating Time 12hr)
Step 3: Method A (With MeOH as solvent): A solution of
100 mg (0.520 mmol) of 2-chloro-3-formyl-6-methyl-
quinoline 2. and 75.5 mg (0.520 mmol) of phenyl
hydrazine hydrochloride 3.in 5 ml of dried MeOH was
stirred at room temperature. The product precipitated
gradually, and the reaction was monitored by TLC for the
disappearance of the reactants, which took about 15 h.
After the reaction is completed, the solution was diluted
with 5 ml of ice-cold water with stirring. The precipitated
yellow solid was collected by filtration, washed with
water and recrystallized from 20 ml of MeOH to get 133
mg (0.471 mmol, 90%)
(Heating time- 15hr)
Step 4: (a) Method-Prepare a solution of -hydroxy
derivative (2 mmol), 4-dimethylaminopyridine (1 mmol)
and acetic anhydride (20 mmol) in chloroform (5 mL).
Stir the solution. Reflux the solution for 3 hours until the
starting material is fully consumed (TLC). Remove the
solvent under low pressure.
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EXPERIMENTAL WORK
Purify the product using chromatography column with

eluent system.
(Heating time- 4 hr)
Step 4: (b) A stirred suspension of hydroxy derivative
(0.70 g; 2.79 mmol) or equivalent, triethylamine (0.4 ml;
2.79 mmol) and a propionyl chloride (0.2 ml; 2.79 mmol)
in dry 1, 4-dioxan (40 ml) was heated to reflux, with the
exclusion of moisture (CaCl2), for 2 h and cooled to room
temperature. The reaction mixture was then poured onto
water (250 ml) and the solid filtered off.
(Heating time -2 hr)
Step 4: (c) A stirred suspension of hydroxy derivative
(0.70 g; 2.79 mmol) or equivalent, triethylamine (0.4 ml;
2.79 mmol) and butyryl chloride (0.2 ml; 2.79 mmol) in
dry 1,4-dioxan (40 ml) was heated to reflux, with the
exclusion of moisture (CaCl2), for 2 h and cooled to room
temperature. The reaction mixture was then poured onto
water (250 ml) and the solid filtered off. (Heating time-
2hr).
Page | 29
RESULT AND DISCUSSION
5.RESULT AND DISCUSSION

5.1 Results of Molecular docking studies:-
Molecular docking has become a standard tool in

computational chemistry for predicting the binding
orientation of small molecule drug candidates with their
protein targets in order to predict the affinity and activity
of the small molecule. Thus , molecule docking plays an
important role in the rational design of drugs .
The search for the best ways is to fit ligand
molecules into protein structure using AUTODOCK
VINA resulted in docking files that contained detailed
records of docking. The obtained log files were read in
ADT ( Auto Dock Tool ) to analyze the results of docking-
Binding affinities that are reported.
The designed Quinoline derivatives are docked well into
the active site of the target protein (PDB ID : 7ALV) using
autodock software. All the designed compound shows
appropriate binding to the target protein. Almost all the
compunds were active with minimum binding affinity are
selected as potent inhibitors of NLPRP3. Standard drug
(MCC950) shows more binding affinity to the selected
protein.
Page | 30
Fig 5.1.1: 3D structure of binding interaction of

ligand(MCC950)
Fig 5.1.2: 2D structure of binding interaction of ligand

(MCC950)
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Page | 34
5.2 Result of ADME Prediction of hit compound

Table 5.2.1: Pharmacokinetic properties (ADME) as
shown in following
Standard Acyl Propionyl Butyryl
Drug derivative derivative derivative
(MCC950)
Molecular
weight 426.46 g/mol 381.81 g/mol 395.84 g/mol 409.87 g/mol
BBB permeant No No No No
CYP2C19 No Yes Yes Yes

inhibitor
CYP2C9 No Yes Yes Yes
inhibitor
CYP2D6 Yes No No No
inhibitor
CYP3A4 Yes No Yes Yes
inhibitor
Bioavailability 0.55 0.55 0.55 0.55
Score
GI absorption High High High High
ESOL Solubility 1.06e-02 5.00e-03 2.59e-03 1.57e-03

(mg/ml) mg/ml mg/ml mg/ml mg/ml
Page | 35
• Pharmacokinetic properties (ADMET) were

predicted using the Swiss ADMET tool.
• Swiss ADME tool were used to calculate the
permeability of the blood-brain barrier (BBB).
The importance of BBB permeation is in affecting
the CNS When drug is administered during
treatment. All compounds showed non-
permeation to the BBB & hence there are no
chances CNS side effects
• For Gastrointestinal Absorption Standard drug
(MCC950) & Acyl, Butyryl, Propionyl, derivative
show higher absorption.
• Acyl, propionyl, and butyryl shows CYP2C19 and
CYP2C9 Inhibitor screening.
• Propionyl and butyryl show CYP2CA4 Inhibitor
screening except Acyl.
• Hence, all designed compounds interact with
cytochrome either as the substrate or as inhibitors.
Page | 36
5.3 Result of Pass prediction:-
Table 5.3.1 An anti-inflammatory study is shown in

the following
PASS Prediction
Compound Activity
Pa Pi
Standard 0,092 0,092 Leukotriene synthesis inhibitor

(MCC950)
0,081 0,080 Anti-inflammatory
Acyl 0,259 0,028 Interleukin agonist

derivative
Propionyl 0,229 0,042 Interleukin agonist

derivative
Butyryl 0,254 0,030 Interleukin agonist

derivative
Page | 37
Anti-inflammatory predictions:-
 The anti-inflammatory activity, Interleukin

agonist, and Leukotriene synthesis inhibitor
results of the PASS prediction suggested that
three-hit compounds, including standard
(MCC950) compound were classified as active
compounds.
 PASS ONLINE software was used to identify
compounds likely to predict anti-inflammatory
activity.
 Three compounds i.e. Acyl, Butyryl, Propionyl
derivative, showed higher anti-inflammatory
activity than Standard drug (MCC950).
Page | 38
5.4 Result of synthesized compound:-
Characteristics of synthesized compounds
Compound 1- (acyl derivative)
Name:-
4-{N-[(E)-(2-chloro-6methylquinoline-
3yl)methylidene]hydrazinecarbonyl}phenylacetate
Molecular weight- 381.82
Appearance – yellow
%Yield- 80%
Melting point - 275-280°C
Solubility- Soluble in alcohol.
RF factor- 0.69
Page | 39
Compound 2 : Propionyl derivative
Name:-
4-{N'-[(E)-(2-chloro-6-methylquinolin-3-
yl)methylidene]hydrazinecarbonyl}phenyl propanoate
Molecular weight- 395.84
Appearance – brownish yellow
% Yield- 70%
Melting point- 280-285°C
Solubility- Soluble in alcohol.
RF factor- 0.38
Page | 40
Compound 3: butyryl derivative
Name:-
4-{N'-[(E)-(2-chloro-6-methylquinolin-3-yl)
methylidene] hydrazine carbonyl} phenyl butanoate
Molecular weight:- 409.87
Appearance:- pitch yellow
% Yield- 70%
Melting point:- 290-300°C
Solubility:- Soluble in alcohol.
RF factor:- 0.25
Page | 41
CONCLUSION
6. CONCLUSION:-
A series of novel quinoline derivatives

containing benzyl were designed and it’s some in silicon
parameters was studied. After comparing the silico study
of designed compound with related literature of similar
molecule, our designed compounds could be potent
NLRP3 inhibitor. From the PASSonline server, it was
predicted that the derivatives have minimum adverse
reaction with NLPR3 inhibitor activity. The designed
compounds violate zero rule from the 'rule of 5' hence it
can be concluded that these derivatives could be orally
active. The ADME study of these compounds reveals that
they are suitable for drug-likeness. These derivatives have
good PPB and GI absorption properties. Also, all
compounds have low cytotoxicity as they do not cross
BBB. All the derivatives show metabolism either by CYP
substrate or inhibitor. The total clearance of all derivatives
was found to be minimum. From docking study, the
derivatives are well docked with lowest minimum binding
energy to the protein. Overall, the studies reveal that
quinoline derivatives show potent inhibitors against
NLRP3 as an anti-inflammatory agent.
Page | 42
FUTURE PROSPECT
7. FUTURE PROSPECT:-
To access the potential threats of
quinoline derivatives, the safety data from sufficient
powered clinical trials are needed; unfortunately, so far,
available data in humans are sparse. Further structure-
activity relationship study is necessary to optimize these
new lead molecules. This will lead is to the development
of novel potential anti-inflammatory agents as NLRP3
inhibitor and will help to eradicate the global burden of
adverse drug.
Moreover, animals will also have to be performed
to evaluate the anti-inflammatory activity. Also, it is
necessary to determine the advantage of these substituted
product-derived drug over traditional therapeutics by
comparing their physicochemical and physiological
properties, cardio protective properties and side effects.
After the overall study, the future prospect is as;
➢ Synthesis of possible derivatives.
➢ Biological evaluation of synthesized compounds.
➢ Looking into the result of present research done so
far, many further ideas can be explored in the
future.
➢ Pharmacophore modelling and QSAR studies.
➢ The compound can be further screened for
immunoassay.
➢ The other alternative short and precise synthetic
route can be worked out to synthesized these
scaffold.
Page | 43
REFERENCES
8. REFERENCES:-
1. Kumar, S., Bawa, S. and Gupta, H., 2009.

Biological activities of quinoline derivatives. Mini
reviews in medicinal chemistry, 9(14), pp.1648-
1654.
2. Wang, X., Shi, J., Li, Z., Li, L., Zhang, R., Bai, Y.,
Li, J., Liang, F. and Tang, Y., 2021. An 8-
Hydroxy-Quinoline Derivative Protects Against
Lipopolysaccharide-Induced Lethality in
Endotoxemia by Inhibiting HMGB1-Mediated
Caspase-11 Signaling. Frontiers in pharmacology,
12, p.1150.
3. Yang, C.Y., Hung, Y.L., Tang, K.W., Wang, S.C.,
Tseng, C.H., Tzeng, C.C., Liu, P.L., Li, C.Y. and
Chen, Y.L., 2019. Discovery of 2-substituted 3-
arylquinoline derivatives as potential anti-
inflammatory agents through inhibition of LPS-
induced inflammatory responses in macrophages.
Molecules, 24(6), p.1162.
4. Tseng, C.H., Tung, C.W., Peng, S.I., Chen, Y.L.,
Tzeng, C.C. and Cheng, C.M., 2018. Discovery of
pyrazolo [4, 3-c] quinolines derivatives as
potential anti-inflammatory agents through
inhibiting of NO production. Molecules, 23(5),
p.1036.
Page | 44
REFERENCES
5. Dai, Z., Chen, X.Y., An, L.Y., Li, C.C., Zhao, N.,
Yang, F., You, S.T., Hou, C.Z., Li, K., Jiang, C.
and You, Q.D., 2020. Development of novel
tetrahydroquinoline inhibitors of NLRP3
inflammasome for potential treatment of DSS-
induced mouse colitis. Journal of Medicinal
Chemistry, 64(1), pp.871-889.
6. Saeedi-Boroujeni, A. Mahmoudian-Sani, M.R.
Nashibi, R. Houshmandfar, S. Tahmaseby
Gandomkari, S. and Khodadadi A., 2021.
Tranilast: a potential anti-Inflammatory and
NLRP3 inflammasome inhibitor drug for COVID-
19. Immunopharmacology and
Immunotoxicology, 43(3), pp.247-258.
7. Chen, X., Wang, N., Zhu, Y., Lu, Y., Liu, X. and
Zheng, J., 2017. The antimalarial chloroquine
suppresses LPS-induced NLRP3 inflammasome
activation and confers protection against murine
endotoxic shock. Mediators of Inflammation,
2017, pp.248-257
Page | 45
P. E. Society’s
Modern College of Pharmacy, Yamunanagar,
Nigdi, Pune-44.
Final Year B-Pharmacy
Academic Year: 2021-22
--------------------------------------------------------------------------
EVALUATION OF DISSERTATION BOOK
Thesis Title: Design, Synthesis and Molecular Docking
of some quinoline derivatives as NLRP3 Inhibitors
Name of the Student: Mr. Dinesh V. Patil
Mr.Shubham M. Patil
Roll Number: 446, 449
Evaluation Table:
Objective(s) Methodology Results Conclusions Total

of the work adopted and and ( 75 )
done (20) Discussions Outcomes
(15)
(20) (20)
Name and Signature of project guide

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“DESIGN, SYNTHESIS AND MOLECULAR

DOCKING OF SOME QUINOLINE DERIVATIVES

Final year B. Pharm Project submitted to

Under the Guidance of

SAVITRIBAI PHULE PUNE UNIVERSITY

Date: Mr. Shubham M. Patil,

Certificate by the Guide

This is to certify that the thesis entitled, “Design,

Date: Dr. Vithhal V.Chopade

Place: Nigdi, PUNE M. Pharm, PhD.

Endorsement by the Principal

This is to certify that the partial research work for the

Date: Dr. P. D. Chaudhari

It is a moment of gratification and pride to look back with

Mr. Dinesh V. Patil,

Sr. Name Page No.

Sr. Title Page.

Sr. No. Title Page.

This suggests that an inflammation does not begin

1.3 NLRP3 Inflammasomes: -

procaspase-1, is by far the best studied and characterised

Various inhibitors of the NLRP3 inflammasome

Quinoline and its fused heterocyclic derivatives, which

tested their biological effects. The current study examines

1.5 Drug design:-

The development of pharmacophore-based and molecular

ligand match. Predicting the manner of protein ligand

1.6 Molecular Docking:-

Molecular docking is one of the most widely used virtual

Fig 1.1: Schematic representation on docking

Mohd Discovery of 2019

Xiaoli Chen et al. The 2017

Fig 3.1: tetra quinoline derivative

Rebecca C. Coli et al discovered that The NLRP3 could

of NLRP3 can be a useful treatment option for

Fig3.2: Structure of MCC950

Fig 3.3: Structure of Tranilast

David harrison et al. discovered that ester moiety act as a

We have designed the drug by combining the amide,

4.1 Design o of Molecule :-

The various steps involved in docking are as follows :

➢ Preparing the Protein:

that would be seen in an electron density map

Table4.1.1: Target Protein 7ALV

Fig 4.1.1: (PDB ID-7ALV) Protein structure of

In the next step, 7ALV_edited.pdb

➢ Preparation of grid: The receptor grid was

➢ Preparation of docking: After preparing the

1) A grid map file, that is generated by

poses with lowest binding affinities and active site amino

. By default , ADT showed the ligand as a ball and stick

Table4.1.2. : Softwares and websites required:

4.2 General scheme of synthesis:-

Step 1: Synthesis of 4-methyl acetanilide

Step 2: Synthesis of 2-chloro 3-formyl 6-methyl quinoline

Step 4: a) Synthesis of acyl derivative

Step 4: b) Synthesis of propionyl derivative

Step 4: c) Synthesis of butyryl derivative

4.3 Procedure of synthesis:-

Step 1: acetic anhydride (10 mL,0.1 mol) & glacial acetic

then POCl3(Phosphorous oxychloride) (7 eq) was added

Purify the product using chromatography column with

5.RESULT AND DISCUSSION