Accepted Manuscript

SYNTHESIS, CHARACTERIZATION, THEORETICAL, ANTI-BACTERI-

AL AND MOLECULAR DOCKING STUDIES OF QUINOLINE BASED
CHALCONES AS A DNA GYRASE INHIBITOR

Muhammad Imran Abdullah, Asif Mahmood, Murtaza Madni, Sara Masood,

Muhammad Kashif

PII: S0045-2068(14)00018-2
DOI: [Link]
Reference: YBIOO 1707

To appear in: Bioorganic Chemistry

Received Date: 19 January 2014

Please cite this article as: M.I. Abdullah, A. Mahmood, M. Madni, S. Masood, M. Kashif, SYNTHESIS,
CHARACTERIZATION, THEORETICAL, ANTI-BACTERIAL AND MOLECULAR DOCKING STUDIES OF
QUINOLINE BASED CHALCONES AS A DNA GYRASE INHIBITOR, Bioorganic Chemistry (2014), doi: http://
[Link]/10.1016/[Link].2014.03.006

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SYNTHESIS, CHARACTERIZATION, THEORETICAL, ANTI-
BACTERIAL AND MOLECULAR DOCKING STUDIES OF
QUINOLINE BASED CHALCONES AS A DNA GYRASE
INHIBITOR.
Muhammad Imran Abdullah,a* Asif Mahmood,b Murtaza Madni,c Sara Masood,d

and Muhammad Kashifa

a
Institute of Chemistry, University of the Punjab, Lahore, Pakistan
b
Department of Chemistry, University of Sargodha, Sargodha, Pakistan
c
Department of Chemistry, Quaid-e-Azam University, Islamabad, Pakistan
d
Sheikh Zayed Medical College&Hospital Rahim Yar Khan Pakistan

*E-mail: [Link]@[Link]

Cell: +92-300-7476871

ABSTRACT: A series of fourteen (A1 – A14) new qunioline based chalcones were synthesized

by condensing 2,7-dichloro-8-methyl-3-formyl quinoline with acetophenone and

acetylthiophenes, and subsequently characterized by IR, NMR and Mass spectroscopy. All the

compounds were screened for antibacterial activities and found potentially active antibacterial

agents. Bioassay, theoretical and dockings studies with DNA gyrase (the enzyme required for

super coiling of DNA of bacteria) results showed that the type and positions of the substituents

seemed to be critical for their antibacterial activities. The bromo and chloro substituted chalcone

displayed high anti-bacterial activity. The A4 and A6 showed high interaction with DNA gyrase,

contributing high free binding energy (ΔG -8.18 and -8.88 Kcal).

KEYWORDS: Quinoline, chalcone, theoretical, acetophenone, anti-bacterial, DNA gyrase.

1
INTRODUCTION
Chalcones are α,β-unsaturated ketones, chalcones constitute an important class of natural

products that serve as the precursor for synthesis of various heterocyclic compounds like

pyrimidines, imidazoles [1], pyrazoles [2], 2-pyrazolines [3,4], and flavonoids [5]. Chalcones,

either natural or synthetic, are known to exhibit various biological activities such as anti-

inflammatory [6-7], antifungal [8-11], antioxidant [12-15], antimalarial [11-19], antituberculosis

[20], analgesic [21], anti-human immunodeficiency virus (HIV) [22], and antitumor activities

[23-24]. Some of them also act as anticancer [25], antiviral [26], and anti-AIDS agents [27].

Quinoline-based chalcones have been reported to possess antimalarial activity [28]. Chalcones as

well as some quinoline derivatives have already been recognized for their antimicrobial [29-30],

and antileishmanial activities [31-33].

Chalcones are considered the precursors of naturally-occurring compounds flavonoids

and isoflavonoids, but their true importance is extended in two branches: (i) The biological

activity associated with them (including anti-inflammatory, antimitotic, anti-leishmanial, anti-

invasive, anti-tuberculosis, anti-fungal, anti-malarial, anti-tumor), and (ii) anti-oxidant properties

[5,34-43].

2
Figure 1: Representation of different active part of newly synthesized drug against bacteria.

In the present study we have incorporated quinoline nucleus in the newly synthesized

anti-bacterial drugs which act as DNA gyrase inhibitor and efflux pumps inhibitor in bacteria

[44, 45]. To increase drug like properties of our compounds (A1 – A14) we have introduced

benzene and thiophene rings which contained high lipophilic groups like bromo, and chloro that

assist the drug molecule to enter into bacterial cell very easily. We have also changed the

position of substituent groups on benzene and thiophene rings in order to find out the substituent

effect on antimicrobial activities (Figure 1).

RESULTS AND DISCUSSION

The precursor 2,7-dichloro-8-methyl-3-formyl quinoline was prepared by the previously reported

method [46]. Synthesis of the title compounds (A1 – A14) were prepared through Claisen–

Schmidt reaction [36,42] by the condensation of formyl quinoline with commercially available

methyl aryl ketones in the presence of sodium hydroxide (Scheme 1 and Chart 1). All the newly

3
synthesized chalcones were in full agreement with the proposed structures. The E-configuration

was confirmed by the spectral data (IR, 1H-NMR and MS). Configuration of the double bond

was ascertained by coupling constants in 1H-NMR.

Scheme 1: Reaction protocol for the synthesis of chalcones (A1 – A14). (a) AcOH, H3PO4, reflux,
4-6 hrs, (b) POCl3, DMF, 80 °C, (c) Methyl Aryl Ketones, NaOH, rt, 2 hrs.

Chart 1: methyl Aryl (Ar) Moiety of Aromatic Ketones (A1 – A14).

4
ANTIBACTERIAL ACTIVITY: Antibacterial activities of the synthetic compounds were

tested against six bacterial species including Escherichia coli (Gram-negative), Klebsiella

aerogenes (Gram-negative), Salmonella typhimurium (Gram-negative), Staphylococcus aureus

(Gram-positive), Streptococcus pyogenes (Gram-positive) and Bacillus subtilis (Gram-positive)

by using method of Minimum Inhibitory Concentration (MIC). A commercial antibacterial

capsule Chloramphenicol and ciprofloxacin were used as a positive control for its comparative

study with all synthetic compounds

The bioactivity results indicate that substituted phenyl derivatives (A2 and A3) are less active

than A4 (IC50= 0.125mg/ml against Staphylococcus aureus, Staphylococcus aureus and

Escherichia coli, IC50=0.5mg/ml against Klebsiella aerogenes , Salmonella typhimurium and

Bacillus subtilis). The only difference between these molecules is the position of bromine on the

benzene ring. The activity is decreased considerably by the introduction of Br group at 3- and 2-

positions (A2 and A3, respectively) of phenyl ring whereas the activity is relatively increased on

moving the Br substituent to 4-position (A4) of phenyl ring. Conversely, replacing the bromo by

chloro (A6), and methoxy at 4-position of phenyl ring (A7) results in decrease activity. This may

be attributed to the greater hydrophobic effect of bromo group, than chloro, and methoxy at 4-

position.

The substituted hetero-aryl derivatives A8, A14 and A10 have shown promising antibacterial

activities against all the bacterial strains. Among the compounds, substituted thiophenyl

derivatives (A11, A12, A13,) exhibit almost equivalent antibacterial activities to that of the

standard (Table 1). Activity decreases considerably by the substitution of methyl on thiophene

ring (A11, A12, and A13). Incorporation of chlorine at 5-position of thiophene ring (A10) enhanced

the activity to a reasonable extent; it is further increased by the incorporation of another bromine

5
 atom at 5-position (A8). However, incorporation of bromo group at 2-positions (A9) decreases

activity. The incorporation of a second methyl group at 2-position (A11) have decreases the

activity than that of mono substituted one (A12, A13). In general, activity is enhanced by the

substitution of aromatic rings with electron withdrawing groups and is suppressed by the

incorporation of electron donating methyl groups. No systematic change has been observed in

antibacterial activities for the rest of the compounds. All the above results are also supported by

theoretically calculated LogP and bioassay studies.

Table 1: Anti-bacterial Activities of the Chalcones (A1 – A14) (IC50 mg/ml)

Compound Bacillus Staphylococ Streptococc Escherichia Klebsiella Salmonella

subtilis cus aureus us pyogenes coli aerogenes typhimurium
A-1 1.0 1.0 1.0 1.0 1.0 1.0
A-2 1.0 1.0 1.0 1.0 1.0 1.0
A-3 1.0 1.0 0.5 1.0 0.5 1.0
A-4 0.5 0.125 0.125 0.125 0.5 0.5
A-5 0.5 0.5 0.5 0.5 0.5 0.5
A-6 0.5 0.125 0.5 0.5 0.125 0.5
A-7 1.0 1.0 1.0 1.0 1.0 1.0
A-8 0.5 0.125 0.125 0.5 0.5 0.125
A-9 1.0 1.0 1.0 1.0 1.0 1.0
A-10 0.125 1.0 0.5 0.5 0.5 0.5
A-11 1.0 1.0 1.0 0.5 1.0 1.0
A-12 1.0 0.5 1.0 1.0 0.5 1.0
A-13 1.0 0.5 1.0 0.5 1.0 0.5
A-14 0.5 0.125 0.125 0.125 1.0 1.0
Chloramphenicol 1.0 1.0 1.0 1.0 1.0 1.0
Ciproflxoacin 1.0 0.5 1.0 0.5 1.0 1.0

DNA gyrase inhibitory assay

To understand the mechanism by which the quinoline based chalcones derivatives induced

the anti-bacterial activity, the inhibitory activity of the selected compound were performed

against the DNA gyrase. The DNA gyrase was isolated from [Link], the result were shown in

6
the Table 2. The compound A4 and A6 with strong anti-bacterial activities powerfully inhibited

the [Link] DNA gyrase with IC50 of 0.14 and 0.16ug/ml respectively. There was good

agreement between the docking experiment and DNA gyrase inhibitory MIC values, indicating

that the inhibition of the bacterial growth is due to the inhibition of DNA gyrase by quinoline

based chalcones. The inhibitory assay of DNA gyrase are good in agreement with docking

experiments.

Table 2: The inhibitory analysis of selected chalcones against DNA gyrase

Compounds [Link] DNA

gyrase IC50 µg/ml
A-4 0.14
A-6 0.16
A-7 5.15
A-8 1.01
A-14 0.95
Ciprofloxacin 0.22

Molecular descriptors-based SAR studies of Chalcones: From the data obtained by the

theoretical evaluation of the ADME properties, one can notice that hybrid compounds A1 – A14

do not break any point of the Lipinski’s rule of five, making them promising leads for drug

candidates [47]. LogP (Table 3) values are compatible with those described as a predictive

indicator of a drug’s capacity for membrane penetration [48]. The field template showed a good

similarity of the most active chalcone A4 with quinoline based drug ciproflaoxcin as shown in

Figure 2.

7
 Figure 2: Representing the similarity of A4 with ciprofloxacin calculated by Field Templater.

For the comprehension of three-dimensional microscopic interactions and binding between a

ligand and a receptor, a detail analysis in SAR is important in drug design and synthesis. A

number of chemical parameters are reported to be responsible for their molecular interactions.

Although many reports on the structure activity relationships based on the biological properties

of chalcones have been the subject of a large number of investigations [49, 50], but the present

study is the first report of its nature on the antibacterial activity of chalcones against different

bacterial strain types of molecular descriptors. Calculations were performed by software

package, Namely ACDlabs, in order to derive a quantitative relation between bactericidal

8
activity and structural properties. The values of the calculated the parameters are given in Table

3.

Molecular descriptors

(1) Octanol-water partition coefficient (log P)

(2) Bioconcentraion Factor(BCF)
(3) Polar surface Area (PSA)

Regarding a correlation of the steric parameters with activity, all chalcones of both set were

found to correlate directly with the activity. The octanol-water partition coefficient (log P) is

representative of steric interactions and in the present study it showed a good correlation with the

bactericidal action of chalcones. A direct correlation of the activity with log P was indicative of

the fact that chalcones with a higher log P are expected to be more active as is reflected in the

activity of halogenated chalcones (Table 1).

The bromo and chloro groups’ containing compounds (A4, A6, A9, and A10) are more active

ones in the series of 14 compounds (Table 3). Their LogP values (6.42, 6.24, 614, and 6.00,

respectively) are higher than the other compounds. As stated earlier in this discussion that the

position of substitutent groups also affects the activity of the compounds e.g A2 (2-bromo) is less

active (LogP = 5.83) than A4 (5-bromo) (LogP = 6.42. This difference is caused by steric

difference in both molecules. Experimental and theoretical data indicates that the halogen group

is somewhat more active against bacteria than methyl group. This difference can be attributed to

the hydrophobic property that bromo is more hydrophobic than chloro and in turn it has greater

binding energy to bind with biomolecule and inhibit their life cycle and growth.

9
Table 3: Calculated Molecular Discriptors

Compounds R (LogP) LogKoc LogBCF No. of [Link] H- PSA

(Koc) (BCF) H- acceptor
Donor
A-1 3,4-OCH3 5.72 4.1(30712.00) 4.1(13040.45) 0 4 48.40
A-2 2,-Br 5.83 4.5(35322.38) 4.2(15854.16) 0 2 29.94
A-3 3-Br 6.14 4.9(77083.59) 4.7(47165.41) 0 2 29.95
A-4 4-Br 6.42 4.6(44596.77) 4.9(74054.95) 0 2 29.96
A-5 2-Br, 4-OCH3 6.12 4.7(32737.98) 4.4(26168.79) 0 3 39.19
A-6 4-Cl 6.24 4.5(20229.20) 4.8(59355.09) 0 2 29.96
A-7 4-OCH3 5.65 4.4(28074.36) 4.1(11502.72) 0 3 39.19
A-8 3-Br 6.14 4.7(51823.04) 4.4(27084.16) 0 2 58.20
A-9 2-Br 5.38 4.3(20229.20) 3.9(7277.05) 0 2 58.20
A-10 5-Cl 6.00 4.6(43889.70) 4.3(21473.55) 0 2 58.20
A-11 2,5-CH3 5.64 4.7(48218.08) 4.4(24448.88) 0 2 29.09
A-12 4-CH3 5.89 4.4(24667.63) 4.0(9600.87) 0 2 29.08
A-13 3-CH3 5.54 4.4(24667.63) 4.0(9600.87) 0 2 58.20
A-14 1,4- 5.65 4.4(26078.77) 4.1(11505.26) 0 4 48.22
benzodioxane

Docking Studies: The docking experiments were performed using the bacterial DNA gyrase

structure with newly synthesized quinoline based chalcones (A1-A14) and with reference

compounds (Ciprofloxacin and Chloramphenicol). The DNA gyryase structure was obtained

from RCSB Portein data base (PDB ID 1Zi0). DNA gyrase is a unique type II topoisomerase

which is responsible for the surper-coiling activity of DNA in bacteria. We used the AutoDock to

determine the docked energy, the free energy of binding (ΔG) and the inhibition constant Ki

compared with experimental biological activity (Table 1 and Table 2). The calculated interaction

energies for (A1-A14) are good in agreement with experimental result which shows that A4, A6,

A8 and A14 are potent inhibitor of DNA gryase as compared to other chalcones and reference

standard (Table 4). The chain residue of DNA gryase to which the most potent inhibitors A4, A6,

A8 and A14 binds are LEU735, GLN788, ASP686, SER734, VAL737, VAL787, ARG739,

VAL685, LEU836 and VAL784 as shown in (Figure 3 and Figure 4).

10
Figure 3: Predicted binding conformations of chalcones A4 and A6 with catalytic portion of

DNA gyrase.

The close contact of quinoline ring of A4 and A6 with ILE736, VAL737 and VAL787 residue

(3.85, 3.69 and 3.62A0) suggested hydrophobic interaction. The nitrogen of quinoline make

interaction through hydrogen bonding with ASP686 and LEU735 residue (3.42 and 3.46A0

respectively). On the other hand –Cl of pyridine ring make interaction with VAL737 and

GLN788 residue (3.37 and 3.20 A0 respectively). The Br group on para position of benzene also

interact with VAl 784 (3.07) and the Cl group on para position of benzene interact with VAL784

and LEU836 (3.40 and 3,59 A0 respectively). These interactions stabilized the most potent

compound A4 and A6 in the active site of DNA gyrase as contributing favorable free binding

energy(ΔG -8.19 and -8.88 Kcal/mol respectively ).

11
 Figure 4: Docking model derived for A4 and A6 with the catalytic portion of DNA gyrase.

Figure 4 predict the 3D model of DNA gyrase complexed with compounds A4 and A6 occupies

the same space which shows a great affinity with DNA gyrase. The result of the docking

experiment support the postulation that compounds A4 and A6 may act as potent inhibitor of the

DNA gyrase. These theoretical results were also supported by the experimental data (Table 2).

Table 4: Summary of calculated binding parameters of chalcones (A1-A14) docked with

DNA gyrase

Compounds Free energy of Docking Ki (Inhibition

binding (ΔG) energy Constant) µM
Kcal/mol Kcal/mol
A-1 -7.09 -7.49 6.37
A-2 -6.77 -7.52 10.84
A-3 -7.04 -8.01 1.24
A-4 -8.19 -9.07 992.57
A-5 -7.31 -7.81 4.36
A-6 -8.88 -9.72 310.99
A-7 -7.22 -8.36 5.12
A-8 -8.05 -8.79 11.91
A-9 -6.62 -7.40 5.96
A-10 -7.75 -8.45 2.29
A-11 -7.09 -8.12 2.37
A-12 -7.56 -8.34 2.88

12
 A-13 -7.23 -8.08 2.79
A-14 -8.00 -8.88 30.98
Ciprofloxacin -6.33 -7.19 23.10
Chloramphenicol -4.20 -6.44 15.66

Spectral data: Spectral data (IR, 1H-NMR and MS) of all the newly synthesized compounds

were found in good agreement with the proposed structures. IR spectra of the compounds A1 –

A14 showed an absorption band at 1659 cm-1, typical of the stretching vibrations of chalcone

moiety. No peaks were found due to starting material aldehydic functionality as impurity. In the
1
H-NMR spectra of the chalcones two very sharp doublets were found around δ 7.61 – 7.63 ppm

for Hα and δ 8.17 – 8.13 ppm for Hβ with J-value 15 – 16 Hz for the trans chalcones. The

molecular ion observed in the mass spectra for all the chalcones confirmed their molecular

masses. The base peak, in the mass spectra of most of the chalcones, was obtained possibly by

the cleavage of Ar─Cl bond. While in bromo and chloro substituted chalcones the base peak is

due to the cleavage of two bonds i.e. CO-phenyl and phenyl─ Cl/Br bonds.

CONCLUSIONS AND OUTLOOK

It is concluded from the above discussion that the chalcones (A4, A5, A6, A8, and A10) are

comparatively more active than (A1, A2, A3, A7, A9, A11, A12, and A13). We have divided these

compounds into four categories for their antibacterial activities and represented in Table 1 i.e.

IC50 = 0.125mg/mL or below as significantly active, 0.5mg/mL as good active, 1.0mg/mL as

moderately active and >1mg/mL as low active. The compounds (A4, A5, A6, A8 A10 and A14) are

found potentially active antibacterial agents. The activity depends on the position of substituent,

the hydrophobic effect of bromo group is greater at 4-position of benzene and thiophene as

exhibited in A4 and A8. On the other hand the hydrophobic effect of bromo group decreases at 2-

13
position of benzene and thiophene as in A2, A3 and A9 respectively. This is also supported by

docking experiment, bioassay the calculated LogP values of studied Compounds. The docking

experiment and bioassay tell us that compounds A4 and A6 are the most potent inhibitor of DNA

gyrase.

Category Compounds
Significant A4, A6, A8
Good A5, A10 A14
Moderate A1, A2, A3, A7, A9
A11, A12, A13 and
A14

EXPERIMENTAL SECTION
General: Melting points were taken on Gallenkamp melting point apparatus. IR spectra were

recorded in KBr pellets on Perkin Elmer infrared spectrophotometer. 1H NMR spectra were

performed in CDCl3 on Brücker/XWIN NMR (300 MHz) and TMS was used as internal standard

(chemical shifts, δ in ppm) unless otherwise specified. Mass spectra were recorded on a Jeol MS

Route instrument. Thin layer chromatography (TLC) was performed with aluminium sheets -

Silica gel 60 F254 purchased from Merck. Purification of synthesized compounds was made by

recrystallization from appropriate solvents. Reagent grade chemicals such as phosphoryl

chloride, acetophenones, N,N-dimethylformamide (Sigma-Aldrich and Alfa Aesar) were used.

Bioassay conditions
Anti-bacterial activity
We have performed the anti-bacterial activity against Escherichia coli (Gram-negative),

Klebsiella aerogenes(Gram-negative), Salmonella typhimurium(Gram-negative), Staphylococcus

14
aureus (Gram-positive), Streptococcus pyogenes (Gram-positive) and Bacillus subtilis (Gram-

positive). We have used MH medium (Mueller-Hinton medium) which composed of 1.5g of beef

extract, 1.7g of starch and casein hydroxylate. All the stock solutions of new synthesized

chalcones were prepared in DMSO. The stock solutions of new synthesized chalcones were used

along with sterilized MH medium. The colorimetric method was used to determine the MIC

(minimum inhibitory concentration) values by using MMT dye.

Enzyme inhibition

We have been used the [Link] [51] method for the purification of [Link] DNA gyrase

enzyme form the crude extract of [Link]. The [Link] was grown in a medium which was

composed of 2g of yeast extract, 1.2g of (NH4)2SO4, 8g of Na2HPO4, 2g of K2H2PO4, 0.2g of

MgSO4, 4.0g of glucose per liter of the distilled water. According to [Link] [59] the

decatenation and supercoiling were performed.

Docking protocol

The docking studies were performed by using the autodock docking software. The structures

of the studied compounds were drawn using ChemBio Ultra 11.0, then energies of all the

compounds were minimized by using Gaussian 09 at B3LYP/6-31(d). The Gasteiger-Huckel

method was used to assign the charges to the ligand. The DNA gyryase structure was obtained

from RCSB Portein data base (PDB ID 1Zi0).In order to prepare the enzymes the hydrogens

atoms were added at a PH range of (6.5-8.1). By the aid of AutoDock tools the salvation

parameters and kollman united atom type chargers were added (Morris, Goodsell et al., 1998).

The autogrid program was used to generate the affibity (gird) maps of 20x20x20 A0 gird points

and 0.375 A0 spacing. (Morris, Goodsell et al., 1998).

15
Methods of Preparation of Precursors for Chalcones
N-acetylation of Substituted Aniline
0.1 mole of substituted aniline, 0.2 mole of glacial acetic acid was added in a 250ml flask. To

this mixture, catalytic amount of phosphoric acid was added and the mixture was refluxed for 5-6

hours. After the completion of the reaction (by TLC monitoring), the mixture was poured in ice-

cold water and stirred well. The crude product was precipitated out at once. These precipitates

were filtered and washed with ice-cold water. The pure product was obtained by recrystallized

from boiling water.

Synthesis of 2-chloro-3-formyl quinoline (Conventional Thermal Method)

Vilsmeier reagent was prepared by adding POCl3 (107.4g, 64.4ml, 0.70 mol) dropwise in DMF

(18.6g, 19.2ml, 0.25 mol) at 0 0C with constant stirring. To this solution acetanilide (0.10 mol)

was added and the mixture was stirred for 15mins at room temperature. Then this mixture was

stirred at 70-80 0C for 16 hrs. After the completion of reaction (TLC monitoring), the mixture

was poured in ice-cold water (500 ml) and stirred vigorously (30mins at 0 0C-10 0C). The 2-

chloro-3-quinolinecarbaldehyde was precipitated out, which was filtered off and washed with

water (200ml), dried and recrystallized from ethanol.

General Method for the Synthesis of quinolinyl Chalcones (A1-A14)

A mixture of formylquinoline (1mmol) and an aromatic ketone (1mmol) in methanol (50 mL)

was stirred at room temperature, followed by dropwise addition of aq. NaOH (4 mL, 10%). The

stirring was continued for 2 h and the reaction mixture was then kept at 0 °C for 24 h.

Subsequently, it was poured onto ice-cold water (200 mL). The precipitates were collected by

filtration, washed with cold water followed by cold MeOH. The resulting chalcones were

recrystallized from CHCl3 and dried in vacuo..

16
(E)-3-(2,7-dichloro-8-methylquinolin-3-yl)-1-(3,4-dimethoxyphenyl)prop-2-en-1-one (A1)

Yield, 85%. M.p. 180-182 °C. IR (KBr, cm-1): 1652 (C=O), 1596 (C=C), 1156 (C=N of

quinoline ring). 1H-NMR (CDCl3) δ: 8.403 ( 1H, s, Ar-H), 8.152 (1H, d, J=15.6Hz, CH of

olefinic bond), 7.698 (1H, dd, J1=2 Hz, J2= 8.4Hz, Ar-H), 7.63 (3H, m, CH of olefinic bond and

Ar-H), 7.564 ( IH ,m, Ar-H ), 6.939 ( 1H, d, J=8.4Hz, Ar-H), 3.972 (6H, s, OCH3), 2.818 (3H, s,

CH3); Calc Mass 402.27 MS, 402.17 [M + H]+ (Molecular ion Peak) 366.25 [M + H]+ -Cl.

(E)-1-(2-bromophenyl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A2)

Yield, 78%. M.p. 198-200 °C. IR (KBr, cm-1): 1659 (C=O), 1598 (C=C), 1152 (C=N of

quinoline ring). 1H-NMR (CDCl3) δ: 8.139 ( 1H, s, Ar-H), 8.057 (2H, d, J=7.6Hz, CH of olefinic

bond and Ar-H), 7.629 (2H, d, J=8.8Hz ,CH of olefinic bond and Ar-H), 7.526 (3H, m, Ar-H),

7.24 ( 1H, s, Ar-H), 2.821 (3H, s, CH3); Calc Mass 421.11 MS, 421.92 [M + H]+ (Molecular ion

Peak) 388.17 [M + 2 + H]+ -Cl.

(E)-1-(3-bromophenyl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A3)

Yield, 80% white Color solid. M.p. 172-175 °C. IR (KBr, cm-1): 1654 (C=O), 1591 (C=C), 1157
(C=N of quinoline ring). 1H-NMR (CDCl3) δ: 8.428 (1H, s, Ar-H ), 8.180 (2H, m, CH of olefinic
bond and Ar-H), 7.96 ( 1H, d, J =7.6Hz, Ar-H), 7.734 (1H, d, J =8Hz, Ar-H), 7.658 (1H, d, J
=8.4Hz, Ar-H), 7.551 ( 2H, m, CH of olefinic bond and Ar-H), 7.406 (IH, t, J =8Hz Ar-H),
2.816 (3H, s, CH3); Calc Mass 421.11 MS, 421.92 [M + H]+ (Molecular ion Peak) 388.17 [M +
2 + H]+ -Cl.
(E)-1-(4-bromophenyl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A4)

Yield, 84% white Color solid. M.p 185-188 °C. IR (KBr, cm-1): 1651 (C=O), 1599 (C=C), 1153
(C=N of quinoline ring). 1H-NMR (CDCl3) δ: 8.408 ( 1H, s, Ar-H), 8.181 (1H, d, J =16Hz, CH
of olefinic bond), 7.906 (2H, d, J =8.8 Hz, Ar-H), 7.653 (3H , m, CH of olefinic bond and Ar-H),

17
7.558 (1H, m, Ar-H), 7.519 (IH, s, Ar-H), 2.814 (3H, s, CH3); Calc Mass 421.11 MS, 421.92
[M + H]+ (Molecular ion Peak) 388.17 [M + 2 + H]+ -Cl.

(Z)-1-(2-bromo-4-methoxyphenyl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one
(A5)

Yield, 84% white Color solid. M.p. 190-192 °C. IR (KBr, cm-1): 1655 (C=O), 1593 (C=C), 1154
(C=N of quinoline ring). 1H-NMR (CDCl3) δ: 8.133 ( 1H, s, Ar-H), 8.048 (2H, d, J =8.8 Hz, CH
of olefin and Ar- H), 7.625 (1H, d, J =8.8 Hz, CH of olefin), 7.554 (1H, d, J =8.4Hz, Ar-H ),
7.240( 1H, s, Ar-H ), 6.965 (2H, d, J =8.8 Hz, Ar-H), 3.877 (3H, s, OCH3), 2.821 (3H, s,CH3 );
Calc Mass 451.14 MS, 451.08 [M + H]+ (Molecular ion Peak) 415.58 [M + H]+ -Cl.

(E)-1-(4-chlorophenyl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A6)

Yield, 75%. M.p. 168-170 °C. IR (KBr, cm-1): 1650 (C=O), 1590 (C=C), 1150 (C=N of
quinoline ring). 1H-NMR (CDCl3) δ: 8.410 ( 1H, s, Ar-H), 8.183 (1H, d, J =15.6Hz, CH of
olefin), 7.987 (2H, d, J =8.4 Hz, Ar-H), 7.645( 1H , d, J =8.8 Hz, Ar-H), 7.530(4H, m, CH of
olefin and Ar-H), 2.818 (3H, s, CH3); Calc Mass 376.66 MS, 376.00 [M + H]+ (Molecular ion
Peak) 340.00 [M + H]+ -Cl.

(E)-3-(2,7-dichloro-8-methylquinolin-3-yl)-1-(4-methoxyphenyl)prop-2-en-1-one (A7)

Yield, 70%. M.p 200-202 °C. IR (KBr, cm-1): 1662 (C=O), 1594 (C=C), 1160 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.402 ( 1H, s, Ar-H ), 8.153 (1H, d, J =16 Hz, CH of olefin), 8.065
(2H, d, J =1.6Hz, Ar-H), 7.629 (2H ,m, CH of olefin and Ar-H), 7.561 (1H, m, Ar-H), 6.995
(2H, d, J =8.4Hz, Ar-H), 3.893(3H, s, OCH3 ), 2.816 (3H, s, CH3); Calc Mass 372.14 MS, 372.08
[M + H]+ (Molecular ion Peak) 336.08 [M + H]+ -Cl.

(E)-1-(5-bromothiophen-2-yl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A8)

Yield, 56%. M.p. 162 °C. IR (KBr, cm-1): 1660 (C=O), 1592 (C=C), 1159 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.389 (1H, s, Ar-H), 8.209 (1H, d, J =15.6Hz, CH of olefin), 7.633
(2H, m, Ar-H), 7.56 (1H, d, J =8.8 Hz, CH of thiazole), 7.393 (1H, d, J =15.6Hz, CH of olefin),
7.171 (1H ,d, J =4Hz, CH of thiophene), 2.816 (3H, s, CH3); Calc Mass 427.14 MS, 428.00 [M
+ 4 + H]+ (Molecular ion Peak) 392.17 [M + 2 + H]+ -Cl.
18
(E)-1-(3-bromothiophen-2-yl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A9)

Yield, 60%. M.p. 170 °C. IR (KBr, cm-1): 1648 (C=O), 1593 (C=C), 1161 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.406 (1H, s, Ar-H ), 8.22 (1H, d, J =15.6Hz, CH of olefin), 7.831
(1H, d, J =15.6Hz, CH of olefin), 7.657(1H, d, J =8.8 Hz, Ar-H ), 7.575 (2H, m, CH of thiazole),
7.164 (1H, d, J =8.8 Hz, Ar-H), 2.814 (3H, s, CH3); Calc Mass 427.14 MS, 427.92 [M + H]+
(Molecular ion Peak) 392.08 [M + H]+ -Cl.

(E)-1-(5-chlorothiophen-3-yl)-3-(2,7-dichloro-8-methylquinolin-3-yl)prop-2-en-1-one (A10)

Yield, 53%. M.p. 148 °C. IR (KBr, cm-1): 1656 (C=O), 1594 (C=C), 1158 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.383 (1H, s, Ar-H ), 8.202 (1H, d, J =15.6Hz, CH of olefin), 7.747
(2H, m, Ar-H), 7.66(1H, m, CH of olefin), 7.425 (1H, d, J =10 Hz, CH of thiophene), 7.024 (1H,
d, J =4Hz, CH of thiophene), 2.812 (3H, s, CH3); Calc Mass 380.95 MS, 382.99 [M + H]+
(Molecular ion Peak) 347.49 [M + H]+ -Cl

(E)-3-(2,7-dichloro-8-methylquinolin-3-yl)-1-(2,5-dimethylthiophen-3-yl)prop-2-en-1-one
(A11)

Yield, 60%. M.p. 158 °C. IR (KBr, cm-1): 1658 (C=O), 1595 (C=C), 1157 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.353 (1H, s, Ar-H), 8.076 (1H, d, J =15.6Hz, CH of olefin), 7.627
(1H, d, J =8.8 Hz, Ar-H), 7.542 (1H, d, J =8.8 Hz, Ar-H ), 7.339 (1H, d, J =16Hz, CH of olefin),
7.096 (1H, s, CH of thiophene), 2.811 (3H, s,CH3), 2.721 (3H, s, CH3), 2.442 (3H, s, CH3); Calc
Mass 376.30 MS, 376.17 [M + H]+ (Molecular ion Peak) 340.17.08 [M + H]+ -Cl.

(E)-3-(2,7-dichloro-8-methylquinolin-3-yl)-1-(4-methylthiophen-2-yl)prop-2-en-1-one(A12)

Yield, 50%. M.p. 140 °C. IR (KBr, cm-1): 1654 (C=O), 1589 (C=C), 1149 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.395 (1H, s, Ar-H), 8.198 (1H, d, J =15.6Hz, CH of olefin), 7.707
(1H, s, CH of thiazole), 7.645 (1H, d, J =8.8 Hz, Ar-H), 7.553 (1H, d, J =8.8 Hz, Ar-H), 7.454
(1H, d, J =15.6 Hz, CH of olefin), 7.315 (1H, s, CH of thiophene), 2.814 (3H, s, CH3), 2.330
(3H, s, CH3); Calc Mass 362.27 MS, 362.17 [M + H]+ (Molecular ion Peak) 326.25 [M + H]+ -
Cl.

(E)-3-(2,7-dichloro-8-methylquinolin-3-yl)-1-(5-methylthiophen-2-yl)prop-2-en-1-one(A13)
19
Yield, 67%. M.p. 130 °C. IR (KBr, cm-1): 1654 (C=O), 1598 (C=C), 1160 (C=N of quinoline
ring). 1H-NMR (CDCl3) δ: 8.380 (1H, s, Ar-H), 8.172 (1H, d, J =15.6 Hz, CH of olefin), 7.715
(1H, d, J =4Hz, CH of thiazole), 7.637 (1H, d, J =8.8 Hz, Ar-H), 7.547(1H, d, J =8.8 Hz, Ar-H ),
7.436 (1H, d, J =15.6Hz, CH of olefin), 6.868 (1H, d, J =4Hz, CH of thiophene), 2.811 (3H, s,
CH3), 2.572 (3H, s, CH3); Calc Mass 362.27 MS, 362.11.08 [M + H]+ (Molecular ion Peak)
364.11 [M + H]+ -Cl.

(E)-3-(2,7-dichloro-8-methylquinolin-3-yl)-1-(2,3-dihydrobenzo[b][1,4]dioxin-5-yl)prop-2-
en-1-one (A14)

Yield, 45%. M.p. 158-160 °C. IR (KBr, cm-1): 1658 (C=O), 1595 (C=C), 1156 (C=N of
quinoline ring). 1H-NMR (CDCl3) δ: 8.391 (1H, s, Ar-H ), 8.147 (1H, d, J =15.6Hz, CH of
olefin), 7.606 (4H, m, CH of olefin and Ar-H ), 7.542 (1H,m, Ar-H ), 6.961 (1H, d, J =8.8 Hz,
Ar-H ), 4.319 (4H, dd, J1=5.2Hz, J2=13.2Hz, CH2 of acetal), 2.811 (3H, s, CH3); Calc Mass
400.25 MS, 400.17 [M + H]+ (Molecular ion Peak) 364.17 [M + H]+ -Cl.

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24
Graphical Abstract

25
 SYNTHESIS, CHARACTERIZATION, THEORETICAL,
ANTI-BACTERIAL STUDIES AND MOLECULAR DOCKING
OF QUINOLINE BASED CHALCONES AS A DNA GYRASE
INHIBITOR.

Highlights
 A series of quinoline based chalcones A1-A14 were synthesized.

 Their anti-bacterial activity by using method of Minimum Inhibitory

Concentration (MIC). Activities were evaluated.

 Tested compounds displayed potential anti-bacterial activity against all bacterial

strain

 Docking studies of chalcones A1-A14 with DNA gyrase were performed

 Better binding affinities were achieved with novel inhibitors than commercial ones.

26