Phytochemistry Letters 54 (2023) 23–27

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Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol

Semi-synthesis of novel 1,4-disubstituted-1,2,3-triazole derivatives of

penicillin G and their antibacterial activity
Kaveh Rasoolijokari a, Peyman Salehi a, *, Atousa Aliahmadi b, Bahareh Heidari a,
Morteza Bararjanian a
a
Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Evin, 1983963113 Tehran, Iran
b
Department of Biology, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, Iran

A R T I C L E I N F O A B S T R A C T

Keywords: A series of 12 novel 1,2,3-triazole penicillin G derivatives were subject to synthesis starting from propargylation
Penicillin G of penicillin G followed by copper(I) catalyzed azide alkyne cycloaddition (CuAAC) click reaction. Also, one-pot
Antibacterial Strecker reaction was applied aiming for the preparation of 5 novel α-amino nitrile triazole tethered penicillin G
CuAAC reaction
derivatives. The derivatives’ chemical structures were confirmed by means of 1H NMR, 13C NMR, FT-IR, and HR-
Click chemistry
MS. The synthesized compounds (5a-5 l and 10a-10e) were evaluated in vitro as antimicrobial agents against
Strecker reaction
Staphylococcus aureus plus Enterococcus faecalis as Gram-positive and Escherichia coli as Gram-negative bacteria,
and the ultimate results were compared with penicillin G. Compounds 5a and 5b revealed the uppermost potency
in inhibiting bacterial growth with the minimum inhibitory concentration (MIC) value of 0.049 µg/ml against
S. aureus. Amongst α-amino nitrile derivatives, 10d and 10e exhibited greater potency in inhibiting bacterial
growth (MIC=0.19 µg/ml) against S. aureus. Moreover, an investigation of minimum bactericidal concentration
(MBC) of 5a-5 l and 10a-10e derivatives put on display the fact that 5b and 5 h were potent to kill the assessed
S. aureus cells in concentration just two fold above that of penicillin G’s.

1. Introduction et al., 2013). Shortly after, however, penicillin resistance turned to be a

Antibiotic microbial resistance (AMR) is a natural defect occurred to
antibiotics through which bacteria evolve to withstand drugs’ action
caused by overuse and/or misuse of drugs and antibiotics. In April 2014,
World Health Organization (WHO) first report on the surveillance of
AMR, exposed us to the reality that the phenomenon has been observed
all around the world (Prestinaci et al., 2015). Resultantly, an
ever-increasing demand for novel and nontoxic antibacterial drugs to
treat bacterial infections has hit pharmaceutical industries. In an effort
to overcome the above mentioned obstacle or as a minimum, to post­
pone the matter for a longer while, faster development of biologically
active antibacterial agents that are new as well as more responsible
utilization of currently available antibiotics are urgently on demand by
the whole world.
β-Lactam antibiotic development was known as a successful con­
trolling agent against bacterial infections during World War II. The
penicillin antibiotics contain a nucleus of 6-aminopenicillanic acid
discovered by Sir Alexander Fleming in 1928 (Piddock, 2012; Sengupta
substantial clinical headache, threatening possible advancements of the
prior decade (Spellberg and Gilbert, 2014).
Despite the emergence of resistance, β-lactams nevertheless remain
the most widely utilized antibiotic class owing to a number of their fine
features namely their potent activity, comparatively high effectiveness,
ease of delivery, low toxicity, and the last but not the least, minimal side
effects (King et al., 2016).
There exist three main routes of bacterial resistance to β-lactam an­
tibiotics: hydrolysis of antibiotic by β-lactamases, development of cell
wall transpeptidases which is resistant to β-lactam antibiotics, and
finally the presence of efflux pumps in Gram-negative as well as Gram-
positive bacteria excreting unknown molecules from bacterial cells
(Wilke et al., 2005).
To overcome drug resistance, the synthesis of analogs of current
drugs plays a crucial role in the design of new candidates and along the
drug discovery process. Heterocyclic compounds have obtained
considerable attention in the field of medicinal chemistry due to their
broad spectrum of their biological activities (Choudhary and Silakari,

* Corresponding author.
E-mail address: p-salehi@sbu.ac.ir (P. Salehi).

https://doi.org/10.1016/j.phytol.2023.01.005
Received 2 July 2022; Received in revised form 24 December 2022; Accepted 4 January 2023
Available online 18 January 2023
1874-3900/© 2023 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.
K. Rasoolijokari et al. Phytochemistry Letters 54 (2023) 23–27

Scheme 1. Synthesis of propargylated penicillin G (3).

Scheme 2. Synthesis of penicillin G triazole derivatives 5a-5l.

Scheme 3. Synthesis of α-amino nitrile 1,2,3-triazole tethered derivatives of penicillin G using Strecker reaction.

2018; Liu et al., 2011; Su et al., 2013). Among N- and S-containing
heterocycles, imidazole, indole, pyridine, quinaxoline, and triazole rings
are especially more appealing (De et al., 2021). The 1,2,3-triazole and its
derivatives are well-known nitrogen-containing aromatic heterocycles
that have received a greater portion of attention due to their diverse
biological activities, such as antitubercular (Gill et al., 2008),
antibacterial (Holla et al., 2005; Slootweg et al., 2014), anti-HIV (da
Silva et al., 2009), antifungal (Aher et al., 2009), and anticancer (Kamal
et al., 2008; Nemati et al., 2020; Queiroz et al., 2019). 1,2,3-Triazole
moiety presents marked stability under oxidative, reductive, and hy­
drolytic conditions. The structural feature of 1,2,3-triazole ring is
capable to mimic various functional groups, justifying its wide

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K. Rasoolijokari et al. Phytochemistry Letters 54 (2023) 23–27

Table 1
Synthesis of 1,2,3-triazole tethered penicillin G derivatives.

1 4a 5a 98 7 4g 5g 60

2 4b 5b 95 8 4h 5h 89

3 4c 5c 88 9 4i 5i 98

4 4d 5d 98 10 4j 5j 90

5 4e 5e 68 11 4k 5k 98

6 4f 5f 64 12 4l 5l 91

utilization as amide bioisostere in the synthesis of new active com­
pounds (Bonandi et al., 2017).
The α-amino nitriles possesses a biological and practical prominence
and this structural motif has been found in the structure of unalike al­
kaloids (Kouznetsov and Galvis, 2018). The Strecker reaction is the first
multi-component procedure for the synthesis of α-amino nitriles via re­
action of an aldehyde, an amine, and either alkaline metal cyanide or
hydrogen cyanide (Strecker, 1850).
Considering the significance of the 1,2,3-triazole heterocycle as the
main component of countless biologically active compounds as well as
in the pursuit of our own interest in the synthesis of its derivatives
(Gharehnaghadeh et al., 2020; Hasanpour et al., 2021; Nami et al.,
2022), hereby we present the synthesis of a library of 12 novel 1,2,3-tri­
azole derivatives as well as a series of 5 novel α-amino nitrile triazole
tethered compounds of penicillin G. In order to evaluate the efficacy of
the synthesized products as antibacterial agents, all compounds were
assessed in vitro against two Gram-positive and a Gram-negative bacte­
ria, and the results have been compared with penicillin G.
2. Results and discussion
2.1. Chemistry
Consequently, for the synthesis of the target triazole-penicillin G
derivatives (5a-5 l), various azides (4a-4 l), in the presence of sodium
ascorbate and Cu(OAc)2 in H2O/DCM/MeOH (1:1:1 v/v) were reacted
with 3 at room temperature (Scheme 2). The results are summarized in
Table 1. To obtain a structurally rich library of novel products, a wide
variety of azides were applied including phenyl azide derivatives with
varied electron-withdrawing substituents such as chlorine (4d), fluorine
(4c), nitro (meta and para-substituted 4a and 4b), aldehyde (4 g), and
carboxylic acid (4e and 4 f) as well as electron-releasing groups such as
methyl (4i), ethyl (4j) and methoxy (4 h), and also benzyl azide (4k)
and 3-fluorobenzyl azide (4 l).
The second part of this investigation corresponded to the synthesis of
akin structures containing an α-amino nitrile moiety connected to the
triazole ring. The Strecker reaction of an aldehyde, an amine, and so­
dium cyanide, conducted in acetic acid as solvent, was employed to
provide compounds 9a-9e (Scheme 3). To obtain a library of novel
products, a variety of amines were used including two aniline de­
rivatives (9a and 9b), benzyl amine (9c) and two amino acid analogs
(valine and phenylalanine methyl esters 9d and 9e).
Finally, the obtained compounds were reacted with penicillin G
propargyl ester (3) to produce the target molecules 10a-10e (Scheme 3)
and the results are summarized in Table 2.

Our strategy for the synthesis of target molecules is presented in
Schemes 1–3.
To achieve triazole derivatives of penicillin G, the first step was
propargylation of its carboxylic acid group. Thus, reaction of penicillin
G with propargyl bromide in DMF, resulted in the formation of the
corresponding ester (3) in high yield (Scheme 1).
2.2. Antibacterial activity evaluation
The present study’s main goal was to prepare a series of novel 1,2,3-
triazole penicillin G derivatives to assess their potency as antibacterial
agents. While penicillins are capable of inhibiting Gram-positive and
many Gram-negative and anaerobic bacteria, penicillin G is effective

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K. Rasoolijokari et al. Phytochemistry Letters 54 (2023) 23–27

Table 2 Determination of the MIC values of 1,4-substituted-1,2,3-triazole

Synthesis of α-amino nitrile derivatives of penicillin G.
1 10a
2 10b
3 10c
derivatives of penicillin G (5a-5 l) showed that compounds 5a and 5b
displayed the highest potency in inhibiting S. aureus growth with the
92 MIC value of 0.049 µg/ml (Table 3). Apparently, presence of a nitro
group on the aryl ring has a positive impact on the antibacterial activity
of the products.
Also, the MIC values of α-amino nitrile compounds attached to the
triazole ring of penicillin G depicted that 10d and 10e possessed greater
potency to inhibit bacterial development (MIC=0.19 µg/ml) than other
compounds (10a, 10b, and 10c) against S. aureus. It seems that presence
92
of an amino acid in the structure of the products, boosts the antibacterial
activity of α-amino nitrile triazolyl compounds.
Also, investigation of MBCs of 5a-5 l and 10a-10e derivatives
revealed that 5b and 5 h were capable of killing assessed S. aureus cells
in concentration just two fold above that of penicillin G (Table 3).
Throughout the present study we evaluated the synthesized compounds
95 against E. coli in an attempt to find new derivatives being capable of
inhibiting this model of Enterobacteriaceae. However, none of the tested
compounds could inhibit this bacterium at concentrations below that of
penicillin G.

4 10d 89
5 10e 85
against Gram positive such as penicillin-susceptible S. aureus, Strepto­
coccus pneumoniae, Streptococcus pyogenes, viridans group streptococci,
Streptococcus bovis, and some Gram-negative organisms such as Neisseria
gonorrhoeae and Neisseria meningitides. Enterobacteria such as E. coli are
more susceptible to ampicillin and cephalosporines than penicillin G
(Yao et al., 2007)
3. Conclusion
In the present study, we did our best to properly report the synthesis
and antibacterial evaluation of several novel penicillin G-derived 1,2,3-
triazole products via CuAAC click chemistry and Strecker reactions in
various yields ranging from good to excellent. All compounds were
characterized and investigated in vitro for their antibacterial activity
against S. aureus and E. faecalis as Gram-positive and E. coli as Gram-
negative bacteria. The results revealed that the compounds exhibited
superior antimicrobial activity against the assessed Gram-positive bac­
teria. The MIC results of the synthesized series put on display the horizon
that there were some promising compounds (5a and 5b as well as 10d
and 10e) that inhibited S. aureus growth and also for eradication of this
bacterial strain (5b and 5 h). Also, E. coli assessed strain was neither
inhibited nor killed by any of the compounds at concentrations below
25 µg/ml.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence

Table 3
Antibacterial evaluations of 1,2,3-triazole penicillin G derivatives.
Entry Product MIC µg/ml Entry Product MIC µg/ml

S. aureus E. faecalis S. aureus E. faecalis

1 Pen G 0.024 0.78 10 5i 0.098 12.5

2 5a 0.049 25 11 5j 0.098 25
3 5b 0.049 25 12 5k 0.19 25
4 5c 0.098 25 13 5l 0.098 25
5 5d 0.19 25 14 10a 0.78 25
6 5e 0.19 25 15 10b 0.39 25
7 5f 0.78 25 16 10c 0.39 25
8 5g 0.39 12.5 17 10d 0.19 12.5
9 5h 0.098 25 18 10e 0.19 12.5

Entry Product MBC µg/ml Entry Product MBC µg/ml S. aureus

S. aureus
1 Pen G 0.39 10 5i 12.5
2 5a 6.25 11 5j 6.25
3 5b 0.78 12 5k 12.5
4 5c < 12.5 13 5l < 12.5
5 5d < 12.5 14 10a < 12.5
6 5e 3.125 15 10b 12.5
7 5f 12.5 16 10c 12.5
8 5g < 12.5 17 10d 6.25
9 5h 0.78 18 10e 12.5

26
K. Rasoolijokari et al.
the work reported in this paper.
Data availability
No data was used for the research described in the article.
Acknowledgments
The authors acknowledge partial support of Research and Technol­
ogy council of Shahid Beheshti University.
Author contributions
The manuscript was written through contributions of all authors. The
final version of the manuscript has received approval by every one of the
authors.
Appendix A. Supporting information
Supplementary data associated with this article can be found in the
online version at doi:10.1016/j.phytol.2023.01.005.
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