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SYNTHESIS AND ANTIBACTERIAL ACTIVITY OF NEW IMIDAZOPYRIDINE

DERIVATIVES BASED ON 2,3-DIAMINOPYRIDINE

Article · January 2025

DOI: 10.47743/achi-2024-2-0013

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DOI: 10.47743/achi-2024-2-0013 ACTA CHEMICA IASI, 32_2, 237-248 (2024)

SYNTHESIS AND ANTIBACTERIAL ACTIVITY

OF NEW IMIDAZOPYRIDINE DERIVATIVES
BASED ON 2,3-DIAMINOPYRIDINE

Oday A. Obaidia, Zeyad Kadhim Oleiwi b*,

Nidhal Hatif Hammoodb, Hussein Ali Al-Bahranic, d

a
Department of Pharmacy, Kufa Technical Institute, Al-Furat Al-Awsat
Technical University, Kufa, Iraq
b
Department of Pharmaceutical Chemistry, Faculty of Pharmacy,
University of Kufa, Najaf, Iraq
c
College of Nursing, University of Al-ameed, Kerbala, Iraq
d
Department of Chemistry. College of Education for Pure Science,
University of Kerbala, Iraq

Abstract: The present research work focuses on the synthesis and evaluation of
antibacterial activity of newly developed imidazopyridine analogues arising from
2,3-aminopyridine substructure. Three new imidazopyridine derivatives have been
synthesized from the reaction of 2,3-aminopyridine with ibuprofen, naproxen and
etodolac employing HCl as both the solvent and the catalyst. The resulting
compounds were identified by using (FTIR) and (¹H-NMR). The antibacterial
activity of these newly synthesized compounds was carried out against a series of
bacteria: Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus &
Streptococcus mutans by microdilution method of 96 well microplates. These
findings revealed that all synthesized compounds possessed pronounced
antimicrobial effects against all the bacterial strains studied. The highest
antimicrobial activity was registered for compound 1-((-[(1H-imidazo
[4,5-b]pyridin-2-yl)methyl)-]-1,8-diethyl-1,3,4,9-tetrahydropyrano[3,4-b]indole
that was most effective toward Escherichia coli. Based on these discoveries, it can
be assumed that the newly imidazopyridine derivatives synthesized might be of
interest for further study, targeting potential antibacterial activity.

Keywords: Imidazopyridine, 2,3-Diaminopyridine, Antibacterial Activity, Synthesis

*
Zeyad Kadhim Oleiwi e-mail: [Link]@[Link]
238 Zeyad Kadhim Oleiwi et al.

Introduction
Substituted pyridines are versatile scaffolds that have various
biological activities: antipyretic/analgesic, antiprotozoal, antibacterial,
antitumor, antifungal, anti-inflammatory, and antiapoptotic.1 In these
compounds, imidazopyridine and its derivatives are the most important
heterocyclic compounds that have attracted great attention in natural
products, medicinal chemistry and synthetic chemistry. Due to the specific
structure of the imidazopyridine ring, it can bind to different enzymes and
receptors through weak interactions, switching on a number of biological and
pharmacological effects.2 Imidazopyridines have been found in many
investigations to display valuable therapeutic effects such as antimicrobial,3
and antibacterial,4 antifungals,5 anticandidal,6,7 kinase inhibitors,8,9
antitubercular agents,10 anti-plasmodial agents11 and anticancer agents,12 In
addition, research on synthesized imidazopyridine derivatives has shown that
they possess anti-inflammatory, antiviral, antiosteoporosis, anti-parasitic and
antihypertensive properties.13,14

Results and Discussion

Synthesis
This study includes the synthesis of new imidazopyridine derivatives
through the reaction of 2,3-diaminopyridine (A) with different carboxylic
acids containing drugs NSAID (Naproxen, Ibuprofen and Etodolac) in the
presence of HCl as solvent and catalyst as shown in Scheme 1. Thus,
2,3-diamino pyridine A (0.01 mole) was refluxed with etodolac, ibuprofen or
naproxen (0.01 mole each) in the presence of 4N hydrochloric acid (HCl) for
6 hours at 100 °C to give the compounds 1-3. Their structures were confirmed
by FTIR and 1H-NMR spectroscopy.
 Synthesis and antibacterial activity of new imidazopyridine derivatives …
N NH 2 HO H
HCl, 100 oC, 6hrs. N N
+ R
R + 2H2O
NH 2 O N
(A)
H
N N
N
O N N
N NH
NH
N H
N
O
(1)
(2)
(3)

Scheme 1. Synthesis of imidazopyridine derivatives.

Antimicrobial activity of compounds 1-3 against E. coli

This study assessed the antibacterial efficacy of three novel
compounds against Escherichia coli (E. coli), a prevalent gram-negative
bacterium, as it is shown in Figure 1. According to the MIC, compound 1
exhibited a limited inhibitory effect on E. coli. (MIC = 906.1 µg/mL) since
the highest MIC, the lowest the antimicrobial activity. However, compound
2 showed a significantly lower MIC value of 171.9 µg/mL indicating a
stronger antimicrobial activity compared to compound 1. On the other hand,
compound 3 demonstrated remarkably low MIC value as the most potent
antimicrobial activity with an MIC of 9.091 µg/mL indicating a highly
effective antimicrobial agent against E. coli.

Figure 1. Antimicrobial activity of compounds 1, 2 and 3 against E. coli.

240 Zeyad Kadhim Oleiwi et al.

Antimicrobial activity of compounds 1-3 against

Klebsiella pneumoniae
In contrast to findings showed in the previous section, the antimicrobial
activities of the three compounds evaluated against Klebsiella pneumoniae,
another gram-negative bacterium showed that compound 1 exhibited the highest
inhibitory effect on Klebsiella pneumoniae (MIC = 20.3 µg/mL) Figure 2.
However, compound 2 showed a significantly higher MIC value of 581.6 µg/mL
indicating a weaker antimicrobial activity compared to compound 1. On the other
hand, compound 3 demonstrated remarkably high MIC value as the least potent
antimicrobial activity with an MIC of 994.2 µg/mL.

Figure 2. Antimicrobial activity of compounds 1, 2 and 3 against Klebsiella pneumoniae.

Antimicrobial activity of compounds 1-3 against

Staphylococcus aureus
When tested against Staphylococcus aureus, a common gram-positive
bacterium compounds 1, 2 and 3 displayed MIC of 843.2, 1270 and
566.4 µg/mL, repectively Figure 3. Although compound 3 showed the lowest
MIC against Staphylococcus aureus which means it has the highest
antimicrobial activity among the three compound against this species, it still
represents a high MIC.
 Synthesis and antibacterial activity of new imidazopyridine derivatives …

Figure 3. Antimicrobial activity of compounds 1, 2, and 3 against Staphylococcus aureus.

Antimicrobial activity of compounds 1-3 against

Streptococcus mutans
The three compound were also tested against Streptococcus mutans,
another gram-positive bacterium. Compounds 1, 2 and 3 showed MIC of
1219, 581.6 and 843.2 µg/mL, respectively Figure 4. Again, although
compound 2 showed the lowest MIC against Streptococcus mutans which
means it has the highest antimicrobial activity among the three compound
against this species, the value still represents a high MIC.

Figure 4. Antimicrobial activity of compounds 1, 2, and 3 against Streptococcus mutans.

242 Zeyad Kadhim Oleiwi et al.

Experimental
All reactions were performed in oven dried glassware. The solvents
and reagents used in the commercial grade were distilled before use. Thin
layer chromatography (TLC) on silica gel GF 254 plates was used in
determining the reaction progress. Melting points were obtained using an
electrothermal melting point apparatus in capillary tubes with the capillary
being open at both ends. ¹H-NMR experiments were recorded at ambient
temperature using Bruker Avance 400 MHz NMR spectrometer. Preparation
of samples involved the dissolving of the compounds in deuterated
chloroform (CDCl₃) with (TMS) as the internal reference. Chemical shifts are
expressed in (ppm) abbreviated as δ.

General Procedure for Synthesis of Compounds 1-315

2,3-Diamino pyridine (A) (0.01 mole) was refluxed with etodolac,
ibuprofen or naproxen (0.01 mole each) in the presence of 4N hydrochloric
acid (HCl) for 6 hours at 100 °C, with a stirring speed of 320 rpm using a
reflux condenser. The reaction progress was monitored by TLC, employing a
chloroform: methanol (9:1) as a mobile phase. After completion, the mixture
was gradually neutralized with 10% NaOH (w/v) until the pH was shifted to
8-9. The resulted solution was further chilled in an ice bath for 5 minutes to
facilitate precipitation. The precipitate was obtained through filtration and
subsequently dried.
Synthesis and spectral data of 2-[1-(4-isobutylphenyl)ethyl]-1H-
-imidazo[4,5-b] pyridine 1 (figure 5)
N H
N

Figure 5. Structure of compound 1 derived from Ibuprofen.

 Synthesis and antibacterial activity of new imidazopyridine derivatives …

It was obtained using the general procedure. A conversion was observed after
6 hours. The resulting product is a white powder, with a melting point of
98°C, yield 71 %. FTIR (KBr), ν(cm-1): N-H (3512), C-H aliphatic (2970-
2850), C-H aromatic (3070), C=C (1590, 1454), C=N (1665), C-N (1340-
1000). 1H NMR (400 MHz, CDCl3) δ(ppm): 12.90 (1H, s, N-H), 8.32 (1H, d,
J = 7.5 Hz, C-Hpyridine), 7.84 (1H, d, J = 7.5 Hz, C-Hpyridine), 7.15 (1H, t,
J = 7.5 Hz, C-Hpyridine), 7.18, (2H, d , J =7.5, Hz, C-Hbenzene), 7.04 (2H, d ,
J =7.5, Hz, C-Hbenzene), 4.16 (1H, q, J = 6.8 Hz, C-H), 2.43 (2H, d, J = 7.0 Hz.
CH2), 1.82 (1H, heptet, J = 7.0 Hz, CH), 1.62 (3H, d, J = 6.8 Hz CH3), 0.87
(6H, d, J = 6.8 Hz, 2 x CH3).
Synthesis and spectral data of (S)-2-[1-(6-methoxynaphthalen-2-
-yl)ethyl]-1H-imidazo[4,5-b]pyridine 2 (figure 6)

H
N

N
O
N

Figure 6. Structure of compound 2 derived from Naproxen.

It was obtained using the general procedure. A conversion was
observed after 6 hours. The resulting product is a faint pink powder, with a
melting point of 95°C, yield 65%. FTIR (KBr), ν(cm-1): N-H (3490), C-H
aliphatic (2990-2860), C-H aromatic (3080), C=C (1587, 1470), C=N (1670),
C-O (1290-990). 1H NMR (400 MHz, CDCl3), δ(ppm): 12.80 (1H, s, N-H),
8.30 (1H, d, J = 7.5 Hz, C-Hpyridine) 7.83 (1H, d, J = 7.5 Hz, C-Hpyridine), 7.78
(1H, d, J = 7.5 Hz, C-Hbenzene), 7.73 (1H, d, J = 7.5 Hz, C-Hbenzene), 7.48 (1H,
d, J = 1.5 Hz, C-Hbenzene), 7.34 (1H, d, J = 1.5 Hz, C-Hbenzene), 7.23 (1H, d ,
J = 1.5 Hz, C-Hbenzene), 7.15 (1H, t, J = 7.5 Hz, C-Hpyridine), 7.06 (1H, d,
J = 7.5 Hz, C-Hbenzene), 4.15 (1H, q, J = 6.8 Hz, CH), 3.81 (3H, s, CH3), 1.68
(3H, d, J = 6.8 Hz, CH3).
244 Zeyad Kadhim Oleiwi et al.

Synthesis and spectral data of 1-[(1H-imidazo[4,5-b]pyridin-2-

-yl)methyl]-1,8-diethyl-1,3,4,9-tetrahydropyrano[3,4-b]indole3
(figure 7)

HN N

H
N

Figure 7. Structure of compound 3 derived from Etodolac.

It was obtained using the general procedure. A conversion was
observed after 6 hours. The resulting product is a brown powder, with a
melting point of 92°C, yield 59%. FTIR (KBr), ν(cm-1): N-H (3515-3490),
C-H aliphatic (2997-2890), C-H aromatic (3087), C=C (1589,1480), C=N
(1680), C-O (1300-995). 1H NMR (400 MHz, CDCl3) δ (ppm): 12.88 (1H, s,
N-H), 11.67 (1H, s, N-Hindole), 8.32 (1H, d, J = 7.5 Hz, C-Hpyridine), 7.84 (1H,
d, J = 7.5 Hz, C-H pyridine), 7.17 (1H, t, J = 7.5, Hz, C-Hindole), 7.15 (1H, t,
J = 7.5 Hz, C-Hpyridine), 7.09 (1H, d, J = 7.5, Hz, C-Hindole), 7.07 (1H, d,
J = 7.5, Hz, C- Hindole), 3.69 (2H, m, CH2-O), 3.02 (2H, s, CH2), 2.71 (2H, q,
J = 8.0 Hz, CH2-CH3), 2.62 (2H, t, J = 7.0 Hz, CH2-CH2 -O), 1.67 (2H, q,
J = 8.0 Hz, CH2-CH3), 1.18 (3H, t, J = 8.0 Hz, CH3-CH2), 0.89 (3H, t,
J = 8.0 Hz, CH3-CH2).
Experimental procedure for antimicrobial testing
preparation of bacterial cultures: Bacterial strains were cultured in Mueller-
Hinton broth, a nutrient-rich medium suitable for supporting bacterial growth.
Microdilution method setup: The antimicrobial activity of the
synthesized compounds was tested using the microdilution method in 96-well
 Synthesis and antibacterial activity of new imidazopyridine derivatives …

microplates. This approach allows for precise measurement of bacterial

inhibition at various concentrations of the test compounds.
Preparation of test compounds: Serial dilutions of the synthesized
compounds were prepared. The concentration range of these dilutions
spanned from 0.5 µg/mL to 1000 µg/mL.
Incubation: The microplates containing the bacterial cultures and test
compounds were incubated for 24 hours at 37°C to allow interaction between
the bacteria and the compounds.
Determination of minimum inhibitory concentration (MIC): MIC
was identified as the lowest concentration of the test compound at which no
visible bacterial growth was observed after the incubation period. The
absence of visible turbidity in the wells indicated effective bacterial
inhibition.

Conclusions
The research paper focused on the synthesis of three novel
imidazopyridine compounds derived from 2,3-diaminopyridine. The
derivatives were tested for their antibacterial activity against several bacterial
strains. The findings showed that all the synthesized compounds exhibited
notable antimicrobial effects, with compound 3 demonstrating the strongest
activity, particularly against Escherichia coli. This study suggests that these
new derivatives are promising candidates for further investigation into their
potential as antibacterial agents. The results highlight the importance of these
compounds for future therapeutic applications.

Acknowledgements
The authors would like to express our sincere gratitude to the Head of the
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Kufa,
for her invaluable support, guidance, and encouragement throughout the course of
this research.
246 Zeyad Kadhim Oleiwi et al.

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