European Journal of Medicinal Chemistry 264 (2024) 116037

Contents lists available at ScienceDirect

European Journal of Medicinal Chemistry

journal homepage: www.elsevier.com/locate/ejmech

Review article

Click chemistry-aided drug discovery: A retrospective and

prospective outlook
Rui Zhao a, b, 1, Junlong Zhu a, b, 1, Xiaoying Jiang a, b, Renren Bai a, b, *
a
School of Pharmacy, Hangzhou Normal University, Hangzhou, 311121, PR China
b
Key Laboratory of Elemene Class Anti-cancer Chinese Medicines, Engineering Laboratory of Development and Application of Traditional Chinese Medicines,
Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, 311121, PR China

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

Keywords: Click chemistry has emerged as a valuable tool for rapid compound synthesis, presenting notable advantages and
Click chemistry convenience in the exploration of potential drug candidates. In particular, in situ click chemistry capitalizes on
In situ click chemistry enzymes as reaction templates, leveraging their favorable conformation to selectively link individual building
Compound library
blocks and generate novel hits. This review comprehensively outlines and introduces the extensive use of click
1,2,3-Triazole
chemistry in compound library construction, and hit and lead discovery, supported by specific research exam­
Drug discovery
ples. Additionally, it discusses the limitations and precautions associated with the application of click chemistry
in drug discovery. Our intention for this review is to contribute to the development of a modular synthetic
approach for the rapid identification of drug candidates.

1. Introduction is frequently applied as a linker of proteolysis targeting chimeras

Researchers have always been trying to find reliable solutions to
break through technological barriers and thus shorten the drug discov­
ery process for decades. Fortunately, the emergence and ongoing
development of artificial intelligence (AI), computer-aided drug design
(CADD), and click chemistry may improve the efficiency and accelerate
the process of drug discovery (Fig. 1) [1–5].
Click chemistry is a combinatorial method initially proposed by
Professor Sharpless for the rapidly synthesis C-X-C atoms [6]. The most
widely used click chemical reaction in organic chemistry, chemical
biology, and biomedical fields is the Husigen 1,3-dipole cycloaddition
reaction (CuAAC) between azides and acetylene. The reaction has
become a classic click chemistry reaction [7–10].
Click chemistry has a wide range of applications in the field of drug
discovery, especially medicinal chemistry. Compared to conventional
radiolabeling strategies, click reactions exhibited many advantages,
including increased radiochemical yield [11], convenience connecting
complex radiotracers [12], and significantly improved availability of
radiopharmaceuticals [13,14]. Moreover, click chemistry also plays a
primary role in protein-lipid modification. It also serves as reasonable
linkers for conjugates or probes [9,15,16]. For example, click chemistry
(PROTACs) [17]. More importantly, large or small compound libraries
can be quickly constructed through click chemistry reactions, providing
a large library for screening active compounds [18]. For a specific drug
target, click chemistry combines two similar or different pharmacophore
groups through a specific skeleton, the triazole heterocyclic ring, to
obtain a compound with high affinity (Fig. 2) [19,20]. This approach
also makes click chemistry a viable lead compound discovery strategy
[21]. If lead compounds or pharmacophores are unavailable for a certain
target like enzyme, in situ click chemistry can be used to directly
generate hits with research value in the binding pocket of the target. The
process of synthesizing and screening enzyme inhibitors can be
streamlined [22].
Undertaking a comprehensive review of the complete applications of
click chemistry in medicinal chemistry is a daunting task requiring
extensive content and ample examples. Due to limitations in space, word
count and journal scope, this review will primarily focus on the signif­
icant aspects of click chemistry in drug discovery, with an emphasis on
medicinal chemistry. Specifically, we summarized its applications in the
generation of compound libraries, as well as the discovery of leads and
hits. Furthermore, we also discussed the challenges and prospects
encountered in the utilization of click chemistry in drug discovery.

* Corresponding author. School of Pharmacy, Hangzhou Normal University, Hangzhou, 311121, PR China.
E-mail address: renrenbai@hznu.edu.cn (R. Bai).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.ejmech.2023.116037
Received 22 October 2023; Received in revised form 20 November 2023; Accepted 8 December 2023
Available online 12 December 2023
0223-5234/© 2023 Elsevier Masson SAS. All rights reserved.
R. Zhao et al. European Journal of Medicinal Chemistry 264 (2024) 116037

Ultimately, this review aims to provide pharmaceutical chemists with a

valuable point of reference for their future research endeavors involving
click chemistry.

2. Rapid construction of compound libraries assisted by click

chemistry

The hit-to-lead stage is a crucial and time-consuming aspect of drug

discovery, also the most expensive [23]. However, the introduction of
CuAAC click chemistry and in situ screening has expedited this process
while enhancing both its efficiency and the quality of drug discovery
[24,25]. This approach offers the advantage of rapidly and efficiently
obtaining lead compounds and testing their activity results. After the
discussion of the structure-activity relationship (SAR), the lead com­
pounds were screened again to form a virtuous circle [24]. This
approach had great potential in the field of medicinal chemistry (Fig. 3)
[26].
Chan’s group reported the preparation of flavonoid dimers by
CuAAC reaction, which was used as the MPR1 inhibitors. Compound 1
(Fig. 4) exhibited significant inhibitory activity and included two
identical flavonoid moieties. Moreover, PEG was used as the linker to
couple two identical moieties, showing potent inhibitory activity. Af­
terward, this group prepared a library of triazole compounds with two
distinct flavonoid fragments. A 300-member MPR1 inhibitors library
was assembled in microtiter plates without any purification, among Fig. 2. Summary of click chemistry application in medicinal chemistry.
which compound 2 has shown a potent inhibitory effect (EC50 = 53 nM)
[27]. click chemistry in 2013. In this work, all library compounds were easily
HDAC3 is a member of the HDAC family and belongs to Type I HDAC prepared by treating nine alkynes with fifty-five azides, and evaluated
(1, 2, 3 and 8). Abnormal increases in HDAC3 are involved in the for inhibitory effect on HDAC3 and two compounds (7 and 8) displayed
development of many diseases, including cancer [28], inflammation potent and selective HDAC3 inhibitory effects [18].
[29], and neurodegenerative diseases [30]. In 2012, Miyata and col­ Previous investigations have shown that quinone-containing frag­
leagues designed a library of HDAC8 inhibitors via click chemistry. ments play an important role in inhibiting Cdc25 phosphatase activity.
Further primary screening at a concentration of 3 μM of the product To find Cdc25 phosphatase inhibitors with strong inhibitory activity and
revealed that compounds 3–6 displayed considerable inhibition on high subtype selectivity, Zhan et al. designed a quinone derivatives li­
HDAC8 with IC50 values of 0.35, 0.18, 0.10, and 0.007 μM, respectively. brary of candidate Cdc25-selective inhibitors, in which two types of
At first, the CuAAC reaction between eight alkynes and fifteen azides alkynes and forty-eight azides were connected by a triazole-containing
provides a 120-member library. Fortunately, without undergoing puri­ linker. Finally, a 96-member Cdc25-selective inhibitors library was
fication, two crude compounds 3 and 4 exhibited potent inhibitory ac­ assembled in microtiter plates using the CuAAC reaction. As shown in
tivity against HDAC8. Subsequently, to prepare HDAC8-selective Fig. 3, compound 9 exerted an obvious inhibitory effect on the Cdc25
inhibitors with more significant activity than the control 4, they subtype with an IC50 value of 0.09 μM [32].
generated a 31-trizole library by click chemistry. Notably, compounds 5 For further structural optimization, this group used click chemistry
and 6 show more potent inhibitory activity than compound 4 at a con­ between the key intermediates alkynes and a set of 37 azides gave a
centration of 0.2 μM. The triazole products were confirmed by TLC and library including quinone fragments. Then, those crude compounds
LCMS [31]. were tested for activity. Notably, compound 10 showed more potent
On the base of the previous investigations, this group further re­ inhibition with an IC50 value of 0.06 ± 0.04 μM, which was equal to the
ported a 504-member library of compound HDAC3 inhibitors through control NSC663284 (IC50 = 0.23 ± 0.01 μM) [33].

Fig. 1. Click chemistry serves as a useful strategy and accelerates the process of hits and leads discovery.

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R. Zhao et al. European Journal of Medicinal Chemistry 264 (2024) 116037

Fig. 3. Compound library rapid generation by click chemistry.

Fig. 4. The chemical structures of hits 1–10 targeting MPR1, HDAC, and Cdc25.

3. Hit discovery aided by in situ click chemistry
Target-guided synthesis (TGS) is a synthesis method of enzyme in­
hibitors based on the selective assembly of complementary functional
groups of biological targets (e.g. enzymes) as templates, mainly divided
into dynamic combinational chemistry (DCC) and dynamic TGS [34].
The application of copper-catalyzed cycloaddition reactions of azides
and alkynes in TGS is named in situ click chemistry [35]. The in situ click
chemistry technique, in contrast to conventional fragment-based drug
design, can use biological targets to chemically construct their inhibitors
in a templated way by creating covalent links between fragments.
Developing novel, extremely effective inhibitors of AChE and CA has
shown in situ click chemistry to be a revolutionary drug discovery
technique to synthesize more effective self-enzyme inhibition under the
physiological activity of enzymes (Fig. 5) [36]. In 2002, Sharpless’s
group first discovered and demonstrated an effective AChE inhibitor

3
R. Zhao et al.
Fig. 5. Discovery of potent “hit” by in situ click chemistry stratergy.
through in situ click chemistry [37].
3.1. Hits targeting acetylcholinesterase
The first target for the application of in situ click chemistry was
AChE.
In 2005, Hartmuth et al. discovered a potent AChE inhibitor by in
situ click chemistry, which confirmed the great potential of in situ click
chemistry in medicinal chemistry (Fig. 5) [22]. Inspired by
AChE-selective double binding sites, Renard’s group designed novel
hupine inhibitor heterodimers. Compound 11 (Fig. 6) showed potent
inhibitory activity against recombinant human-AChE (rh-AChE) ligand,
with an IC50 value of 0.43 nM. Notably, the synthesis of enzyme in­
hibitors was thermodynamically driven to control the regional selec­
tivity (syn or anti) of Huisgen reactions, leading to the successful
identification of effective enzyme inhibitors [38]. Akbarzadeh and his
colleagues discovered a series of novel 1,2,3-triazole derivatives by in
situ click chemistry. Among these, compound 12 demonstrated obvious
inhibition against AChE (IC50 = 7.31 μM). The results of molecular
docking showed that the planar structure of 1,2,3-triazole was closely
Fig. 6. The chemical structures of hits 11–23 targeting AChE and COX.
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European Journal of Medicinal Chemistry 264 (2024) 116037
related to the activity [39].
3.2. Hits targeting cyclooxygenase
COX is a heme protein located in the cell membrane, existing in two
distinct subtypes: constitutive COX-1 and inducible COX-2. COX-2 plays
a critical role in promoting inflammation.
Wuest et al. reported a series of COX-2 inhibitor derivatives, which
were prepared by treatment azide and alkyne groups at the COX-2
binding site by in situ click chemistry. Two potent COX-2 inhibitory
and selective compounds, 13 (triacoxib, IC50 = 90 nM) and 14 (IC50 =
50 nM), were successfully identified, which exhibited anti-inflammatory
activity similar to the control (celecoxib, IC50 = 70 nM). In the
carrageenan-induced rat paw edema model, compounds 13 (ED50 =
0.44 mg/kg) and 14 (ED50 = 0.12 mg/kg) showed potent anti-
inflammatory activity, superior to celecoxib (ED50 = 10.8 mg/kg)
[40]. Similarly, the same group reported radiotracer [18F]triacoxib (15)
for PET imaging of COX-2 by in situ click chemistry. The
anti-inflammatory activity of compound 15 was similar to [18F]pyr­
icoxib both in vitro and in vivo. In HCA-7 tumor-bearing mice model,
R. Zhao et al.
compound 15 was found to bind specifically to COX-2, confirming that
compound 15 was an excellent radiotracer [41].
Based on the structure of COX-2 inhibitors, it was found that most of
the compounds contained five-membered heterocyclic rings. Wuest
et al. synthesized five 1,4-diaryl-1,2,3-triazole derivatives by Cu(I)
catalyzing the cycloaddition reaction between 1-azido-4-methane sul­
fonyl benzene and phenylacetylene containing different p-substituents.
Among these, phenyl substituted derivative 16, fluorophenyl substituted
derivative 17, and chlorophenyl derivative 18 exerted promising ac­
tivities, with IC50 values ranging from 0.15 to 0.21 μM. More impor­
tantly, the substitution of 1,2,3-triazole of the central cyclopene portion
can reduce the lipophilicity of the compound and thus serve as a tracer
of in vivo radioactivity by reducing non-specific binding in vivo [42].
Kharbanda’s group also identified a series of 1,2,3-triazole-contain­
ing compounds as inhibitors of COX through in situ click chemistry.
Treatment of 2-mercaptobenzooxazolyl compounds and various
substituted aromatic azides gave a library which compounds containing
a double heterocyclic ring structure. Further biological evaluation
proved that compounds 19, 20, 21, 22, and 23 significantly reduced
inflammation and the absence of gastric ulceration. The result indicated
that they demonstrated potential anti-inflammatory medication prop­
erties. Furthermore, compounds 19 and 21 had a higher COX-2 selec­
tivity (IC50 = 3.8 μM and 2.8 μM, SI = 64.79 and 66.47, respectively)
[43].
3.3. Hits targeting HIV-1
HIV-1 protease is a target for treating acquired immunodeficiency
syndrome (AIDS). Inhibiting HIV protease activity disrupts HIV’s ability
to replicate and infect.
Using in situ click chemistry, Whiting et al. synthesized a triazole
compound 24 (Fig. 7), which used to be an inhibitor of HIV-1-Pr and its
mutants (IC50 = 6 nM, Ki = 1.7 nM) [44]. However, single-target drug
has the disadvantage of easy drug resistance. To address this issue, the
application of dual-target or multi-target provides a new idea for the
discovery of anti-HIV drugs. Brik et al. retained the core structure of
hydroxyethylamine in HIV-1-Pr marketed drugs (e.g. Amprenavir [45]
and Nelfinavir [46]) and used it to generate novel inhibitors. The
promising compounds 25–28 were obtained from a 100-member library
that showed potent inhibitory activity against HIV-1-Pr at the concen­
tration of 10 nM. Notably, the inhibition of compounds 25 and 27 on
Fig. 7. The chemical structures of hits 24–29 targeting HIV-1.
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European Journal of Medicinal Chemistry 264 (2024) 116037
HIV-1-Pr, G48V, V82F, and V82A were as low as nanomolar (25: IC50 =
6 ± 0.5 nM, 19 ± 1 nM, 39 ± 1 nM, 46 ± 2 nM, 27: IC50 = 13 ± 0.5 nM,
24 ± 1 nM, 17 ± 1 nM, 52 ± 2 nM, respectively) [47].
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) occupy a
large proportion in the treatment of HIV disease due to their high
antiviral ability. Targeting HIV-1 reverse transcriptase (RT), Kang’s
group screened out 22 compounds that displayed markedly suppressed
HIV NNRTI by in situ click chemical. Within this series, compounds
comprising 1,2,3-triazole exhibited strong activity (EC50 = 3.28–10.4
nM) superior to the control NVP (EC50 = 163 nM). In particular, com­
pound 29 demonstrated the best inhibition against HIV-1 NNRTI (IC50
= 3.28 nM) and the lowest cytotoxicity (CC50 > 210 μM) [48].
3.4. Hits targeting abelson tyrosine kinase
The earliest Bcr-abelson (Abl) tyrosine kinase inhibitors (TKIs) were
used for the treatment of chronic myelogenous leukemia and acute
lymphoblastic leukemia. However, due to the existence of drug resis­
tance and mutants, the development of new TKIs is of great clinical
significance.
The CuAAC reaction between azide and acetylene allowed Peruz­
zotti’s group to generate a small candidate library, which can bind to the
active position site of Abl. Within the series, compound 30 (Fig. 8)
exhibited inhibitory activity against K-562 cell line and tyrosine kinase
with IC50 values of 0.89 μM and 0.9 ± 0.1 μM, respectively [49].
Kalesh et al. designed a library of candidate Abl inhibitors containing
about 344 compounds using in situ click chemistry by treating ADP-
alkynes and azides. The SAR study of the library showed that the
short-chain C-2 position of azides demonstrated remarkably inhibitory
activity. The previous result showed that the piperazine fragment of
imatinib did not show a significant contribution to the inhibition of Abl.
Consequently, based on the structural information of imatinib and the
primary screening of products, 90 triazole compounds were synthesized,
containing the core structure of the imatinib. 11 compounds exhibited
effective inhibition against Abl over Src kinase. Compounds 31 and 32
displayed remarked inhibition against Abl (IC50 = 0.70 μM and 1.12 μM,
respectively), while the inhibition of the parent of compounds 31 and 32
with IC50 values of 1.36 μM and 2.73 μM, respectively [50].
R. Zhao et al.
Fig. 8. The chemical structures of hits 30–46 targeting Abl, CA, Chi, and OGA.
3.5. Hits targeting carbonic anhydrase
CA is a zinc-containing enzyme catalyzing the mutual conversion of
bicarbonate and carbon dioxide. Abnormal elevation of CA has been
observed in a variety of diseases, including tumors and AD.
Kugler et al. utilized in situ click chemistry to design five inhibitors
targeting hCA-II. Among these, compound 33 exerted potent inhibitory
activity (Ki = 2.5 ± 1.4 nM and SI = 4). Notably, the Ki of compound 33
was higher than the parent (20-fold) [51].
Sulfonamide and glycoconjugates were identified as pharmaco­
phores for CA inhibitors. Poulsen’s group designed a series of potential
CA inhibitors. 12 compounds presented potent inhibition against bCA-II
(Kds = 0.2–7.1 nM). Notably, compounds also showed higher affinity
due to the higher selection of the enzyme [52]. Wilkinson et al. syn­
thesized a series of glycoconjugate benzene sulfonamide compounds
using in situ click chemistry, which treated the carbohydrate azides and
arylsulfonamide alkynyl. Within the series, compounds 34 and 35 dis­
played potent suppressed hCA-II isozyme activity (Ki = 7.5 nM and 2.3
nM, respectively), superior to the parent alkyne (Ki = 5.1 nM) [53].
Based on these results, Michaela et al. developed a PET [18F] mo­
lecular imaging probe compound 36 using in situ click chemistry, which
can confirm the expression of CA-II in vivo [54]. Additionally, an inte­
grated microfluidic device for large-scale in situ click chemistry can
remarkably decrease reagent consumption and screening time [55].
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European Journal of Medicinal Chemistry 264 (2024) 116037
3.6. Hits targeting chitinase
Chitinase (Chi) is the main component of fungal cell walls, and Chi
inhibitors have the potential of bactericidal, insecticidal, and asthmatic
in chemotherapy.
Argifin (37) is a cyclic pentapeptide natural product, which was
proved to be an effective Chi inhibitor (SmChiA: IC50 = 0.025 μM;
SmChiB: IC50 = 6.4 μM). In particular, Nw-methylcarbamoyl-L-arginine
scaffold was considered as the key moiety of Argifin, which can be used
as a model to obtain more potent inhibitory activity than Argifin [56].
Derivative 38 displayed potent inhibitory activity against SmChiB with
an IC50 value of 0.58 ± 0.044 μM. Subsequently, using in situ click
chemistry, Hirose et al. constructed a 1,2,3-triazole candidates library.
Compound 39 was prepared by compound 38 and alkyne comprising
dibenzylamide analog and displayed significant Chi inhibitory against
SmChiB (IC50 = 0.022 μM), superior to the azide precursor. Further
X-ray crystallography and the control assay without the presence of Chi
showed that SmChiB can promote the formation of compound 39 [56,
57].
The CuAAC reaction was chosen as the linking method to couple
erythronolide A oxime alkyne with azide comprising N-methyl­
carbamoylguanidinyl fragment to generate compound 40 at C3 position
by Sunazuka’s et al. Simultaneously, compounds with C5 and C9 sub­
stitution positions were synthesized by the same procedure. The results
indicated that compounds with C3, C5, and C9 substitution exhibited
remarkably Chi inhibitory activity against SmChiB, with IC50 values of
R. Zhao et al.
3.3 μM, 11.9 μM, and 24.1 μM, respectively. Notably, C3 substitution
induced potent inhibitory activity [58].
3.7. Hits targeting O-GlcNAc
O-GlcNAc (OGA) plays an important role in many basic cellular
processes, and its dysregulation has been linked to the causes of car­
diovascular diseases, type 2 diabetes, cancer, and neurological diseases.
Li et al. reported an efficient synthetic method for the preparation of
a series of 1,2,3-triazole candidates using in situ click chemistry. Further
inhibitory investigation of human OGA revealed that most of the com­
pounds showed effective inhibitory activity against OGlcNAcase. In
particular, compound 41 demonstrated the best potency and lowest
toxicity, with an inhibition of 81.3 % (Ki = 185.6 μM). Notably, the
triazole fragment of compound 41 enhanced interaction with biological
targets via hydrogen bonding and dipole-dipole [59].
By use of in situ click chemistry, a series of glucopyranoside triazole
derivatives library was constructed by Carvalho’s group, which were
prepared from sugar azides and terminal alkynes. Within the series,
compounds 42–44 exhibited significant inhibitory and selective activity
against OGA, with IC50 values of 0.50 ± 0.02 μM, 0.52 ± 0.01 μM, and
0.72 ± 0.02 μM, respectively. Moreover, compounds were shown to be
nontoxic at the concentration of 10 μM. Based on the structure infor­
mation, the presence of heteroatoms (N or O) and the two carbons bridge
between the aromatic ring and five-membered heterocyclic ring may be
the key pharmacophore to the increased activity [60].
Zhang et al. used in situ click chemistry to generate a triazole library
and screened for novel O-GlcNAc transferase (OGT)-selective inhibitors,
among which compounds 45 and 46 showed potent inhibitory against
OGT on COS-7 cells in vitro (IC50 = 66.7 ± 0.8 μM and 139 ± 14 μM,
respectively). Their deacetylate precursors displayed more significant
cytotoxicity in vitro. Besides, they showed no effect on the viability of
normal cells. It was noteworthy that the inhibitory activity of compound
45 increased more than 60-fold comparable with the parent alkynes and
azides, which was comparable to positive drug BZX in vivo [61].
Fig. 9. The chemical structures of anti-AD leads 47–55.
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European Journal of Medicinal Chemistry 264 (2024) 116037
4. Click chemistry-aided drug discovery
4.1. Anti-Alzheimer’s disease leads
AD is a neurodegenerative disease commonly observed in the
elderly. The possible pathogenesis is downregulation of acetylcholine
(ACh) level, activation of oxidative stress, deposition of beta-amyloid
protein (Aβ), and hyperphosphorylation of tan protein. AChE in­
hibitors (tacrine) are widely accepted by researchers for the treatment of
AD. Coumarin exerts a wide range of biological activities, such as anti-
inflammatory, anti-tumor, and anti-AD [62]. Studies have revealed
that coumarin can inhibit Aβ-protein aggregation, oxidative stress, and
ACh degradation. Oxidative damage and neurotoxicity created by H2O2
are considered as most important factors controlling the progress of
neurodegenerative disorder [63].
Foroumadi et al. recently reported that coumarin-lipoic acid conju­
gate 47 (Fig. 9) exerted AChE-selective inhibitory activity with an IC50
of 16.4 μM, and exhibited neuroprotective effect against H2O2-induced
oxidative in PC12 cells and Aβ1-42-induced cytotoxicity in SH-SY5Y cells
[64]. Saeedi and co-workers synthesized a series of 1,2,3-triazole-chro­
menone carboxamides derivatives by click chemistry. Among these
compounds, compound 48 was demonstrated to be a potent AChE and
beta-secretase 1 (BACE1) inhibitor with IC50 values of 1.8 μM and 21.13
μM, respectively. Compound 48 also displayed significant neuro­
protectivity and showed good metal-chelating properties to Cu2+ and
Zn2+ ions [65].
Based on the CuAAC reaction between five alkynes and nine
tryptamine-derived azides, a series of derivatives were prepared.
Further biological evaluation proved that compound 49 exhibited
potent AChE inhibitory activity (IC50 = 18 nM). Interestingly, the linker
triazole moiety increased hAChE activity [66]. Muńoz-Torrero et al.
synthesized a series of huprine derivatives by click chemistry with
cholinesterase (ChE) inhibitor huprine Y and capasaicin, targeting
oxidative stress-related diseases and neuroinflammation diseases.
Among the hybrids, compound 50 exerted potent ChE inhibitory activity
and antioxidant activity, with hAChE and hBChE inhibition of 1.06 nM
and 7.3 nM, respectively. Additionally, compound 50 attained a higher
brain concentration superior to the reference drug donepezil [67].
R. Zhao et al.
Carvalho’s group reported a series of aryl-1,2,3-triazyl benzylpiperidine
inhibitors. Further investigation revealed that compound 51 presented
significant inhibition (IC50 = 0.17 nM) and selective (>58,000-fold) of
hBChE compared to donepezil (IC50 = 9.14 μM) without cytotoxicity
[68]. Khoobi et al. prepared a series of multi-target directed ligand
(MTDL) compounds based on chromones and lipoic acid. Results indi­
cated that compound 52 exhibited inhibitory activity against BChE
(IC50 = 7.55 μM) and potent neuroprotection activity by remarkably
decreasing reactive oxygen species (ROS). As reported before, amid and
disulfide bonds of chelating sites lead to antioxidant effects of compound
52 [69].
Miri and co-workers reported that iminochromene-2H-carboxamide
53 showed high selective BACE1 inhibitory activity and neuroprotective
activity, with IC50 values of 2.2 μM and 2.2 μM, respectively, superior to
the positive control (caffeic acid, IC50 = 75.8 μM) [70].
Xie’s group prepared a series of dual-target coumarin derivatives
against AD. The most potent compound 54 markedly inhibited the iron
chelation activity (pFe3+ = 17.4) and anti-monoamine oxidase-B (MAO-
B) activity (IC50 = 0.68 μM) [71].
Vajragupta’s group used click chemistry to prepare a series of
ascorbic derivatives through an ascorbic core and tryptoline or phenolic
moieties. Investigation on sodium-dependent vitamin C transporter-2
(SVCT2) inhibitory activity and targeting BBB permeation of all ascor­
bic derivatives revealed that compound 55 might interact with the
Fig. 10. The chemical structures of anti-inflammatory leads 56–66.
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European Journal of Medicinal Chemistry 264 (2024) 116037
transporter SVCT2 to cross the blood-brain barrier (BBB) and enter the
brain. Additionally, 55 showed moderate amyloid aggregation inhibi­
tory comparable to the control group (71.79 ± 1.42 % inhibition at a
concentration of 100 μM) and markedly antioxidant effect (IC50 = 72.26
μM). Moreover, the expression of COX-2 and iNOS genes was effectively
reduced by compound 55 [21].
4.2. Anti-inflammatory leads
A series of thioquinazolinone molecules were prepared from 2-Mer­
capto-3-substituted phenyl-quinazlin-4(3H)-ones intermediates and ar­
omatic azides using click chemistry. Compound 56 (Fig. 10)
demonstrated a significant inhibitory effect against COX-1 and COX-2
(IC50 = 2.97 μM and 0.11 μM, respectively, SI = 27) and moderate
inhibitory activity against 15-LOX (IC50 = 7.26 μM). Further monocyte-
to-macrophage differentiation inhibitory tests revealed that compound
56 exhibited a potent effect in comparison to diclofenac (IC50 = 0.11 μM
and 0.8 μM, respectively). Additionally, in vivo Formalin-induced rat
paw edema evaluation showed that compound 56 demonstrated
doubled inhibition compared to celecoxib [72].
Compound 57, prepared by Kharbanda et al., was a 2-mercaptoben­
zoxazole derivative containing 1,2,3-triazoles. Compound 57 displayed
significant inhibition on COX-2 (IC50 = 3.8 μM and SI = 64.79) and the
level of Nitric Oxide (NO). In particular, 57 decreased the expression of
R. Zhao et al.
COX-2 genes (0.94-fold) and exhibited effective gastric mucosa toler­
ance. In particular, it showed potent anti-inflammatory activity with
66.66 % inhibition [43].
Han’s group synthesized a series of hybridizable compounds based
on tryptamine and paeonol by click chemistry. Compound 58 was
demonstrated to be a potent generation inhibitor for NO and STAT-3 in a
dose-dependent manner in an LPS-induced BV2 cell inflammatory
model. More importantly, compound 58 exerted lower toxicity associ­
ated with the free phenolic hydroxyl moiety of peaonol [73].
Haider et al. reported a novel anti-inflammation compound 59 based
on piperine. Anti-inflammation evaluation revealed that compound 59
showed potent in vivo anti-inflammatory activity in a carrageenan-
induced rat paw edema model (80.40 % inhibition) and decreased
TNF-α level in RAW 264.7 cell lines in vitro (73.73 % inhibition), which
was superior to the control piperine (54.72 % and 49.67 % inhibition,
respectively). Simultaneously, compound 59 demonstrated remarked
analgesic and ulcerogenic activity, without cytotoxicity [74].
Serafini’s group synthesized a series of biphenyl triazoles based on
the structure of Synta66 for the treatment of acute pancreatitis. Com­
pound 60, with a 3-carboxyphenyl key moiety, showed a potential
inhibitory activity (87.8 ± 2.9 % inhibition) comparable to Synta66. For
further structural optimization obtained compound 61, displaying
effective inhibitory in HEK cells with an IC50 value of 851 ± 54 nM.
Moreover, compound 61 did not show cytotoxicity at the concentration
of 60 μM and high aqueous solubility. Surprisingly, the 1,2,3-triazole
fragment increased the targeting of the compound [75].
Compound 62 was a dual steric/bitopic agent, synthesized by Man­
era et al. Compound 62 inhibited the production of pro-inflammatory
media IL-6 release and increased the expression of IL-10. The anti-
inflammation activity and neuroprotection of compound 62 were
confirmed in HMC3 cells and a mouse model of nociceptive behavior. In
particular, it was not cytotoxic in microglia [76].
Ghanim’s group designed a series of anti-inflammation candidate
inhibitors based on ibuprofen (Ibu) and indomethacin triazole
Fig. 11. The chemical structures of anti-virus leads 67–79.
9
European Journal of Medicinal Chemistry 264 (2024) 116037
conjugates. These favorable findings indicated that Ibu conjugates
demonstrated potent effects, superior to indomethacin conjugates.
Among these Ibu-triazole conjugates, compounds 63–66 showed sig­
nificant anti-inflammatory, peripheral analgesic, and central analgesic
effects. Noticeably, compounds 63, 64, and 66 were potent and selective
inhibitors of COX-2 (IC50 = 0.743 ± 0.02 μM, 12.35 ± 0.35 μM, and
4.467 ± 0.13 μM, respectively) with the SI values ranged from 2.15 to
23.10 [77].
4.3. Anti-virus leads
A series of 1,2,3-triazole glycosides comprising benzimidazole,
benzoxazole, and benzotriazole complexes were prepared through click
chemistry by Ali’s group. All the compounds were evaluated for antiviral
activity of H1N1, H5N1wild, H5N1V116A, and H5N1N295S. Compound 67
(Fig. 11) displayed a potent antiviral activity with IC50 values of 1.64,
2.25, 1.65, and 55.09 μM, respectively. It was also suggested that
compound 67 was a potent and selective H1N1 inhibitor with an SI
value of 184.1. In further investigation of plaque reduction in vitro and
mice models, compound 67 showed potent antiviral activity. Further­
more, the antiviral activity and safe profiles of compound 67 were su­
perior to the reference drug oseltamivir [78].
Kataev’s group reported a series of 1,2,3-triazolyl nucleoside ana­
logues using CuAAC reaction. The promising compound 68 demon­
strated a significant anti-H1N1 activity (IC50 = 15 μM) and effectively
selective index (SI = 5) but showed moderate cytotoxicity (CC50 = 79
μM). This effect could depend on the polymerase acidic protein of RNA-
dependent RNA polymerase (RdRp). Notably, 1,2,3-triazole nucleoside
derivatives containing 6-menthyluracil and luinazoline-2,4-dione exer­
ted significant antiviral effects, while uridine and thymidine derivatives
demonstrated lower activity against the influenza virus [79].
In 2022, this group further synthesized a series of 1,2,3-triazolyl
nucleoside analogues and their 5-phosphorylated derivatives. The
promising compound 69 showed potent and selective inhibition against
R. Zhao et al.
the H1N1 influenza virus (IC50 = 25 μM, SI = 21), while its parent
compound absence of activity. The result also indicated that 5-triphos­
phate derivative demonstrated significant anti-H1V1 activity,
compared to the nucleoside analogues [80].
He et al. prepared a series of 1,2,3-triazole rupestonic aid derivatives
through click chemistry. Further in vivo biological evaluation proved
that compound 70 demonstrated a significant antiviral effect against
influenza A virus H1N1 (IC50 = 2.82 ± 0.36 μg/mL) and compound 71
showed a potent antiviral effect against influenza B (IC50 = 1.71 μg/mL),
which were superior to the reference drugs oseltamivir (IC50 = 2.81 μg/
mL) and ribavirin (IC50 = 7.84 ± 4.41 μg/mL) [81].
Yu et al. successfully obtained a series of oleanolic acid (OA)-
saccharide complexes by click chemistry, connected by a triazole-
containing linker. All productions were employed for the anti-
influenza assay, and compound 72 showed a significant inhibitory ef­
fect against A/WSN/33 virus (IC50 = 5.47 μM) and displayed a wide
range of biological activities against OSV-p resistant viruses. Moreover,
compound 72 did not show cytotoxicity at the concentration of 100 μM
[82].
Liu et al. designed and synthesized a series of dihydro-alkylthio-
benzyl-oxopyrimidines (S-DABOs) derivatives incorporating 1,2,3-tria­
zole by click chemistry. Among these, compound 73 (EC50 = 3.22 μM)
showed the best activity against HIV-1 in comparison to reference drug
3 TC (EC50 = 2.24 μM). In terms of chemical structures, methyl and
sulfonamide moieties of compound 73 were associated with anti-HIV-1
activity [83].
Rana’s group synthesized a series of 1,2,3-triazoles based on the
structure of RN-18-based viral infectivity factor (VIF). It was noticed
that the 1,4- disubstituede-1,2,3-triazole analogue showed the most
significant anti-HIV activity, with an IC50 value of 1.2 μM. Further
structural modifications provided compound 74, displaying potently
Fig. 12. The chemical structures of anti-infective leads 80–87.
10
European Journal of Medicinal Chemistry 264 (2024) 116037
suppressed HIV-1 (IC50 = 10 nM, H9 cell line), which was superior to the
original lead molecule [84].
Zhan et al. used CuAAC reaction to prepare 1,2,3-triazole-containing
phenylalanine derivatives, among which one potent anti-HIV-1 inhibitor
75 was identified. Compound 75 exhibited significant anti-HIV-1 NL4-3
virus activity with an EC50 value of 4.33 μM (SI > 13.33), comparable to
the original lead compound PF-74. Further surface plasmon resonance
assay showed that compound 75 show potent activity via inhibiting the
early and late stages replication of HIV-1 [85].
Compound 76 was a diarylpyrimidine derivative reported by Liu’s
group, showing promising and selective inhibitory activity against wild-
type HIV-1 strain (IC50 = 0.013 μM, SI > 1000). Additionally, it also
displayed higher water solubility and lower cytotoxicity [86].
Based on the structure of dUY11, a series of 1,2,3-triazole derivatives
were prepared by Korshun’s group, which converse the linker of the
triple bond to 1,2,3-triazole ring. Among those, compounds 77 and 78
showed significant activities against tick-borne encephalitis virus
(TBEV), with EC50 values of 0.031 ± 0.013 and 0.023 ± 0.012 μM,
respectively. In particular, the perylene residue-contained triazolyl
heterocycle was then selected as an antiviral drug fragment for further
study [87].
Aouad et al. synthesized phenylpyrazolone derivatives consisting of
triazole ring and lipophilic aromatic terminals. Within the series, com­
pound 79 exhibited effective inhibitory activity against SARS-CoV-2
Main protease (IC50 = 3.16 ± 1.2 μM) and Vero E6 cells (IC50 = 22.1
μM). Furthermore, compound 79 displayed essentially lower cytotoxic
and higher antiviral effects than the positive drug GC-376 (IC50 = 12.85
μM) at the concentration of 10 μM [88].
R. Zhao et al.
4.4. Anti-infective leads
Zhang et al. reported a series of novel compounds, with 1,2,3-tria­
zoles linked to (2R,3S)-2-(2,4-difluorophenyl)-3-menthyl-1-(1H-1,2,4-
triazol-1-yl)pent-4-yn-2-ol moiety. The promising compound 80
(Fig. 12) demonstrated in vitro potent antifungal activity against
C. albicans SC5314, C. neoformans H99, and A. fumigatus 7544 (MIC =
0.0313 μM, 0.0625 μM, and 1.0 μg/mL, respectively). It also exhibited
potent broad-spectrum and protection in vivo. Moreover, it showed low
cytotoxicity and good safety [89].
Bhattacharya et al. designed a series of novel compounds with 1,2,3-
triazoles linked to glycoconjugate moieties for further antifungal
investigation. Compound 81 displayed excellent antifungal activity
against A.fumigatus, with an IC50 value of 5.42 μM and an MIC value of
10.86 μM. Additionally, compound 81 was not cytotoxic on an un­
transformed cell line with a CC50 value of 43.46 μM and an SI value of
8.01 [90].
Shen et al. chose click chemistry as a linking method to prepare
triazole-piperidine side chains derivatives The result indicated that most
of the derivatives demonstrated broad-spectrum antifungal activity. The
promising compound 82 showed remarkably antifungal activity in vitro,
superior to the control fluconazole and itraconazole, with MIC80 values
from 0.0125 to 2 μg/mL, against Candida albicans, Candida parapsilosis,
and Cryptococcus neoformans strains. It was noteworthy that compound
82 showed potent antifungal activity due to the triazole ring being
associated with the coordination bond and hydrophobic interaction of
CACYP51 [91].
1-alkyl-1H-1,2,3-triazole-4-carboxylic acids (ATCs) incorporating
1,2,3-triazole were prepared based on the structure of cis-2-dodecenoic
acid (BDSF) by Wang’s group. Compound 83 exerted a potent antifungal
activity effect against C. albicans yeast-cell growth and germ-tube for­
mation in vitro (95 % inhibition at the concentration of 200 μM and
length remarkably reduced from 71.9 to 53.3 μM, respectively). The
1,2,3-triazole hybridized structure increases the antifungal properties of
BDSF analogues [92].
Kumar et al. designed a library of antibacterial candidates in which
C12-sphinganine and 1,2,3-triazole analogues were connected by a
triazole-containing linker. The most active compound 84 demonstrated
potent antibacterial activity against C. albicans MTCC 227, C. albicans
854 (MIC = 3.9 μg/ml and 3.9 μg/ml, respectively), and potent sup­
pression on biofilm formation against C. albicans MTCC 227, staphylo­
coccus aureus MTCC 96 (IC50 = 1.9 μg/ml and 2.9 μg/ml, respectively)
[93].
A small library of novel 1,2,3-triazoles bearing β-D-ribofuranosyl
were synthesized by Prasad’s group. Within this series, compound 85
exhibited potent inhibitory activity against DNA gyrase at the concen­
tration of 0.66 μM (IC50 = 0.8 μM) and InhA enzyme at the concentration
of 5 μM. Furthermore, it showed good safety and selectivity compared to
streptomycin in the human THP-1 cell line [94].
Kumar et al. designed a series of chalcone triazole derivatives using
click chemistry. The promising compound 86 showed significant anti­
bacterial activity against E.coli, superior to the positive drug fluconazole
(MIC = 0.0032 μmol/mL and 0.0102 μmol/mL, respectively). Notice­
ably, molecular docking showed that its potent antibacterial effect
depended on the DNA topoisomerase inhibition of the chalcone triazole
structure [95].
The pleuromutilin complexes comprising 1,2,3-triazole fragments
were prepared through click chemistry by Jin’s group. Among which,
compound 87 demonstrated potent antibacterial activity against
multidrug-resistant Staphylococcus aureus (MRSA) (MIC = 0.25 μg/
mL). In the thigh infection and MRSA infection mouse model, compound
87 further showed effective amelioration compared to the standard drug
tiamulin [96].
11
European Journal of Medicinal Chemistry 264 (2024) 116037
4.5. Anti-cancer leads
Xu et al. applied click chemistry to couple 1,2,3-triazole fragments
with different C6 substituted quinazoline scaffolds antiproliferative
evaluation revealed that compound 88 (Fig. 13) showed significant
antitumor activity against human lung cancer A549 and colorectal
cancer HCT116 cells in vitro, with IC50 values of 1.18 μM and 0.36 μM,
respectively. Besides, compound 88 also exerted a broad-spectrum
antiproliferative activity against various cell lines. Results also showed
compound 88 inhibited tumor growth by tethering RNF186 with P62
[97].
He et al. designed a calothrixin A (CAA) compound library, prepared
from various sugar/polyhydroxy azides and CAA using click chemistry.
The most active compound 89 displayed potent antiproliferation activ­
ity against A357 and HCT116 cell lines, superior to the control CPT and
VP-16, with IC50 values of 20 nM, 50 nM, 660 nM, and 280 nM,
respectively. Additionally, the results also showed that compound 89
enhanced antitumor activity and water solubility compared to the
parent compound CAA [98].
Zhang et al. generated 31 triazole compounds processing mel­
ampomagnolide B (MMB) fragments via click chemistry. The promising
compound 90 exhibited potent inhibition on the growth and migration
against HCT116 cell line, superior to the parent compound MMB (IC50 =
0.43 μM and 4.93 μM, respectively). Additionally, it also demonstrated
effective antitumor activity against Bel704, PANC1, A549, and U87 cell
lines, while did not significantly affect the viability of normal cells [99].
He et al. identified a novel series of indoleamine 2,3-dioxygenase-1
(IDO1) and indoleamine 2,3-dioxygenase-2 (IDO2)-selective inhibitors
by using click chemistry, generating the promising candidate compound
91. It exhibited significant antitumor activity against IDO1 and IDO2 in
vitro, with IC50 values of 28 nM and 144 nM, respectively. In the CT26
xenograft tumor mouse nude model, compound 91 exhibited potent
suppression of tumor growth (TGI = 69.7 %) and was nontoxic [100].
Bhadra et al. prepared a series of quinazolinone derivatives incor­
porating triazole fragments for further antiproliferative investigation.
Among which, compound 92 presented significant anticancer against
HeLa cell line (IC50 = 5.94 μM and SI = 43.09) and moderate activity
over MCF-7, K562, and HEK cell lines (IC50 = 8.0 μM, 20.16 μM, and
256 μM, respectively). Notably, compound 92 exhibited an improved
antitumor activity due to the triazole ring interacting with Zn2+-medi­
ated HDACs [101].
Zhu’s group synthesized a series of HDAC8-selective inhibitors based
on PCI-34051 and 1,2,3-triazole rings. Within the series, compound 93
displayed considerable inhibition on A549 cells (IC50 = 9.55 μM)
through reducing proliferation. Furthermore, it also remarkably
decreased HDAC8 protein levels in A549 cells (DC50 = 0.59 ± 0.06 μM).
In particular, co-treatment with compound 93 and irradiation (IR)
demonstrated a significant antiproliferative effect (IC50 = 6.04 μM) su­
perior to either compound 93 or PCI-34051 alone [102].
The synthesis of a library of 1,2,3-triazole derivatives was conducted
to find potent and selective antitumor agents by Raić-Malić et al. Among
these, compound 94 showed more potent and selective inhibition
against non-small cell lung cancer A549 cells, with an IC50 value of 70
nM, which was superior to the 5-FU (IC50 = 2.80 μM). Besides, com­
pound 94 was not cytotoxic in a submicromolar range and suppressed
the TGM2, CDK9, SK1, and p38 MAPK protein kinases [103]. In 2019,
the same group synthesized a series of benzimidazo[1,2-α]quinoline
complexes with 1,2,3-triazole. Compound 95 exhibited potent anti­
proliferative activity against HCT116 and H460 cell lines (IC50 = 0.5 μM
and 0.7 μM, respectively) and significant selection over HaCaT cells
(IC50 = 3.5 μM and SI > 7) [104]. Subsequently, In 2019, Raić-Malić
et al. generated a library of anticancer compounds, namely 4-substituted
6-(1,2,3-triazolyl)-2,3-dibenzyl-L-ascorbic acid derivatives. Among
these, compound 96 demonstrated a significant inhibitory effect against
breast adenocarcinoma (MCF-7) cells (IC50 = 0.08 μM) through the
HIF-1α signaling pathway. Besides, it also showed the best lipophilic
R. Zhao et al.
Fig. 13. The chemical structures of anti-cancer leads 88–103.
(clog P = 5.25) property and selectivity against tumor cells [105].
Additionally, compound 97, one of the coumarin analogues comprising
the 1,2,3-triazole group, was identified. It exhibited the greatest potency
and high selectivity against Hepatocellular carcinoma (HepG2) cells
with an IC50 value of 0.9 μM and an SI value of 50. Besides, it also
moderately suppressed over other tumor cell lines with IC50 values
ranging from 17.48 to 45.33 [106].
The imidazo[2,1-b]thiazole complexes were prepared through click
chemistry reported by Kamal’s group. The most potent compound 98
significantly inhibited the A549 and MCF-7 cell lines (IC50 = 0.778 μM
and 1.013 μM, respectively), superior to the control group (IC50 = 1.57
12
European Journal of Medicinal Chemistry 264 (2024) 116037
μM). Further biological evaluation proved that the antiproliferative ac­
tivity of compound 98 depended on disrupting the microtubule orga­
nization [107]. Subsequently, a series of podophyllotoxin derivatives
with 4β-amidotriazole were synthesized. The further results showed that
compound 99 potently suppressed human tumor cell proliferation
against DU-145, Hela, HepG2, and MCF-7 cell lines (IC50 = 0.7 μM, 0.78
μM, 0.78 μM, and 0.97 μM, respectively) [108].
Zhao et al. designed a library of novel triazole candidates against
dehydrogenase (hDHODH), containing 1,4-benzoquinone fragments.
Compound 100, with a diphenyl skeleton, displayed remarkable inhi­
bition on hDHODH with an IC50 value of 4.5 nM. It also demonstrated
R. Zhao et al.
significant proliferation inhibition over human cancer cell lines with
IC50 values ranging from 0.02 to 9.61 μM. Furthermore, nanocrystalline
samples of compound 100 exhibited a higher safety profile and anti­
tumor activity in vivo [109].
Crook’s group reported a series of MMB derivatives with 1,2,3-tria­
zole rings. Compound 101 showed significant antitumor activity
against the RXF393 renal cancer cell line (GI50 = 20 nM). Besides, it also
suppressed the growth of colon cancer, melanoma, renal cancer, and
breast cancer sub-panels [110].
To discover new MLL1-WDR5 inhibitors, Gao et al. provide a triazole
library containing fragments of phenyl-triazole. All derivatives were
evaluated for antiproliferation activity. Compound 102 exhibited potent
and selective inhibitory against MV4-11 and MOLM-13 cell lines (IC50 =
8.0 μM and 9.9 μM, respectively), with little impact on normal cells
(IC50 > 100 μM). Moreover, it also demonstrated remarkably binding
affinity (IC50 = 8.6 ± 1.3 nM, Kd = 11.6 ± 4.0 nM), proper solubility,
and permeability. The further in vivo results showed that compound 102
displayed potent antitumor activity via reducing the level of H3K4me
and the expression of HOXA9 and Meis1 genes [111].
The 1,2,3-triazole-tethered dehydroabietic acid derivatives were
discovered through click chemistry by Xu’s group. The most selective
compound 103 displayed potent antitumor activity against MDA-MB-
231 cells (IC50 = 0.7 μM), superior to positive drug 5-FU. Furthermore, it
also exhibited a moderate antiproliferative effect on various human
tumor cells with IC50 values ranging from 0.6 to 1.8 μM, including
MCF7, HCT116, PC-3, and SKOV-3 cell lines. Notably, the SAR study
indicated that the potent antiproliferative activity was due to the pres­
ence of 1,2,3-triazole fragment and its C-4 substituents [112].
5. Challenges of click chemistry-aided drug discovery
Click chemistry has demonstrated significant advantages in the rapid
synthesis of compounds, offering considerable convenience in the search
Fig. 14. Sources and clinical applications of triazole heterocycles.
13
European Journal of Medicinal Chemistry 264 (2024) 116037
for potential drug candidates (Fig. 14). However, the application of click
chemistry in drug discovery also presents several challenges and cannot
be considered as a universal solution. When generating a large number
of compounds at a fast pace, it is unclear whether click chemistry en­
sures the desired drug-like properties in the resulting compounds.
Firstly, it is important to note that all click chemistry reactions yield
products with a triazole core structure, leading to significant enhance­
ment of the lipophilicity and consequently reducing the solubility of the
compounds. Poor solubility, a key negative determinant of drug-like
properties, can compromise bioavailability and hinder the further
development of a compound. Additionally, a review of synthesized
compounds reveals that the majority of click chemistry-assisted prod­
ucts possess larger structures, which negatively impacts their drug-like
characteristics and further dissolution, penetration, and absorption.
Lastly, despite the decades since its discovery, click chemistry has pri­
marily aided researchers in identifying multiple hits and leads for spe­
cific targets, but few marketed drugs were successfully discovered by
click chemistry. Consequently, the application of click chemistry in
medicinal chemistry and drug discovery presents both advantages and
disadvantages, along with numerous challenges. Well understanding the
project objectives in utilizing click chemistry is therefore crucial.
6. Conclusion and perspectives
In traditional drug discovery, the design and synthesis of a large
number of compounds typically entail significant time, manpower, and
financial resources, leading to lengthy drug development cycles. How­
ever, the utilization of click chemistry in compound discovery has
considerably reduced the time needed for synthesis. Click chemistry
offers several advantages, including resistance to oxygen and water,
high yields, regioselectivity, and stereoselectivity. These attributes
enable researchers to rapidly prepare target compounds and build both
small and large compound libraries. Additionally, in situ click chemistry
exploits enzymes as reaction templates, capitalizing on their favorable
conformation in physiologically active conditions. This approach
selectively connects individual building blocks to synthesize novel in­
hibitors of the enzyme.
Nevertheless, when employing click chemistry in drug discovery,
careful and deep consideration must be given to balancing and opti­
mizing both biological activity and drug-like properties. Such optimi­
zation ensures that the synthesized active compounds possess desirable
physical and chemical characteristics. Only when this equilibrium is
attained can the compound exhibit favorable dissolution and absorp­
tion, thereby warranting further development. Furthermore, as addi­
tional types of click chemistry reactions are discovered, researchers will
be empowered to swiftly prepare compounds with diverse and intricate
structures, surpassing those solely containing triazoles. This will expand
the chemical space of drug-like compounds and significantly enhance
the potential for identifying promising hits and leads in drug discovery.
CRediT authorship contribution statement
Rui Zhao: Data curation, Investigation, Writing – original draft.
Junlong Zhu: Data curation, Resources, Writing – original draft.
Xiaoying Jiang: Writing – review & editing, Funding acquisition.
Renren Bai: Conceptualization, Data curation, Funding acquisition,
Supervision, Writing – original draft, Writing – review & editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
R. Zhao et al. European Journal of Medicinal Chemistry 264 (2024) 116037

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