Microwave-assisted synthesis of 2-styrylquinoline-4-carboxylic acid derivatives to improve the toxic effect against Leishmania (Leishmania) amazonensis

Article abstract of DOI:10.1002/jhet.4217

The identification of new compounds is urgent to develop safe and efficacious candidates for leishmaniasis treatment, especially from natural products as a potential source of active molecules against neglected tropical parasite diseases. Inspired by the efficacious quinoline alkaloid microbial effects, we have previously reported the synthesis and biological activity of 2-phenylquinoline-4-carboxylic acids and poly-substituted quinolines against parasites. In this work, a series of eighteen 2-styryl-4-quinolinecarboxylic acids were synthesized under microwave irradiation settings obtaining from good to excellent yields (60%-90%), shorter reaction times (2 minutes), and eco-friendly experimental conditions. All these products were evaluated against infective forms of Leishmania (Leishmania) amazonensis, such as promastigotes and intracellular amastigotes, based on cytotoxicity assays, including host macrophage infection assays. Compounds 4 and 5 possessing a 2-chloro or 4-chlorostyryl moiety, respectively, were considered the most promising antileishmanial agents due to the parasite killing effect in intracellular forms inside infected macrophages. Thus, our results revealed that the 2-styryl-4-quinolinecarboxylic acid backbone structure was essential for the activity against intracellular pathogens like L. (L.) amazonensis.

Full text of DOI:10.1002/jhet.4217

Received: 17 August 2020
DOI: 10.1002/jhet.4217
Revised: 16 December 2020
Accepted: 23 December 2020
A R T I C L E
Microwave-assisted synthesis of 2-styrylquinoline-
4-carboxylic acid derivatives to improve the toxic effect
against Leishmania (Leishmania) amazonensis
Ayelen Luczywo1 | Ismael Pretto Sauter2 | Thalita Camêlo da Silva Ferreira2
Mauro Cortez2 | Gustavo P. Romanelli3 | Gabriel Sathicq3 | Silvia E. Asís1
|
1Facultad de Farmacia y Bioquímica,
The identification of new compounds is urgent to develop safe and efficacious
candidates for leishmaniasis treatment, especially from natural products as a
potential source of active molecules against neglected tropical parasite dis-
eases. Inspired by the efficacious quinoline alkaloid microbial effects, we have
previously reported the synthesis and biological activity of 2-phenylquinoline-
4-carboxylic acids and poly-substituted quinolines against parasites. In this
work, a series of eighteen 2-styryl-4-quinolinecarboxylic acids were synthesized
under microwave irradiation settings obtaining from good to excellent yields
(60%-90%), shorter reaction times (2 minutes), and eco-friendly experimental
conditions. All these products were evaluated against infective forms of Leish-
mania (Leishmania) amazonensis, such as promastigotes and intracellular
amastigotes, based on cytotoxicity assays, including host macrophage infection
assays. Compounds 4 and 5 possessing a 2-chloro or 4-chlorostyryl moiety,
respectively, were considered the most promising antileishmanial agents due
to the parasite killing effect in intracellular forms inside infected macrophages.
Thus, our results revealed that the 2-styryl-4-quinolinecarboxylic acid back-
bone structure was essential for the activity against intracellular pathogens like
L. (L.) amazonensis.
Departamento de Química Orgánica,
Universidad de Buenos Aires, Buenos
Aires, Argentina
2Laboratório de Imunobiologia da
~
Interaçao Leishmania-macrófagos,
Departamento de Parasitologia, Instituto
de Ciências Biomédicas, Universidade de
~
~
Sao Paulo, Sao Paulo, Brazil
3Centro de Investigación y Desarrollo en
Ciencias Aplicadas “Dr. Jorge J. Ronco”
(CINDECA-CONICET-CCT-La Plata),
Universidad Nacional de La Plata, La
Plata, Argentina
Correspondence
Mauro Cortez, Laboratório de
~
Imunobiologia da Interaçao Leishmania-
macrófagos, Departamento de
Parasitologia, Instituto de Ciências
~
Biomédicas, Universidade de Sao Paulo,
Av. Prof. Lineu Prestes, 1374, CEP
~
05508-000 Sao Paulo, Brazil.
Email: mcortez@usp.br
Silvia E. Asís, Facultad de Farmacia y
Bioquímica, Departamento de Química
Orgánica, Universidad de Buenos Aires,
Junín 956, C1113AAD Buenos Aires,
Argentina.
Email: elizabet@ffyb.uba.ar
Funding information
Conselho Nacional de Desenvolvimento
Científico e Tecnológico, Grant/Award
~
Number: 443816/2014-0; Fundaçao de
~
Amparo à Pesquisa do Estado de Sao
Paulo, Grant/Award Number:
2012/24105-3; Secretaria de Ciencia y
Tecnica, Universidad de Buenos Aires,
Grant/Award Number:
20020170200206BA
822
© 2020 Wiley Periodicals LLC.
wileyonlinelibrary.com/journal/jhet
J Heterocyclic Chem. 2021;58:822–832.

LUCZYWO ET AL.
823
1 | INTRODUCTION
extracted from the filamentous bacteria Streptomyces
nodosus, and paromomycin obtained from Streptomyces
rimosus.[12] Moreover, reports identifying natural prod-
ucts with antileishmanial activity are always frequent. A
series of 2-substituted quinolines isolated from Galipea
longiflora K. Krause (family Rutaceae), a Bolivian tree
traditionally used to cure ulcerations of CL by the native
Chimanes tribe, was first reported by Fournet et al.[13]
Pure quinoline alkaloids, such as 2-n-propylquinoline,
2-n-pentylquinoline, 4-methoxy-2-phenylquinoline, and
Leishmaniasis is a neglected vector-transmitted tropical
disease, affecting 12 million people around the world,
with 350 million at risk of infection in at least 98 coun-
tries and territories.[1,2] The disease caused by a proto-
zoan parasites species of the genus Leishmania, which is
transmitted by sandflies, presents a broad spectrum of
clinical manifestations, where three principal ones stand
out: cutaneous leishmaniasis (CL), mucocutaneous leish-
maniasis (MCL), and visceral leishmaniasis (VL).[3,4]
Known also as kala-azar, VL is fatal if left untreated in
over 95% of cases. Most of the human cases occur in Bra-
zil, India, East Africa, and South-East Asia, with an esti-
mated 50 000 to 90 000 new cases occurring worldwide
each year, out of which only about 25% to 45% are
reported to the World Health Organization (WHO). CL is
the most common form of leishmaniasis and causes skin
lesions, mainly ulcers, on exposed parts of the body,
whereas MCL leads to the partial or total destruction of
mucous membrane tissues such as nose, mouth, and
throat. About 95% of CL/MCL cases occur in the Ame-
ricas (mainly in Brazil, Peru, Colombia, and Mexico), the
Mediterranean Basin, the Middle East, and Central Asia,
with annually estimated 600 000 to 1 million new cases
worldwide.[2]
The pharmacological treatment for leishmaniasis is
limited, relying primarily on pentavalent antimonials,
amphotericin B (under deoxycholate or liposomal formu-
lations), and miltefosine.[5,6] The WHO has made signifi-
cant efforts by promoting equitable access to health
services and essential drug treatment. Significant achieve-
ments include reducing the price of two of the five existing
medicines for VL by 90% for liposomal amphotericin B
and by 60% for meglumine antimoniate, and a new agree-
ment until 2021 for the donation of 445 000 vials of liposo-
mal amphotericin B injection to treat more than 50 000
VL cases in eligible endemic countries.[2] However, these
few chemotherapeutic options are mostly unsatisfactory
not only by the high costs but also due to low efficacy,
poor safety, the emergence of parasite resistance, among
other factors related to treatment (such as administration
route or length of treatment) that are unsuitable for the
socioeconomic reality of affected populations.[6–11] There-
fore, the identification of new compounds is urgent to
develop safe and efficacious candidates for leishmaniasis
treatment, especially from natural products as a potential
source of active molecules against neglected tropical para-
site diseases.[3] In this context, new and established
pharmacophores are being identified based on synthetic
and natural product chemistry.
2-(3,4-methylenedioxyphenylethyl)-quinoline,
demon-
strated promising activity against Leishmania donovani
(species related to VL) in vitro, and activity by oral route
in the in vivo evaluation.[14] This discovery encouraged
other authors to develop C2-substituted quinolines as
antileishmanial agents.[15,16] Noteworthy, the synthesis of
diverse C2-aryl quinoline derivatives with activity against
Leishmania (Vianna) braziliensis (species related to CL
and MCL), and their possible mechanism of action were
reported.[17]
In addition to their recognized activity as antimalarials,
many synthetic quinoline derivatives are antibacterial, anti-
fungal, antitumor,[18] and antimycobacterial agents.[19,20]
We have previously reported the microwave-assisted syn-
thesis of substituted 2-phenylquinoline-4-carboxylic
acids[21] and poly-substituted quinolines,[22] both series
showing antiparasitic activities. Among them, we found a
hit compound with antimalarial and antichagasic (Chagas
diseases caused by the protozoan Trypanosoma cruzi)
activities and four derivatives with moderate activity
against Leishmania (Leishmania) infantum chagasi and
L. donovani (Figure 1). Furthermore, 2-methyl and 2-aryl-
4-quinolinecarboxylic
acids[23]
and
4-amino-2-sty-
rylquinolines[24] with antileishmanial activity were recently
reported. All these findings prompted us to investigate the
influence on the antileishmanial activity of the vinyl-bridge
insertion between the 2-aryl ring and the quinoline core of
these new 4-quinolinecarboxylic acids.
2 | RESULTS AND DISCUSSION
2.1 | Synthesis and characterization of
2-styryl-4-quinolinecarboxylic acid
Herein, we present the microwave-assisted reaction of
2-methyl-4-quinolinecarboxylic acid (i) and arylaldehydes
(ii) under trifluoroacetic acid (TFA) catalysis (Scheme 1),
to obtain eighteen 2-styryl-4-quinolinecarboxylic acids
(iii, 1-18). All these products were evaluated against
promastigotes and intracellular amastigotes of L. (L.)
amazonensis, based on cytotoxicity assays, including host
macrophages.
Leishmaniasis chemotherapy has been historically
associated with natural products, such as amphotericin B

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LUCZYWO ET AL.
SCHEME 1 Microwave-assisted (MW) synthesis from
2-methyl-4-quinolinecarboxylic acid (i) and arylaldehydes (ii) under
trifluoroacetic acid (TFA) catalysis, to obtain 2-arylvinyl-
4-quinolinecarboxylic acids (iii, 1-18 derivatives)
FIGURE 1 2-methyl and 2-aryl-4-quinolinecarboxylic acids
(two compounds each) previously shown to possess in vitro
antileishmanial activity
coefficient (logP), topological polar surface area (TPSA),
the number of hydrogen-bond donors (HBD) and accep-
tors (HBA), and the number of rotatable bonds for all
synthesized compounds (iii) were estimated using the
online prediction tools from the Molinspiration server.[31]
The starting material (i) was prepared employing the
Pfitzinger reaction from isatin and ketone,[25] then it was
subjected to Knoevenagel condensation to provide the
2-styryl-4-quinolinecarboxylic acids (iii) in good to excel-
lent yields (60%-90%) and short reaction times
2.2 | In vitro activity against L. (L.)
amazonensis and bone marrow
macrophages
1
(2 minutes). The H NMR spectra showed that the com-
pounds (iii) are E stereoisomers owing to the value of
16.0 to 16.6 Hz for the double bond hydrogen atoms cou-
pling constant J. Moreover, the IR spectra contained
absorption bands at 960 to 968 cm−1, typical of stretch
vibrations of the CH=CH bond in the trans form.[26,27]
Derivatives 1 to 3 and 13 were prepared under thermal
heating in acetic anhydride,[26,27] whereas compound
6 was obtained from isatin.[28] In the literature, only their
melting point values and the 1H NMR spectrum for com-
pound 13 were found.[27] In this work, all the yields are
higher than the previously reported values, including 5,
7, and 14 to 17.[9,25] To evaluate the method, the EcoScale
Calculator Index was determined.[29] This procedure
receives a significantly excellent score of 77 (score from
0 to 100) based on economic and ecological parameters.
Most of the selected arylaldehydes (ii) possess system-
atic changes at positions 2 to 5 of the phenyl-ring to
determine the effect of substitution with atoms or moie-
ties, bearing different electronic, hydrophobic, and steric
properties on antileishmanial potency and cytotoxicity.
In this way, 2-fluoro, 2-nitro, 2-chloro, 4-methoxyphenyl
groups, and their position isomers were the main
targeted compounds.[21,30] The octanol-water partition
The inhibitory effect of the different derivative com-
pounds on L. (L.) amazonensis proliferation (strain IFLA/
BR/67/PH8) was in vitro assayed using the proliferative
biological stage promastigote in culture media. The inhib-
itory concentration of parasite growth was determined by
3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium
bromide (MTT) assay[32] after 24, 48, and 72 hours of the
proliferation assays in the presence of the compounds,
and analyzed statistically by Student t test in Prism 5.0
software. All compounds were tested at 29.3 μM, single
dose (IC50, previously determined), and each test was car-
ried out in duplicate in two independent experiments.
The treatment with compounds 1 to 6 and 14 decreased
parasite viability (Figure 2), so they were selected for fur-
ther investigation of their inhibitory activity in the intra-
cellular parasite proliferative-stage amastigotes, by a
biological model previously reported.[33,34]
To perform the assay of the derivatives inhibitory
activity against amastigotes, bone marrow macrophages
(BMMs) were infected with promastigotes for 2 hours,
washed, and incubated in media for 24 hours to obtain

LUCZYWO ET AL.
825
intracellular amastigotes. After incubation, amastigotes-
infected BMMs were treated with different derivatives. A
33% to 40% decrease in the number of parasites per cell
was observed in compounds 2, 4, and 5 (Figure 3A),
although the compounds did not affect the percentage of
infected cells (Figure 3B). In the representative images of
amastigote-infected BMMs (Figure 4), in the untreated
sample (Figure 4A) it is possible to visualize two or more
parasites inside of the rounded vacuoles (parasitophorous
vacuoles, PV) in the BMMs, similarly to the derivatives
without effects (compounds 1, Figure 4C; 3, Figure 4E; 6,
Figure 4H; and 14, Figure 4I) whereas in the treatment
with inhibitory effect (compounds 2, Figure 4D; 4,
Figure 4F; and 5, Figure 4G) individual amastigotes were
visualized inside PV (at similar for the inhibitory control
such as amphotericin, Figure 4B). From these results,
compounds 4 and 5 were considered the most promising
antileishmanial agents.
2.3 | SAR analysis, cytotoxicity assay,
and possible target enzyme
The molecular properties of the active compounds 1 to
6 and 14 were calculated (Table 1). Strikingly, it is worthy
to note that only the Log P values for the active com-
pounds 4 and 5 (*) exceed Lipinski's rule of five,[35] the
other derivative compounds falling within the rule.
Lipinski's rule states that, in general, an orally active
drug has no more than one violation of the criteria.
Moreover, 3D molecular structures were created using
Molinspiration Galaxy 3D generator and were examined
in various display modes, including visualization of sur-
face properties, such as molecular lipophilicity potential
(MLP) and polar surface area (PSA). Compounds 4, 5,
and 14 exhibited similar MLP and shape, where the
(substituted) phenyl moiety is perpendicular to the
quinoline-ring. For compounds 1, 2, and 6, the phenyl
significantly approaches the quinoline plane and exhibits
a hydrophilic region (Figure 5).
When the effects of 2-arylvinyl-4-quinolinecarboxylic
acid derivatives on their proliferative (growth%) inhibi-
tion activity were compared in both biological forms of
L. (L.) amazonensis, derivative compounds 1 and 6, which
showed the best inhibitory results on promastigotes, lost
any effect on the intracellular amastigotes (as well as
compound 14). In contrast, derivative compounds 2, 4,
and 5 kept inhibiting the intracellular stage. This infor-
mation is critical since all these derivative molecules
need to cross several membranes (host plasmatic mem-
brane, PV membrane, and parasite surface membrane)
until reaching the parasite inside the host cell. It could
mean that derivatives 1, 6, and 14 were incapable of pen-
etrating the membrane (1 and 6 possess lower Log
FIGURE 2 Inhibitory effect of 2-arylvinyl-
4-quinolinecarboxylic acids derivatives on the proliferation of L. (L.)
amazonensis promastigotes (29.3 μM, single dose). Amp,
amphotericin B; CTL, control sample; DMSO, dimethyl sulfoxide;
Milt, Miltefosine. P-value: ***P < .001; **P .001-.01
FIGURE 3 Inhibitory effect of 2-arylvinyl-4-quinolinecarboxylic acid derivatives on the proliferation of L. (L.) amazonensis intracellular
amastigotes (29.3 μM, single dose). Bone marrow macrophage (BMM)-containing intracellular amastigotes and treated with the compounds
1 to 6, and 14. A, The number of intracellular parasites per 100 infected BMMs. B, Percentage of amastigotes infection. Control: untreated
sample. Amp, amphotericin B. P-value: ***P < .001; **P .001-.01

826
LUCZYWO ET AL.
FIGURE 4 Representative images of amastigote-infected bone marrow macrophages (BMMs) untreated (A, Control) or treated as
indicated (B, amphotericin B; C, Compound 1; D, Compound 2; E, Compound 3; F, Compound 4; G, Compound 5; H, Compound 6; I,
Compound 14). After incubation, the samples were processed by fluorescence staining to recognize the nuclei of macrophages and parasites
(PI, propidium iodide), including an overlay of the contrast phase (phase). Arrows indicate the selected crop for zooming (at the bottom left
of the image). Scale bar: 10 μm [Colour figure can be viewed at wileyonlinelibrary.com]
TABLE 1 Molecular properties of the active products (iii) 1 to 6, and 14 calculated from Molinspiration server
iii
1
LogPa
4.31
TPSAb
96.02
96.02
96.02
50.19
50.19
79.65
50.19
Number of atomsc
MWd
320.30
320.30
320.30
309.75
309.75
321.33
275.30
Number of ONe
Number of OHNHf
Ro5g
24
24
24
22
22
24
21
6
6
6
3
3
5
3
1
1
1
1
1
2
1
0
0
0
1
1
0
0
2
4.52
3
4.54
4
5.03*
5.26*
3.92
5
6
14
4.58
aLogP octanol-water partition coefficient;
bTPSA, topological polar surface area;
cNumber of nonhydrogen atoms;
dMolecular weight;
eNumber of hydrogen-bond acceptors (O and N atoms);
fNumber of hydrogen-bond donors (OH and NH groups);
gNumber of rule of five violations.
*LogP outside Lipinski's rule of five.
P values compared to the others), or these compounds
could be unstable and easily degraded by host factors
(acidic intravacuolar environment, degradation by host
enzymes, among others). Although these synthetic com-
pounds seem to be quite stable,[25] differences in the final
target molecules of the derivative compounds, perhaps in

LUCZYWO ET AL.
827
FIGURE 5 Visualization of
molecular lipophilicity potential
(MLP) for representative 3D
molecular structures, to see which
parts of the surface are hydrophobic
(encoded by violet and blue colors)
and which hydrophilic (orange and
red) [Colour figure can be viewed at
wileyonlinelibrary.com]
the presence or quantities, might differ in the different
biological stages, an important feature when comparing
in the Leishmania spp. drug discovery field.[36,37] Because
the more suitable biological form to test new compounds
against Leishmania spp. is amastigotes,[38,39] derivative
compounds 4 and 5 were found to be promising as
antileishmanial molecules, according to this SAR
analysis.
A further cytotoxicity assay was performed, includ-
ing some compounds in RAW cells, a macrophage-like
cell linage, due to the impossibility to work with mice.
As visualized in Figure S1, similar results of viability
were obtained with trypan blue staining assay compar-
ing to the control and amphotericin B, for 72 hours.
The compounds 5, 6, and 14 with leishmanicidal effect
are not toxic for the host cells. However, MTT assays
showed an effect with the compound 2, which could be
a metabolic effect in macrophages owing to the nitro
group.
As a possible candidate affected by 2-arylvinyl-
4-quinolinecarboxylic acid derivatives, the flavoenzyme
dihydroorotate dehydrogenase (DHODH) is a validated
target for the inhibitory effect against Leishmania spp., as
well as an antiparasite drug in the research field of new
drug discovery.[40] DHODHs catalyze the stereoselective
oxidation of (S)-dihydroorotate (DHO) to orotate (ORO)
in the fourth reaction of the de novo pyrimidine biosyn-
thetic pathway, been divided into Class 1 and Class
2 according to sequence similarity, subcellular location,
and preference for the substrate.[41] Class 1 DHODHs are
cytosolic enzymes, present in Leishmania genus and
T. cruzi, with a few examples of specific inhibitors being
mentioned in the literature.[42,43]
Noteworthy, L. donovani DHODH has explicitly been
proposed as a target for 2-aryl-4-quinolinecarboxylic
acids.[23] By aligning the 3D structures of T. cruzi,
L. major, and human DHODH active sites, it can be seen
that, despite the substitution of some residues, the profile
of interactions for molecular recognition is highly con-
served between the two classes.[44] Furthermore, human
DHODH inhibitors included brequinar (synthetic
4-quinolinecarboxylic acid) and several series of
4-quinolinecarboxylic acid derivatives,[45] also with ant-
iviral activity,[46] strongly suggesting the active com-
pounds 4 and 5 as Leishmania DHODH inhibitors, based
on these reported data.
3 | CONCLUSION
Eighteen (E) 2-styryl-4-quinolinecarboxylic acids were
prepared in good to excellent yields, short reaction
times, and simple work-up procedure under eco-
friendly conditions. Seven of these derivatives are novel
structures and for the remaining known compounds,
this work provides additional spectroscopic data as well
as an improved synthetic method with higher yields
than reported. It was shown that the 2-styryl-
4-quinolinecarboxylic acid backbone is essential for the
activity against L. (L.) amazonensis. The compounds 5,
6, and 14 with leishmanicidal effect are not toxic for the
host cells. The SAR studies indicate that chlorine atoms
attached at the 2-styryl moiety increase such activity,
for this reason, compounds 4 and 5 are attractive candi-
dates for hit-to-lead development, maybe as potential
DHODH inhibitors.

828
LUCZYWO ET AL.
4 | EXPERIMENTAL SECTION
4.1 | General
4.1.3 | (E)-2-(3-nitrostyryl)quinoline-
4-carboxylic acid 2
Yield 86%, pale yellow powder, mp > 320ꢀC (lit. 316ꢀC-
317ꢀC).[26] 1H NMR (600 MHz, DMSO-d6) δ 8.66 (d,
J = 8.5 Hz, 1H, H3), 8.62 (s, 1H, H18), 8.31 (s, 1H, H9),
8.28 (d, J = 7.8 Hz, 1H, H6), 8.20 (dd, J = 8.1; 1.6 Hz, 1H,
H16), 8.11 (d, J = 8.4 Hz, 1H, H14), 8.08 (d, J = 16.4 Hz,
1H, H12), 7.85 (t, J = 7.6 Hz, 1H, H15), 7.81 (d,
J = 16.4 Hz, 1H, H11), 7.75 (t, J = 7.9 Hz, 1H, H1), 7.69
(t, J = 7.6 Hz, 1H, H2). 13C NMR (151 MHz, DMSO-d6) δ
168.0, 155.3, 148.9, 148.9, 138.5, 133.8, 132.9, 131.3, 130.8,
130.65, 129.9, 128.2, 126.0, 124.2, 123.6, 122.3, 121.3. IR
(cm−1): ν 3438, 3064, 1734, 1593, 1530, 1367, 955, 734.
Chemical Formula: C18H12N2O4.
The structures of the synthesized compounds were
1
established through their H and 13C-NMR, MS, and IR
spectra. Melting points were determined in a capillary
Electrothermal 9100 SERIES-Digital apparatus and are
uncorrected. 1H and 13C-NMR spectra were obtained
using a Bruker 600 spectrometer. The operating frequen-
cies for protons and carbons were 600 and 151 MHz,
respectively. The chemical shifts (δ) were given in ppm.
IR spectra were recorded on an FT Bruker from KBr
disks. Mass spectra were measured on MS/DSQ II
Thermo Scientific DPC. Elemental analysis data for syn-
thesized compounds were consistent with calculated
values. Analytical time of reaction completion (TLCs)
was performed on DC-Alufolien Kieselgel 60 F254 Merck.
Microwave-assisted reactions were carried out in a Glass
vial G10, Anton Paar Monowave Series (Serial Number:
81920884, Instrument Software Version: 4.10.9376.7).
4.1.4 | (E)-2-(4-nitrostyryl)quinoline-
4-carboxylic acid 3
Yield 89%, pale yellow powder, mp > 320ꢀC (lit. 324ꢀC-
325ꢀC).[26] 1H NMR (600 MHz, DMSO-d6) δ 10.18 (s, 1H,
H24), 8.66 (d, J = 8.5 Hz, 1H, H3), 8.43 (d, J = 8.6 Hz,
1H), 8.34 (s, 1H, H9), 8.29 (d, J = 8.71 Hz, 1H), 8.15 (d,
J = 16.6 Hz, 1H, H12), 8.09 to 8.04 (m, 3H), 7.86 (dt,
Jo = 8.2, Jp = 0.7 Hz, 1H, H1), 7.82 (d, J = 16.4 Hz, 1H,
H11), 7.71 (dt, Jo = 8.0, Jp = 0.7 Hz, 1H, H2). 13C NMR
(151 MHz, DMSO-d6) δ 192.8, 167.9, 155.0, 148.7, 147.5,
143.3, 140.5, 137.7, 133.1, 131.1, 130.9, 129.8, 128.8, 128.5,
126.0, 124.7, 124.5, 124.2, 121.5. IR (cm−1): ν 3438, 3064,
1734, 1593, 1530, 1367, 955, 734. MS: 320.1 (M+). Chemi-
cal Formula: C18H12N2O4.
4.1.1 | General procedure for the
synthesis of 2-arylvinyl-
4-quinolinecarboxylic acids iii 1 to 18
A neat mixture of 0.25 mmol (0.05 g) of 2-methyl-
4-quinolinecarboxylic acid and 0.25 mmol of the
corresponding arylaldehyde with 0.1 mL TFA was sub-
jected to MW irradiation at 170ꢀC and 850 W. The mix-
ture was cooled to room temperature to give a solid
product which was then triturated from EtOH; TLC is
2 minutes.
4.1.5 | (E)-2-(2-chlorostyryl)quinoline-
4-carboxylic acid 4
4.1.2 | (E)-2-(2-nitrostyryl)quinoline-
4-carboxylic acid 1
Yield 73%, pale yellow powder, mp 312.4ꢀC-312.6ꢀC. 1H
NMR (600 MHz, DMSO-d6) δ 8.66 (d, J = 8.5 Hz, 1H,
H3), 8.20 (s, 1H, H9), 8.18 (d, J = 16.1 Hz, 1H, H12),
8.12 (d, J = 8.4 Hz, 1H, H6), 8.03 (dd, J = 7.6, 1.3 Hz,
1H, H17), 7.83 (dt, J = 8.1, 1.1 Hz, 1H, H2), 7.69 (dt,
J = 8.1, 1.1 Hz, 1H, H1), 7.66 (d, J = 16.1 Hz, 1H, H11),
Yield 77%, pale yellow powder, mp > 300ꢀC (lit. 293ꢀC-
294ꢀC).[26] 1H NMR (600 MHz, DMSO-d6) δ 14.04 (s, 1H,
H24), 8.67 (d, J = 8.2 Hz, 1H, H3), 8.24 (s, 1H, H9), 8.16
(d, J = 16.2 Hz, 1H, H12), 8.11 (d, J = 8.01 Hz, 1H, H6),
8.10 (t, J = 7.1 Hz, 1H, H16), 8.07 (dd, J = 8.1; 0.9 Hz,
1H,H17), 7.85 (td, J = 8.3; 1.1 Hz, 1H, H15), 7.82 (t,
J = 7.5 Hz, 1H, H1), 7.71 (td, J = 8.2; 1.1 Hz, 1H, H2),
7.64 (td, J = 8.3; 0.9 Hz, 1H, H14), 7.63 (d, J = 16.01 Hz,
1H, H11). 13C NMR (151 MHz, DMSO-d6) δ 167.9, 154.9,
148.9, 148.8, 137.6, 134.1, 132.9, 131.4, 130.7, 130.1, 130.0,
129.6, 129.1, 128.4, 125.9, 125.1, 124.3, 121.7. IR (cm−1): ν
3469, 3063, 3032, 1734, 1593, 1523, 1375, 969, 733. Chem-
ical Formula: C18H12N2O4.
7.56 (dd,
J = 7.8,0.9 Hz, 1H, H14), 7.44 (dt,
J = 7.2,1.1 Hz, 1H, H15), 7.40 (td, J = 7.6, 1.5 Hz, 1H,
H16). 13C NMR (151 MHz, DMSO-d6) δ 167.9, 155.1,
148.9, 137.5, 134.2, 133.5, 131.3, 130.8, 130.7, 130.4,
130.02, 129.9, 128.2, 128.1, 128.0, 125.9, 124.2, 122.0. IR
(cm−1): ν 3485, 3050, 1734, 1593, 1390, 953, 875. MS:
309.8 (M+). Calcd. Analysis for C18H12ClNO2: C,
69.80; H, 3.90; N, 4.52. Found: C, 69.72; H,
4.05; N, 4. 49.

LUCZYWO ET AL.
829
4.1.6 | (E)-2-(4-hydroxy-3-methoxystyryl)
quinoline-4-carboxylic acid 6
4.1.9 | (E)-2-(2,3-difluorostyryl)
quinoline-4-carboxylic acid 10
1
Yield 90%, orange powder, mp 314ꢀC-316ꢀC (lit. 282ꢀC-
283ꢀC).[28] 1H NMR (600 MHz, DMSO-d6) δ 13.91 (s,
1H, H22), 8.61 (dd, J = 8.5; 0.6 Hz, 1H, H3), 8.21 (s, 1H,
H9), 8.05 (d, J = 8.3 Hz, 1H, H6), 7.82 (d, J = 16.3 Hz,
1H, H12), 7.80 (td, J = 8.3; 1.2 Hz, 1H, H1), 7.65 (td,
J = 8.2; 1.2 Hz, 1H, H2), 7.41 (d, J = 16.2 Hz, 1H, H11),
7.39 (d, J = 2.0 Hz, 1H, H18), 7.18 (dd, J = 8.2;2.0 Hz,
1H, H14), 6.85 (d, J = 8.03 Hz, 1H, H15), 3.87 (s, 3H,
OCH3). 13C NMR (151 MHz, DMSO-d6) δ 168.1, 156.3,
148.5, 148.4, 137.1, 135.9, 130.5, 129.5, 128.1, 127.5,
125.9, 125.3, 123.7, 122.1, 121.0, 120.9, 116.1, 111.1,
56.1. IR (cm−1): ν 3453, 3078, 2984, 2945, 1641, 1594,
1390, 1281, 1345, 954, 875. Chemical Formula:
C19H15NO4.
Yield 72%, yellow powder, mp 289.7ꢀC-290.2ꢀC. H NMR
(600 MHz, DMSO-d6) δ 14.00 (s, 1H, H23), 8.64 (dd,
J = 8.5;0.7 Hz, 1H, H3), 8.25 (s, 1H,H9), 8.12 (dd, J = 8.4;
0.5 Hz, 1H, H6), 7.98 (d, J = 16.3 Hz, 1H,H12), 7.84 (dt,
J = 7.0;1.5 Hz, 1H, H2), 7.77 (dt, J = 7.0;1.5 Hz, 1H, H1),
7.72 (d, J = 16.3 Hz, 1H, H11), 7.69 (dt, J = 8.3;1.2 Hz,
1H, H14), 7.45 (dc, J = 8.2;1.2 Hz, 1H, H15), 7.45 (dc,
J = 8.0;1.1 Hz, 1H, H16). 13C NMR (151 MHz, DMSO-d6)
δ 167.9, 167.9, 160.3, 155.0, 151.5, 149.7, 149.3, 149.2,
148.9, 148.5, 147.7, 137.7, 136.4, 132.3, 132.3, 130.7, 130.1,
130.0, 129., 128.3, 127.6, 126.6, 126.5, 125.9, 125.8, 125.7,
125.6, 125.5, 124.8, 124.2, 123.9, 123.4, 123.1, 121.7, 117.8,
117.7. IR (cm−1): ν 3420, 3060, 1656, 1485, 1250, 976, 780.
Calcd. Analysis for C18H11F2NO2: C, 69.45; H, 3.56; N,
4.50. Found: C, 69.37; H, 3.62; N, 4.41.
4.1.7 | (E)-2-(3,5-difluorostyryl)
quinoline-4-carboxylic acid 8
4.1.10 | (E)-2-(2,6-difluorostyryl)
quinoline-4-carboxylic acid 11
1
Yield 82%, yellow powder, mp 319.3ꢀC-319.9ꢀC. H NMR
1
(600 MHz, DMSO-d6) δ 8.66 (d, J = 8.5 Hz, 1H, H3), 8.20
(s, 1H, H9), 8.17 (d, J = 16.1 Hz, 1H, H12), 8.12 (d,
J = 8.5 Hz, 1H, H6), 8.03 (d, J = 7.7 Hz, 1H, H14), 7.83 (t,
J = 7.6, 1H, H2), 7.69 (td, J = 8.5, 2.8 Hz, 1H, H1), 7.65
(d, J = 16.1 Hz, 1H, H11), 7.55 (d, J = 8.9 Hz, 1H, H18),
7.46 to 7.40 (m, 1H, H16). 13C NMR (151 MHz, DMSO-
d6) δ (151 MHz, DMSO) δ 167.9, 164.0, 162.3, 155.2,
148.9, 140.6, 140.5, 137.3, 132.9, 131.3, 130.7, 129.9, 128.3,
125.9, 124.2, 121.4, 110.7, 104.5, 104.3, 104.2. IR (cm−1): ν
3438, 3078, 3031, 1640, 1593, 1328, 1125, 984, 859. Calcd.
Analysis for C18H11F2NO2: C, 69.45; H, 3.56; N, 4.50.
Found: C, 69.38; H, 3.63; N, 4.43.
Yield 60%, yellow powder, mp 284.6ꢀC-285.0ꢀC. H NMR
(600 MHz, DMSO-d6) δ 8.63 (d, J = 8.5 Hz, 1H, H3), 8.22 (s,
1H, H9), 8.13 (d, J = 8.4 Hz, 1H, H6), 7.90 (d, J = 16.5 Hz,
1H, H12), 7.84 (t, J = 7.6 Hz, 1H, H2), 7.71 (t, J = 8.3, 1H,
H1), 7.68 (d, J = 16.5 Hz, 1H, H11), 7.47 (m, 1H, H17), 7.24
(t, J = 8.6 Hz, 2H, H15,H16). 13C NMR (151 MHz, DMSO-
d6) δ 167.9, 161.9, 160.2, 155.0, 148.8, 137.7, 134.3, 131.0,
130.7, 130.1, 128.3, 125.9, 124.3, 121.8, 120.9, 113.6, 112.8.
IR (cm−1): ν 3437, 3078, 1711, 1633, 1484, 1256, 1000,
984, 781. Calcd. Analysis for C18H11F2NO2: C, 69.45; H,
3.56; N, 4.50. Found: C, 69.39; H, 3.60; N, 4.44.
4.1.11 | (E)-2-(2,5-difluorostyryl)
quinoline-4-carboxylic acid 12
4.1.8 | (E)-2-(3,4-difluorostyryl)
quinoline-4-carboxylic acid 9
Yield 67%, yellow powder, mp 299ꢀC-300ꢀC. 1H NMR
(600 MHz, DMSO-d6) δ 14.0 (s, 1H, H23), 8.65 (dd,
J = 8.0, 0.6 Hz, 1H, H3), 8.21 (s, 1H, H9), 8.12 (d,
J = 8.3 Hz, 1H, H6), 7.94 (d, J = 16.3 Hz, 1H, H12), 7.85
(td, J = 3.7 Hz, 1H, H14), 7.83 (td, J = 8.3, 7.0 Hz, H1),
7.74 (d, J = 16.3 Hz, 1H, H11), 7.69 (ddd, J = 8.2,
7.0,1.0 Hz, 1H, H2), 7.38 (td, J = 9.0, 4.5 Hz, 1H, H17),
7.27 (ddd, J = 9.0, 7.4, 3.5 Hz, 1H, H16). 13C NMR
(151 MHz, DMSO-d6) δ (151 MHz, DMSO) δ 167.9, 159.7,
158.2, 157.7, 156.1, 155.0, 148.9, 137.6, 132.2, 132.1, 130.7,
130.0, 128.3, 125.9, 125.7, 124.3, 121.7, 117.4, 114.5. IR
(cm−1): ν 3420, 3060, 1656, 1485, 1250, 969, 780. Calcd.
Analysis for C18H11F2NO2: C, 69.45; H, 3.56; N, 4.50.
Found: C, 69.40; H, 3.59; N, 4.48.
1
Yield 84%, yellow powder, mp 309.0ꢀC-309.6ꢀC. H NMR
(600 MHz, DMSO-d6) δ 8.64 (d, J = 7.9 Hz, 1H, H3), 8.23
(s, 1H, H9), 8.08 (d, J = 8.1 Hz, 1H, H6), 7.91 (ddd,
J = 7.9, 1.7 Hz, 1H, H14), 7.87 (d, J = 16.3 Hz, 1H, H12),
7.82 (ddd, J = 8.3, 6.8, 1.1 Hz, 1H, H2), 7.67 (ddd, J = 8.2,
6.9, 1.1 Hz, 1H, H1), 7.61 (m, 1H, H18), 7.59 (d,
J = 16.3 Hz, 1H, H11), 7.49 (dt, J = 10.3, 8.6 Hz, 1H,
H15). 13C NMR (151 MHz, DMSO-d6) δ 168.0, 155.4,
150.8, 150.8, 149.5, 149.5, 149.3, 148.9, 148.9, 137.3, 134.5,
133.0, 130.6, 129.8, 128.1, 125.9, 125.2, 124.0, 121.2, 118.4,
118.3, 116.2, 116.1. MS: 311.1 (M+). Calcd. Analysis for
C18H11F2NO2: C, 69.45; H, 3.56; N, 4.50. Found: C,
69.39; H, 3.65; N, 4.43.

830
LUCZYWO ET AL.
4.1.12 | (E)-2-(3-fluorostyryl)quinoline-
4-carboxylic acid 13
promastigotes were incubated with MTT (3-[4,5-dimethyl-
2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide; Sigma-
Aldrich) and the optical density was determined in a plate
reader (POLARstar Omega, BMG Labtech) with a refer-
ence wavelength of 690 nm and a test wavelength of
595 nm. Results were expressed as the mean percentage
viability of treated parasites compared with untreated par-
asites (Control and DMSO control). Positive control treat-
ments were performed by incubating the parasite with
2 μM of amphotericin B (Sigma-Aldrich) or 30 μM
miltefosine (Sigma-Aldrich).
Yield 89%, yellow powder, mp 299.3ꢀC-299.9ꢀC (lit.
265ꢀC-266ꢀC).[27] 1H NMR (600 MHz, DMSO-d6) δ 8.64
(d, J = 8.4 Hz, 1H, H3), 8.25 (s, 1H, H9), 8.09 (d,
J = 8.4 Hz, 1H, H6), 7.91 (d, J = 16.3 Hz, 1H, H12), 7.83
(dt, J = 8.4, 1.2 Hz, 1H,), 7.70 to 7.59 (m, 3H), 7.64 (d,
J = 16.3 Hz, 1H, H11), 7.48 (c, J = 7.8 Hz, 1H, H15), 7.20
(dt, J = 8.5, 2.4 Hz, 1H). 13C NMR (151 MHz, DMSO-d6)
δ 168.0, 163.8, 162.2, 155.5, 148.9, 139.2, 139.2, 137.4,
133.9, 133.9, 131.2, 130.6, 129.9, 128.1, 125.9, 124.3, 121.3,
115.9, 113.9. Chemical Formula: C18H12FNO2.
4.4 | Intracellular multiplication of
parasites in treated bone marrow
macrophages
4.1.13 | (E)-2-(4-bromo-
3,5-dimethoxystyryl)quinoline-4-carboxylic
acid 18
BMMs were prepared as previously described. Briefly,
BMMs from female mice were obtained after 7 days of
differentiation of bone marrow in RPMI media (Gibco)
supplemented with 20% (v/v) FBS and 20% (v/v) of L-929
cell supernatant. For macrophage infection, 8 × 104
(cytotoxic assay) or 2 × 105 (fluorescence assay) BMMs
were plated in 96 well or 24 well dishes, respectively,
24 hours prior to experiments.
For the cytotoxic assay, BMMs were incubated with
the compounds for 24, 48, and 72 hours, and the viability
was accessed using MTT assay as described before.
For fluorescence assay, BMMs were plated on glass cov-
erslips in dishes. Promastigotes were added at an MOI = 5
in RPMI supplemented with 10% (v/v) FBS and 2% (v/v) of
L-929 cell supernatant for 2 hours at 34ꢀC. BMMs were
then washed three times with PBS and incubated for
24 hours. Later, the BMMs were incubated for 48 hours at
34ꢀC in the presence of the compounds 1, 2, 3, 4, 5, 6, and
14. Coverslips were then fixed with methanol 100%. The
number of intracellular parasites was quantified by cou-
nting the total number of macrophages, infected macro-
phages, and the total number of intracellular parasites. The
results were expressed as a number of parasites per number
of 100 infected BMMs or the percentage of infected cells.
Control treatment was performed by incubating the differ-
ent forms of the parasite with Amphotericin B.
1
Yield 60%, orange powder, mp 279ꢀC-280ꢀC. H NMR
(600 MHz, DMSO-d6) δ 8.68 (d, J = 8.4 Hz, 1H, H3), 7.95
(m, 2H, H9;H6), 7.81 (d, J = 16.2 Hz, 1H, H12), 7.70 (d,
J = 7.2 Hz, 1H, H2), 7.66 (d, J = 16.2 Hz, 1H, H11), 7.50
(t, J = 7.4, Hz, 1H, H1), 7.18 (s, 2H, H14;H18), 3.94 (s,
6H, H24;H26). 13C NMR (151 MHz, DMSO-d6) δ 169.3,
157.2, 155.5, 148.9, 137.7, 133.6, 130.6, 129.5, 129.1, 127.8,
126.1, 125.2, 118.7, 104.5, 100.4, 57.0 (C24;C26). IR
(cm−1): ν 3422, 3062, 2969, 1641, 1609, 1516, 1328, 1297,
984, 781. Calcd. Analysis for C20H16BrNO4: C, 57.99; H,
3.89; N, 3.38. Found: C, 57.94; H, 3.92; N, 3.34.
4.2 | Biological assays
4.2.1 | L. (L.) amazonensis culture
L. (L.) amazonensis (IFLA/BR/67/PH8) were propagated
as promastigotes at 26ꢀC in M199 media supplemented
with
5%
penicillin/streptomycin,
0.1%
hemin
(25 mg mL−1 in 0.1 N NaOH), 10 mM adenine, and 10%
FBS, pH 7.4. In the experiments, 7 days cultures (station-
ary phase) were used. Parasites were washed three times
in PBS before use in experiments.
For fluorescence assay, Methanol-fixed cells were
blocked with Saponine/BSA/TBS (0.1%) and incubated
with 10 μg mL−1 of Propidium Iodide (Sigma-Aldrich) for
1 hour. Images were acquired by fluorescence microscopy
DMI6000B/AF6000 (Leica) coupled to a digital camera
system (DFC 365 FX).
4.3 | Compounds activity against
promastigotes assessed by MTT assay
The compounds' activity was determined by incubating
L. (L.) amazonensis promastigotes in the presence of dif-
ferent compounds (29.3 μM—EC50, previously deter-
mined). The number of viable cells was determined as
described. Briefly, after 24, 48, and 72 hours of treatment,
ACKNOWLEDGMENTS
Fundings were received from Universidad de Buenos
Aires (UBACyT 20020170200206BA), and Unión

LUCZYWO ET AL.
831
Ramanathan, P. Bose, M. V. S. Rao, S. Gupta, S. K. Puri, D.
Launay, D. Martin, Eur. J. Med. Chem. 2013, 69, 527.
[17] D. Bompart, J. Núñez-Durán, D. Rodríguez, V. V. Kouznetsov,
C. M. Meléndez, F. Gómez, F. Sojo, G. Arvelo, A. Visbal, X.
Alvarez, Y. G.-M. Serrano-Martín, Bioorg. Med. Chem. 2013,
21, 4426.
[18] N. A. Liberto, J. Baptista Simões, S. de Paiva Silva, C. J. da
Silva, L. V. Modolo, A. de Fátima, L. M. Silva, M. Derita, S.
Zacchino, O. M. Portilla Zuñiga, G. P. Romanelli, S. A.
Fernandes, Bioorg. Med. Chem. 2017, 25, 1153.
[19] C. Nava-Zuazo, S. Estrada-Soto, J. Guerrero-Alvarez, I. León-
Rivera, G. M. Molina-Salinas, S. Said-Fernández, M. J. Chan-
Bacab, R. Cedillo-Rivera, R. Moo-Puc, G. Mirón-López, G.
Navarrete-Vazquez, Bioorg. Med. Chem. 2010, 18, 6398.
[20] G. C. Muscia, J. P. Carnevale, A. Luczywo, M. V. Peláez, A. R.
O'Toole, G. Y. Buldain, J. J. Casal, S. E. Asís, Arab. J. Chem.
2019, 12, 932.
Iberoamericana de Universidades (UIU) “Discovery of
New Antiparasitic Agents” (Proposal 1054, 1st and 2nd
Research Collaboration Fund: 2017-2018 and 2019-2020).
Also, this work was funded by Brazilian grants (Mauro
Cortez) from the FAPESP (grant 2012/24105-3), CNPq
(grant 443816/2014-0), and CAPES (Finance Code #001).
DATA AVAILABILITY STATEMENT
Data openly available in a public repository that issues
datasets with DOIs.
ORCID
Silvia E. Asís https://orcid.org/0000-0002-3978-8638
REFERENCES AND NOTES
[21] G. C. Muscia, J. P. Carnevale, M. Mariela Bollini, S. E. Asís,
J. Heterocyclic Chem. 2008, 45, 611.
[22] G. C. Muscia, M. Bollini, J. P. Carnevale, A. M. Bruno, S. E.
Asís, Tetrahedron Lett. 2006, 47, 8811.
[23] M.A.S. Abdelwahid, T. Elsaman, M.S. Mohamed, S.A. Latif,
M. Moawia. M.M. Mukhtar, M.A. Mohamed. J. Chem. 2019,
2019, 9.
[24] M. Staderini, M. Piquero, M. A. Abengózar, M. Nachér-
Vázquez, G. Romanelli, P. López-Alvarado, L. Rivas, M. L.
Bolognesi, J. C. Menéndez, Eur. J. Med. Chem. 2019, 171, 38.
[25] G. C. Muscia, S. E. Asís, G. Y. Buldain, Med. Chem. 2017,
13, 448.
[1] J. Alvar, I. D. Velez, C. Bern, M. Herrero, P. Desjeux, J. Cano,
J. Jannin, M. den Boer, the WHO Leishmaniasis Control
Team, PLoS One 2012, 7, e35671.
[2] WHO, Weekly Epidemiological Record (Relevé Épidémiologique
Hebdomadaire), Vol. 91, WHO, Geneve, Switzerland 2016,
p. 285. https://apps.who.int/iris/handle/10665/251908.
[3] I. Kevric, M. A. Cappel, J. Keeling, Dermatol. Clin. 2015,
33, 579.
[4] WHO, Technical Report Series Control of the Leishmaniases,
World Health Organization, Geneva, Switzerland 2010.
[5] S. L. Croft, K. Seifert, V. Yardley, Indian J. Med. Res. 2006,
123, 399.
[26] A. E. Lipkin, Z. P. Bespalova, Pharm. Chem. J. 1970, 4, 23.
[27] A. N. Dubrovin, A. I. Mikhalev, S. V. Ukhov, A. G. Goldshtein,
V. V. Novikova, T. F. Odegova, R. R. Makhmudov, Pharm.
Chem. J. 2015, 49, 309.
[6] S. R. B. Uliana, C. T. Trinconi, A. C. Coelho, Parasitology
2018, 145, 464.
[7] A. Hefnawy, M. Berg, J. C. Dujardin, G. De Muylder, Trends
Parasitol. 2017, 33, 162.
[28] H. John, J. Prakt. Chem. 1927, 117, 214.
[29] K. Van Aken, L. Strekowski, L. Patiny, J. Beilstein, Org. Chem.
2006, 3, 2.
[30] J.C. Sloop, J. Chem. (Hindawi) 2017, 2017, 15.
[31] ©Molinspiration Cheminformatics homepage, http://www.
molinspiration.com (accessed: May 2020).
[32] F. V. Cabral, M. T. Pelegrino, I. P. Sauter, A. B. Seabra, M.
Cortez, M. S. Ribeiro, Nitric Oxide 2019, 93, 25.
[33] M. Cortez, C. Huynh, M. C. Fernandes, K. A. Kennedy, A.
Aderem, N. W. Andrews, Cell Host Microbe. 2011, 9, 463.
[34] I. P. Sauter, K. G. Madrid, J. B. de Assis, A. Sá-Nunes, A. C.
Torrecilhas, D. I. Staquicini, R. Pasqualini, W. Arap, M.
Cortez, JCI Insight 2019, 4, 1, 18
[35] C. A. Lipinski, F. Lombardo, B. W. Dominy, P. J. Feeney, Adv.
Drug Delivery Rev. 2001, 46, 3.
[36] J. L. Siqueira-Neto, S. Moon, J. Jang, G. Yang, C. Lee, H. K.
Moon, E. Chatelain, A. Genovesio, J. Cechetto, L. H. Freitas-
Junior, PLoS Neglected Trop. Dis. 2012, 6, e1671.
[8] B. Purkait, A. Kumar, N. Nandi, A. H. Sardar, S. Das, S.
Kumar, K. Pandey, V. Ravidas, M. Kumar, T. De, D. Singh, P.
Das, Antimicrob. Agents Chemother. 2012, 56, 1031.
[9] S. Rijal, B. Ostyn, S. Uranw, K. Rai, N. R. Bhattarai, T. P. C.
Dorlo, J. H. Beijnen, M. Vanaerschot, S. Decuypere, S. S.
Dhakal, M. L. Das, P. Karki, R. Singh, M. Boelaert, J. C.
Dujardin, B. P. Koirala, Clin. Infect. Dis. 2013, 56, 153.
[10] S. Srivastava, J. Mishra, A. K. Gupta, A. Singh, P. Shankar, S.
Singh, Parasit. Vectors 2017, 10, 49.
[11] S. Sundar, T. K. Jha, C. P. Thakur, J. Engel, H. Sindermann, C.
Fischer, K. Junge, A. Bryceson, J. Berman, N. Engl. J. Med.
2002, 347, 1739.
[12] I. A. Rodrigues, A. M. Mazotto, V. Cardoso, R. L. Alves,
A. C. F. Amaral, J. R. de Andrade Silva, A. S. Pinheiro, A. B.
Vermelho, Mediators Inflamm. 2015, 2015, 835910.
[13] A. Fournet, A. A. Barrios, V. Muñoz, R. Hocquemiller, A.
Cavé, J. Bruneton, Antimicrob. Agents Chemother. 1993,
37, 859.
[37] M. De Rycker, I. Hallyburton, J. Thomas, L. Campbell, S.
Wyllie, D. Joshi, S. Cameron, I. H. Gilbert, P. G. Wyatt, J. A.
Frearson, A. H. Fairlamb, D. W. Gray, Antimicrob. Agents
Chemother. 2013, 57, 2913.
[38] M. Vermeersch, R. I. da Luz, K. Toté, J. P. Timmermans, P.
Cos, L. Maes, Antimicrob. Agents Chemother. 2009, 53, 3855.
[39] L. M. Alcântara, T. C. S. Ferreira, F. R. Gadelha, D. C. Miguel,
Int. J. Parasitol. Drugs Drug Resist. 2018, 8, 430.
[14] A. Fournet, J. C. Gantier, A. Gautheret, L. Leysalles, M. H.
Munos, J. Mayrargue, H. Moskowitz, A. Cavé, R.
Hocquemiller, J. Antimicrob. Chemother. 1994, 33, 537.
[15] N. Campos Vieira, C. Herrenknecht, J. Vacus, A. Fournet, C.
Bories, B. Figadere, L. Salmen Espindola, P. M. Loiseau,
Biomed. Pharmacother. 2008, 62, 684.
[16] V. S. Gopinath, J. Pinjari, R. T. Dere, A. Verma, P.
Vishwakarma, R. Shivahare, M. Moger, P. S. Kumar Goud, V.

832
LUCZYWO ET AL.
[40] D. Boschi, A. C. Pippione, S. Sainas, M. L. Lolli, Eur. J. Med.
Chem. 2019, 183, 111681.
[41] R. A. G. Reis, F. A. Calil, P. R. Feliciano, M. P. Pinheiro, M. C.
Nonato, Arch. Biochem. Biophys. 2017, 632, 175.
[42] R. Ochoa, S. J. Watowich, A. Flórez, C. V. Mesa, S. M.
Robledo, C. Muskus, J. Comput.-Aided Mol. Des. 2016, 30, 541.
[43] L. A. Chibli, T. J. Schmidt, M. C. Nonato, F. A. Calil, F. B. Da
Costa, Eur. J. Med. Chem. 2018, 5, 852.
[44] J. Cheleski, J. R. Rocha, M. P. Pinheiro, H. J. Wiggers, B. F.
Albérico, A. B. F. da Silva, M. C. Nonato, C. A. Montanari,
Eur. J. Med. Chem. 2010, 45, 5899.
SUPPORTING INFORMATION
Additional supporting information may be found online
in the Supporting Information section at the end of this
article.
How to cite this article: Luczywo A, Sauter IP,
da Silva Ferreira TC, et al. Microwave-assisted
synthesis of 2-styrylquinoline-4-carboxylic acid
derivatives to improve the toxic effect against
Leishmania (Leishmania) amazonensis.
J Heterocyclic Chem. 2021;58:822–832. https://doi.
org/10.1002/jhet.4217
[45] J. T. Madak, C. R. Cuthbertson, Y. Miyata, S. Tamura, E. M.
Petrunak, J. A. Stuckey, Y. Han, M. He, D. Sun, H. D.
Showalter, N. Neamati, J. Med. Chem. 2018, 61, 5162.
[46] T. T. H. Hajalsiddig, A. E. M. Saeed, Eur. J. Chem. 2019, 10, 45.