7-Chloroquinolinotriazoles: Synthesis by the azide-alkyne cycloaddition click chemistry, antimalarial activity, cytotoxicity and SAR studies

Article abstract of DOI:10.1016/j.ejmech.2013.11.022

Twenty-seven 7-chloroquinolinotriazole derivatives with different substituents in the triazole moiety were synthesized via copper-catalyzed cycloaddition (CuAAC) click chemistry between 4-azido-7-chloroquinoline and several alkynes. All the synthetic compounds were evaluated for their in vitro activity against Plasmodium falciparum (W2) and cytotoxicity to Hep G2A16 cells. All the products disclosed low cytotoxicity (CC₅₀ > 100 μM) and five of them have shown moderate antimalarial activity (IC₅₀ from 9.6 to 40.9 μM). As chloroquine analogs it was expected that these compounds might inhibit the heme polymerization and SAR studies were performed aiming to explain their antimalarial profile. New structural variations can be designed on the basis of the results obtained.

Full text of DOI:10.1016/j.ejmech.2013.11.022

Accepted Manuscript
7-Chloroquinolinotriazoles: synthesis by the azide-alkyne cycloaddition click
chemistry, antimalarial activity, cytotoxicity and SAR studies
Guilherme R. Pereira, Geraldo Célio Brandão, Lucas M. Arantes, Háliton A. de
Oliveira Júnior, Renata Cristina de Paula, Maria Fernanda A. do Nascimento, Fábio
M. dos Santos, Ramon K. da Rocha, Júlio César D. Lopes, Alaíde Braga de Oliveira
PII:
S0223-5234(13)00764-2
10.1016/j.ejmech.2013.11.022
EJMECH 6568
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 19 April 2013
Revised Date: 8 November 2013
Accepted Date: 20 November 2013
Please cite this article as: G.R. Pereira, G.C. Brandão, L.M. Arantes, H.A. de Oliveira Júnior, R.C.d.
Paula, M.F.A. do Nascimento, F.M. dos Santos, R.K. da Rocha, J.C.D. Lopes, A.B.d. Oliveira, 7-
Chloroquinolinotriazoles: synthesis by the azide-alkyne cycloaddition click chemistry, antimalarial
activity, cytotoxicity and SAR studies, European Journal of Medicinal Chemistry (2013), doi: 10.1016/
j.ejmech.2013.11.022.
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ACCEPTED MANUSCRIPT
7
-Chloroquinolinotriazoles: synthesis by the azide-alkyne cycloaddition click chemistry, antimalarial
activity, cytotoxicity and SAR studies
Pereira, G. P.; Brandão, G. C.; Arantes, L. M.; Oliveira, H.; Paula, R. C.; Nascimento, M. F. A.; Santos, F. M.; Rocha, R. K.; Lopes, J. C. D.;
Oliveira, A. B.

ACCEPTED MANUSCRIPT
7
-Chloroquinolinotriazoles: synthesis by the azide-
alkyne cycloaddition click chemistry, antimalarial
activity, cytotoxicity and SAR studies
a
b
c
Guilherme R. Pereira , Geraldo Célio Brandão , Lucas M. Arantes , Háliton A. de
a
a
a
Oliveira Júnior , Renata Cristina de Paula , Maria Fernanda A. do Nascimento , Fábio
M. dos Santos , Ramon K. da Rocha , Júlio César D. Lopes , Alaíde Braga de
d
c
c
a*
Oliveira .
a
Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de
Minas Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, CEP 31270-901, Belo Horizonte,
MG, Brazil
b
Faculdade de Farmácia, UFOP, Rua Costa Sena 171, CEP 35400-000, Ouro Preto, MG,
Brazil
c
Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas
Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, CEP 31270-901, Belo Horizonte, MG,
Brazil
d
Programa de Pós-Graduação em Bioinformática, Instituto de Ciências Biológicas, Universidade
Federal de Minas Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, CEP 31270-901, Belo
Horizonte, MG, Brazil.
*
Corresponding author
Tel. +55 31 3409 6950 Fax: +55 31 3441 5575
E-mails: alaidebraga@terra.com.br
alaide.braga@pq.cnpq.br

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Abstract
Twenty-seven 7-chloroquinolinotriazole derivatives with different substituents in
the triazole moiety were synthesized via copper-catalyzed cycloaddition
(CuAAC) click chemistry between 4-azido-7-chloroquinoline and several
alkynes. All the synthetic compounds were evaluated for their in vitro activity
against Plasmodium falciparum (W2) and cytotoxicity to Hep G2A16 cells. All
the products disclosed low cytotoxicity (CC50 > 100 µM) and five of them have
shown moderate antimalarial activity (IC50 from 9.6 to 40.9 µM). As chloroquine
analogs it was expected that these compounds might inhibit the heme
polymerization and SAR studies were performed aiming to explain their
antimalarial profile. New structural variations can be designed on the basis of
the results obtained.
Keywords:
7-Chloroquinolinotriazoles; Quinolines; Triazoles, Click chemistry; Antimalarial
activity; Plasmodium falciparum; Cytotoxicity in HepG2
1. Introduction
Malaria remains one of the most serious problems in public heath around
the world, despite being one of the oldest diseases. It is estimated that around
00 million people are infected in subtropical and tropical countries and 2.5
5
million deaths occur annually. In Brazil, it is restricted to the Amazonia area.
Malaria is an infectious disease, caused by protozoa parasites from the genus
Plasmodium that is transmitted by mosquitoes of the genus Anopheles. P.
falciparum is responsible for the most lethal form of malaria [1]. Chloroquine (1)
was the most widely and effective antimalarial clinically used drug but parasite
resistance led to its substitution by artemisinin and its semi-synthetic derivatives
(artemether, artesunate). However, emergence of P. falciparum resistance to
these drugs is a serious cause of concern. To prevent the selection of resistant
parasites, WHO has recommended the combination of artemisinins with
traditional antimalarial drugs such as lumefantrine, amodiaquine and mefloquine
and ACT (Artemisinin Combination Therapy) formulations are presently adopted
in most endemic countries [2, 3]. Therefore, new drugs to treat human malaria

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are urgently needed. Synthesis of molecular hybrids containing different
moieties which are representatives of known or putative antimalarial
compounds is presently being extensively exploited. Recently, the synthesis of
1
,2,3-triazoles by a process known as Cu-mediated click chemistry [4] has been
explored to combine different molecules affording new analogues of chloroquine
5], chalcones [6], naphthoquinones [7] and several other hybrid antimalarial
[
molecules have been synthesized [8-14]. The aim of the present work was to
synthesize new compounds with a 7-chloroquinolinotriazole basic structure
supporting different side chains linked to the triazole moiety, via click chemistry.
Considering chloroquine (1) as the basic antimalarial moiety, we have planned
the substitution of its N-alkyl chain by a 1,2,3-triazole moiety supporting different
substituents at position C-4. The final products could be represented by three
general structures A, B and C, as shown in Scheme 1. The three types would
be obtained by a click reaction between 4-azido-7-chloroquinoline and an
alkyne via copper-catalyzed cycloaddition reactions (CuAAC) [4] (Scheme 1).
INSERT SCHEME 1
2. Synthetic Chemistry
4-Azido-7-chloroquinoline (3), a key intermediate, was prepared from 4,7-
dichloroquinoline (2) that is commercially available. The click reactions were
carried out according to the methodology described by Sharpless and co-
workers [4] for the Cu(I)-catalyzed azide-alkyne 1,3-dipolar cycloaddition
(
CuAAC) that is presently one of most widely used for the preparation of 1,2,3-
triazole hybrids [7-11]. The reactions were carried out in dichloromethane or
acetonitrile and water, in the presence of CuSO .5H O and sodium ascorbate,
4
2
stirring at room temperature, for 2-24 hours. The crude reaction products were
generally purified by preparative TLC or column chromatography. The yields for
the several click reactions reported here were in the range of 49 to 81%, except
in two cases that were of 28 and 31% (Table 1).
The simplest route, leading to derivatives of group A, was carried out using
commercially available alkynes (Scheme 2). Compounds 4 – 11 were prepared
in high yield reactions (49 to 77%) (Table 1). It is interesting to emphasize the
applicability of this process to the synthesis of 7-chloroquinolinotriazole

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derivatives with different classes of side chains such as alkyl, aryl and alcohol
groups, as described here.
INSERT SCHEME 2
Ester analogs, representing type B 7-chloroquinolinotriazole derivatives,
were obtained by reaction of propargyl alcohol (12) and 1-pentynol (13) with
different benzoyl chlorides leading to a total of 14 new compounds (14-27)
(
Scheme 3).
INSERT SCHEME 3
Based on chloroquine (1) structure, it seemed interesting to synthesize a
series of 7-chloroquinolinotriazoles containing a methylamine side chain with a
unity between the two nitrogen atoms, although in quite different situations,
C
4
such as a C aliphatic chain, as in chloroquine (1), and an aromatic triazole ring.
4
The methylamine group in the designed molecules would favor their action in
the acidic parasite vacuole by inhibition of the heme polymerization similarly to
known antimalarial 4-aminoquinolines, including chloroquine (1) itself [3]
(
Scheme 4). The necessary propargylamine moiety was easily formed from
bromopropine (28) by simple nucleophilic displacement, and it was further used
without isolation in the click reaction with 4-azido-7-chloroquinoline (3) in a one-
pot-process that afforded a total of five type C 7-chloroquinolinotriazoles with an
amine side chain (29-33).
INSERT SCHEME 4
The structure of the quinolinotriazole hybrids were assigned on the
1
13
basis of spectrometric data including HRMS-ESI-IT-TOF, IR, H and C NMR.
3. Biological assays
Continuous cultures of Plasmodium falciparum
P. falciparum W2 clone, which is chloroquine-resistant and mefloquine-
sensitive [15], was kept in a continuous culture at 37 ºC in human erythrocytes
using the candle jar method [15, 16]. The antimalarial effect of the compounds
was measured by the HRP2 assay [17,18]. Parasites were kept in complete

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culture medium (RPMI) containing sodium bicarbonate, 21mM), D-glucose
(11mM), HEPES (25mM), hypoxanthine (300µM) and gentamicin (40µg/ml) that
was supplemented with 10% human plasma on culture dishes, the culture
medium being changed daily. All experiments were performed in duplicate; the
compounds were tested in triplicate at each concentration. The cultures with
predominantly ring-stage parasites were concentrated by sorbitol-
synchronization [19]. A suspension of red blood cells with 0.05% parasitemia
and 1.5% hematocrit was distributed in a 96-well microtiter plate (180 µl/well).
The parasite growth was evaluated by the ELISA immunoenzymatic anti-HRP2
test, as summarized below.
Evaluation of the in vitro antimalarial activity by the HRP2 assay
The HRP2 test was performed as previously described [17, 18]. Briefly,
cultures of P. falciparum (1.5% hematocrit; 0.05% parasitemia) were placed in
96 well microplates with the test compounds and controls at different
concentrations, and were incubated for 48 h under the same culture conditions,
as described above. After 24 h incubation, the content of six wells
corresponding to medium and no test sample controls were harvested and
frozen in microtubes, as to allow subtracting the average value obtained from
these wells from the other wells for excluding the background value (production
of HRP2 during the first 24 h of incubation). After a total of 48 h incubation, the
plates were frozen and thawed twice for total erythrocyte lyses and 100 µl/well
of the material was placed in another plate for the ELISA test. This plate was
pre-coated overnight at 4 °C with 1 mg/ml of the pri mary antibody anti-HRP2
(
MPFM-55A ICLLABs) and then the content was discarded, replaced by the
blocking solution (PBS/BSA 2% 200 µl/well), incubated for 2 h, and finally the
content was discarded. The hemolyzed cultures were transferred to the ELISA
pre-coated plate, incubated (1 h, room temperature), discarded, incubated for 1
h with 0.05 mg/ml of the secondary antibody (MPFG55P-ICLLAB; 100 µl/well),
then incubated with 100 µl /well of TMB chromogen (15 min at room
temperature) in the dark. The reaction was stopped with 50 ml/L of 1M sulfuric
acid and the absorbance was read at 450nm in a spectrophotometer
®
(
Infinite 200 PRO, Tecan). The results were evaluated with the software
MicrocalOrigin 8.5 for determination of the dose-response curves plotted with

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sigmoidal fit. The 50% inhibitory concentration growth of the parasites (IC50
)
was determined by comparison with controls with standard drug and without
drugs.
Cytotoxicity evaluation in human hepatome cell cultures – Hep G2A16
The hepatome cells Hep G2A16 were maintained at 37ºC, 5% CO in 75-
2
2
®
cm sterile culture flasks (Corning ) with RPMI 1640 culture medium
supplemented with 5% FBS, penicillin (10 U/ml), and streptomycin (100 g/ml),
with changes of medium twice a week. The cells were maintained in weekly
passages (at 1:3 dilutions in sterile culture flasks) and grown to 80% [20]. They
were used for experiments after being trypsinized (0.05% trypsin/0.5 mM EDTA)
and plated on 96 well microplates [21]. When confluent, the monolayers were
trypsinized, washed, counted, diluted in complete medium, distributed in 96-well
3
microplates (4 x 10 cells/well), then incubated for another 24 h at 37ºC. The
test samples and controls were diluted to a final concentration of 0.02% DMSO
in culture medium to yield four concentrations in serial dilutions starting at 1,000
mg/ml. After a period of 24 h incubation at 37 °C , 1 8µl of MTT solution (5mg/ml
in PBS ) were added to each well, followed by another 1h30min incubation at
37 °C .The supernatant was then removed, and 180µl of DMSO were added to
each well. The culture plates were read in a spectrophotometer with a 570 nm
filter [22]. The minimum cytotoxicity concentration was determined as described
with slight modifications. Each test was performed in duplicate; the
concentration that killed 50% of the cells (CC ) were determined [23]. The
50
selectivity index (SI) for the antimalarial activity was then calculated based on
the rate between CC50 and IC50 for the in vitro activity against P. falciparum as
described [24]. Most of the compounds showed SI >10 and could be considered
non-toxic [25].
4. Results and discussion
The twenty-seven 7-chloroquinolinotriazole synthetic compounds were
evaluated for their in vitro antimalarial activity against P. falciparum W2 strain
that is chloroquine resistant and mefloquine-sensitive. Table 1 shows the values
of IC for the in vitro antimalarial assay (HRP2 method), the CC values for the
5
0
50
cytotoxicity (Hep G2A16 cells) and the SI for these compounds. All the

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compounds disclosed low cytotoxicity with CC50 > 100 µM, some of them
reaching values > 3,000 µM. Five of them showed moderate antimalarial
activity with IC < 50 µM, in the range of 9.6 to 40.9 µM. SI values varied from
5
0
2.2 to 71.1; about half of them were close to 20 as consequence of high CC50
values (low cytotoxicity).
TABLE 1
The most active 7-chloroquinolinotriazole derivative was compound 8 (type A)
with an IC50 of 9.6 µM and a reasonable SI (15.7). Interestingly, the other active
compounds, 30 (IC50 27.4 µM), 4 (IC50 40.3 µM) and 33 (IC50 40.5 µM) have a
nitrogen or an oxygen atom in the end of the triazole side chain. However,
compound 29 was approximately sixfold less potent (IC50 > 166.0 µM) than the
closely related compound 30. The only structural differences between 29 and
30 are the substituents on the amino group of the triazole side chain, diethyl
and dimethyl, respectively. It is surprising that such a small structural difference
could have a remarkable effect in the antimalarial potency of these compounds.
Considering that these quinolinotriazole derivatives are putative inhibitors of
heme polymerization, it sounds reasonable to suppose that the observed effect
is a consequence of side chain variation of these triazole derivatives. Indeed,
such effect has been previously shown for a systematic variation of the
branching and basicity of the side chain of chloroquine in a series of 7-chloro-4-
aminoquinolines [26,27]. On the other hand, in the case of the alcohol derivative
30, the decreased activity of the corresponding ester 19 (IC50 >123.1 µM) is a
clear demonstration of the importance of a free O-H group for the antimalarial
activity. Besides, all the benzoyl ester derivatives, except 27, disclosed low
activity with IC50 >100 µM) and, therefore, the expectation that they could act as
pro-drugs affording the supposed more active alcohols after hydrolysis was not
a favorable one. The effect of a side chain hydroxyl group in the antimalarial
activity of this series of 7-chloroquinolinotriazole derivatives is evidenced when
comparing the piperidylamine 31 (IC50 >152.8 µM) and the corresponding
hydroxy derivative 33 (IC50 40.9 µM) that is about four times more potent.
As pointed out, it was expected that the synthesized compounds, as
chloroquine (1) analogs, could inhibit the heme polymerization that is involved in
the parasite-mediated hemoglobin digestion. This process, the heme

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detoxification, leads to hemozoin formation (malarial pigment) that takes place
in the digestive or food vacuole of intra-erythrocytic parasites and is an
established drug target. The food vacuole is an acidic compartment (pH 5.0-5.4)
where chloroquine (1) is trapped and concentrated by protonation following its
active uptake by parasite transporter(s), and then binding to a parasite specific
receptor [28]. Many other antimalarials target the food vacuole indicating the
importance of this organelle and its various functions to the survival of the
parasite [29].
The acid-base character of quinoline derivatives should be taken into
account for its putative action in the food vacuole and the pKa values of the
presently described 7-chloroquinolinotriazole derivatives were calculated by the
Marvin 5.11 (2012) Chemaxon’s Calculator Plugins [30] (Table 2).
TABLE 2
A significative decrease of pKa was shown for the quinoline ring nitrogen
of these quinolinotriazoles in comparison with chloroquine: from 7.3 for
chloroquine (1) to 5.6 to 5.8 for the triazole derivatives. This difference can be
explained by the low resonance contribution of the triazole ring, due to its
aromatic character, to the stabilization of the protonated form of the quinoline
nitrogen. As a consequence of the pKa decrease for the quinoline nitrogen of
the quinolinotriazole derivatives, in comparison to chloroquine (1), it would be
expected that its protonation would be largely decreased in the digestive
vacuole with pH 5.0-5.4. It is known that the chloroquine−hematin interaction is
largely a function of its quinoline moiety, but inhibition of hemozoin formation
and parasite growth is also influenced by the 4-amino side chain [31].
Therefore, taken together, the lower basicity (pKa) of these nitrogens in the
quinolinotriazoles, in comparison with those in the corresponding positions in
the chloroquine (1) structure, might affect the antimalarial effect of the
quinolinotriazole derivatives described here.
Linear correlation analyses between the calculated pKa and
experimentally determined IC50 values described here disclose an excellent
2
correlation (R =0.993) for the active quinolinotriazoles 8, 27, 30 and 33 (IC50
<
50 µM) while, when chloroquine (1) is inserted in the series, a slight degradation
2
is registered (R =0.985). These results are in agreement with the recognized

ACCEPTED MANUSCRIPT
relevance of the quinoline nitrogen protonation for the antimalarial activity of
chloroquine and 4-aminoquinolines [32]. Therefore it is clear that when a given
pH value is considered, compounds with lower pKa will have a smaller
population of the acidic form (protonated species) than its conjugated basic
form resulting in lower antimalarial activity, as can be seen in Figure 1 that
depicts the correlation of the calculated pKa versus pIC50. The active
quinolinotriazole 4 (IC50 = 41 µM) does not fit to the curve of Figure 1 what can
be explained by its differentiated structure showing at C4 a strong electron
donor p-N(Me
compounds of this series. In this situation, compound 4 shows a high pKa
5.81), close to that one of compound 8, the most active (lowest IC50) of this
2
)-phenyl group, instead of alkyl groups, as in the other active
(
series but with a marked decrease of the antimalarial activity.
FIGURE 1
Aiming to understand how charge distribution could affect the antimalarial
effect of the quinolinotriazoles, semi-empirical calculations were also
undertaken for generation of the most stable conformers of the most stable
microspecies in pH 5. Such a pH is being considered to assuring protonation of
the quinoline nitrogen in each of the compounds and proximity to the pH of the
digestive vacuole. Initially, the major microspecies at pH 5 was calculated by
Chemaxon’s Calculator Plugins [30] and then submitted to the OMEGA2
program from OpenEye [33] to generate a set of stable conformers for each
compound. All conformations of each compound were submitted to
MOPAC2012 [34] for minimization and charge distribution calculations with AM1
method [35]. Vibrational frequencies were calculated to ensure that all
conformations are true minima, and only the most stable one was considered to
charge distribution comparisons. The partial charge calculations performed at
semi-empirical level with AM1 method show that the total charge in the
quinoline ring of the quinolinotriazoles becomes more positive in relation to
chloroquine (1) (Table 2). Therefore, the triazole ring effect on the quinoline
system can be considered just as an inductive electron attracting group when
compared with the amino group of chloroquine (1). Again, the analyses of the
partial charges indicate that compound 4 shows a distinct behavior in
comparison with other quinolinotriazole derivatives. The total charge in both

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quinoline (QN) and triazole rings are less pronounced, being the quinoline ring
(QN) less positive and the triazole ring (TR) less negative what is coherent with
the lower antimalarial activity of compound 4 in the group of the five active
quinolinotriazoles . These results would suggest that not only the protonation of
the quinoline ring is important to the antimalarial activity, but that, for the
quinolinotriazole derivatives, the negative charge in the triazole ring could
interfere in its interaction with heme. Considering that the main target for
chloroquine analogs is the ferroprotoporphyrin IX [36], a high electron density in
the triazole ring (TR) could also contribute to the activity. These distinct effects
of compound 4 might even lead to speculate on an alteration in the mechanism
of action, concerning the detailed mechanism of the chemical interaction of this
compound with the ferroprotoporphyrin IX for inhibition of the heme
polymerization.
It is interesting to note the remarkable resemblance of the potential
pharmacophores of compounds 4 and 8 with methylene blue (MB), a synthetic
thiazine dye that shows good antimalarial activity. Recently, its in vitro activity
against resistant strains of P. falciparum was reported and disclosed that it is 20
to 50 fold more active than chloroquine (1) [37, 38]. A close look to the potential
pharmacophores of compounds 4, 8 and MB was undertaken by the Discover
Studio Visualizer [39]. The following reference groups were foccused: a) the
quaternary nitrogen (positive center), the unprotonated nitrogen of the p-
N(Me
hydrogen bond acceptor of the MB structure, at pH 5, b) the protonated
quinoline nitrogen (positive center), the unprotonated nitrogen of the p-N(Me )-
phenyl group (negative center) and the N - triazole nitrogen as a hydrogen bond
acceptor of compound 4, and 8) the protonated quinoline nitrogen (positive
center), the oxygen of the triazole side chain (negative center) and the N
2
)-phenyl moiety (negative center) and the endocyclic nitrogen as a
2
1
1
-
triazole nitrogen as a hydrogen bond acceptor of compound 8. The model
structures (Figure 2) show that the putative pharmacophores disclose similar
positions in these three compounds, including distances and angles, as can be
seen in their schematic representation in Figure 3. It is interesting to notice that,
in both cases, the aromatic ring position is conserved as well. Might we ascribe
the higher potency of quinolinotriazoles 4 and 8 to their closer structural

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resemblance to MB? It should be noticed that in this case, compound 4,
showing a more extended conjugation, planarity and rigidity, would be expected
to be more potent. However compound 8 is about four times more active than
compound 4. Therefore the combination of a 1,2,3-triazole moiety with the 7-
chloroquinoline unity brings in a new hydrogen acceptor pharmacophore in
which the distance between the positive center and the hydrogen acceptor is
6.2 in these triazole derivatives and is comparable to that one of 5.6 A for MB.
FIGURE 2
FIGURE 3
Finally, the SAR analyses described in the present study has shown that
the click reaction to afford triazole derivatives should still be exploited to
produce a diverse structural series of 7-chloroquinolinotriazoles with stronger
4
electron donating group at position C and a spacer between the quinoline and
the triazole group. Indeed, very recently, the synthesis of a series of 1H-1,2,3-
triazole-tethered-7-chloroquinoline-isatin conjugates with or without an alkyl
chain and its in vitro antimalarial evaluation against W2- chloroquine resistant
strain of P. falciparum demonstrated the applicability of our SAR studies. The
activity profiles showed dependence on the presence or absence of an alkyl
chain between the 7-chloroquinoline ring and the triazole moiety, as well as the
length of this alkyl chain and the substituent at the C-5 position of the isatin ring.
IC values < 5 µM, in the range of 3.85 to 1.21 µM, are reported for the most
5
0
active compounds, all of them with the spacer alkyl chain between the 7-
chloroquinoline ring and the triazole moiety, while compounds without this
spacer are less active [40]. However, the IC50 have not been exactly determined
(IC50 > 5 µM) not allowing an appreciation of the structural effect in this series.
5. Conclusion
In the present work reports the synthesis of twenty-seven 7-
chloroquinolinotriazole derivatives in which a 1,2,3-triazole ring supporting
different substituents in position C is directly linked by the N to the C
quinoline position. Evaluation of the antimalarial activity and cytotoxicity of three
4
1
4

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different structural series disclosed that the most promising compounds have
some structural similarities such as a heteroatom in the triazole side chain. The
facility to obtain these compounds via click chemistry by the copper-catalyzed
cycloaddition (CuAAC) reaction can be an advantageous strategy for SAR
investigation. The moderate to low activity of the 7-chloroquinolinotriazoles we
have synthesized was explained by the low resonance contribution of the
triazole ring to stabilize the acidic form of the quinoline ring what decreases the
pKa of the quinoline nitrogen that is known to be important for the heme
detoxification in the malaria parasite. Good correlations were observed between
pIC50 and pKa, and semi-empirical calculations confirmed the low resonance
effect of the triazole ring on the quinoline ring that could be foreseen. However,
the extension of this effect on the antimalarial activity of quinolines was not
estimated. On the other hand, the analyses of pharmacophores described in the
present study has shown that the click reaction to afford triazole derivatives
should still be exploited to produce a
diverse structural series of 7-
chloroquinolinotriazoles with an stronger electron donating group at position C
4
and a spacer between the quinoline and the triazole group. The significance of
the triazole side chain for the antimalarial activity was also appreciated. Taking
into account the results of the present study the synthesis of promising new
similar derivatives is on progress.
6. Experimental section
6.1. General
Chemicals and reagents were purchased from commercial suppliers and
used as received unless noted otherwise. Reactions were monitored by thin
layer chromatography (TLC) on precoated 0.2 mm silica gel 60 F254 (Merck)
plates and visualized in several ways with an ultraviolet light source at 254nm,
by spraying with Hanissam reagent (ceric ammonium molibidate-CAM),
anisaldehyde acid, Dragendorff or iodine chamber. All reactions were performed
in standard dry glassware without inert atmosphere. Evaporation and
concentration were done at standard rotavapor Büchi using vacuum pump.
Thin-layer chromatography (TLC) was carried out with silica gel 60 with
fluorescent indicator (e.g., Silica Gel. F-254 or IB-F, Merck) and activated
previously activated with heating at 100 °C overnigh t and visualized by UV light

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or Dragendorff indicator. Melting points (mps) were measured with a Fisher-
1
13
Jones melting point apparatus and are uncorrected. H and C NMR spectra
were measured on the Bruker. Advance DPX 200 with FT analysis. Chemical
shifts
δ
(ppm) are reported in SiMe
4
. Coupling constants (J) are given in hertz.
or DMSO-d . Splitting patterns have been
Used deuterated solvent were CDCl
3
6
designated as follows: s = singlet; bs = broad singlet; d = doublet; t = triplet; q =
quartet; qt = quintuplet; td = triplet doublet; m = multiplet. Protons signals
attribution was done for each structural moiety of the molecules and the
following abbreviations were used: Cq = chloroquinoline, Tr = triazole, Ch =
cyclohexane, Ph = phenyl, Np = naphthalene, To = tolyl, An = aniline, iPr =
isopropyl, Pr = propyl, Ea = ethylamine, Ma= methylamine, Mo = Morpholine,
Pm = pyrimidine, Bz = benzoate, Po = Pentynol. Infrared spectra were recorded
on FT-IR, Spectrum One, Perkin-Elmer, with system ATR and are reported in
-
1
wave number (cm ). High-resolution EI mass spectra were recorded on a
Shimadzu LCMS instrument with direct injection. Samples were dissolved in
methanol-formic acid 0.1 % solution.
7. Materials
4,7-Dichloroquinoline, benzoyl chloride, 4-methylbenzoyl chloride, 4-
methoxylbenzoyl chloride, 4-(trifluoromethyl)benzoyl chloride, 4-fluorobenzoyl
chloride, 4-tert-butylbenzoyl chloride, 4-ethylbenzoyl chloride, 2-ethylbenzoyl
chloride,
2,3,5-trifluoro-4-methoxybenzoyl
chloride,
2-ethynyl-6-
methoxynaphthalene; 4-ethynyltoluene; 1-ethynyl-4-fluorobenzene; 1-ethynyl-4-
fluorobenzene; 4-Ethynyl-N,N-dimethylaniline; ciclehexylacethylene; 2-methyl-3-
butyn-2-ol; phenylacethylene, molecular sieves A4, triethylamine, piperidin-4-ol,
pent-4-yn-1-ol, prop-2-yn-1-ol, 3-bromoprop-1-yne, RPMI 1640 medium, sodium
bicarbonate, D-glucose, HEPES, hypoxanthine, gentamicin, D-sorbitol, PBS,
BSA, TMB, FBS, penicillin, streptomycin, tripsin/EDTA, DMSO were obtained
®
from Sigma-Aldrich USA, Ltd. Copper sulphate pentahydrate was obtained
®
®
from Reagen , ascorbic acid was obtained from Synth , MPFG-55P and
®
MPFM-55A antibodies were purchased from ICLLABS , sulfuric acid, sodium
®
bicarbonate and sodium sulphate were obtained from FMaia , MTT,
®
dimethylamine, piperidine and diethylamine from Merck , and were used
without further purification, glassware from Hialoquímica Ltda.

ACCEPTED MANUSCRIPT
8
. Synthesis and characterization
.1.1. 4-Azido-7-chloro-quinoline (3) [41]
8
To a solution of 4,7-dichloroquinoline (2) (2.0 g, 10 mmol) in 5 mL anhydrous
DMF and molecular sieves A4, sodium azide (1.3 g, 20 mmol) was added in
°
one portion at room temperature, and the resulting mixture stirred at 85 C for 8
h, when TLC indicated reaction completion. The reaction mixture was then
allowed to cool to room temperature and then it was diluted with 100 mL
CH Cl , washed with water (3x 40 mL), dried over anhydrous Na SO , and
2
2
2
4
evaporated to dryness. The resulting product residue was purified by small
column chromatography eluted with CH
2
Cl
2
/Hexane mixture 1:1 to yield the final
°
pure product as colorless, needle-like crystals (1.8 g, 91%); mp 115 C (from
-
1
1
CH
NMR (200 MHz, CDCl
Hz, H ), 7.88 (1H, d, J = 8.8 Hz, H
2
Cl
2
/Hex); Rf (EtOAc/Hex 3:7) 0.29; IR νmax (neat)/cm 3036, 2118, 1608. H
) : Cq 8.76 (1H, d, J = 5.0 Hz, H ), 8.00 (1H, d, J = 2.0
), 7.49 (1H, dd, J = 2.0 Hz, J = 8.8 Hz, H
). C NMR (50 MHz, CDCl3) 151.33, 149.59,
46.30, 136.57, 128.23, 127.52, 123.77, 119.91, 108.73.
3
δ
2
8
5
6
)
1
3
7
.13 (1H, d, J = 5.0 Hz, H
3
δ
1
General methodology for route A. 4-Azido-7-chloroquinoline (3) and
commercial alkyne was dissolved in CH Cl (2 mL), followed by addition of
CuSO .5H O (0.3 equivalents) and an aqueous solution of sodium ascorbate
0.6 equivalents) (2 mL) freshly prepared. The reaction mixture was left over
2
2
4
2
(
night and was stopped when it was completed as shown in TLC. Work up of
the reaction mixture with CH Cl , water (3x 10 mL), dried over Na SO and final
2
2
2
4
purification by preparative TLC or column chromatography.
8.1.2. 4-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)-N,N-dimethylaniline (4)
-1
Yield: 76%; mp 202.5-205.3 °C; IR ( νmax, cm ): 3129, 3056, 2995, 2970, 2938,
2
1
8
842, 1632, 1613, 1593, 1557, 1504, 1477, 1450, 1446, 1433, 1394, 1372,
352, 1265, 1235, 1206, 1187, 1164, 1118, 1080, 1024, 953, 900, 857, 815,
1
04, 770, 731, 716, 677. H NMR (200 MHz, CDCl ): Cq
δ
9.04 (d, 1H, J = 4.6
3
Hz, H
2
); 8.23 (d, 1H, J = 1.8 Hz, H
8
); 8.13 (d, 1H, J = 9.2 Hz, H ); 7.57 (dd, 1H,
5
J = 2.0 Hz, J= 9.2 Hz, H ); 7.51 (d, 1H, J = 4.6 Hz, H ); Tr 8.10 (s, 1H, H ); An
6
3
5
7.79 (d, 2H, J = 8.6 Hz, H2,
H
6
); 6.68 (d, 2H, J = 8.6 Hz, H3,
H
5
); 3.01 (s, 6H, 2x -
3 5
CH ). HRMS-ESI-IT-TOF: m/z cal C19H17ClN (M+H) 350.1172, found 350.1106;

ACCEPTED MANUSCRIPT
1
3
C NMR (50 MHz, CDCl
3
): δ 151.59; 150.95; 150.41; 149.24; 141,31; 136.97;
129.45; 129.08; 127.12; 125.12; 120.86; 119.63; 117.50; 115.93; 112.58, 40.54.
HRMS-ESI-IT-TOF: m/z calculated C19
H
17ClN
5
(M+H) 350.1172, found
350.1106; HRMS-ESI-IT-TOF: m/z cal C H ClN Na (M+H+2) 352.1143, found
19 16 5
352.1108. HRMS-ESI-IT-TOF: m/z calculated C19
H
16ClN
5
Na (M+Na) 372.0992,
found 372.0957
8.1.37-Chloro-4-(4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl)quinoline (5)
-
1
Yield: 58%; mp 165.1-168.5 ºC; IR (νmax, cm ): 3140, 3056, 2923, 2852, 1609,
1
1
δ
591, 1561, 1504, 1493, 1455, 1438, 1420, 1342, 1301, 1232, 1218, 1159,
1
030, 973, 952, 879, 840, 824, 795, 769, 677. H NMR (200 MHz, DMSO-d
6
):
); 8.14 (d, 1H, J =
); 7.72 (dd, 1H, J = 1.6 Hz, J= 9.0 Hz,
); Ph 7.82 (dd, 2H, J = 5.6 Hz, J = 8.2 Hz, H2, ); 7.30 (t,
162.96; 150.54; 147.84;
Cq 9.12 (d, 1H, J = 4.2 Hz, H
.0 Hz, H ); 7.85 (d, 1H, J = 4.6 Hz, H
); Tr 9.22 (s, 1H, H
2 8
); 8.21 (d, 1H, J = 1.6 Hz, H
9
5
3
H
6
5
H
6
1
3
2
1
1
H, J = 8.6 Hz, H3,
H
5
). C NMR (50 MHz, DMSO-d
6
):
δ
44.71; 138,69; 138.84; 127.22; 126.47; 126.07; 125.91; 124.64; 123.90;
21.63; 118.41; 114.92; 114.48, 114.05 (t, 2C, J = 21.5 Hz). HRMS-ESI-IT-
9 4
TOF: m/z cal C17H ClFN (M-H) 323.0499, found 323.0521. HRMS-ESI-IT-TOF:
m/z cal C H ClFN (M+H+2) 327.0627, found 327.0503
17
9
4
8.1.4. 7-Chloro-4-(4-p-tolyl-1H-1,2,3-triazol-1-yl)quinoline (6)
-
1
Yield: 77%; mp 1163.9-165.7 ºC; IR (νmax, cm ): 3112, 3048, 2919, 2853, 1611,
1594, 1559, 1497, 1448, 1430, 1375, 1343, 1300, 1234, 1192, 1128, 1084,
1
1
027, 1017, 955, 911, 875, 849, 812, 767, 722, 678. H NMR (200 MHz,
CDCl
3
):
H, J = 9.0 Hz, H
.0 Hz, H ); Tr 9.29 (s, 1H, H
δ
Cq 9.04 (d, 1H, J = 4.2 Hz, H
); 7.58 (d, 1H, J = 4.2 Hz, H
); To 7.82 (d, 2H, J = 7.6 Hz, H2, H
6
2
); 8.21 (d, 1H, J = 3.8 Hz, H
); 7.72 (dd, 1H, J = 3.8 Hz, J =
); 7.26 (d, 2H,
151.59;
8
); 8.05 (d,
1
9
5
3
6
5
1
3
J = 7.6 Hz, H H ); 2.34 (s, 3H, -CH ). C NMR (50 MHz, DMSO-d6):
δ
3
,
5
3
1
50.40; 148.80; 141.17; 139,08; 139.00; 137.09; 129.95; 129.59; 129.17;
26.86; 126.07; 124.92; 120.98; 120.87; 120.82; 116.07, 21.56. HRMS-ESI-IT-
1
TOF: m/z calculated C17
H10ClN
4
(M-H) 305.0593, found 321.0912. HRMS-ESI-
IT-TOF: m/z cal C17
H
10ClN
4
(M+H) 323.0877, found 323.0821.

ACCEPTED MANUSCRIPT
8.1.5. . 7-Chloro-4-(4-phenyl-1H-1,2,3-triazol-1-yl)quinoline (7)
-1
Yield: 59%; mp 160.3-162.9 ºC; IR (νmax, cm ): 3144, 3127, 3053, 1608, 1593,
1560, 1502, 1484, 1453, 1433, 1316, 1301, 1234, 1212, 1187, 1155, 1081,
1
1
033, 1021, 954, 920, 877, 833, 820, 811, 770, 691. H NMR (200 MHz,
DMSO-d
6
):
.8 Hz, H ); 7.90 (d, 1H, J = 4.4 Hz, H
H, H ); Ph 7.99 (d, 2H, J = 7.2 Hz, H
): 152.41; 149.51; 147.24; 140.45; 135,51; 129.86;
29.15; 129.05; 128.63; 128.18; 125.66; 125.64; 123.67; 120.19; 116.86.
δ
Cq 9.15 (d, 1H, J = 4.4 Hz, H
); 7.75 (d, 1H, J = 9.0, H
,H ); 7.54-7.40 (m, 3H, H
2
); 8.21 (bs, 1H, H
8
); 8.06 (d, 1H, J =
8
1
5
3
6
); Tr 9.29 (s,
13
5
2
6
3
,H
4
,H
5
).
C
NMR (50 MHz, DMSO-d
6
δ
1
HRMS-ESI-IT-TOF: m/z calculated
C
17
H
10ClN
10ClN
10ClN
4
(M-1) 305.0593, found
4
(M+Na) 329.0570, found
3
3
3
05.0712. HRMS-ESI-IT-TOF: m/z cal C17
H
29.0368. HRMS-ESI-IT-TOF: m/z cal C17
31.0362.
H
4
(M+Na+2) 331.0540, found
.
8.1.6. 3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propan-1-ol (8)
-1
Yield: 75%; mp 107.0-112.1 °C; IR ( νmax, cm ): 3321, 3079, 3037, 1608, 1578,
1
1
8
566, 1557, 1492, 1417, 1373, 1352, 1301, 1278, 1200, 1206, 1146, 1071,
1
012, 963, 896, 880, 840, 819, 777, 769, 671. H NMR (200 MHz, CDCl
3
): Cq
); 7.45
); 3.05
d, 2H, J = 7.2 Hz, H ); 7.78 (m, 2H, J = 6.0 Hz, J = 7.2 Hz, H ). C NMR (50
δ
.98(d, 1H, J = 4.6 Hz, H
2
); 8.10 (s, 1H, H
8 5
); 7.46 (d, 1H, J = 9.0 Hz, H
(d, 2H, J = 4.6 Hz, H
3
); Tr 8.05 (s, 1H, H
5
); Pr 3.87 (d, 2H, J = 6.0 Hz, H
1
1
3
(
3
2
3
MHz, CDCl ): δ 151.08; 149.45; 148.28; 140.67; 136,37; 128.87; 128.22;
1
24.51; 125.12; 122.92; 120.03; 115.60; 60.98, 31.80, 21.73. HRMS-ESI-IT-
TOF: m/z calculated C14 ONa (M+Na) 311.0675, found 311.0687.
HRMS-ESI-IT-TOF: m/z cal C14 ONa (M+Na+2) 313.0646, found
H
13ClN
4
H
13ClN
4
313.0957.
8.1.7. 2-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propan-2-ol (9)
-1
Yield: 49%; mp 143.5-145.6 °C; IR ( νmax, cm ): 3438, 3160, 3037, 2972, 2925,
1613, 1592, 1559, 1505, 1449, 1435, 1362, 1314, 1237, 1176, 1128, 1101,
1
1
033, 962, 925, 875, 855, 826, 816, 808, 775, 726, 684, 671. H NMR (200

ACCEPTED MANUSCRIPT
MHz, CDCl
3
):
H, J = 9.2 Hz); 7.48-7.41 (m, 2H); 1.75 (s, 6H). C NMR (50 MHz, CDCl
156.78; 151.37; 150.08; 141.08; 136,94; 129.40; 128.86; 124.79; 121.53;
4
20.55; 115.99; 68.71; 30.68. HRMS-ESI-IT-TOF: m/z calculated C14H14ClN O
δ
8.94 (bs, 1H); 8.21 (bs, 1H, J = 3.8 Hz
8
); 8.00 (bs, 1H); 7.92 (d,
1
3
1
δ
3
):
1
(
M+H) 289.07779, found 289.0855; HRMS-ESI-IT-TOF: m/z calculated
ONa (M+Na) 311.0676, found 311.062
14 4
C H13ClN
8.1.8. 7-Chloro-4-(4-cyclohexyl-1H-1,2,3-triazol-1-yl)quinoline (10)
-
1
Yield: 64%; mp 115.9-120.3 ºC; IR (νmax, cm ): 3127, 3047, 2924, 2849, 1610,
1
8
4
1
593, 1560, 1504, 1449, 1438, 1348, 1310, 1244, 1115, 1046, 1018, 922, 875,
1
34, 822, 813, 767, 672. H NMR (200 MHz, DMSO-d
6
):
δ
Cq 9.09 (d, 1H, J =
.2 Hz, H ); 8.21 (d, 1H, J = 1.4 Hz, H ); 8.06 (d, 1H, J = 9.2 Hz, H ); 7.76 (d,
2
8
5
H, J = 4.2 Hz, H
3
); 7.75 (dd, 1H, J = 1.4 Hz, J = 9.2, H
6
5
); Tr 8.54 (s, 1H, H ),
1
3
Ch 1.90-1.80 (m, 1H, H
50 MHz, DMSO-d ): 153.10; 152.31; 149.47; 140.62; 135,33; 128.84; 128.10;
25.73; 122.94; 120.19; 116.58; 34.59; 32.39; 25.65; 25.60. HRMS-ESI-IT-TOF:
1
); 1.78-1.30 (m, 10H, H14, H15, H16, H17, H18). C NMR
(
6
δ
1
m/z calculated C17
H17ClN
4
Na (M+Na) 335.1039, found 335.1236. HRMS-ESI-IT-
TOF: m/z cal C17
H17ClN
4
Na (M+Na+2) 337.1010, found 337.1045.
8
.1.9. 7-Chloro-4-(4-(6-methoxynaphthalen-2-yl)-1H-1,2,3-triazol-1-yl)quinoline
11)
Yield: 56%; mp 229.8-232.9 ºC; IR (νmax, cm ): 3129, 3056, 2995, 2938, 1632,
(
-
1
1613, 1593, 1557, 1504, 1477, 1446, 1433, 1394, 1265, 1235, 1206, 1164,
1
1
118, 1080, 1024, 953, 900, 877, 857, 815, 804, 770, 677 H NMR (200 MHz,
1
DMSO-d
6
):
δ
H NMR (200 MHz, DMSO-d
); 8.19 (d, 1H, J = 9.2 Hz, H
); Tr 9.37 (s, 1H, H ); Np 8.20 (d, 1H, J = 2.0 Hz, H
); 7.90 (dd, 1H, J = 2.2 Hz, J = 9.2 Hz, H ); 7.38 (d, 1H, J = 2.4,
); 7.38 (dd, 1H, J = 2.4, J = 9.2, H ); 3.90 (s, 3H, -OCH
): 157.77; 152.51; 149.54; 147.44; 140,97; 140.53; 138.86; 135.51;
34.30; 129.10; 128.56; 128.21; 127.65; 125.83; 125.79; 125.04; 124.25;
20.19; 119.41; 119.30; 55.34. HRMS-ESI-IT-TOF: m/z calculated C22
6
): Cq 9.19 (d, 1H, J = 4.6 Hz, H
); 8.07 (d, 1H, J = 9.2 Hz, H ); 7.98-
); 7.98-7.91
2
);
8
.49 (bs, 1H, H
91 (m, 1H, H
, H
8
5
6
7
3
5
2
(m, 2H, H
4
7
8
1
3
H
7
5
3
). C NMR (50 MHz,
DMSO-d
6
δ
1
1
4
H16ClN O

ACCEPTED MANUSCRIPT
(
(
M+1) 387.1012, found 387.0430. HRMS-ESI-IT-TOF: m/z cal C22
M+H+2) 389.0983, found 389.0929
4
H16ClN O
General methodology for route B. To a solution of amine (8 mmol) in 4 mL of
acetonitrile, 3-bromoprop-1-yne (2 mmol) was added in drops at room
temperature, followed by sodium bicarbonate (8 mmol) and the resulting mixture
was stirred at room temperature overnight. The next step was carried in the
same reaction pot. First, 4-azido-7-chloro-quinoline (1 mmol) dissolved in
enough CH
prepared aqueous solution of sodium ascorbate, made using ascorbic acid (53
mg, 0.3 mmol), NaHCO (25 mg, 0.3 mmol) and CuSO .5H O (0.3 mmol). The
reaction mixture was stirred for 24 hours, then 30 mL CH Cl was added,
washed with water (3x 40 mL), the organic layer was dried over anhydrous
Na SO , and evaporated to dryness. The resulting residue was purified by
preparative TLC to afford the final compound.
2 2
Cl was added to the reaction, followed by a 2 mL of a freshly
3
4
2
2
2
2
4
8
.1.10.
ethylethanamine (29)
Yield: 40%; yellow oil; IR (νmax,cm ): 3053, 2969, 2933, 2873, 2815, 1611,
N-((1-(7-chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)methyl)-N-
-1
1594, 1561, 1504, 1454, 1437, 1372, 1345, 1299, 1229, 1188, 1159, 1112,
1
1
070, 1034, 1001, 954, 877, 813, 769, 733, 665, 671.m H NMR (200 MHz,
CDCl
3
): Cq
δ
9.04 (d, 1H, J = 4.2 Hz, H
2
); 8.22 (s, 1H, H
8
); 8.03 (d, 1H, J = 9.2
); Tr 8.01 (s, 1H,
Hz, H
5
); 7.58 (d, 1H, J = 9.2 Hz, H
6
); 7.50 (d, 1H, J = 4.2 Hz, H
3
H ); Ea 3.92 (s, 2H, -CH ); 2.63 (q, 4H, J = 7.0 Hz, 2 x -CH ); 1.14 (t, 6H, J =
5
2
2
1
3
7
1
1
3
3
.0 Hz, 2 x -CH
3
). C NMR (50 MHz, CDCl
3
):
δ
151.51; 150.31; 146.56;
41.21; 136,94; 129.47; 129.05; 124.88; 124.46; 120.71; 116.00; 47.78; 47.10;
1.96. HRMS-ESI-IT-TOF: m/z calculated C16 (M+H) 316.1328, found
(M+H+2) 318.1299, found
H
19ClN
5
16.1075; HRMS-ESI-IT-TOF: m/z cal C16
H
19ClN
5
18.1054.
8.1.11.
1-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)-N,N-
dimethylmethanamine (30)
Yield: 53%; yellow oil; IR (νmax, cm ): 3398, 2947, 2869, 2828, 2782, 2647,
611, 1595, 1563, 1505, 1456, 1439, 1371, 1348, 1303, 1235, 1189, 1174,
-
1
1

ACCEPTED MANUSCRIPT
1
1
115, 1045, 1023, 1000, 956, 878, 849, 818, 762, 733, 672. H NMR (200 MHz,
CDCl ): ): Cq 9.03 (d, 1H, J = 4.4 Hz, H ); 8.28 (s, 1H, H ); 8.10 (d, 1H, J =
.0 Hz, H ); 7.59 (d, 1H, J = 4.4 Hz, H ); 7.56 (d,1H, J= 9.0 Hz, H ); Tr 8.15 (s,
H, H ); Ma 3.76 (s, 2H, -CH ); 2.63 (s, 6H, 2 x –CH ). C NMR (50 MHz,
3
δ
2
5
9
8
3
6
1
3
1
5
2
3
3
CDCl ): δ 150.81; 149.12; 144.83; 140.08; 135,55; 128.26; 127.80; 124.21;
1
19.64; 115.32; 53.16; 44.35. HRMS-ESI-IT-TOF: m/z calculated C14
M+H) 288.1016, found 288.2605; HRMS-ESI-IT-TOF: m/z cal C14
M+H) 288.1016, found 288.0975. HRMS-ESI-IT-TOF: m/z cal C14
H
14ClN
14ClN
5
(
(
(
H
5
H
13ClN
5
Na
M+H+2) 290.0986, found 290.0972. HRMS-ESI-IT-TOF: m/z calculated
Na (M+Na) 310.0835, found 310.0788.
14 5
C H13ClN
8.1.12. 4-((1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)methyl)morpholine (31)
-1
Yield: 62%; mp 157.0-161.2 °C ; IR ( νmax,cm ): 3150, 3063, 2991, 2857, 2804,
2
1
8
4
608, 1594, 1562, 1504, 1453, 1437, 1339, 1319, 1310, 1285, 1263, 1240,
225, 1201, 1140, 1112, 1071, 1035, 1012, 1004, 956, 915, 879, 849, 867, 847,
1
24, 813, 772, 733, 698, 670 H NMR (200 MHz, CDCl ):
δ
Cq 8.90 (d, 1H, J =
3
.4 Hz, H
2
); 8.06 (s, 1H, H
8
); 7.87 (d, 1H, J = 9.2 Hz, H
5
); 7.40 (d, 1H, J = 9.2
Hz, H ); 7.37 (d, 1H, J = 4.4 Hz, H ); Tr 7.95 (s, 1H, H ) Mo 3.71 (s, 2H, -CH );
6
3
5 ;
2
1
3
3
.63 (s, 4H, 2 x –CH
2
O); 2.51 (s, 4H, 2 x –CH
2
N). C NMR (50 MHz, CDCl
3
):
δ
151.28; 149.28; 145.01; 140.78; 136,62; 129.19; 128.77; 124.58; 120.35;
15.79; 66.72; 53.42; 53.42. HRMS-ESI-IT-TOF: m/z cal C16 O (M+H)
30.1121, found 330.0890. HRMS-ESI-IT-TOF: m/z cal C H ClN O (M+H+2)
1
3
3
H17ClN
5
1
6
17
5
32.1092, found 332.0845.
8.1.13. 7-Chloro-4-((4-(piperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)quinoline (32)
-1
Yield: 55%; mp 105.4-108.7 °C; IR ( νmax, cm ): 3150, 3068, 2941, 2855, 2823,
2
1
7
8
780, 2609, 1596, 1560, 1504, 1449, 1435, 1373, 1317, 1301, 1252, 1228,
199, 1183, 1156, 1109, 1069, 1033, 1010, 956, 884, 877, 858, 819, 809, 788,
1
68, 670 H NMR (200 MHz, CDCl
3
):
); 7.58 (d, 1H, J = 9.0 Hz, H
); Pm 3.83 (s, 2H, -CH ); 2.56 (s, 4H, 2 x –CH
2
δ
Cq 9.06 (bs, 1H, H
); 7.37 (d, 1H, J = 2.8
O);
.63 (s, 4H, 2 x –CH ); 1.49 (s, 2H, –CH N). C NMR (50 MHz, CDCl3):
2 5
); 8.23 (s, 1H, H );
.03 (d, 1H, J = 9.0 Hz, H
); Tr 8.04 (s, 1H, H
8
6
Hz, H
3
5
2
1
3
1
2
2

ACCEPTED MANUSCRIPT
δ
151.34; 150.05; 145.69; 140.94; 136,63; 129.18; 128.79; 124.74; 124.51;
20.45; 115.82; 66.72; 54.48; 25,79; 23.99. HRMS-ESI-IT-TOF: m/z calculated
1
17 5
C H19ClN (M+1) 328.1328, found 328.1076. HRMS-ESI-IT-TOF: m/z cal
C H ClN (M+H+2) 330.1299, found 330.1048.
1
7
19
5
8
.1.14.
33)
Yield: 31%; mp 158.0-163.3 °C ; IR ( νmax,cm ): 3151, 2991, 2935, 2857, 2830,
1-((1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)methyl)piperidin-4-ol
(
-1
2
1
8
804, 2608, 1594, 1562, 1504, 1454, 1437, 1369, 1339, 1319, 1310, 1264,
240, 1225, 1201, 1140, 1112, 1071, 1035, 1013, 1004, 956, 915, 880, 867,
1
47, 824, 813, 772, 738, 671, 654. H NMR (200 MHz, CDCl
3
):
); 7.37 (d, 1H, J = 9.0 Hz, H
); 7.10 (dd, 1H, J = 1.6 Hz, J = 9.0 Hz H ); Tr 8.03 (s, 1H,
H ); 3.04 (s, 2H, -CH N); Pm 2.16-2.10 (m, 2H, 2x –CHN); 1.55-1.51 (m, 2H, 2x
δ
Cq 8.45 (d, J
=
4.6 Hz, 1H, H
2
); 7.56 (d, J = 1.6 Hz, 1H, H
8
5
); 7.16
(d, 1H, J = 4.6 Hz, H
3
6
5
2
–
CHN); 1.09-1.04 (m, 2H,2x –CH); 0.77-0.72 (m, 2H,2x –CH). Note: signal for
CHOH in the pyperimidyl cycle was probably superimposed to the solvente
–
1
3
signal. C NMR (50 MHz, CDCl
3
): δ 152.40; 149.46; 144.67; 140.54; 135,41;
128.98; 128.15; 126.07; 125.69; 120.29; 116.91; 66.17; 52.42; 50.69; 34.27.
HRMS-ESI-IT-TOF: m/z calculated C17
H
10ClN (M-H) 305.0593, found
4
3
3
3
05.0712. HRMS-ESI-IT-TOF: m/z cal C17
H
10ClN (M+H) 344.1278, found
4
44.1268. HRMS-ESI-IT-TOF: m/z cal C H ClN (M+H+2) 346.1249, found
1
7
10
4
46.1265.
General methodology for route C. To a solution of acid chloride (30.00 mmol)
in 4 mL CH Cl , pent-4-yn-1-ol (1.3 g, 20 mmol) was added in one portion at
2
2
room temperature, then trietylamine (19.40 mmol, 2,58 mL) was added in drops
and the final mixture was stirred overnight, where upon TLC indicated reaction
completion. The crude was diluted with 20 mL CH
0 mL), HCl 0,1M (1x 10 mL) and sodium hydroxide 0,1M (1x 10 mL) then dried
over anhydrous Na SO , and evaporated to dryness. The resulting product
2 2
Cl , washed with water (3x
4
2
4
residue was used in the next step without further purification.
In the next step, 4-azido-7-chloroquinoline (102 mg, 0.5mmol) and an alkyne
ester (0.5 mmol) were dissolved in CH Cl (2 mL), added CuSO .5H O (38 mg,
2
2
4
2

ACCEPTED MANUSCRIPT
0.15 mmol) and a freshly prepared solution of sodium ascorbate, made using
ascorbic acid (53 mg, 0.3 mmol) and NaHCO
water. The resulting mixture was stirred overnight, the reaction was completed
by TLC and final compound was elaborated with CH Cl , water (3x 10 mL);
dried over Na SO and purified by preparative TLC or chromatography column.
.1.15.1-Pent-4-ynyl-benzoate (34)
3
(25 mg, 0.3 mmol) in 2 mL of
2
2
2
4
8
-1
IR (νmax, cm ): 3298, 3064, 2916, 2119, 1790, 1714, 1601, 1584, 1491, 1451,
1
1
388, 1353, 1314, 1268, 1212, 1175, 1113, 1070, 1027, 997, 707, 686 H NMR
(
200 MHz, CDCl
3
):
); Po 4.31 (t, J = 6.2 Hz, 2H, H
); 1.21 (pseudo q, J = 6.2 Hz, J = 7.4 Hz, 2H, H
δ
Bz 7.94 (d, J = 7.4 Hz, 2H, H2, ); 7.58-7.42 (m, 3H, H
H
6
3
,
H
H
δ
4
3
, H
5
1
); 2.75 (bs, 1H, H ); 2.31 (t, J = 7.4 Hz, 2H,
5
13
2
3
). C NMR (50 MHz, CDCl ):
165.70; 133.11; 130.30; 129.16; 128,55; 83.30; 71.30; 63.34; 27.23; 14.73.
8.1.16. Pent-4-ynyl 4-methylbenzoate (35)
-
1
IR (νmax, cm ): 3297, 2959, 2120, 1784, 1712, 1611, 1577, 1509, 1446, 1408,
1
1
387, 1269, 1221, 1176, 1108, 1032, 1020, 1004, 840, 751, 690. H NMR (200
3 6
MHz, CDCl ): δ Bz 7.92 (d, J = 7,8 Hz, 2H, H2, H ); 7.21 (d, J = 7.8 Hz, 2H, H3,
H ); 2.42-2.33 (m, 3H, -CH ); Po 4.40 (t, J = 6.2 Hz, 2H, H ); 2.42-2.33 (m, 3H,
5
3
1
1
3
H
3
, H
5
); 2.04-1.94 (m, 2H, H
2
). C NMR (50 MHz, CDCl
3
):
δ
166.55; 143.61;
129.61; 129.09; 127.50; 83.10; 69.19; 63.30; 27.74; 21.64; 15.38.
8.1.17. Pent-4-ynyl 4-tert-butylbenzoate (36)
-1
IR (νmax, cm ): 3300, 2963, 2906, 2870, 2121, 1785, 1716, 1608, 1572, 1506,
1463, 1409, 1389, 1364, 1314, 1271, 1224, 1187, 1178, 1117, 1039, 997, 854,
1
7
75, 705 H NMR (200 MHz, CDCl
3
):
δ
Bz 7.98 (d, J = 8.6 Hz, 2H, H2,
H
6
); 7.44
); Po 4.41 (t, J = 6.4 Hz, 2H, H );
). C NMR (50
(
d, J = 8.6 Hz, 2H, H3,
H
5
); 1.34 (s, 9H, 3 x CH
3
1
13
2
.36 (td, J = 2.6 Hz, J = 7.0 Hz, 2H, H
):
5.30; 31.14; 27.78; 15.38.
3
2 5
); 2.05-1.97 (m, 3H, H , H
MHz, CDCl
3
δ 166.46; 156.57; 129.61; 127.51; 125,36; 83.09; 69.21; 63.25;
3
8.1.18. Pent-4-ynyl 4-etyl-benzoate (37)
-1
IR (νmax, cm ): 3300, 2966, 2119, 1784, 1713, 1610, 1575, 1509, 1460, 1415,
1
1
387, 1269, 1219, 1177, 1108, 1032, 1020, 1000, 852, 760, 701. H NMR (200
3 6
MHz, CDCl ): δ Bz 7.81 (d, J = 8.4 Hz, 2H, H2, H ); 7.10 (d, J = 8.4 Hz, 2H, H3,

ACCEPTED MANUSCRIPT
H
5
); 2.55 (q, J = 7.8 Hz, 2H, CH
.0 Hz, 2H, H ); 2.97-2.90 (m, 2H, H
MHz, CDCl ): 166.39; 149.70; 130.68; 129.65; 128,35; 127,68; 83.01; 69.15;
2
); 1.10 (t, J = 7.8 Hz, 3H, CH
3
); Po 4.33 (t, J =
1
3
6
1
3
); 2.25-2.15 (m, 3H, H , H
2
5
). C NMR (50
3
δ
63.21; 29.02; 27.70; 15.21.
8.1.19. Pent-4-ynyl 4-methoxybenzoate (38)
-1
IR (νmax, cm ): 3295, 2961, 2120, 1778, 1707, 1605, 1580, 1510, 1461, 1420,
1
1
387, 1316, 1272, 1251, 1221, 1165, 1114, 1102, 1025, 995, 846, 769, 695. H
1
H NMR (200 MHz, CDCl
3
):
); 3.68 (s, 3H, -OCH
); 2.20-2.15 (m, 3H, H
δ
Bz 7.37 (d, J = 9.0 Hz, 2H, H2,
H
6
); 6.73 (d, J =
); 2.96-
). C NMR (50 MHz, CDCl ):
9
2
δ
.0 Hz, 2H, H3,
H
5
3
); Po 4.30 (t, J = 6.0 Hz, 2H, H
1
1
3
.89 (m, 2H, H
3
2
, H
5
3
166.27; 163.42; 131.64; 122.68; 113,66; 83.21; 69.19; 63.23; 55.46; 27.82;
5.43.
1
8.1.20. Pent-4-ynyl 4-(trifluoromethyl)benzoate (39)
-1
IR (νmax, cm ): 3309, 2963, 1720, 1586, 1514, 1412, 1323, 1272, 1166, 1100,
1
1
064, 1017, 863, 820, 775, 752, 703, 689. H NMR (200 MHz, CDCl ):
δ
Bz
3
8.16 (d, J = 7.0 Hz, 2H, H2,
H
6
); 7.74 (d, J = 7.0 Hz, 2H, H3,
H
5); Po 4.47 (t, J =
13
6
.0 Hz, 2H, H ); 2.43-2.40 (m, 3H, H , H ); 2.05-1.99 (m, 2H, H ).
C NMR (50
1
5
3
2
MHz, CDCl
3
):
δ
163.45; 134.54; 132.21; 132.05; 128,71; 123,88 (q, 1C, J = 11.5
Hz); 81.66; 68.94; 62.69; 26.08; 13.74.
8.1.21. 3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propyl-benzoate (14)
-1
Yield: 82%; mp 35.2-42.7 ºC; IR (νmax, cm ): 3132, 3059, 2933, 2895, 1713,
1609, 1602, 1584, 1571, 1504, 1491, 1450, 1432, 1314, 1272, 1251, 1238,
1
1
173, 1112, 1069, 1038, 1025, 977, 960, 874, 824, 795, 774, 685, 660 H NMR
(
200 MHz, CDCl
3
):
); 7.55-7.36 (m, 2H, H
); 3.07 (t, 2H, J = 7.2 Hz, H
); Bz 8.00 (d, J = 7.0 Hz, 2H, H
NMR (50 MHz, CDCl ): 166.49; 151.33; 150.01; 149.96; 147.70; 140,94;
36.63; 131.77; 129.46; 129.18; 128.74; 128.36; 124.74; 122.93; 120.46;
15.81, 63.93, 28.18, 22.24.HRMS-ESI-IT-TOF: m/z cal C21 (M+H)
(M+H+2)
δ
Cq 9.03 (bs, 1H, H
, H ); Tr 7.87 (s, 1H, H
); 2.37 (pseudo q, 2H, J = 6.0 Hz, J = 7.4
2
); 8.21 (s, 1H, H
8
); 7.98 (d, 1H, J = 9.2
Hz, 1H, H
5
3
6
5
); Pr 4.47 (t, 2H, J =
6
.2 Hz, H
1
3
13
Hz, H
2
2
, H
6
); 7.55-7.36 (m, 3H, H
3
, H
4
, H
5
).
C
3
δ
1
1
3
3
H
18ClN
4 2
O
93.1118, found 393.1330. HRMS-ESI-IT-TOF: m/z cal C21
H
18ClN O
4 2
95.1293, found 393.1089.

ACCEPTED MANUSCRIPT
8.1.22. 3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propyl-4-ethylbenzoate
(
15)
Yield: 60%; mp 94.7-97.2 ºC; IR (νmax,cm ): 3134, 3094, 3049, 2969, 2918,
-
1
1
1
6
708, 1611, 1590, 1561, 1505, 1458, 1437, 1342, 1325, 1306, 1283, 1245,
228, 1179, 1114, 1068, 1042, 1021, 956, 918, 873, 852, 821, 810, 773, 747,
1
97, 683. H NMR (200 MHz, CDCl
3
):
); 7.29 (bs, 1H, H
); 7.10 (d, 2H, J = 8.4 Hz, H
); 1.10 (t, 3H, J = 7.6 Hz, CH ); Pr 4.33 (t, 2H, J = 6.0 Hz, H
H, J = 7.2 Hz, H ), 2.20 (m, 2H, H
δ
Cq 8.87 (bs, 1H, H
); Tr 8.05 (s, 1H, H
, H ); 2.54 (q, 2H, J = 7.6
); 2.93 (t,
166.59;
2
); 7.84-7.79 (m, 1H,
H
5
, H
d, 2H, J = 8.4 Hz, H3,
Hz, CH
8
); 7.40 (d, 1H, J = 9 Hz, H
6
3
5
); Bz 7.81
(
H
5
2
6
2
3
1
1
3
2
1
1
1
4
4
3
2
). C NMR (50 MHz, CDCl
3
):
δ
51.32; 150.06; 149.90; 147.79; 140,95; 136.65; 129.64; 129.19; 128.82;
27.92; 127.54; 124.79; 122.95; 120.49; 115.83, 63.78, 28.90, 28.25, 22.33,
5.22. HRMS-ESI-IT-TOF: m/z calculated C H ClN O (M+H) 421.1431, found
2
3
21
4
2
21.1355. HRMS-ESI-IT-TOF: m/z cal C23
23.1371.
H
21ClN
4
O
2
(M+H+2) 423.1431, found
8.1.23.
3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propyl-4-tert-butyl-
benzoate (16)
Yield: 67%; mp 91.3-94.2 °C ; IR ( νmax, cm ): 3134, 3047, 2921, 1704, 1611,
-
1
1591, 1561, 1505, 1480, 1451, 1429, 1354, 1304, 1275, 1255, 1233, 1177,
1
1
121, 1110, 1034, 1016, 995, 956, 918, 880, 872, 818, 750, 689, 670. H NMR
(
200 MHz, CDCl
3
):
); 7.47-7.32 (m, 2H, H
, H
t, 2H, J = 6.2 Hz, H
δ
Cq 8.91 (bs, 1H, H
2
); 8.10 (d, J = 2.0 Hz, 1H, H
); Tr 7.90-7.86 (m, 1H, H ); Bz 7.88
, H ); 1.24 (s, 9H, 3xCH ); Pr 4.39
); 2.99 (t, 2H, J = 7.2 Hz, H ); 2.26 (pseudo q, 2H, J = 6.2
): 166.47; 156.64; 151.29;
49.96; 147.73; 140,90; 136.57; 129.35; 129.10; 128.72; 127.24; 125.32;
24.75; 122.92; 120.39; 115.74, 63.72, 34.98, 31.02, 28.19, 22.27. HRMS-ESI-
8
); 7.90-
7
.86 (m, 1H, H
5
3
, H
6
5
(d, 2H, J = 8.2, H
3
5
); ); 7.34 (d, J = 8.2, H
2
6
3
(
1
3
1
3
Hz, J = 7.2 Hz, H
2
). C NMR (50 MHz, CDCl
3
δ
1
1
IT-TOF: m/z calculated C H ClN O Na (M+Na) 471.1563, found 471.1610.
2
5
25
4
2
HRMS-ESI-IT-TOF: m/z cal C25
73.1482.
H
4 2
25ClN O Na (M+Na+2) 473.1534, found
4

ACCEPTED MANUSCRIPT
8
.1.24.
benzoate (17)
Yield: 81%; mp 94.2-97.9 °C ; IR ( νmax, cm ): 3131, 3069, 3009, 2933, 1698,
3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propyl-4-methoxy-
-
1
1
1
7
605, 1579, 1561, 1510, 1480, 1451, 1431, 1376, 1356, 1299, 1276, 1256,
233, 1198, 1165, 1120, 1100, 1029, 995, 968, 951, 881, 874, 843, 818, 786,
1
66, 694, 671. H NMR (200 MHz, CDCl
3
):
); 7.30-7.27 (m, 2H, H3,
Bz 7.88-7.80 (m, 2H, H3, ); 6.73 (d, 2H, J = 8.6 Hz, H2,
OCH3); Pr 4.33 (t, J = 5.6 Hz, 2H, H ); 2.93 (t, 2H, J = 6.8 Hz, H
δ
Cq 8.85 (bs, 1H, H
). Tr 7.88-7.80 (m, 1H, H
); 3.68 (s, 3H,
); 2.18 (pseudo
166.04; 163.19;
51.19; 149.83; 147.63; 140,73; 136.36; 131.32; 128.91; 128.58; 124.67;
22.83; 122.25; 120.22; 115.58, 113.44; 63.52, 55.24; 28.11, 22.17. HRMS-
(M+1) 423.1223, found 423.1204.
HRMS-ESI-IT-TOF: m/z cal C H ClN O (M+H+2) 425.1194, found 425.1187.
2
); 8.02 (s, 1H,
H
8
); 7.88-7.80 (m, 1H, H
5
H
6
5
);
H
5
H
6
1
3
1
3
2 3
q, 2H, J = 5.6 Hz, J = 6.8 Hz, H ). C NMR (50 MHz, CDCl ): δ
1
1
4 3
ESI-IT-TOF: m/z calculated C22H19ClN O
22
19
4
3
8
.1.25. 3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propyl-4-trifluorometoxy
benzoate (18)
Yield: 75%; mp 112.5-116.4 °C; IR ( νmax, cm ): 3130, 3089, 3051, 2963, 2931,
-
-1
1
1
8
1
1
712, 1611, 1590, 1561, 1505, 1480, 1450, 1437, 1412, 1376, 1325, 1310,
285, 1250, 1165, 1135, 1124, 1101, 1064, 1039, 1017, 996, 954, 917, 878,
1
62, 853, 819, 808, 773, 704, 682. H NMR (200 MHz, CDCl
3
):
δ
Cq 8.98 (bs,
H, H ); 8.15-8.09 (m, 1H, H ); 7.95 (d, J = 9.0 Hz, 1H, H ); 7.51(d, J = 9.0 Hz,
2
8
5
H, H
6
); 7.40 (d, 1H, J = 4.0 Hz, H
3
); Bz 7.96 (d, 2H, J = 8.2, H
2
, H
6
); 7.64 (d, J
=
8.2 Hz, 2H, H3,
t, 2H, J = 7.2 Hz, H
50 MHz, CDCl3):
H
5
); Tr 7.86 (s, 1H, H
); 2.30 (pseudo q, 2H, J = 6.2 Hz, J = 7.2 Hz, H
165.39; 151.39; 150.14; 147.70; 141.06; 136.87; 134.83;
5
); Pr; 4.47 (t, 2H, J = 6.2 Hz, H ); 3.03
1
1
3
(
(
3
2
). C NMR
δ
1
33.83 (d, 1C, J = 38.6 Hz); 130.03; 129.77; 129.36; 128.90; 128.72; 124.77,
23.04 (d, 1C, J = 245.1 Hz), 122.93, 115.90, 64.62, 28.27, 22.30. HRMS-ESI-
1
IT-TOF: m/z calculated C22
HRMS-ESI-IT-TOF: m/z cal C22
85.0624.
H
16ClF
3
N
4
O
2
Na (M+Na) 483.0811, found 483.0529.
H
16ClF N O Na (M+Na+2) 485.0782, found
3 4 2
4

ACCEPTED MANUSCRIPT
8.1.26. 3-(1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)propyl-4-trifluorometoxy
-
benzoate (19)
Yield: 68%; mp 81.0-83.9 °C ; IR ( νmax, cm ): 3150, 3066, 2968, 2952, 2905,
-
1
2
1
7
870, 1700, 1609, 1596, 1560, 1507, 1478, 1451, 1429, 1359, 1278, 1254,
228, 1189, 1126, 1114, 1028, 1014, 993, 962, 950, 882, 872, 852, 819, 812,
1
69, 733, 704, 671. H NMR (200 MHz, CDCl
3
):
); 7.86 (d, 1H, J = 2.0 Hz, H
); 7.31 (d, 1H, J = 4.6 Hz, H ); Tr 7.85 (s, 1H, H
d, 2H, J = 8.2 Hz, H3, ); 7.09 (d, J = 8.2 Hz, 2H, H2, ); 2.27 (s, 3H, CH
δ
Cq 8.87 (d, 1H, J = 4.6 Hz,
); 7.41 (dd, 1H, J =
); Bz 7.80
); Pr
H
2
); 8.05 (d, 1H, J = 2.0 Hz, H
8
5
2.0 Hz, J = 8.8 Hz, H
6
3
5
(
H
5
H
6
3
4
6
1
1
.35 (t, 2H, J = 6.2 Hz, H
1
); 2.96 (t, 2H, J = 7.4 Hz, H
3
); 2.21 (pseudo q, 2H, J =
1
3
.2 Hz, J = 7.4 Hz, H
2
). C NMR (50 MHz, CDCl
3
):
δ
165.45; 151.22; 149.89;
47.66; 143.59; 140,80; 136.47; 129.40; 128.99; 128.98; 128.63; 127.21;
24.71; 122.87; 120.30; 115.65, 63.70, 28.13, 22.22, 21.52. HRMS-ESI-IT-TOF:
m/z cal C H ClN O (M+H) 407.1275, found 407.1539.HRMS-ESI-IT-TOF: m/z
2
2
20
4
2
4 2
cal C22H20ClN O (M+H+2) 409.1245, found 409.0144.
Route C (small chain)- To a solution of acid chloride (12.95 mmol) in 4 mL
CH Cl , prop-2-yn-1-ol (0.48 g, 8,63 mmol) was added in one portion at room
2
2
temperature, then trietylamine (19.40 mmol, 2,58 mL) was added in drops and
the final mixture was stirred overnight, where upon TLC indicated reaction
completion. The crude was diluted with 20 mL CH
0 mL), HCl 0,1M (1x 10 mL) and sodium hydroxide 0.1M (1x 10 mL) then dried
over anhydrous Na SO , and evaporated to dryness. The resulting product
2 2
Cl , washed with water (3x
4
2
4
residue was used in the next step without further purification.
In the next step, 4-azido-7-chloroquinoline (102 mg, 0.5mmol) and ester alkyne
(
0.5 mmol) were dissolved in CH
mmol) and a freshly prepared solution of sodium ascorbate, made using
ascorbic acid (53 mg, 0.3 mmol) and NaHCO (25 mg, 0.3 mmol) in 2 mL of
water. The resulting mixture was stirred overnight, the reaction was completed
by TLC and final compound was elaborated with CH Cl , water (3x 10 mL);
dried over Na SO and purified by preparatory TLC.
2 2 4 2
Cl (2 mL), added CuSO .5H O (38 mg, 0,15
3
2
2
2
4

ACCEPTED MANUSCRIPT
8.1.27. Prop-2-ynyl benzoate (40)
-1
IR (νmax, cm ): 3295, 2946, 2129, 1786, 1719, 1600, 1584, 1492, 1451, 1435,
1
7
4
369, 1315, 1262, 1211, 1175, 1105, 1095, 1069, 1026, 979, 925, 872, 804,
1
1
77, 706, 671. H NMR (200 MHz, CDCl
.76 (d, J = 1.4 Hz, 2H, H ); 2.43 (bs, 1H, H
); 7.36-7,23 (m, 3H, H3,
3
):
δ
H NMR (200 MHz, CDCl
).Bz 7.90 (d, J = 7.6 Hz, 2H, H2,
). C NMR (50 MHz, CDCl ): 165.61; 133.26;
3
): δ Pr
1
3
13
H
6
H
4
, H
5
3
δ
129.71; 129.34; 128,38; 77.86; 75.17; 52.37.
8.1.28. Prop-2-ynyl 4-methylbenzoate (41)
-
1
IR (νmax, cm ): 3250, 2979, 2947, 2739, 2603, 2497, 2124, 1774, 1712, 1609,
1508, 1475, 1444, 1397, 1383, 1366, 1267, 1222, 1209, 1171, 1120, 1099,
1
1
035, 1015, 981, 924, 851, 840, 824, 807, 752, 730, 682. H NMR (200 MHz,
CDCl3):
d, J = 8.2 Hz, 2H, H2,
NMR (50 MHz, CDCl
δ
Pr 4.88 (d, J = 2.4 Hz, 2H, H ); 2.52 (t, J = 2.4 Hz, 1H, H ); Bz 7.94
3
1
1
3
(
H
):
6
); 7.20 (d, J = 8.2 Hz, 2H, H3,
H
5
3
); 2.38 (s, 3H, -CH ). C
3
δ 165.27; 144.11; 129.17; 128.99; 126,17; 77.88; 75.03;
52.28; 21.82.
8.1.29. Prop-2-ynyl 4-tert-butylbenzoate (42)
-
1
IR (νmax, cm ): 3297, 2964, 2906, 2871, 2129, 1786, 1721, 1608, 1571, 1505,
1463, 1434, 1409, 1366, 1314, 1299, 1266, 1224, 1187, 1112, 1093, 1041,
1
1
1
016, 983, 925, 853, 874, 773, 706, 673. H NMR (200 MHz, CDCl
): Pr 4.91(d, J = 2.4 Hz, 2H, H ); 2.52 (t, J = 2.4 Hz, 1H, H
Bz 8.00 (d, J = 8.6 Hz, 2H, H2, ); 7.45 (d, J = 8.6 Hz, 2H, H3, ); 1.35 (s, 9H,
). C NMR (50 MHz, CDCl ): 165.76; 157.09; 129.71; 126.67; 125,47;
7.98; 74.92; 53.58; 52.27; 31.09.
3
):
δ
H NMR
(200 MHz, CDCl
3
δ
3
1
);
H
6
H
5
1
3
3x -CH
3
3
δ
7
8.1.30. Prop-2-ynyl 4-ethyllbenzoate (43)
-1
IR (νmax, cm ): 3296, 2968, 2129, 1720, 1689, 1610, 1574, 1509, 1433, 1417,
1
1
369, 1310, 1263, 1177, 1095, 1051, 1019, 981, 925, 852, 701, 675 H NMR
(200 MHz, CDCl
3
):
δ
Pr 4.88 (d, J = 2.4 Hz, 2H, H
1
); 2.52 (t, J = 2.4 Hz, 1H, H
3
);
Bz 7.94 (d, J = 8.0 Hz, 2H, H2,
H
6
); 7.23 (d, J = 8.0 Hz, 2H, H3,
H
5
); 2.65 ( q, J =
1
3
7.6 Hz, 3H, -CH
2 3 3
); 1.22 (t, J = 7.6 Hz, 2H, -CH ). C NMR (50 MHz, CDCl
):
δ
165.80; 150.27; 129.94; 128.42; 127,98; 77.91; 74.92; 52.26; 28.96; 15.18.

ACCEPTED MANUSCRIPT
8.1.31. Prop-2-ynyl 2-ethyllbenzoate (44)
-1
IR (νmax, cm ): 3295, 2972, 2130, 1785, 1720, 1601, 1575, 1487, 1448, 1366,
1
1
290, 1252, 1229, 1198, 1140, 1129, 1077, 1056, 977, 751, 708, 661. H NMR
(
200 MHz, CDCl
Bz 7.94 (dd, J = 0.8 Hz, J = 7.4 Hz, 1H, H
J = 2.0 Hz, J = 5.6 Hz, 1H, H ); 2.98 (q, J = 7.6 Hz, 2H, CH
Hz, 3H, CH ). C NMR (50 MHz, CDCl ): 166.67; 146.50; 132.51; 131.44;
3
):
δ
Pr 4.88 (d, J = 2.4 Hz, 2H, H
); 7.45-7.38 (m, 2H, H
); 1.26 (t, J = 7.6
1
); 2.52 (t, J = 2.4 Hz, 1H, H
3
);
6
5
, H ); 7.25 (dd,
4
3
2
13
δ
3
3
130.85; 130,35; 125,82; 77.87; 75.01; 52.23; 27.62; 15.94.
8.1.32. Prop-2-ynyl 2,4,5-trifluoro-3-methoxybenzoate (45)
-1
IR (νmax, cm ): 3300, 3078, 2954, 2847, 2132, 1804, 1724, 1620, 1508, 1473,
1435, 1377, 1351, 1276, 1213, 1185, 1100, 1054, 982, 956, 932, 910, 874, 780,
1
7
63, 733, 684. H NMR (200 MHz, CDCl ):
δ
Pr 4.94 (d, J = 2.4 Hz, 2H, H1);
3
13
2
.58 (t, J = 2.4 Hz, 1H, H ); Bz 7.56-7,474 (m, 1H, H ); 4.10 (s, 3H, -OCH ). C
3 6 3
NMR (50 MHz, CDCl3):
δ 161.87; 155.23; 149.53(m, 1C); 144.66 (m, 1C);
139.00 (m, 1C); 113.80 (m, 1C); 112.43(d, J = 20.9 Hz, 1C); 77.11; 75.72;
62.42; 53.20.
8.1.33. Prop-2-ynyl 4-(trifluoromethyl)benzoate (46)
-1
IR (νmax, cm ): 3308, 2950, 2131, 1728, 1619, 1587, 1514, 1437, 1412, 1371,
1323, 1265, 1166, 1126, 1095, 1064, 1016, 979, 924, 861, 820, 790, 773, 752,
1
7
03, 689. H NMR (200 MHz, CDCl
3
):
); Bz 8.16 (d, J = 8.2 Hz, 2H, H2,
). C NMR (50 MHz, CDCl ): 164.62; 134.55 (q, J = 32.5 Hz, 1C); 132.75;
δ
Pr 4.97 (d, J = 2.0 Hz, 2H, H
1
); 2.54
(
bs, 1H, H
3
6
H ); 7.69 (d, J = 8.2 Hz, 2H, H3,
1
3
H
5
3
δ
130.28; 123,69 (d, J = 271.0 Hz, 1C); 120,98; 744.40; 75.57; 53.01.
8.1.34. Prop-2-ynyl 4-methoxybenzoate (47)
-1
IR (νmax, cm ): 3238, 2952, 2118, 1705, 1604, 1578, 1511, 1465, 1428, 1369,
1
1
254, 1167, 1095, 1026, 1008, 982, 922, 844, 768, 726, 694. H NMR (200
MHz, CDCl
3
):
δ
Pr 4.89 (d, J = 2.4 Hz, 2H, H
); 6.92 (d, J = 9.0 Hz, 2H, H3,
163.83; 132.48; 132.06; 121.91; 113.86;
1
); 2.53 (t, J = 2.4 Hz, 1H, H
3
); Bz
8.02 (d, J = 9.0 Hz, 2H, H2,
H
6
H
5
); 3.86 (s, 3H, -
1
3
OCH ). C NMR (50 MHz, CDCl ):
δ
3
3
77.88; 75.03; 55.61; 52.34.
8.1.35. Prop-2-ynyl 4-chloro-benzoate (48)

ACCEPTED MANUSCRIPT
1
H NMR (200 MHz, CDCl
H, H ); Bz 7.99 (d, J = 8.6 Hz, 2H, H2,
NMR (50 MHz, CDCl ): 165.05; 139.98; 131.33; 128.95; 127,97; 77.87; 75.43;
3
):
δ
Pr 4.91(d, J = 2.4 Hz, 2H, H
1
); 2.54 (t, J = 2.4 Hz,
13
1
3
H
6
); 7.42 (d, J = 8.6 Hz, 2H, H3,
5
H ). C
3
δ
52.05.
8.1.36. (1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)methyl-4-ethylbenzoate
(
20)
-1
Yield: 67%; mp 95.7-102.1 °C ; IR ( νmax, cm ): 3155, 2964, 2930, 1707, 1609,
1
1
δ
594, 1562, 1507, 1455, 1417, 1343, 1299, 1273, 1246, 1182, 1112, 1040,
1
022, 982, 955, 882, 875, 853, 825, 766, 701, 656. H NMR (200 MHz, CDCl
3
):
); 7.98 (d, J = 9.2
); 7.49 (d, 1H, J = 4.6 Hz,
);
Cq 9.04 (d, J = 4.6 Hz, 1H, H
2
); 8.21 (d, J = 1.8 Hz, 1H, H
8
Hz, 1H, H
); 7.59 (dd, J = 1.8 Hz, J= 9.0 Hz, 1H, H
5 6
H
3
); Tr 8.23 (s, 1H, H
5
); 5.62 (s, 2H, CH
2
); Bz 7.98 (d, J = 8.2 Hz, 2H, H2,
H
6
7
7
1
1
.26 (d, 2H, J = 8.0 Hz, H H ); 2.94 (q, 2H, J = 7.6 Hz, CH ); 1.24 (t, 3H, J =
3,
5
2
1
3
.6 Hz, CH
3
). C NMR (50 MHz, CDCl
3
):
δ
166.65; 151.48; 150.52; 150.26;
44.09; 140,90; 137.02; 130.05; 129.59; 129.07; 128.14; 127.04; 126.02;
24.69; 120.62; 116.17, 57.75, 29.09, 15.33. HRMS-ESI-IT-TOF: m/z calculated
C H ClN O (M+H) 393.1118, found 393.0648. HRMS-ESI-IT-TOF: m/z cal
2
1
18
4
2
C
21
H18ClN
4
O
2
(M+H+2) 395.1089, found 395.1322.
8
.1.37. (1-(7-Chloroquinolin-4-yl)-1H-1,2,3-triazol-4-yl)methyl-2-ethylbenzoate
21)
Yield: 70%; mp 84.1-87.3 °C ; IR ( νmax, cm ): 3138, 3095, 2985, 2972, 1718,
(
-
1
1608, 1591, 1560, 1505, 1484, 1451, 1439, 1320, 1293, 1257, 1245, 1229,
1
1
142, 1115, 1088, 1038, 954, 877, 872, 845, 820, 811, 752, 709, 672, 653. H
NMR (200 MHz, CDCl
3
):
); 7.94 (d, J = 9.2 Hz, 1H, H
.46 (d, J = 4.6 Hz, 1H, H ); Tr 8.25 (s, 1H, H
7.2 Hz, 1H, H ); 7.26 (d, J = 7.2 Hz, 2H, H
.99 (q, J = 7.4 Hz, 2H, CH
δ
Cq 9.05 (d, J = 4.6 Hz, 1H, H
); 7.50 (dd, J = 2.0 Hz, J= 9.0 Hz, 1H, H
); 5.61 (s, 2H, CH ); Bz 7.92 (t, J
); 7.40 -7.27 (m, 2H, H , H ) ;
). C NMR (50 MHz,
2
); 8.21 (d, J = 2.0 Hz,
1H, H
8
5
6
);
7
3
5
2
=
6
4
3
5
1
3
2
2
); 1.22 (t, 3H, J = 7.4 Hz, CH
3
CDCl3):
δ 167.35; 151.43; 150.52; 146.48; 143,94; 140,82; 136.95; 132.91;
1
30.88; 130.41; 129.51; 128.98; 128.50; 125.87; 125.86; 124.56; 120.54;
16.13, 57.67, 27.64, 15.96. HRMS-ESI-IT-TOF: m/z calculated C H ClN O
2
1
2
1
18
4