Structural Chemistry

https://doi.org/10.1007/s11224-020-01512-0

ORIGINAL RESEARCH

Synthesis, antibacterial evaluation and molecular docking studies

of novel series of acridone- 1,2,3-triazole derivatives
Mohammed Aarjane 1 & Siham Slassi 1 & Bouchra Tazi 2 & Mohamed Maouloua 3 & Amina Amine 1

Received: 29 May 2019 / Accepted: 21 February 2020

# Springer Science+Business Media, LLC, part of Springer Nature 2020

Abstract
Development of new drugs with antibacterial potency is an important solution to overcome drug-resistance problems.
In the goal to develop novel structure with antibacterial potency, we designed and synthesized novel acridone
derivatives bearing triazole nucleus. The novel synthesized compounds were tested for their in vitro antibacterial
activity against four bacterial human pathogenic strains. The compound 4h displayed significant antibacterial activ-
ities against Staphylcoccus aureus (MRSA) with MIC = 19.6 μg/mL. The synthesized compounds were subjected for
docking study to understand the interaction of our compounds and the dihydropteroate synthase (DHPS) in
methicillin-resistant Staphylcoccus aureus (MRSA).

Keywords Synthesis . Acridone . Triazole . Antibacterial activity . Docking

Introduction attractive for synthesis since they possess diverse pharma-

cological properties such as antifungal [4], antibacterial
Antibiotic resistance is currently considered as major [5], anti-malarial [6], antiviral [7], antitumoral [8], and
public health problem [1]. Therefore, the development anti-inflammatory [9] activities. Moreover, the synthesis
of new compounds with antimicrobial activity is impor- of 1,2,3-triazole via click chemistry by 1,3-dipolar cyclo-
tant errands of this century. One of the researched ther- addition of substituted azides and alkynes in the presence
apeutic targets in the field of antibacterial discovery is of Cu(I) gives regioselective 1,4-disubstituted 1,2,3-tri-
dihydrofolate reductase (DHFR), an essential enzyme azole with good yields.
whose role is to regenerate folic acid into its reduced On the other hand, acridone and acridine derivatives are
form tetrahydrofolate; this enzyme represents an attrac- known for many years for their biological activities such as
tive antibacterial target [2]. antitumor [10], antiviral [11], anti-inflammatory [12], antima-
Triazole is an important heterocyclic skeleton with larial [13], and antimicrobial bioactivity [14] . Numerous
large biological activity. 1,2,3-triazoles are important class acridone compounds that have antibacterial activity have been
in the triazoles series [3]; these compounds continue to be developed, including some heterocyclic such as triazole, thia-
zole, and 1,3,4-oxadiazole nucleus [15–17] . Several methods
Electronic supplementary material The online version of this article are reported for the preparation of the acridone ring, among
(https://doi.org/10.1007/s11224-020-01512-0) contains supplementary them are Ullmann reaction which was considered as the most
material, which is available to authorized users.
efficient [18]; it consist the condensation of o-halobenzoic
acid with aromatic amine in the presence of Cu followed by
* Mohammed Aarjane
aarjane.mohammed@gmail.com intramolecular cyclization using sulfuric acid.
In a view of the significant bio-potential of 1,2,3-triazole
1 and acridone nucleus and in order to develop novel bioactive
LCBAE, Faculty of Science, University Moulay Ismail, Zitoune,
11201 Meknes, Morocco therapeutic agents, we focused our work on the preparation
2 and evaluation of novel acridone bearing 1,2,3-triazole nucle-
Department of Basic Sciences, National School of Agriculture,
Meknes, Morocco us as antibacterial agents against four bacteria. The synthe-
3 sized compounds had led to some important structure with
Medical Microbiology Laboratory, Mohamed V. Hospital,
Meknes, Morocco interesting antibacterial activity.
 Struct Chem

Materials and methods Synthesis

Materials Synthesis of acridone from 2-(phenylamino)benzoic acid

All materials were purchased from commercial suppliers. IR A mixture of aniline (0.65 g, 7 mmol), o-bromobenzoic acid
spectra were recorded using JASCO FT-IR 4100 spectropho- (1 g, 5 mmol), anhydrous potassium carbonate (0.82 g,
tometer. Mass spectrometric measurements were recorded 6 mmol), 0.1 g of copper powder, and 0.04 g of copper oxide
using SHIMADZU 8040 LC/MS/MS. The 1H, 13C NMR was refluxed for 4 h in 15 mL of amyl alcohol. The amyl
spectra was recorded with Bruker Avance 300 at 25 °C. alcohol was evaporated then poured into (100 mL) hot water,
cooled to room temperature, acidification with HCl lead to 1
as a white precipitate which was then recrystallized from eth-
Determination of minimum inhibitory concentration anol. The N-phenylanthranilic acid (1 g, 4.7 mmol) was taken
in 4 mL of concentrated sulfuric acid and heated on water bath
The synthesized compounds were tested for their antibac- for 3 h. Reaction mixture was added to hot water and the
terial activity by the disk diffusion method; the active resulting precipitates were filtered to get acridone. The sample
compounds were subjected to the determination of the of acridone 2 was recrystallized from acetic acid. Yield 70%,
MIC, using the broth microdilution method. The micro- Rf 0.71, mp > 330 C. IR (KBr) 3275 (N–H), 3084 (C–H aro-
organisms utilized for the test were Escherichia coli, matic), 1640 (C=O), 1570 (C=C). 1H NMR (300 MHz,
Staphylococcus aureus, Pseudomonas putida, and [D6]DMSO, 25 °C, TMS): δ 11.74 (s, 1H, NH), 8.23 (dd,
Klebsiella pneumonia. They were collected from clinical J = 8.1, 1.2 Hz, 2H, H1-H8), 7.70 (td, J = 8.4, 1.5 Hz, 2H,
isolates. Bacterial inoculums were prepared by H3-H6), 7.53 (d, J = 8.1 Hz, 2H, H4-H5), 7.23 (td, J = 8.4,
subculturing microorganisms into MHB at 37 °C for 1.5 Hz, 2H, H 2-H 7 ). 13 C NMR (75 MHz, [D 6 ]DMSO,
18 h and were diluted to approximately 106 CFU mL−1. 25 °C, TMS) δ: 177.29, 141.35, 133.94, 126.46, 121.49,
Initial solution with concentration 0.5 mg/mL of the 120.91, 117.80.
compounds (4a-h) were prepared in DMF; further serial
dilutions were made in the microplates, and 100 μl of Synthesis of 10-(prop-2-yn-1-yl)acridone (3)
MHB containing each test microorganism were added to
the microplate [19–21], then incubated at 36 °C for 24 h. To a mixture of acridone 2 (0.5 g, 2.5 mmol), sodium hydride
After incubation, 20 μL of TTC (0.04 mg/mL) were (45 mg, 3.6 mmol) and TBAB (0.5 g, 2 mmol) in DMF
added to each microplate. The color changes of TTC (3 mL), propargyl bromide (0.38 g, 3.6 mmol) was added
from colorless to red were accepted as microbial growth and the mixture was stirred at 85 °C for 3 h. After which, it
[22]. A negative control was prepared using the inocu- was poured into crushed ice and the white yellow formed
lum and DMF without derivative, whereas chloramphen- precipitate was recrystallized from ethanol-DMF.
icol was used as positive control. White solid; yield 74%, Rf 0.84, mp = 178 °C. IR (KBr):
3206, 3008, 2913, 1639, 1600, 1495 cm −1 . 1 H NMR
(300 MHz, [D6]DMSO, 25 °C, TMS): δ 8.34 (d, 2H, H1-
Molecular docking studies H8), 7.85–7.81 (m, 4H, H3-H6-H4-H5), 7.38–7.32 (m, 2H,
H2-H7), 5.34 (s, 2H, CH2), 2.48 (s, 1H, CH); 13C NMR
Molecular docking of the compounds (4a-h) into the (75 MHz, [D6]DMSO, 25 °C, TMS) δ 177.08, 141.70,
S. aureus DHFR complex structure (PDB code, 2W9S) 134.76, 134.46, 127.18, 127.00, 122.37, 122.20, 117.44,
[23] was carried out using Molecular Operating 116.35, 79.12, 76.16, 36.
Environment (MOE-Dock 2015.10) software [24]. The
Discovery Studio (version 4.5) was used for graphical General procedure for the synthesis
visualization. The structure of 2W9S in docking study of acridon-1,2,3-triazole derivatives (4a-h)
was downloaded from Protein Data Bank. The 3D struc-
tures of compounds (4a-h) were constructed using A mixture of 10-(prop-2-yn-1-yl)acridone (0.1 g, 0.42 mmol),
Chem ultra 12.0 software and were energetically mini- azide (0.63 mmol), copper sulfate (21 mg, 0.081 mmol), and
mized, then the 3D structure of compounds (4a-h) was sodium ascorbate (33 mg, 0.16 mmol) in DMF (5 mL) was
placed in database. For protein preparation, the hydro- stirred at room temperature for 12 h. Then the mixture was
gen atoms were added; water and impurities were re- diluted with water, poured onto ice, and the precipitate was
moved. Different types of interactions between protein filtered off, washed with cold water, and purified by flash
and the docked compounds (4a-h) were analyzed using chromatography on silica gel using hexane/diethyl ether
Discovery Studio. (1:4) to afford desire product.
Struct Chem

Ethyl 2-{4-[(9-oxoacridin-10-yl)methyl] 7.2 Hz, 2H, H1’,H4’), 7.36 (d, J = 7.2 Hz, 4H,
-1,2,3-triazol-1-yl}acetate (4a) H2,H7,H2’,H3’), 5.87 (s, 2H, CH2), 2.36(s, 3H, CH3). 13C
NMR (75 MHz, [D6]DMSO, 25 °C, TMS):177.07, 144.10,
Yellow solid; yield 80%, Rf 0.73 (hexane:diethyl ether = 1:6), 142.51, 138.85, 134.61, 130.85, 127.14, 122.42, 121.94,
mp > 300 °C. IR (KBr): 3132, 2986, 2914, 1745, 1634, 1600, 120.52, 116.74, 42.39, 20.96. MS (ESI) for C23H18N4O
1493, 1227 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, [M + 1]+, calcd 367.17, found 367.15.
TMS): δ 8.36 (dd, J = 8.1, 1.5 Hz,2H, H1-H8), 8.10 (s, 1H,
triazole), 7,95 (d, J = 8,7 Hz, 2H, H4, H5), 7.80 (td, J = 15.6, Ethyl 2-{4-[(2-methyl-9-oxoacridin-10-yl)methyl]
6.9 1.5 Hz, 2H, H3, H6), 7.33 (t, J = 7.5 Hz, 2H, H2,H7), 5.83 -1,2,3-triazol-1-yl}acetate (4e)
(s, 2H, CH2), 5.31(s, 2H, CH2), 4,11 (q, J = 7.2 Hz, 2H, CH2),
1,13 (t, J = 7.2 Hz, 3H,CH3). 1 3 C NMR (75 MHz, Yellow solid; yield: 84%, Rf 0.77 (hexane:diethyl ether = 1:6),
[D6]DMSO, 25 °C, TMS): 177.07, 167.53, 143.01, 142.23, mp > 300 °C. IR (KBr): 3130, 2981, 1746, 1635, 1602, 1496,
134.65, 127.13, 125.25, 122.20, 122.00, 116.74, 61.89, 50.86, 1228 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, TMS):
41.94, 14.34. MS (ESI) for C20H18N4O3 [M + 1]+, calcd 8.34 (d, J = 7.8 Hz,1H, H1), 8.13 (s,1H, H8), 8.05 (s,1H, tri-
363,13, found 363.14. azole), 7,93–7.85 (m, 3H, H3,H4, H5), 7.63 (d, J = 8.1 Hz,
1H, H6), 7.30 (t, J = 7.5 Hz, 1H, H2), 5.80 (s, 2H, CH2),
{4-[(9-oxoacridin-10-yl)methyl]-1,2,3-triazol-1-yl}acetic acid 5.29(s, 2H, CH2), 4,11 (q, J = 7.2 Hz, 2H, CH2), 2.41 (s,3H,
(4b) CH3), 1,13 (t, J = 7.2 Hz, 3H,CH3). 13C NMR (75 MHz,
[D6]DMSO, 25 °C, TMS): 176.91, 167.47, 143.07, 142.09,
Yellow solid; yield 82%, Rf 0.62 (hexane:diethyl ether = 1:4), 140.36, 135.92, 134.42, 131.21, 127.15, 126.35, 125.16,
mp > 300 °C. IR (KBr): 3400, 3120, 2970, 1700, 1635, 1601, 122.13, 121.69, 116.73, 116.55, 61.87, 50.87, 41.87, 20.63,
1497, 1236 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, 14.32. MS (ESI) for C21H20N4O3 [M + 1]+, calcd 377.15,
TMS): 10.82 (s, 1H, OH), 8.36 (dd, J = 8.0, 1.1 Hz,2H, H1- found 377.16.
H8), 8.11 (s, 1H, triazole), 7,96 (d, J = 8,7 Hz, 2H, H4, H5),
7.80 (td, J = 15.6, 6.9 1.8 Hz, 2H, H3, H6), 7.33 (t, J = 8.1 Hz, {4-[(2-methyl-9-oxoacridin-10-yl)methyl]
2H, H2,H7), 5.81 (s, 2H, CH2), 5.19(s, 2H, CH2). 13C NMR -1,2,3-triazol-1-yl}acetic acid (4f)
(75 MHz, [D6]DMSO, 25 °C, TMS): 177.08, 168.91, 142.79,
142.23, 134.66, 127.12, 125.21, 122.16, 122.00, 116.78, Yellow solid; yield: 81%, Rf 0.65 (hexane:diethyl ether = 1:4),
51.08, 41.93. MS (ESI) for C18H14N4O3 [M + 1]+, calcd mp > 300 °C. IR (KBr): 3400, 3118, 2976, 1700, 1635, 1601,
335.10, found 335.12. 1490, 1237 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C,
TMS): 10.85 (s,1H, OH), 8.34 (d, J = 7.8 Hz,1H, H1), 8.12
10-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)acridin-9(10H)-one (s,1H, H8), 8.06 (s,1H, triazole), 7,94–7.75 (m, 3H, H3,H4,
(4c) H5), 7.64 (d, J = 8.1 Hz, 1H, H6), 7.30 (t, J = 7.5 Hz, 1H, H2),
5.78 (s, 2H, CH2), 5.13(s, 2H, CH2), 2.40(s,3H, CH3). 13C
White solid; yield: 82%, Rf 0.69 (hexane:diethyl ether = 1:4), NMR (75 MHz, [D6]DMSO, 25 °C, TMS): 176.89, 167.55,
mp > 300 °C. IR (KBr): 3111, 1638, 1614, 1593, 1502, 142.07, 140.34, 135.96, 134.46, 131.20, 127.13, 126.32,
1255 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, TMS): 125.19, 122.06, 121.70, 116.80, 116.62, 50.87, 41.84, 20.66.
8,71(s, 1H, triazole), 8.42 (d, J = 7.2 Hz,2H, H1-H8), 7,98 (d, MS (ESI) for C19H16N4O3 [M + 1]+, calcd 349.13, found
J = 8,1 Hz, 2H, H4, H5), 7.81–7.62 (m, 4H, H3, H6, H1’,H4’), 349.13.
7.35 (t, J = 7.2 Hz, 5H, H2,H7,H2’,H3’,H4’), 5.87 (s, 2H,
CH2). 13C NMR (75 MHz, [D6]DMSO, 25 °C, 2-methyl-10-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)
TMS):177.12, 144.07, 142.57, 138.75, 134.61, 130.85, acridin-9(10H)-one (4 g)
127.24, 122.32, 121.91, 120.42, 116.44, 42.49. MS (ESI) for
C22H16N4O [M + 1]+, calcd 353.13, found 353.12. White solid; yield: 80%, Rf 0.69 (hexane:diethyl ether = 1:4),
mp > 300 °C. IR (KBr): 3110, 3054, 1636, 1614, 1601, 1500
10-{[1-(4-methylphenyl)-1,2,3-triazol-4-yl] 1266 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, TMS):
methyl}acridin-9-one (4d) 8,71(s, 1H, triazole), 8.32 (d, J = 7.2 Hz,2H, H1), 8.14 (s, 1H,
H8), 7,94–7.78 (m, 2H, H4, H5), 7.71–7.62 (m, 4H, H3, H6,
Yellow solid; yield 90%, Rf 0.70 (hexane:diethyl ether = 1:4), H1’,H5’), 7.37 (d, J = 7.2 Hz, 5H, H2,H2’,H3’, H4’), 5.82 (s,
mp > 300 °C. IR (KBr): 3110, 3060, 1637, 1617, 1598, 1504, 2H, CH2), 2,38 (s, 3H, CH3). 13 C NMR (75 MHz,
1265 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, TMS): [D6]DMSO, 25 °C, TMS): 177.01, 144.29, 142.13, 140.10,
8,72(s, 1H, triazole), 8.40 (d, J = 7.2 Hz,2H, H1-H8), 7,98 (d, 138.74, 136.12, 134.57,134.12, 131.18, 130.21, 127.22,
J = 8,1 Hz, 2H, H4, H5), 7.82 (t, 2H, H3, H6), 7.73 (d, J = 126.35, 122.30, 121.13, 120.71, 120.37, 116.92, 116.84,
 Struct Chem

Scheme 1 Synthesis of acridone linked to 1,2,3-triazole derivatives 4

42.36, 20.86. MS (ESI) for C23H18N4O [M + 1]+, calcd (75 MHz, [D 6 ]DMSO, 25 °C, TMS): 177.02, 144.23,
367.14, found 367.15. 142.23, 140.50, 138.81, 136.00, 134.67,134.52, 131.18,
130.61, 127.12, 126.33, 122.15, 121.83, 121.71, 120.37,
116.82, 116.64, 42.16, 20.99, 20.68. MS (ESI) for
2-methyl-10-{[1-(4-methylphenyl)-1,2,3-triazol-4-yl] C24H20N4O [M + 1]+, calcd 381.17, found 381.16.
methyl}acridin-9-one (4 h)

Yellow solid; yield: 87%, Rf 0.70 (hexane:diethyl ether = 1:4),

Results and discussion
mp > 300 °C. IR (KBr): 3112, 3063, 1638, 1616, 1600, 1502
1267 cm−1. 1H NMR (300 MHz, [D6]DMSO, 25 °C, TMS):
Synthesis
8,72(s, 1H, triazole), 8.35 (d, J = 7.2 Hz,2H, H1), 8.14 (s, 1H,
H8), 7,94–7.78 (m, 2H, H4, H5), 7.71–7.62 (m, 4H, H3, H6,
The synthesis of novel acridone bearing 1,2,3-triazole com-
H1’,H4’), 7.36 (d, J = 7.2 Hz, 3H, H2,H2’,H3’), 5.83 (s, 2H,
pounds was performed as described in (Scheme 1). Initially
CH2), 2,41 (s, 3H, CH3), 2.32(s, 3H, CH3). 13C NMR
acridone 2 was synthesized by Ullmann condensation reaction
Table 1 Synthesized compounds (4a-h) from alkynes and azides of o-bromobenzoic acid with various aniline derivatives in the
presence of potassium carbonate and copper in isoamyl alco-
Entry Compounds R1 R2 Yield (%) hol at reflux to produce 2-arylamino benzoic acids 1 [25],
which was cyclized with sulfuric acid in water bath at 85 °C
Conventional MW
for 4 h to give compounds 2. The propargylation of acridone
1 4a H CH2COOEt 74 80 was attempted by refluxing excess of propargyl bromide with
2 4b – CH2COOH 70 82 acridone derivatives in DMF using sodium hydride.
3 4c – Ph 70 82 We synthesized various azides by utilizing literature proto-
4 4d – p-CH3-C6H4 75 90 cols [9, 26]. Alkyl azides were prepared by refluxing alkyl
5 4e CH3 CH2COOEt 76 84 halides with NaN3 in DMF at 80–90 °C; 4-Azidotoluene and
6 4f – CH2COOH 69 81 azidobenzene were prepared from p-toluidine and aniline
7 4g – Ph 75 80 using diazotization followed by adding NaN3.
8 4h – p-CH3-C6H4 73 87 1,3-diploar cycloaddition was performed with 10-(prop-2-
yn-1-yl)acridone derivatives (1 equiv) (3) and aromatic,
Struct Chem

Fig. 1 The NMR 2D HMBC of compound 4a

aliphatic azides (1.5 equiv) in the presence of copper sulfate (4a-h) were obtained with good yields (Scheme 1). In the goal
(0.1 equiv), and sodium ascorbate (0.2 equiv). The reaction to obtain these 1,2,3-triazoles with excellent yields and shorter
was conducted at room temperature in DMF for 12 h. Then the reaction times under mild reaction conditions, we focused our
mixture was poured into ice water; the precipitate was filtered attention to microwave-assisted synthesis. The model reaction
off and purified by flash chromatography using hexane/ was examined in various solvents with controlled MW powers
diethyl ether (1:4); the novel 1,4-disubstituted 1,2,3-triazoles (100–300 W). The best result was obtained in DMF as solvent
at 200 W. The desired products of 4a–h were obtained in
yields ranging from 85 to 98%. The reaction times under
Table 2 Antibacterial data for the synthesized compounds (4a-h)

compounds S. aureus E.coli K. pneumoniae P. putida

2a 122.8 133.4 137.9 156.3

2b 118.4 124.2 130.4 145.5
4a 90.91 122.81 122.81 122.81
4b 56.60 122.81 137.93 122.81
4c 46.60 112.36 138.32 126.12
4d 38.46 90.91 107.14 107.14
4e 56.60 74.07 74.07 115.04
4f 56.60 74.07 56.60 107.14
4g 41.32 66.36 81.32 119.36
4h 19.61 56.60 90.91 122.81
Chloramphenicol 11.65 22.41 15.38 37.03
DMF – – – – Fig. 2 Redocking pose and RMSD value of 0.9 Å (blue = docked, gray =
original)
 Struct Chem

Table 3 Docking score,

intramolecular hydrogen bonds Compounds Docking score C=O—ALA7 (d in Å) Hydrophobic interactions
(C=O⋯Ala7), and hydrophobic
interactions of the synthesized 4a 6.74 – Ile 50, Ile 31, Ile 14, Leu 20, Leu 54, and Leu 28
compounds 4b 6.42 – Ile 31, Ile 14, and Leu 20
4c 6.80 C=O---ALA7 (2.49 Å) Ile 50, Ile 31, Ile 14, and Leu 20
4d 6.98 C=O---ALA7 (2.50 Å) Ile 50, Ile 31, Ile 14, and Leu 20
4e 6.75 – Ile 50, Ile 31, Ile 5, and Phe 92
4f 6.58 – Ile 50, Ile 31, Ile 5, Leu 20, and Phe 92
4g 6.98 C=O---ALA7 (2.55 Å) Ile 50, Ile 14, and Leu 20
4h 7.04 C=O---ALA7 (2.55 Å) Ile 5 and Phe 29

microwave irradiation are more acceptable than those of con- supported by 13C and DEPT NMR spectra, which showed all
ventional methods (Table 1). the expected carbon signals corresponding to acridone–
The structure of the 1,2,3-triazoles were demonstrated by triazole derivatives, essentially the aromatic carbons resonat-
their spectral data. The IR spectra of compounds (4a-h) ing at δC 125.2–143.0 ppm corresponding to triazole ring and
showed characteristic absorption bands in the region of signals of CH2 carbons resonated at δC 50.8–41-9 ppm.
1746–1700 cm−1 (C=O) due to the presence of ester and car- The regioselectivity of the cycloaddition reaction under
boxylic acid groups, and characteristic bands of 1,2,3-triazole microwave irradiation was confirmed by using 1 H and 13C
ring was detected in the range 1500–1300 cm−1 which corre- long-range NMR experiments (HMBC). The selective forma-
sponds to N=N & C=C bonds. However, the desperation of tion of 1,4-disubstituted 1,2,3-triazoles was confirmed by the
the vibration bonds of alkyne group in the region of 2110 cm−1 3-bond HMBC correlations of the methylene protons H1a (δ
and 3210 cm−1 confirmed the formation of compounds. 5.83 ppm) and H4a (δ 5.31 ppm) with the carbon of triazole
The 1H NMR spectra of compounds 4a–h showed a singlet C3a (δ 125 ppm). Also, we observe that the signal of the HTz (δ
at δH 8.10–8.72 ppm attributable to the proton H3’ of the 8.10 ppm) correlate with the C3a (δ 125 ppm) (Fig. 1).
triazole moiety and signals in the aromatic region δH 8.40–
7.31 ppm relative to the aromatic protons. In addition, the Antibacterial activity
compounds 4a-d showed signals at δ H 5.83–5.13 ppm
corresponded to –CH2– groups attached to triazole ring, and The antibacterial activity of the compounds (4a-h) was tested
the compounds 4e-f present singlet peak that appeared at δH in vitro against one gram-positive bacteria Staphyloccocus
2.3 ppm corresponded to CH3. These structures were further aureus and three gram-negative bacteria Pseudomonas putida,

Fig. 3 The binding conformations and ligand interactions of the compound 4h at the active site of S. aureus DHFR
Struct Chem

Fig. 4 The binding conformations and ligand interactions of the compound 4b at the active site of S. aureus DHFR

Klebsiella pneumonia, and Escherichia coli. Primary screen- compounds 4e-h show moderate activity against
ing test of the antibacterial effect of the compounds elicited as Escherichia coli and Klebsiella pneumonia with the
inhibition zones of the growth of the tested bacteria. MIC values between 56.6–74.0 μg/mL. The low anti-
Therefore, the MIC was determined for synthesized bacterial activity was observed against Pseudomonas
compounds. putida with MIC value 107.1 μg/mL. The results indi-
According to the antibacterial activity results cate that the substitution of the acridone ring at position
(Table 2), all compounds (4a-h) were found to have 2 with methyl increase the antibacterial activity. In the
good antibacterial activity against Staphylococcus methyl-substituted series, the antibacterial activity of the
aureus, the compound 4h was found to be the best compound 4e showed higher activity against gram-
active derivatives with MIC = 19.6 μg/mL. The tested negative bacteria. Moreover, there is a development in

Fig. 5 Structure-activity
relationship (SAR) of the
acridone-1,2,3-triazole skeleton
 Struct Chem

antibacterial activity after the substitution of acridone These docking results indicate that the compound 4h played
ring with triazole ring for all bacteria. The results of a significant role in binding with the active site of S. aureus
the antibacterial activity revealed that the compounds DHFR receptor, and this may explain its high antibacterial
(4a-h) have good antibacterial activity against gram- activity. Furthermore, the study shows that all compounds
positive bacteria compared with gram-negative bacteria. with high potency for antibacterial activity present a hydrogen
bond interaction with Ala7 and hydrophobic interactions into
Molecular docking studies S. aureus DHFR active site.
The results of the antibacterial activity and docking studies
The antibacterial potency of the compounds (4a-h) was sub- revealed that the substitution of the acridone ring with methyl
jected for docking study to understand the interaction of our group and the acridone-1,2,3-triazole skeleton by aromatic
compounds and methicillin-resistant Staphylcoccus aureus moiety increases the antibacterial activity against S. aureus;
(MRSA). The dihydrofolate reductase (DHFR) is an impor- this could be explained by the hydrophobic interactions in
tant enzyme whose role is to regenerate folic acid into reduced S. aureus DHFR active site (Fig. 5).
form of tetrahydrofolate [27]. The dihydrofolate reductase
(DHFR) is an interesting target for antibacterial agents in sev-
eral studies [2, 28, 29]. Crystal structure of S. aureus DHFR Conclusion
(PDB ID: 2W9S) with trimethoprim as co-crystallized ligands
and NADPH with 1.80 Å resolution was retrieved from The present work reports synthesis of novel heterocyclic com-
Protein Data Bank. In order to evaluate and validate the bind- pounds combining the 1,2,3-triazole and acridone rings.
ing prediction of docking protocol, the co-crystallized ligand Efficient microwave-assisted synthesis of novel acridone
TOP was redocked into the active site of S. aureus DHFR bearing 1,2,3-triazole compounds has been carried out effec-
protein (2W9S). The evaluation of the precision of the tively with short time reaction and in good yields. The com-
docking protocol was based on the RMSD parameter. The pounds (4a-h) were investigated for their in vitro antibacterial
RMSD was calculated on the basis of the difference between activity against Escherichia coli, Klebsiella pneumoniae,
redocking and original poses using the MOE software. The Pseudomonas putida, and Staphyloccocus aureus, indicating
results show the redocked ligand superimposed on the co- that the introduction of 1,2,3-triazole ring on the acridone ring
crystallized one (Fig. 2) with a low RMSD value of 0.9 Å. increases the antibacterial activity. The docking results
The analysis of the binding site revealed that the synthe- showed that compounds with high antibacterial potency
sized ligands is stabilized by several favorable interactions showed hydrogen bond and hydrophobic interactions with
including polar, hydrogen bond, hydrophobic, and Van der S. aureus DHFR active site.
Waals interactions (Table 3); the binding site pocket contains
the following amino acids: Ala 7, Ile 31, Ile 14, Leu 20, Ile 50,
Ile 5, Phe 92, Ser 49, Leu 28, Tyr 98, Thr 111, Gly 93, Thr 46,
Gly 94, Gly 15, Trp 22, Val 6, His 30, and Asp 27.
According to molecular docking analysis, 4h showed a
docking score value of 7.04, reflecting that 4h could bind with References
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