Discovery of a quinoline-based phenyl sulfone derivative as an antitrypanosomal agent

Article abstract of DOI:10.1016/j.bmcl.2018.03.039

A series of natural products-based phenyl sulfone derivative and their property-based analogues were investigated as potential growth inhibitors of Trypanosoma brucei. Trypanosoma brucei is a kinetoplastid protozoan parasite that causes trypanosomiasis. In this work, we found that nopol- and quinoline-based phenyl sulfone derivative were the most active and selective for T. brucei, and they were not reactive towards the active thiol of T. brucei's cysteine protease rhodesain. A thiol reactive variant of the quinoline-based phenyl sulfone was subsequently investigated and found to be a moderate inhibitor of rhodesain. The quinoline-based compound that is not reactive towards rhodesain can serve a template for phenotypic-based lead discovery while its thiol-active congener can serve as template for structure-based investigation of new antitrypanosomal agents.

Full text of DOI:10.1016/j.bmcl.2018.03.039

Accepted Manuscript
Discovery of a Quinoline-Based Phenyl Sulfone Derivative as an Antitrypano-
somal Agent.
Huaisheng Zhang, Jasmine Collins, Rogers Nyamwihura, Shelbi Ware, Marcel
Kaiser, Ifedayo Victor Ogungbe
PII:
S0960-894X(18)30233-6
https://doi.org/10.1016/j.bmcl.2018.03.039
BMCL 25693
DOI:
Reference:
Bioorganic & Medicinal Chemistry Letters
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Revised Date:
Accepted Date:
22 January 2018
6 March 2018
15 March 2018
Please cite this article as: Zhang, H., Collins, J., Nyamwihura, R., Ware, S., Kaiser, M., Ogungbe, I.V., Discovery
of a Quinoline-Based Phenyl Sulfone Derivative as an Antitrypanosomal Agent., Bioorganic & Medicinal Chemistry
Letters (2018), doi: https://doi.org/10.1016/j.bmcl.2018.03.039
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Discovery of a Quinoline-Based Phenyl Sulfone
Derivative as an Antitrypanosomal Agent.
Huaisheng Zhang, Jasmine Collins, Rogers Nyamwihura, Shelbi Ware, Marcel Kaiser, Ifedayo Victor Ogungbe

Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com
Discovery of a Quinoline-Based Phenyl Sulfone Derivative as an Antitrypanosomal
Agent.
Huaisheng Zhanga, Jasmine Collinsa, Rogers Nyamwihuraa, Shelbi Warea, Marcel Kaiserb,c, Ifedayo Victor
Ogungbea*
aDepartment of Chemistry, Jackson State University, Jackson, MS, 39217, USA
bDepartment of Medical Parasitology & Infection Biology, Swiss Tropical and Public Health Institute, 4051 Basel, Switzerland
cUniversity of Basel, Petersplatz 1, 4003 Basel, Switzerland
ARTICLE INFO
ABSTRACT
Article history:
Received
Revised
Accepted
Available online
A series of natural products-based phenyl sulfone derivative and their property-based analogues
were investigated as potential growth inhibitors of Trypanosoma brucei. Trypanosoma brucei is
a kinetoplastid protozoan parasite that causes trypanosomiasis. In this work, we found that
nopol- and quinoline-based phenyl sulfone derivative were the most active and selective for T.
brucei, and they were not reactive towards the active thiol of T. brucei’s cysteine protease
rhodesain. A thiol reactive variant of the quinoline-based phenyl sulfone was subsequently
investigated and found to be a moderate inhibitor of rhodesain. The quinoline-based compound
that is not reactive towards rhodesain can serve a template for phenotypic-based lead discovery
while its thiol-active congener can serve as template for structure-based investigation of new
antitrypanosomal agents.
Keywords:
Trypanosoma brucei
Cysteine protease
Natural products
Quinoline
2009 Elsevier Ltd. All rights reserved.
Human African Trypanosomiasis (HAT), one of the neglected
tropical diseases (NTDs), caused by protozoan Trypanosoma
brucei is a declining public health problem on the African
continent due to a gradual decrease in the number of reported
cases in the past few years. It is most prevalent, at the moment,
in the Democratic Republic of the Congo. Historically, the lack
of adequate and rapid diagnostic tools as well as lack of effective,
safe, and accessible medicines to treat HAT resulted in the death
of hundreds of thousands of people. Despite the decrease in
reported case, the lack of good network of primary healthcare
facilities in most rural and remote places on the continent as well
as the possibility of continuous transmission of the parasite from
animal reservoirs to humans, make the disease a continuous
threat to millions of people.1-4 Discovery and development of
effective oral drugs remains a key objective in combating the
have been widely explored in anti-infective drug discovery. Most
anti-infective agents are natural products-based/inspired.8
However, due to the complexity and scarcity of most active
agents, follow-up studies are usually difficult and rarely pursued
in NTDs drug discovery.
The compounds described in this work were synthesized as
outlined in Schemes 1 and 2. For compounds 5 to 25, allyl
phenyl sulfone (1) was reacted with bromine to obtain the 1,2-
dibromide (2), in good yield (93%). This was followed by
dehalogenation of the vicinal dibromide with sodium carbonate
in diethyl ether to obtain (E)-((3-bromoprop-1-en-1-
yl)sulfonyl)benzene (3). Compounds 4a-c were obtained via
etherification
reaction
between
the
appropriate
4-
hydroxyphenylacetic acids and 3 in ethanol, using potassium
hydroxide and sodium iodide. Compounds 4a-c were then used to
synthesize the corresponding amides (5-10) and esters (11-25)
using CDI or DCC and DMAP as coupling reagents.9-13 Detailed
synthesis and compound characterization data are provided as
supporting information.
The compounds were subsequently tested for their ability to
inhibit the growth of T. brucei in vitro.14 The parasites were
exposed to the compounds for 48 hours. Most of the compounds
displayed selective but moderate growth inhibitory activity
against T. brucei when compared with mammalian cells (Hep
G2).15 Compounds derived from 8-aminoquinoline (9), (1R)-
nopol (15, 24), 6-bromo-2-naphthol (16), (+) fenchol (21) and 4-
disease. In this regard,
a
promising drug candidate,
nitroimidazole fexinidazole, is in the approval stages for the
treatment of human African trypanosomiasis. It would be the first
approved oral medicine to treat human African trypanosomiasis
in several decades. Fexinidazole is also being investigated as a
potential treatment for Chagas Disease.5,6 Despite these recent
gains, the drug development pipeline for HAT is sparse and there
is need for continued investment and investigation into new
chemical entities that can be developed as treatments and/or as
prophylactic agents against the disease. Many plant-derived
natural products have been reported as antiprotozoal agents. See
review by Schmidt and colleagues.7 In addition, natural products
benzylphenol
(23)
were

n/a
0.008 ± 0.0003
Podophyllotoxin
the most active (Table 1). The 8-aminoquinoline-based
compound (9), being the most selective, was evaluated for in vivo
antitrypanosomal activity. Two groups of T. brucei (STIB795)-
infected mice were treated for
4
consecutive days
intraperitoneally with 50 mg/kg/day and 100 mg/kg/day of 9,
respectively.16 The infected mice were positive for parasites 24
hours posttreatment, suggesting that compound 9 lack in vivo
efficacy. Several generations of aminoquinoline-based
compounds have found clinical use in the treatment of malaria
but not in the treatment of trypanosomiasis.17 This is perhaps due
to the unique mechanism of action of aminoquinolines in
plasmodium-infected cells. However, there are increasing reports
of quinoline-based growth inhibitors of trypanosomes, although,
the mechanism of action of the quinoline-based compounds have
not been deciphered.18-21
Scheme 1. Synthesis of target compounds 5-25.
Table 1. The antitrypanosomal activities of compounds 5-27.
R1 R2
R3
T. brucei
Hep G2
IC50
IC50
H
H
H
H
5
6
The presence of the vinyl sulfone moiety in 5-25 suggests that
they are potential covalent inhibitors of trypanosoma cysteine
proteases. Compounds 5-25 were then tested for inhibitory
activity against the major cathepsin L protease in T. brucei,
rhodesain.
Rhodesain is a validated drug target and it is known to be
essential for the survival and infectivity of the parasite. Its role in
the ability of the parasite to proliferate has been extensively
investigated.22-24 None of the compounds displayed noteworthy
inhibition of the protease at 20 µM. The inactivity of the
compounds may be because of the proximity of the vinylic
Michael acceptor to the phenoxide oxygen in 5-25. It is also
possible that the compounds are just unable to adopt favorable
orientation at the active site of the protease. Nevertheless, a
quinoline-based thiol reactive structural variant of 9 was
synthesized and tested for protease inhibition, and for
trypanocidal activity. Compound 27 was synthesized from boc-
11.72 ± 0.83
10.77 ± 0.31
>20
>20
H
H
H
H
7
8
9
>20
>20
>20
>20
H
H
0.76 ± 0.11
>80
H
H
H
H
10
11
5.45 ± 0.20
7.16 ± 0.42
5.62 ± 0.65
>20
>20
>20
>20
>20
H
H
H
H
12
13
protected
(E)-5-phenyl-1-(phenylsulfonyl)pent-1-en-3-amine
(26). Compound 26 was a generous gift from Prof. J Love
(University of British Columbia), and it was reported by Kiemele
and co-workers in 2016.25 Compound 27 was able to completely
inactivate rhodesain at 20 µM after 1 hour of incubation with
estimated IC50 value of 800 nM, and a Kinact/Ki value of 99 M-1s-1
(Figure 1).26
H
H
14
>20
>20
>80
H
H
H
H
15
16
2.01 ± 0.12
2.18 ± 0.25
>80
H
H
H
H
H
H
17
18
19
6.04 ± 0.03
>20
>20
>20
>20
>20
Scheme 2. Synthesis of target compound 27.
H
H
H
H
20
21
5
5.63 ± 0.61
4.04 ± 0.01
>20
N
o i n h i b i t o r
0 . 0 6 µ M
0 . 1 2 µ M
0 . 2 4 µ M
0 . 9 7 µ M
3 . 9 µ M
4
3
2
11.9 ± 1.03
H
H
H
H
H
F
22
23
24
25
27
>20
>20
>80
>80
1.47 ± 0.40
3.04 ± 0.07
7.03 ± 0.17
Cl
H
0
1 0 0 0
2 0 0 0
3 0 0 0
4 0 0 0
5 0 0 0
T
i m
e
( s )
>20
>80
Figure 1. Pseudo-first order inhibition plots for compound 27.
5.97 ± 0.12
n/a
0.004 ± 0.001
Suramin

2a
2b
P1
P2
Figure 2. a. The superposition of docked compound 27 (Yellow) and D1R (Blue) at the active site of rhodesain; b. The interaction
plot of compound 27 with active site residues.
Chandenier, J.; Desoubeaux, G. Emerg Infect Dis 2016,
It has a moderate antitrypanosomal activity with an IC50 value of
5.97 uM. Compound 27 was also tested for inhibitory activity
against human cathepsin L, but it was inactive (from 0.1 µM to
125 µM).26 Crystallographic investigation of rhodesain-inhibitor
(27) complex has been attempted but it has not been successful.
However, it is still being pursued. In order to understand the
interactions responsible for the inhibitory activity of 27 on
rhodesain, template docking was used.27 Compound 27 was
docked in the previously reported crystal structure of rhodesain
using the bound ligand (D1R) as template.28 Three of the top five
docking poses suggests that the vinyl group is in the vicinity of
active thiol (Cys25), while the homophenyl moiety occupies the
P1 site, and the quinoline moiety occupies the P2 site (Figure 2a
and 2b). The phenyl sulfone moiety is predicted to have steric
interactions with Gln19 and His162 while the quinolyl motif
have steric interactions with Met68. Quite noticeable is the empty
P3 site.
22, 935–937.
3.
4.
5.
Mukadi, P.; Lejon, V.; Barbé, B.; Gillet, P.; Nyembo, C.;
Lukuka, A.; Likwela, J.; Lumbala, C.; Mbaruku, J.;
Vander Veken, W.; Mumba, D.; Lutumba, P.; Muyembe,
J.-J.; Jacobs, J. PLOS ONE 2016, 11, e0146450.
Lumbala, C.; Simarro, P. P.; Cecchi, G.; Paone, M.;
Franco, J. R.; Kande Betu Ku Mesu, V.; Makabuza, J.;
Diarra, A.; Chansy, S.; Priotto, G.; Mattioli, R. C.; Jannin,
J. G. Int J Health Geogr 2015, 14.
Nagle, A.; Khare, S.; Kumar, A.; Supek, F.; Buchynskyy,
A.; Mathison, C.; Chennamaneni, N.; Pendem, N.;
Buckner, F.; Gelb, M.; Molteni. V. Chem Rev. 2014, 114,
11332-11333.
In conclusion, a series of phenyl sulfone natural products-based
compounds were synthesized and evaluated as potential
antitrypanosomal agents. Quinoline- and nopol-based
compounds, 9 and 15, were the most active and selective. The
quinoline-based compound (9) can serve a template for
phenotypic-based lead discovery and the thiol-active compound
(27) may serve as template for structure-based investigation of
new covalent antitrypanosomal agents.
6.
7.
Kaiser, M.; Bray, M. A.; Cal, M.; Bourdin, T. B.;
Torreele, E.; Brun, R. Antimicrob Agents Chemother
2011, 55, 5602.
Schmidt, T.J.; Khalid, S.A.; Romanha, A.J.; Alves,
T.MA.; Biavatti, M.W.; Brun, R; Da Costa, F.B.a; de
Castro, S.L.; Ferreira, V.F.; de Lacerda, M.V.G.; Lago,
J.H.G.; Leon, L.L.; Lopes, N.P.; Das Neves Amorin, R.C;
Niehues, M.; Ogungbe, I.V.; et al. Curr Med Chem 2012,
19, 2128-2175.
Acknowledgments: This work was carried out in part by
resources made available by the US National Institutes of Health
(SC2GM109782, SC3GM122629, and G12MD007581). JC and
SW were supported by NSF-RISE (HRD-1547836). We thank
Prof. Jennifer Love (University of British Columbia) for the gift
of compound 26.
8.
a. Newman, D.J.; Cragg, G.M.; Snader, K.M. J Nat Prod
2003, 66, 1022-1037.; b. Newman, D.J.; Cragg, G.M. J
Nat Prod 2007, 70, 461-477.; c. Newman, D.J.; Cragg,
G.M. J Nat Prod 2012, 75, 311-335.
9.
Zindel, J.; de Meijere A. Synthesis 1994, 3, 190-194.
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Ishizaki, M.; Yamada, M.; Watanabe, S.I.; Hoshino, O.;
Nishitani, K.; Hayashida, M.; Tanaka, A.; Hara, H.
Tetrahedron 2004, 60, 7973-7981.
References and Notes
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WHO. Investing to Overcome the Global Impact of
Neglected Tropical Diseases. Third WHO Report on
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11.
12.
Neises, B.; Steglich, W. Angew Chem Int Ed 1978, 17,
522-524.
2.
De Kyvon, M.-A. L.-C.; Maakaroun-Vermesse, Z.;
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M.; De Toffol, B.; Paris, L.; Bernard, L.; Goudeau, A.;
Kamiński, K.; Rapacz, A.; Łuszczki, J.J.; Latacz, G.;
Obniska, J.; Kieć-Kononowicz, K.; Filipek, B. Bioorgan
Med Chem 2015, 23, 2548-2561.

18.
19.
20.
21.
22.
Ramírez–Prada, J.; Robledo, S.M.; Vélez, I.D.; del Pilar
Crespo, M.; Quiroga, J.; Abonia, R.; Montoya, A.; Svetaz,
L.; Zacchino, S.; Insuasty, B. Eur J Med Chem 2017, 131,
237-254.
13.
14.
Lafrance D, Bowles P, Leeman K, Rafka R. Org Lett
2011, 13, 2322 - 2325.
Trypanosoma brucei assay: The growth inhibitory activity
of the compounds was evaluated using the Alamar blue
assay. Bloodstream forms of T. brucei brucei (strain 427)
cultured in HMI-9-medium supplemented with 10% FBS,
Nefertiti, A.; Batista, M.; Da Silva, P.; Batista, D.; Da
Silva, C.; Peres, R.; Torres-Santos, E.; Cunha-Junior, E.;
Holt, E.; Boykin, D.W.; Brun, R. Antimicrob Agents Ch
2017, AAC-01936.
10%
Serum
plus
(SAFC),
0.05
mM
mM
bathocuproinesulfonate, 1.5 mM L-cysteine,
1
hypoxanthine, 0.2 mM β-mercaptoethanol, 0.16 mM
thymidine, 1 mM pyruvate, and 0.0125% Tween 80 were
dispensed into sterile 96-well plates at 5 X 103 cells/well,
and treated with compounds for 48 hours. The compounds
were prepared in DMSO and were tested in triplicates
with a total assay volume of 100 µL. Next, Alamar blue
(20 µL) was added and the plate was incubated at 37°C
Carvalho, L., Martínez-García, M., Pérez-Victoria, I.,
Manzano, J.I., Yardley, V., Gamarro, F. and Pérez-
Victoria, J.M. Antimicrob Agents Ch 2015, 59, 6151-
6160.
Di Pietro, O.; Vicente-García, E.; Taylor, M.C.;
Berenguer, D.; Viayna, E.; Lanzoni, A.; Sola, I.; Sayago,
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105, 120-137.
for
4
hours. Immediately following incubation,
fluorescence signals were read (λex 530 nm, λem 590nm).
IC50 values were determined by testing compounds in a
dose range of 0.3 – 50 µM. Suramin was used as positive
control.
a. Scory, S.; Caffrey, C. R.; Stierhof, Y. D.; Ruppel, A.;
Steverding, D. Exp. Parasitol 1999, 91, 327-333.; b.
Troeberg, L.; Morty, R. E.; Pike, R. N.; Lonsdale-Eccles,
J. D.; Palmer, J. T.; McKerrow, J. H.; Coetzer, T. H. Exp.
Parasitol 1999, 91, 349-355.; c. Steverding, D.; Sexton,
D. W.; Wang, X.; Gehrke, S. S.; Wagner, G. K.; Caffrey,
C. R. Int. J. Parasitol 2012, 42, 481-488.
15.
Cytotoxicity Assay: Human hepatocarcinoma cell line
(Hep G2, CRL-11997™) was used for cytotoxicity
studies. The cells were grown in complete medium
(DMEM: F12 containing L-glutamine and sodium
bicarbonate, 10% FBS, 1% penicillin/streptomycin,
0.0125% Tween 80) incubated at 37°C in a 5% CO2
environment. Once 80-90% confluent, the cells were
washed with phosphate-buffered saline (PBS), treated
with 0.25% (w/v) of trypsin/EDTA, counted and
suspended in fresh complete media. Into 96-well plates,
198 µL of 5 x 105 cells/mL were seeded and incubated for
about 24 hours. Cells were treated with the compounds
prepared in DMSO for 72 hours. After 72 hours, the old
cell medium is removed and fresh DMEM:F12 medium
containing MTT (5 mg/mL in PBS) was added to the
cells, and incubated for 1.5 hour. The MTT-containing
medium was gently removed and replaced with DMSO
(200 μL/well). Lyzed cells were then repeatedly
resuspended in DMSO using multichannel pipette in order
to allow the formazan crystals to dissolve. Plates were
incubated for 10 minutes and absorbance were measured
at 550 nm. All compounds were tested in triplicates. SDS
(10%) was used as assay positive control and
podophyllotoxin was used as cytotoxicity control.
23.
24.
Ettari, R.; Tamborini L.; Angelo I. C.; Micale N.; Pinto
A.; De Micheli C.; Conti P. J Med Chem. 2013, 56, 5637.
Mott, B.; Ferreira, R.; Simeonov, A.; Jadhav, A.; Ang, K.;
Leister, W.; Shen, M.; Silveira, J.; Doyle, P.; Arkin, M.;
McKerrow, J.; Inglese, J.; Austin, C.; Thomas, C.;
Shoichet, B.; Maloney, D. J. Med. Chem. 2010, 53, 52.
25.
26.
Kiemele, E.R.; Wathier, M.; Bichler, P.; Love, J.A. Total
synthesis of K777: Successful application of transition-
metal-catalyzed alkyne hydrothiolation toward the
modular synthesis of a potent cysteine protease inhibitor.
Org lett 2016, 18, 492-495.
a. T. brucei cathepsin L (rhodesain) was expressed in
Pichia pastoris. The inhibition assays were carried out in
50 mM sodium acetate (assay buffer) pH 5.5, containing 2
mM DTT and 0.01% Triton X-100. The enzyme (5 µL,
final concentration of 0.8 nM) and test compound (5 µL,
final concentration of 20 μM) mixtures were pre-
incubated for 1 hour, followed by the addition of 100 μL
of the substrate, Z-Phe-Arg-AMC (10 µM), in sodium
acetate buffer pH 5.5. Fluorescence (RFU/sec) resulting
from proteolytic cleavage of the substrate was monitored
at 25 °C (λex 355 nm and λem 460 nm) on a PolarStar
Omega plate reader (BMG LABTECH, Germany). E-64
(10 µM) was used as positive control (100% inhibition).
For time dependent inhibition studies, rhodesain-
compound assay mixture includes 30 µL of compound 27
in DMSO, 30 µL of rhodesain in assay buffer, and 180 µL
of assay buffer. Aliquots (10 µL) of rhodesain-27 mixture
were assayed every 10 minutes using Z-Phe-Arg-AMC
(10 µM) as substrate. Fluorescence (RFU/sec) resulting
from proteolytic cleavage of the substrate was monitored
for 7 minutes as described above. Compound 27 was
assayed at the following concentrations: 0, 0.06, 0.12,
0.24, 0.97, and 3.9 µM.; b. Cathepsin L (CatL) from
human liver (Millipore Sigma) was used for protease
16.
In vivo assay: T. b. brucei STIB795-Luc model was used
for in vivo studies. The studies were conducted at the
Swiss Tropical and Public Health Institute (Basel,
Switzerland). They were approved by the veterinary office
of the Canton Basel-Stadt, Switzerland under license
number 2813. Four female NMRI mice were used per
experimental group. A control group and two treatment
groups (50 mg/kg/day and 100 mg/kg/day). Each mouse
was inoculated intraperitoneally (ip) with 1 x 104
bloodstream forms of STIB795-Luc. Treatments were
administered ip in water (plus 10% DMSO) 3 days post-
infection. All mice were monitored for parasitemia by live
imaging. Moribund mice were euthanized after detection
of parasitemia.
17.
Parhizgar, A.R.; Tahghighi, A. Iran J Med Sci 2017, 42,
115.

selectivity studies. Human CatL (0.58 nM) was assayed in
400 mM sodium acetate (100 µL) pH 5.5, containing 8
mM DTT, 4 mM EDTA, and 0.01% Triton X-100. Human
CatL and 27 were pre-incubated for 1 hour in assay
buffer, followed by the addition of 100 μL of the
substrate, Z-Phe-Arg-AMC (20 µM), in sodium acetate
buffer and the plate was read as described above.
27.
Molecular structure of compound 27 was built using
SPARTAN ’10 for Windows. Its geometries were
optimized using the MMFF 94 force field. The docking
simulations were carried out using the MolDock docking
algorithm of the Molegro Virtual Docker in the template
docking mode. The X-ray crystal structure (PDB ID:
2P7U) of rhodesain was used for the docking calculation.
The bound inhibitor in 2P7U was used as template
molecule. A docking sphere (15 Å radius) was placed on
the binding site (X = -9.38; Y = 1.66; Z = 10.38) in order
to allow different orientations of the ligand to be searched.
The docking algorithm was set at maximum iterations of
1500 with a simplex evolution population size of 50, and a
minimum of 50 runs. The 2D representations of
rhodesain-27 complex was prepared using MOE 2016.
28.
Kerr, I.D.; Lee, J.H.; Farady, C.J.; Marion, R.; Rickert,
M.; Sajid, M.; Pandey, K.C.; Caffrey, C.R.; Legac, J.;
Hansell, E.; McKerrow, J.H. J Bio Chem 2009, 284,
25697-25703.