4-Aminoquinoline-chalcone/-N-acetylpyrazoline conjugates: Synthesis and antiplasmodial evaluation

Article abstract of DOI:10.1016/j.ejmech.2017.07.041

1H-1,2,3-triazole linked 4-aminoquinoline-chalcone/-N-acetylpyrazoline conjugates were synthesized and evaluated against cultured chloroquine (CQ) resistant strain. Antiplasmodial activities of the synthesized conjugates revealed dependence of activity on

Full text of DOI:10.1016/j.ejmech.2017.07.041

Accepted Manuscript
4-Aminoquinoline-chalcone/-N-acetylpyrazoline conjugates: Synthesis and
antiplasmodial evaluation
Sumit Kumar, Anu Saini, Jiri Gut, Philip J. Rosenthal, Raghu Raj, Vipan Kumar
PII:
S0223-5234(17)30563-9
10.1016/j.ejmech.2017.07.041
EJMECH 9604
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 9 June 2017
Revised Date: 19 July 2017
Accepted Date: 21 July 2017
Please cite this article as: S. Kumar, A. Saini, J. Gut, P.J. Rosenthal, R. Raj, V. Kumar, 4-
Aminoquinoline-chalcone/-N-acetylpyrazoline conjugates: Synthesis and antiplasmodial evaluation,
European Journal of Medicinal Chemistry (2017), doi: 10.1016/j.ejmech.2017.07.041.
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ACCEPTED MANUSCRIPT
4-Aminoquinoline-Chalcone/-N-Acetylpyrazoline Conjugates:
Synthesis and antiplasmodial Evaluation
Sumit Kumar,a Anu Saini,a Jiri Gut,b Philip J. Rosenthal,b Raghu Raj*c, Vipan
Kumar*a
a Department of Chemistry, Guru Nanak Dev University, Amritsar-143005, India
b Department of Medicine, University of California, San Francisco, CA, USA
c Department of Chemistry, DAV College, Amritsar-143001, India
*Corresponding Author. Tel: +91183 2553377 (R.R.), +91 183 2258802-09*3320 (V.K.); Fax: +91 183 2258819-20, Email address:
raghusharma4@gmail.com (R. Raj), vipan_org@yahoo.com (V. Kumar)
Synthesis and in vitro antiplasmodial evaluation of 4-aminoquinoline-chalcone/N-
acetylpyrazoline conjugates against W2 (CQ-R) strain of P. falciparum

ACCEPTED MANUSCRIPT
4-Aminoquinoline-Chalcone/-N-Acetylpyrazoline Conjugates:
Synthesis and antiplasmodial Evaluation
Sumit Kumar,a Anu Saini,a Jiri Gut,b Philip J. Rosenthal,b Raghu Raj*c, Vipan
Kumar*a
a Department of Chemistry, Guru Nanak Dev University, Amritsar-143005, India
b Department of Medicine, University of California, San Francisco, CA, USA
c Department of Chemistry, DAV College, Amritsar-143001, India
*Corresponding Author. Tel: +91183 2553377 (R.R.), +91 183 2258802-09*3320 (V.K.); Fax: +91 183 2258819-20, Email address:
raghusharma4@gmail.com (R. Raj), vipan_org@yahoo.com (V. Kumar)
Abstract: 1H-1,2,3-triazole linked 4-aminoquinoline-chalcone/-N-acetylpyrazoline conjugates were synthesized
and evaluated against cultured chloroquine (CQ) resistant strain. Antiplasmodial activities of the synthesized
conjugates revealed dependence of activity on the length of the alkyl chain as well as on the presence of
methoxy substituents on ring A/ring B of the chalcone. The most potent and non-cytotoxic conjugate showed
comparable antiplasmodial activity with that of CQ, with an IC50 value of 53.7 nM.
Key Words: 4-Aminoquinoline - Chalcone - N-Acetylpyrazoline - Click Chemistry - Antiplasmodial
1. Introduction
Despite substantial progress since 2000 in the fight against malaria, the disease continues to
be the most lethal parasitic infection. According to the latest World Health Organization
(WHO) update, malaria was responsible for 212 million illnesses and 4,29,000 deaths in
2015, with children and pregnant women the most affected groups [1]. Among the five
plasmodial species that infect humans, P. falciparum is most virulent and prevalent form,
accounting for >90% of malarial deaths [2]. CQ was the mainstay for treatment of malaria
[3-6], but it is no longer used to treat falciparum malaria in most areas due to widespread
resistance. WHO now recommends fixed dose artemisinin-based combination therapies
(ACTs) to treat uncomplicated malaria [7-10]. However, emergence of resistance to
artemisinins and frequent failures after therapy with some ACTs in Cambodia and nearby
countries has provided strong impetus to discover new antimalarial agents [1,10].
4-Aminoquinoline-hybridization involving covalent fusion of two or more
pharmacophoric fragments to yield a single hybrid molecule with improved efficacy
compared to the parent drugs has emerged as a promising strategy in antimalarial
chemotherapy [11-14]. The potential of quinoline hybridization is well exemplified by
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trioxaferroquines, artemisinin-quinine, and clotrimazole-quinoline conjugates, all of which
demonstrate potent antimalarial activity [15].
Chalcones, belonging to the flavanoid family of natural products, have been a source
of motivation to synthetic medicinal chemists because of their synthetic accessibility and
diverse biological activities [16-18]. The antiplasmodial potential of chalcones has been
attributed to their ability to inhibit plasmodial aspartic proteases, cysteine proteases, cyclin-
dependent protein kinases and new permeation pathways [19], but mechanisms of action are
uncertain. 4-Aminoquinoline-chalcone conjugates, prepared either via click chemistry or
aromatic nucleophilic substitution, have shown good antimalarial activities against CQ-
resistant strains of P. falciparum [20]. Pyrazoline is another well-known pharmacophore
found in many natural products. It exhibits a wide range of biological activities, including
antimicrobial, antitubercular, antitumor, anticancer and antiplasmodial actions [21-25].
Recent work from our lab described the synthesis and antiplasmodial activities of
amide-tethered 4-aminoquinoline-chalcone/4-aminoquinoline-ferrocenyl chalcone conjugates
[26]. Structure-activity relationship (SAR) studies revealed methoxy substituents on ring B of
the chalcones along with longer alkyl chains, introduced as spacers between two
pharmacophores, to be essential for good activities. Inspired by these results and in
continuation [27], the present manuscript describes the synthesis and antiplasmodial
evaluation of 1H-1,2,3-triazole-tethered 4-aminoquinoline-chalcone/ N-acetylpyrazoline
conjugates (Figure 1) along with their antiplasmodial evaluation against P. falciparum. The
insertion of a 1H-1,2,3-triazole core in the present series of conjugates is based on its well
established stability under acidic, basic, reductive and oxidative conditions, along with
additional favourable properties including capability of hydrogen bonding and rigidity in
binding with biomolecular targets [28].
2. Result and Discussion
2.1 Synthetic Chemistry
The synthetic approach involved an initial treatment of 4,7-dichloroquinoline 1 with
ethanolamine or 1-propanol amine at 120 °C to afford another precursor 2. The treatment of 2
with methane sulphonyl chloride in dry DCM at 0 °C for 2 h, followed by its reaction with
sodium azide at 120 °C in dry DMF for 12 h afforded 3, as depicted in Scheme 1 [29].
The precursors’ acetylenic chalcones 6 and their corresponding pyrazolines 7 were
prepared via an initial propargylation of substituted-4-hydroxy acetophenones 4 to yield the
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corresponding O-propargylated acetophenones 5. Aldol condensation of 5 with substituted
benzaldehydes in the presence of KOH led to the formation of O-propargylated chalcones 6.
The treatment of 6 with hydrazine hydrate in acetic acid at 120°C led to the formation of
corresponding pyrazolines 7 (Scheme 2).
Cu-promoted azide-alkyne cycloaddition reaction of 6 with (2-azido-ethyl/propyl)-(7-
chloro-quinolin-4-yl)-amine 3 in the presence of sodium ascorbate at room temperature
afforded the desired 1H-1,2,3-triazole tethered 4-aminoquinoline-chalcone conjugates 8 in
good to excellent yields. A similar approach, involving the reaction between 3 and 7 in
ethanol: water (85:15) mixture, was employed for the preparation of 1H-1,2,3-triazole-
tethered 4-aminoquinoline-pyrazoline conjugates 9 (Scheme 3).
The structure to the synthesized conjugates was assigned on the basis of spectral data
and analytical evidence. For example, 9p showed a molecular ion peak [M]+ at 699.2561 and
1
[M+2] at 701.2579 in its high resolution mass spectrum (HRMS), while its H NMR
spectrum exhibited the presence of a singlet at δ 2.43 due to the presence of an acetyl group,
a singlet at δ 5.19 due to presence of two methylene proton and a singlet at δ 8.94
corresponding to a triazole ring proton, along with characteristic absorptions due to quinoline
ring protons. The presence of the required number of carbons in the 13C NMR spectrum of 9p
along with characteristic absorption due to a carbonyl carbon at δ 167.9 further confirmed the
assigned structure.
2.2 In vitro Antiplasmodial activity
The synthesized conjugates were assayed for their antiplasmodial efficacy against the
chloroquine-resistant W2 strain of P. falciparum (Table 1). Although, the synthesized
compounds were not as active as artemisinin (ART); some of the conjugates exhibited
comparable or better activities than CQ. A careful analysis of Table 1 revealed the
dependence of activity on the length of the alkyl chain introduced as a spacer along with the
nature of substituents at ring A/ ring B of the chalcones. Comparing the activity data among
ethyl chain linked 4-aminoquinoline-chalcone conjugates viz. 8a-h revealed a reduction in
antiplasmodial efficacy with the introduction of a –OCH3 substituent at ring A irrespective of
the nature of substituents at ring B. The conjugates 8a-d (R=H, ring A) exhibited IC50 values
ranging from 114.4-172.1 nM. Increasing the chain length from ethyl to propyl (8i-p) resulted
in considerable loss in antiplasmodial activities, except in the case 8o, which displayed an
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IC50 value of 357 nM. The introduction of a pyrazoline ring substantially improved the
antiplasmodial activity of the synthesized conjugates. Comparing the activities among ethyl
chain linked 4-aminoquinoline-pyrazoline conjugates, 9a-h revealed improved activity with
the introduction of a –OCH3 substituent at ring A of the chalcone, as evident with conjugates
9f and 9g, exhibiting IC50 values of 58.7 nM and 53.7 nM, respectively. Similar comparison
among conjugates 9i-p, with a propyl linker, revealed reduction in antiplasmodial activities;
except for 9m, with an IC50 value of 77nM. Cytotoxicity of few of the most potent
compounds viz. 8b, 8c, 9b, 9f, 9g and 9m were assessed against mammalian HeLa cells and
the results are enlisted in Table 2. As evident the compounds were non-cytotoxic and
exhibited a good selectivity index.
In conclusion, a series of 4-aminoquinoline-chalcones/N-acetyl-pyrazolines were
synthesized via Cu-promoted azide-alkyne cycloaddition reaction and evaluated against
chloroquine-resistant P. falciparum. SAR studies revealed the conjugates to have comparable
antiplasmodial activities with that of chloroquine. 4-aminoquinoline-pyrazoline conjugate 9g,
with an optimum combination of a –OCH3 substituent at ring A and 3,4-dimethoxy
substituents at ring B proved to be most potent and non-cytotoxic, with an IC50 value of 53.7
nM.
3. Experimental Section
1H NMR spectra were recorded in deuterochloroform and DMSO-d6 with a Jeol 400 (400
MHz) and Bruker 500 (500 MHz) spectrometer using TMS as an internal standard. Chemical
shift values are expressed as parts per million downfield from TMS and J values are in hertz.
Splitting patterns are indicated as s: singlet, d: doublet, t: triplet, m: multiplet, dd: double
13
doublet, ddd: doublet of a doublet of a doublet, and br: broad peak. C NMR spectra were
recorded on a Bruker 400 (100 MHz) and Bruker 500 (125 MHz) spectrometer in
dimethylsulfoxide using TMS as internal standard. High resolution mass spectra were
recorded on a Bruker-micrOTOF-Q II spectrometer.
3.1 General procedure for the synthesis of conjugates 8 or 9: To the stirred solution of
(2-Azido-ethyl/propyl)-(7-chloro-quinolin-4-yl)-amine 3 (1 mmol) and propargylated
substituted chalones 6 (1 mmol) or propargylated substituted pyrazolines 7 (1 mmol) in
ethanol: water (85:15) mixture, was added CuSO4.5H2O (0.055 mmol) and sodium ascorbate
(0.143 mmol). The reaction mixture was allowed to stir at room temperature for 7-8 h, with
progress monitored via TLC. After the usual workup in DCM and water, combined organic
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layers were collected, dried over anhydrous Na2SO4 and evaporated under reduced pressure
to yield crude product which was purified by column chromatography over 60-120 silica gel
using 95:5 (CHCl3: MeOH) mixture.
3.1.1
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-phenyl-propenone (8a): Yield 85%; Light yellow Solid; m.p. 140-141 °C; 1H NMR
(DMSO-d6, 400 MHz): δ 3.50-3.52 (m, 2H, -CH2), 4.48-4.50 (m, 2H, -CH2), 5.21 (s, 2H, -
OCH2), 6.42 (d, J = 5.6 Hz, 1H, H2), 7.10 (m, 2H, ArH), 7.33 (d, J = 9.2 Hz, 1H, H4), 7.42-
7.46 (d, J = 8.8 Hz, 2H, ArH), 7.51 (d, J = 15.6 Hz, 1H, olefinic-H), 7.62 (m, 3H, ArH),7.76
(s, 1H, -NH exchangeable with D2O), 7.81 (d, J = 15.6 Hz, 1H, olefinic-H), 7.89 (s, 1H, H5),
7.95 (d, J = 9.2 Hz, 1H, H3), 8.07 (d, J = 8.4 Hz, 2H, ArH), 8.22 (s, 1H, H6), 8.42 (d, J = 5.6
13
Hz, 1H, H1); C NMR (DMSO-d6, 100 MHz): 48.5, 62.0, 69.9, 99.2, 107.2, 115.4, 117.6,
120.9, 124.1, 125.2, 125.8, 126.4, 128.3, 130.7, 131.9, 134.6, 140.9, 142.6, 144.2, 146.5,
150.2, 151.6, 152.3, 162.1, 188.0; HRMS: Calcd for C29H24ClN5O2 [M]+509.1619 and [M+2]
511.1619, found 509.1610 and 511.1605; Anal. Calcd (%) for: C, 68.30; H, 4.74; N, 13.73,
found: C, 68.43; H, 4.67; N, 13.65.
3.1.2
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-(4-methoxy-phenyl)-propenone (8b):Yield 82%; Light Yellow Solid; m.p. 179-180
°C; 1H NMR (CDCl3, 400 MHz): δ 3.65-3.67 (m, 2H, -CH2-),3.70 (s, 3H, -OCH3), 4.69-4.71
(m, 2H, -CH2), 5.20 (s, 2H, -OCH2), 6.45 (d, J = 5.6 Hz, 1H, H2), 7.02 (d, J = 8.8 Hz, 2H,
ArH), 7.32 (d, J = 9.2 Hz, 1H, H4), 7.42 (d, J = 8.4 Hz, 2H, ArH), 7.54 (d, J = 15.6 Hz, 1H,
olefinic-H), 7.69 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 15.6 Hz, 1H, olefinic-H), 7.88 (s, 1H,
H5), 7.98 (s, 1H, -NH exchangeable with D2O), 8.05 (d, J = 9.2 Hz, 1H, H3), 8.17 (d, J = 8.8
Hz, 2H, ArH), 8.25 (s, 1H, H6), 8.46 (d, J = 5.6 Hz, 1H, H1); 13C NMR (DMSO-d6, 100
MHz): 48.8, 55.2, 62.0, 70.5, 99.0, 107.4, 115.7, 117.5, 121.0, 124.3, 125.4, 126.0, 126.8,
130.7, 131.4, 134.8, 140.6, 142.9, 144.4, 146.3, 150.1, 151.8, 152.2, 157.5, 162.0, 188.3;
HRMS: Calcd for C30H26ClN5O3 [M]+539.1724 and [M+2] 541.1724, found 539.1733 and
541.1720; Anal. Calcd (%) for: C, 66.72; H, 4.85; N, 12.97, found: C, 66.62; H, 4.77; N,
12.84.
3.1.3
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-(3,4-dimethoxy-phenyl)-propenone (8c): Yield 90%; Light yellow Solid; m.p. 162-
1
163 °C; H NMR (DMSO-d6, 500 MHz): δ 3.70 (s, 3H, -OCH3), 3.77-3.79 (m, 2H, -
CH2),3.82 (s, 3H, -OCH3), 4.52-4.55 (m, 2H, -CH2), 5.19 (s, 2H, -OCH2), 6.44 (d, J = 6.0 Hz,
1H, H2), 6.49 (dd, J = 2.0, 8.0 Hz, 1H, ArH), 6.57 (d, J = 2.5 Hz, 1H, ArH), 6.80 (d, J = 8.5
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Hz, 1H, ArH), 7.18 (d, J = 8.5 Hz, 2H, ArH),7.52 (d, J = 15.6 Hz, 1H, olefinic-H), 7.59 (dd, J
= 1.5, 9.0 Hz, 1H, H4), 7.73 (d, J = 8.5 Hz, 2H, ArH), 7.83 (d, J = 15.6 Hz, 1H, olefinic-H),
7.85 (s, 1H, -NH exchangeable with D2O), 8.09 (s, 1H, H5), 8.20 (d, J = 9.0 Hz, 1H, H3), 8.34
13
(s, 1H, H6), 8.46 (d, J = 4.5 Hz, 1H, H1); C NMR (DMSO-d6, 125 MHz): 48.0, 56.4, 60.6,
62.1, 79.3, 99.2, 107.1, 115.4, 117.5,121.2, 123.9, 124.8, 125.1, 126.0, 126.6, 129.8, 131.6,
135.0, 140.2, 142.7, 144.0, 147.4, 150.6, 151.1, 151.8, 152.9, 153.6, 162.5, 188.1; HRMS:
Calcd for C31H28ClN5O4 [M]+569.1830 and [M+2] 571.1830, found 569.1836 and 571.1842;
Anal. Calcd (%) for: C, 65.32; H, 4.95; N, 12.29, found: C, 65.45; H, 5.09; N, 12.24.
3.1.4
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-(3,4,5-trimethoxy-phenyl)-propenone (8d): Yield 87%; Light yellow Solid; m.p.
152-153 °C; 1H NMR (DMSO-d6, 400 MHz): δ 3.74 (s, 3H, -OCH3), 3.77 (s, 6H, 2×-OCH3),
3.81-3.83 (m, 2H, -CH2), 4.74-4.76 (m, 2H, -CH2), 5.20 (s, 2H, -OCH2),6.40 (s, 2H, ArH),
6.46(d, J = 6.0 Hz, 1H, H2), 6.99 (d, J = 9.2 Hz, 2H, ArH), 7.32 (dd, J = 2.0, 8.4 Hz, 1H, H4),
7.51 (d, J = 15.6 Hz, 1H, olefinic-H), 7.58 (d, J = 8.8 Hz, 2H, ArH), 7.72 (s, 1H, H5), 7.79 (d,
13
J = 15.6 Hz, 1H, olefinic-H), 7.85 (m, 2H, H3+ H6), 8.33 (d, J = 5.6 Hz, 1H, H1); C NMR
(DMSO-d6, 100 MHz): 47.9, 55.1, 55.4, 60.3, 61.8, 68.8, 98.9, 104.6, 115.0, 117.2, 121.7,
123.5, 124.5, 125.3, 126.0, 126.6, 128.6, 134.9, 140.1, 142.5, 144.0, 147.5, 150.3, 154.8,
156.9, 159.7, 161.0, 187.2; HRMS: Calcd for C32H30ClN5O5 [M]+599.1935 and [M+2]
601.1935, found 599.1928 and 601.1944; Anal. Calcd (%) for: C, 64.05; H, 5.04; N, 11.67,
found: C, 63.95; H, 5.16; N, 11.59.
3.1.5
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-phenyl-propenone (8e): Yield 90%; Light yellow Solid; m.p. 143-144 °C;
1H NMR (DMSO-d6, 400 MHz): δ 3.71-3.74 (m, 5H, -CH2+ -OCH3), 4.76-4.78 (m, 2H, -
CH2), 5.17 (s, 2H, -OCH2), 6.54 (d, J = 6.0 Hz, 1H, H2), 6.93 (m, 2H, ArH), 7.18 (d, J = 8.0
Hz, 1H, ArH), 7.31 (d, J = 8.8 Hz, 1H, H4), 7.53-7.61 (m, 6H, ArH+ olefinic-H), 7.82 (d, J =
15.6 Hz, 1H, olefinic-H), 7.85 (s, 1H, H5), 8.01 (d, J = 8.8 Hz, 1H, H3), 8.15 (s, 1H, H6), 8.40
(d, J = 6.0 Hz, 1H, H1); 13C NMR (DMSO-d6, 125 MHz): 48.1, 55.1, 61.5, 79.0, 99.1, 106.8,
111.2, 112.5, 117.1, 120.5, 123.2, 124.3, 125.3, 125.7, 126.4, 128.0, 131.4, 135.0, 139.9,
142.2, 143.7, 147.5, 149.8, 150.1, 151.6, 152.8, 161.7, 188.6; HRMS: Calcd for
C30H26ClN5O3 [M]+539.1724 and [M+2] 541.1724, found 539.1731 and 541.1712; Anal.
Calcd (%) for: C, 66.72; H, 4.85; N, 12.97, found: C, 66.62; H, 4.71; N, 12.90.
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3.1.6
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-(4-methoxy-phenyl)-propenone (8f): Yield 83%; Light Yellow Solid; m.p.
182-183 °C; 1H NMR (CDCl3, 400 MHz): δ 3.71 (s, 3H, -OCH3), 3.74 (s, 3H, -OCH3), 3.81-
3.84 (m, 2H, -CH2), 4.59-4.61 (m, 2H, -CH2), 5.20 (s, 2H, -OCH2), 6.47 (d, J = 5.6 Hz, 1H,
H2), 6.80 (d, J = 8.8 Hz, 2H, ArH), 7.21 (d, J = 8.p Hz, 1H, ArH), 7.32 (d, J = 9.2 Hz, 1H,
H4), 7.54-7.62 (m, 5H, 4ArH+olefinic-H), 7.79 (d, J = 15.6 Hz, 1H, olefinic-H), 7.90 (s, 1H,
13
H5), 8.04 (d, J = 9.2 Hz, 1H, H3), 8.20 (s, 1H, H6), 8.42 (d, J = 5.6 Hz, 1H, H1); C NMR
(DMSO-d6, 125 MHz): 48.4, 55.1, 55.6, 62.0, 79.1, 98.8, 106.2, 111.5, 112.7, 117.3, 120.6,
123.4, 124.2, 125.1, 125.7, 126.0, 128.9, 131.0, 135.3, 140.2, 142.4, 143.9, 147.6, 149.7,
150.2, 151.8, 157.4, 162.1, 188.2; HRMS: Calcd for C31H28ClN5O3 [M]+569.1830 and [M+2]
571.1830, found 569.1840 and 571.1837; Anal. Calcd (%) for: C, 65.32; H, 4.95; N, 12.29,
found: C, 65.46; H, 4.87; N, 12.43.
3.1.7
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-(3,4-dimethoxy-phenyl)-propenone (8g): Yield 79%; Light yellow Solid;
1
m.p. 155-156 °C; H NMR (DMSO-d6, 500 MHz): δ 3.82-3.86 (m, 11H, -CH2+3×-OCH3),
4.70-4.72 (m, 2H, -CH2), 5.25 (s, 2H, -OCH2), 6.66 (d, J = 5.6 Hz, 1H, H2), 7.02 (d, J = 8.5
Hz, 1H, ArH), 7.29 (d, J = 8.5 Hz, 1H, ArH), 7.40 (d, J = 9.0 Hz, 1H, H4), 7.53-7.60 (m, 3H,
ArH), 7.68 (d, J = 15.5 Hz, 1H, olefinic-H), 7.81-7.91 (m, 3H, olefinic-H+H3+ArH), 8.02 (s,
1H, H5), 8.27-8.33 (m, 2H, H6+H1); 13C NMR (DMSO-d6, 125 MHz): 48.3, 56.0, 56.1, 56.2,
62.1, 79.6, 99.7, 104.7, 111.3, 111.4, 112.0, 112.6, 119.9, 123.5, 124.1, 124.8, 125.5, 125.8,
128.1, 131.5, 135.1, 139.8, 142.6, 144.0, 149.3, 149.4, 151.5, 151.8, 152.1, 155.0, 162.7,
187.7; HRMS: Calcd for C32H30ClN5O5 [M]+599.1935 and [M+2] 601.1935, found 599.1942
and 601.1948; Anal. Calcd (%) for: C, 64.05; H, 5.04; N, 11.67, found: C, 64.21; H, 5.11; N,
11.55.
3.1.8
1-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-(3,4,5-trimethoxy-phenyl)-propenone (8h): Yield 90%; Light yellow
1
Solid; m.p. 148-149 °C; H NMR (DMSO-d6, 400 MHz): δ 3.71-3.79 (m, 14H, 4×-OCH3+ -
CH2), 4.83-4.85 (m, 2H, -CH2), 5.22 (s, 2H, -OCH2), 6.33 (s, 2H, ArH), 6.70 (d, J = 6.0 Hz,
1H, H2), 7.12 (d, J = 8.0 Hz, 1H, ArH), 7.43 (d, J = 9.2 Hz, 1H, H4), 7.47-7.52 (m, 2H, ArH),
7.65 (d, J = 15.6 Hz, 1H, olefinic-H), 7.82 (d, J = 15.6 Hz, 1H, olefinic-H), 7.85 (s, 1H, H5),
7.99 (d, J = 9.2 Hz, 1H, H3), 8.24 (s, 1H, H6), 8.41 (d, J = 6.0 Hz, 1H, H1); 13C NMR
(DMSO-d6, 125 MHz): 47.4, 55.2, 55.4, 55.5, 60.5, 61.0, 79.6, 99.1, 106.3, 111.4, 112.9,
116.8, 120.1, 124.0, 124.6, 125.4, 125.7, 126.3, 127.9, 131.4, 135.3, 139.7, 142.5, 143.6,
7

ACCEPTED MANUSCRIPT
147.5, 150.3, 154.0, 156.4, 159.5, 162.6, 187.8; HRMS: Calcd for C33H32ClN5O6
[M]+629.2041 and [M+2] 631.2041, found 629.2036 and 631.2049; Anal. Calcd (%) for: C,
62.90; H, 5.12; N, 11.11, found: C, 62.95; H, 5.01; N, 11.04.
3.1.9
1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-phenyl-propenone (8i): Yield 89%; Light yellow Solid; m.p. 161-162 °C; 1H NMR
(DMSO-d6, 400 MHz): δ 2.25-2.28 (m, 2H, -CH2), 3.55-3.57 (m, 2H, -CH2), 4.64-4.66 (m,
2H, -CH2), 5.17 (s, 2H, -OCH2), 6.39 (d, J = 5.2 Hz, 1H, H2), 7.18 (m, 2H, 2ArH), 7.31 (d, J
= 8.8 Hz, 1H, H4), 7.49 (d, J = 8.8 Hz, 2H, ArH),7.57 (d, J = 15.6 Hz, 1H, olefinic-H), 7.68-
7.70 (m, 3H, ArH), 7.85 (d, J = 15.6 Hz, 1H, olefinic-H), 7.93 (s, 1H, H5), 7.97 (d, J = 8.8Hz,
13
1H, H3), 8.06 (d, J = 8.8 Hz, 2H, ArH), 8.19 (s, 1H, H6), 8.41 (d, J = 5.6 Hz, 1H, H1); C
NMR (DMSO-d6, 100 MHz): 28.6, 48.5, 61.7, 70.3, 99.0, 107.3, 114.9, 117.4, 121.1, 124.1,
125.3, 125.8, 126.5, 128.6, 130.7, 131.9, 134.3, 140.7, 142.5, 144.6, 146.4, 150.0, 151.5,
152.5, 162.3, 188.1; HRMS: Calcd for C30H26ClN5O2 [M]+523.1775 and [M+2] 525.1775,
found 523.1768 and 525.1784; Anal. Calcd (%) for: C, 68.76; H, 5.00; N, 13.36, found: C,
68.87; H, 5.08; N, 13.50.
3.1.10
1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-(4-methoxy-phenyl)-propenone (8j): Yield 86%; Light Yellow Solid; m.p. 175-176
1
°C; H NMR (CDCl3, 400 MHz): δ 2.29-2.32 (m, 2H, -CH2), 3.70-3.73 (m, 5H, -CH2+-
OCH3), 4.75-4.77 (m, 2H, -CH2), 5.21 (s, 2H, -OCH2), 6.47 (d, J = 5.6 Hz, 1H, H2), 7.07 (d, J
= 8.8 Hz, 2H, ArH), 7.39 (d, J = 9.2 Hz, 1H, H4), 7.51 (d, J = 8.8 Hz, 2H, ArH), 7.54 (d, J =
15.6 Hz, 1H, olefinic-H), 7.78 (d, J = 8.8 Hz, 2H, ArH), 7.83 (d, J = 15.6 Hz, 1H, olefinic-
H),7.97 (s, 1H, H5), 8.02 (s, 1H, -NH exchangeable with D2O), 8.14 (d, J = 9.2 Hz, 1H, H3),
8.22 (d, J = 8.8 Hz, 2H, ArH), 8.31(s, 1H, H6), 8.44 (d, J = 5.6 Hz, 1H, H1); 13C NMR
(DMSO-d6, 100 MHz): 29.1, 48.1, 55.2, 61.5, 79.0, 99.0, 107.1, 115.6, 117.3, 121.0, 124.3,
125.7, 126.4, 126.7, 130.4, 131.4, 135.2, 140.8, 142.9, 145.1, 146.3, 150.4, 152.2, 152.6,
157.4, 162.3, 187.9; HRMS: Calcd for C31H28ClN5O3 [M]+553.1881 and [M+2] 555.1881,
found 553.1871 and 555.1887; Anal. Calcd (%) for: C, 67.20; H, 5.09; N, 12.64, found: C,
67.31; H, 4.97; N, 12.74.
3.1.11
1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-(3,4-dimethoxy-phenyl)-propenone (8k): Yield 93%; Light yellow Solid; m.p. 146-
1
147 °C; H NMR (DMSO-d6, 400 MHz): δ 2.25-2.28 (m, 2H, -CH2), 3.69 (s, 3H, -OCH3),
3.77-3.79 (m, 2H, -CH2), 3.82 (s, 3H, -OCH3), 4.52-4.55 (m, 2H, -CH2), 5.19 (s, 2H, -OCH2),
8

ACCEPTED MANUSCRIPT
6.44 (d, J = 6.0 Hz, 1H, H2), 6.49 (dd, J = 2.0, 8.0 Hz, 1H, ArH), 6.57 (d, J = 2.5 Hz, 1H,
ArH), 6.80 (d, J = 8.5 Hz, 1H, ArH), 7.18 (d, J = 8.5 Hz, 2H, ArH), 7.59 (dd, J = 1.5, 9.0 Hz,
1H, H4), 7.73 (d, J = 8.5 Hz, 2H, ArH), 7.85 (s, 1H, -NH exchangeable with D2O), 8.20 (d, J
= 9.0 Hz, 1H, H3), 8.29 (s, 1H, H5), 8.34 (s, 1H, H6), 8.46 (d, J = 4.5 Hz, 1H, H1); 13C NMR
(DMSO-d6, 125 MHz): 28.6, 48.2, 56.1, 60.1, 62.0, 78.7, 98.8, 107.0, 115.5, 117.7, 121.4,
124.0, 124.6, 125.1, 126.1, 126.8, 130.6, 131.8, 135.2, 140.4, 142.5, 144.3, 147.7, 150.4,
151.6, 152.0, 153.1, 153.9, 162.4, 188.0; HRMS: Calcd for C32H30ClN5O4 [M]+583.1986 and
[M+2] 585.1986, found 583.1976 and 585.1992; Anal. Calcd (%) for: C, 65.80; H, 5.18; N,
11.99, found: C, 65.89; H, 5.10; N, 12.05.
3.1.12
1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-3-(3,4,5-trimethoxy-phenyl)-propenone (8l): Yield 86%; Light yellow Solid; m.p.
173-174 °C; 1H NMR (DMSO-d6, 500 MHz): δ 2.24-2.27 (m, 2H, -CH2), 3.62-3.64 (m, 2H, -
CH2), 3.72 (s, 3H, -OCH3), 3.87 (s, 6H, 2×-OCH3), 4.54-4.56 (m, 2H, -CH2), 5.29 (s, 2H, -
OCH2), 6.53 (d, J = 6.0 Hz, 1H, H2), 7.20-7.22 (m, 4H, ArH), 7.54 (dd, J = 2.0, 9.0 Hz, 1H,
H4), 7.67 (d, J = 15.5 Hz, 1H, olefinic-H), 7.82-7.83 (m, 2H, H5+-NH exchangeable with
D2O), 7.89 (d, J = 15.5 Hz, 1H, olefinic-H), 7.93 (d, J = 8.5 Hz, 1H, H3), 8.19 (d, J = 9.0 Hz,
13
2H, ArH), 8.33 (s, 1H, H6), 8.43 (d, J = 4.5 Hz, 1H, H1); C NMR (DMSO-d6, 125 MHz):
28.8, 47.8, 56.1, 56.6, 60.6, 61.9, 70.2, 99.1, 106.9, 115.1, 117.4, 121.6, 124.9, 125.2, 125.3,
126.0, 130.8, 131.3, 134.9, 140.0, 142.6, 144.1, 147.0, 150.3, 151.6, 152.8, 153.5, 162.3,
187.7; HRMS: Calcd for C33H32ClN5O5 [M]+613.2092 and [M+2] 615.2092, found 613.2085
and 615.2098; Anal. Calcd (%) for: C, 64.54; H, 5.25; N, 11.40, found: C, 64.64; H, 5.33; N,
11.47.
3.1.13 1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-phenyl-propenone (8m): Yield 79%; Light yellow Solid; m.p. 157-158
1
°C; H NMR (DMSO-d6, 400 MHz): δ 2.35-2.38 (m, 2H, -CH2), 3.72 (s, 3H, -OCH3), 3.82-
3.84 (m, 2H, -CH2), 4.66-4.68 (m, 2H, -CH2), 5.21 (s, 2H, -OCH2), 6.38 (d, J = 5.6 Hz, 1H,
H2), 6.87 (m, 2H, ArH), 7.09 (d, J = 8.0 Hz, 1H, ArH), 7.40 (d, J = 9.2 Hz, 1H, H4), 7.49-
7.56 (m, 6H, 5ArH+olefinic-H), 7.82 (d, J = 15.6 Hz, 1H, olefinic-H), 7.93 (s, 1H, H5), 7.98
(d, J = 9.2 Hz, 1H, H3), 8.20 (s, 1H, H6), 8.43 (d, J = 5.6 Hz, 1H, H1); 13C NMR (DMSO-d6,
125 MHz): 28.2, 48.5, 55.2, 62.0, 70.5, 99.1, 106.0, 111.2, 112.6, 118.1, 119.7, 123.4, 124.5,
125.2, 125.6, 126.8, 128.1, 131.3, 135.2, 139.7, 142.1, 143.6, 147.4, 149.9, 150.0, 151.7,
152.9, 161.8, 188.2; HRMS: Calcd for C31H28ClN5O3 [M]+553.1881 and [M+2] 555.1881,
9

ACCEPTED MANUSCRIPT
found 553.1872 and 555.1893; Anal. Calcd (%) for: C, 67.20; H, 5.09; N, 12.64, found: C,
67.30; H, 5.22; N, 12.57.
3.1.4 1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-(4-methoxy-phenyl)-propenone (8n): Yield 92%; Light Yellow Solid;
1
m.p. 160-161 °C; H NMR (CDCl3, 400 MHz): δ 2.21-2.24 (m, 2H, -CH2-), 3.70 (s, 3H, -
OCH3), 3.76 (s, 3H, -OCH3), 3.83-3.85 (m, 2H, -CH2), 4.72-4.75 (m, 2H, -CH2), 5.18 (s, 2H,
-OCH2), 6.49 (d, J = 5.6 Hz, 1H, H2), 7.09-7.12 (m, 3H, ArH), 7.41 (d, J = 9.2 Hz, 1H, H4),
7.45-7.51 (m, 4H, ArH), 7.58 (d, J = 15.6 Hz, 1H, olefinic-H), 7.83 (d, J = 15.6 Hz, 1H,
olefinic-H), 7.89 (s, 1H, H5), 8.11 (d, J = 9.2 Hz, 1H, H3), 8.17 (s, 1H, H6), 8.41 (d, J = 5.6
Hz, 1H, H1); 13C NMR (DMSO-d6, 125 MHz): 28.0, 47.5, 55.2, 55.5, 62.1, 79.2, 98.9, 105.7,
111.3, 111.8, 118.3, 119.8, 123.1, 124.0, 125.4, 125.6, 126.2, 128.7, 131.1, 135.4, 140.6,
142.1, 143.7, 147.5, 149.8, 150.0, 152.4, 157.0, 161.8, 188.3; HRMS: Calcd for
C32H30ClN5O4 [M]+583.1986 and [M+2] 585.1986, found 583.1991 and 558.1979; Anal.
Calcd (%) for: C, 65.80; H, 5.18; N, 11.99, found: C, 65.89; H, 5.27; N, 11.84.
3.1.15 1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-(3,4-dimethoxy-phenyl)-propenone (8o): Yield 89%; Light yellow Solid;
m.p. 144-145 °C; 1H NMR (DMSO-d6, 400 MHz): δ 2.26-2.29 (m, 2H, -CH2), 3.62-3.64 (m,
2H, -OCH2), 3.71 (s, 3H, -OCH3), 3.75 (s, 6H, 2×-OCH3), 4.84-4.87 (m, 2H, -CH2), 5.22 (s,
2H, -OCH2), 6.39 (d, J = 8.0 Hz, 1H, ArH), 6.54 (d, J = 5.6 Hz, 1H, H2), 6.91 (d, J = 8.4 Hz,
1H, ArH), 7.21 (d, J = 8.0 Hz, 1H, ArH), 7.42 (d, J = 9.0 Hz, 1H, H4), 7.51-7.56 (m, 3H,
ArH), 7.62 (d, J = 15.6 Hz, 1H, olefinic-H), 7.81 (d, J = 15.6 Hz, 1H, olefinic-H), 7.94 (s,
1H, H5), 8.08 (d, J = 9.2 Hz, 1H, H3), 8.19 (s, 1H, H6), 8.29 (d, J = 5.6 Hz, 1H, H1); 13C NMR
(DMSO-d6, 125 MHz): 27.8, 48.0, 55.1, 55.4, 60.2, 62.3, 79.1, 99.5, 104.4, 111.2, 111.5,
114.6, 120.5, 123.1, 124.0, 124.5, 125.3, 125.5, 125.1, 127.9, 131.2, 135.0, 139.6, 142.4,
143.7, 147.2, 149.0, 149.2, 151.2, 151.6, 154.4, 162.1, 188.0; HRMS: Calcd for
C33H32ClN5O5 [M]+ 613.2092 and [M+2] 615.2092, found 613.2085 and 615.2083; Anal.
Calcd (%) for: C, 64.54; H, 5.25; N, 11.40, found: C, 64.61; H, 5.42; N, 11.33.
3.1.16 1-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-3-(3,4,5-trimethoxy-phenyl)-propenone (8p): Yield 86%; Light yellow
1
Solid; m.p. 169-170 °C; H NMR (DMSO-d6, 400 MHz): δ 2.27-2.30 (m, 2H, -CH2), 3.73-
3.80 (m, 14H, 4×-OCH3+ -CH2), 4.59-4.61 (m, 2H, -CH2), 5.16 (s, 2H, -OCH2), 6.36 (s, 2H,
ArH), 6.58 (d, J = 5.6 Hz, 1H, H2), 7.15 (d, J = 8.0 Hz, 1H, ArH), 7.38 (d, J = 8.8 Hz, 1H,
10

ACCEPTED MANUSCRIPT
H4), 7.42-4.46 (m, 2H, ArH), 7.57 (d, J = 15.6 Hz, 1H, olefinic-H), 7.85 (d, J = 15.6 Hz, 1H,
olefinic-H), 7.90 (s, 1H, H5), 8.10 (d, J = 8.8 Hz, 1H, H3), 8.22 (s, 1H, H6), 8.46 (d, J = 5.6
13
Hz, 1H, H1); C NMR (DMSO-d6, 100 MHz): 28.4, 47.5, 55.0, 55.1, 55.5, 60.9, 62.4, 78.6,
99.0, 105.1, 111.4, 112.6, 117.3, 119.1, 123.3, 124.2, 125.5, 125.9, 126.1, 128.2, 131.6,
135.7, 139.9, 142.4, 143.6, 147.5, 150.2, 153.6, 156.9, 159.3, 162.4, 187.7; HRMS: Calcd for
C34H34ClN5O6 [M]+643.2198 and [M+2] 645.2198, found 643.2187 and 645.2204; Anal.
Calcd (%) for: C, 63.40; H, 5.32; N, 10.87, found: C, 63.45; H, 5.21; N, 10.98.
3.1.17 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-phenyl-4,5-dihydro-pyrazol-1-yl]-ethanone (9a): Yield 94%; Light yellow Solid;
1
m.p. 181-182 °C; H NMR (DMSO-d6, 500 MHz): δ 2.41 (s, 3H, -CH3), 2.95 (dd, J = 4.0,
17.5 Hz, 1H, H7), 3.51 (dd, J = 11.5, 17.5 Hz, 1H, H8), 3.81-3.83 (m, 2H, -CH2), 4.72-4.74
(m, 2H, -CH2), 5.22 (s, 2H, -OCH2), 5.59 (dd, J = 4.5, 11.5 Hz, 1H, H9), 6.41 (d, J = 5.5 Hz,
1H, H2), 7.13 (m, 2H, ArH), 7.37 (d, J = 9.0 Hz, 1H, H4), 7.52-7.57 (m, 5H, ArH), 7.88 (s,
1H, H5), 7.92 (d, J = 9.2 Hz, 1H, H3), 8.11 (d, J = 8.5 Hz, 2H, ArH), 8.23 (s, 1H, H6), 8.40 (d,
J = 5.5 Hz, 1H, H1); 13C NMR (DMSO-d6, 125MHz): 21.6, 40.1, 42.5, 47.8,62.1, 79.0, 99.1,
107.7, 115.5, 117.4, 121.1, 123.5, 124.6, 125.4, 125.8, 126.0, 126.8, 128.4, 135.8, 139.7,
143.4, 146.9, 150.1, 151.6, 154.9, 160.5, 167.9; HRMS: Calcd for C31H28ClN7O2
[M]+565.1993 and [M+2] 567.1993, found 565.2004 and 567.1987; Anal. Calcd (%) for: C,
65.78; H, 4.99; N, 17.32, found: C, 65.90; H, 4.94; N, 17.20.
3.1.18 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-(4-methoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9b):Yield 82%; Light
Yellow Solid; m.p. 189-190 °C; 1H NMR (CDCl3, 500 MHz): δ 2.27 (s, 3H, -CH3), 2.91 (dd,
J = 4.0, 17.5 Hz, 1H, H7), 3.48 (dd, J = 11.5, 17.5 Hz, 1H, H8), 3.73 (s, 3H, -OCH3), 3.76-
3.79 (m, 2H, -CH2), 4.78-4.80 (m, 2H, -CH2), 5.19 (s, 2H, -OCH2), 5.56 (dd, J = 4.5, 11.5 Hz,
1H, H9), 6.45 (d, J = 5.5 Hz, 1H, H2), 7.05 (d, J = 8.5 Hz, 2H, ArH), 7.31(d, J = 9.0 Hz, 1H,
H4), 7.41 (d, J = 8.5 Hz, 2H, ArH), 7.73 (d, J = 8.5 Hz, 2H, ArH), 7.92 (s, 1H, H5), 8.00 (s,
1H, -NH exchangeable with D2O), 8.09 (d, J = 9.0 Hz, 1H, H3), 8.19 (d, J = 8.5 Hz, 2H,
13
ArH), 8.30 (s, 1H, H6), 8.41 (d, J = 5.5 Hz, 1H, H1); C NMR (DMSO-d6, 125 MHz): 22.1,
39.8, 42.0, 48.6, 55.1, 62.4, 78.5, 99.3, 107.7, 115.6, 117.8, 121.2, 123.9, 124.4, 125.2, 126.0,
126.5, 128.4, 134.8, 139.3, 143.1, 147.3, 150.2, 151.4, 154.8, 157.5, 160.0, 168.3; HRMS:
Calcd for C32H30ClN7O3 [M]+595.2099 and [M+2] 597.2099, found 595.2091 and 597.2086;
Anal. Calcd (%) for: C, 64.48; H, 5.07; N, 16.45, found: C, 64.38; H, 5.19; N, 16.52.
11

ACCEPTED MANUSCRIPT
3.1.19 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-(3,4-dimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9c): Yield 72%; Light
1
yellow Solid; m.p. 183-184 °C; H NMR (DMSO-d6, 500 MHz): δ 2.30 (s, 3H, -CH3), 2.91
(dd, J = 4.0, 17.5 Hz, 1H, H7), 3.68-3.74 (m, 4H, -OCH3+ H8), 3.79 (s, 3H, -OCH3), 3.83-
3.85 (m, 2H, -CH2), 4.67-4.70 (m, 2H, -CH2), 5.17 (s, 2H, -OCH2), 5.56 (dd, J = 4.5, 11.5 Hz,
1H, H9), 6.43 (dd, J = 2.0, 8.0 Hz, 1H, ArH), 6.58 (d, J = 2.5 Hz, 1H, ArH), 6.60 (d, J = 5.5
Hz, 1H, H2), 6.78 (d, J = 8.5 Hz, 1H, ArH), 7.05 (m, 2H, ArH), 7.53 (dd, J = 1.5, 9.0 Hz, 1H,
H4), 7.67 (d, J = 8.5 Hz, 2H, ArH), 7.88 (s, 1H, -NH exchangeable with D2O), 8.21 (d, J =
13
9.0 Hz, 1H, H3), 8.27 (s, 1H, H5), 8.32 (s, 1H, H6), 8.43 (d, J = 4.5 Hz, 1H, H1); C NMR
(DMSO-d6, 125 MHz): 22.1, 41.7, 42.9, 48.3, 55.6, 56.0, 61.6, 79.6, 99.3, 104.9, 115.3,
117.5, 122.2, 124.6, 124.7, 125.3, 125.6, 126.5, 128.6, 131.0, 134.9, 138.3, 142.8, 147.3,
150.5, 151.4, 154.7, 157.2, 159.9, 160.2, 167.4; HRMS: Calcd for C33H32ClN7O4
[M]+625.2204 and [M+2] 627.2204, found 625.2212 and 627.2208; Anal. Calcd (%) for: C,
63.30; H, 5.15; N, 15.66, found: C, 63.39; H, 5.28; N, 15.54.
3.1.20 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-(3,4,5-trimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9d): Yield 81%;
Light yellow Solid; m.p. 166-167 °C; 1H NMR (DMSO-d6, 400 MHz): δ 2.41 (s, 3H, -CH3),
3.07 (dd, J = 4.8, 17.6 Hz, 1H, H7), 3.67 (dd, J = 12.0, 18.0 Hz, 1H, H8), 3.76 (s, 3H, -OCH3),
3.77 (s, 6H, 2×-OCH3), 3.91-3.93 (m, 2H, -CH2), 4.75-4.77 (m, 2H, -CH2), 5.17 (s, 2H, -
OCH2), 5.47 (dd, J = 4.4, 11.6 Hz, 1H, H9), 6.34 (d, J = 6.0 Hz, 1H, H2), 6.38 (s, 2H, ArH),
6.94 (d, J = 9.2 Hz, 2H, ArH), 7.30 (dd, J = 2.0, 8.4 Hz, 1H, H4), 7.62 (d, J = 8.8 Hz, 2H,
ArH), 7.74 (s, 1H, H5), 7.89 (m, 2H, H3+ H6), 8.36 (d, J = 5.6 Hz, 1H, H1); 13C NMR
(DMSO-d6, 125MHz): 22.0, 42.7, 43.1, 48.7, 56.1, 60.2, 60.8, 61.8, 79.5, 98.5, 102.3, 114.9,
116.7, 122.4, 124.2, 124.6, 126.5, 128.3, 136.6, 137.8, 144.0, 147.0, 150.5, 151.5, 153.6,
153.8, 156.5, 159.8, 161.0, 169.0; HRMS: Calcd for C34H34ClN7O5 [M]+655.2310 and [M+2]
657.2310, found 655.2318 and 657.2302; Anal. Calcd (%) for: C, 62.24; H, 5.22; N, 14.94,
found: C, 62.36; H, 5.19; N, 15.24.
3.1.21 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-5-phenyl-4,5-dihydro-pyrazol-1-yl]-ethanone
(9e): Yield 90%; Light
1
yellow Solid; m.p. 199-200 °C; H NMR (DMSO-d6, 400 MHz): δ 2.41 (s, 3H, -CH3), 3.01
(dd, J = 4.0, 17.6 Hz, 1H, H7), 3.51 (dd, J = 11.6, 18.0 Hz, 1H, H8), 3.73 (s, 3H, -OCH3),
3.79-3.81 (m, 2H, -CH2), 4.81-4.83 (m, 2H, -CH2), 5.25 (s, 2H, -OCH2), 5.46 (dd, J = 4.0,
11.6 Hz, 1H, H9), 6.45 (d, J = 5.6 Hz, 1H, H2), 6.88-6.91 (m, 2H, ArH), 7.10 (d, J = 8.0 Hz,
12

ACCEPTED MANUSCRIPT
1H, ArH), 7.33 (d, J = 9.2 Hz, 1H, H4), 7.51-7.56 (m, 5H, ArH), 7.90 (s, 1H, H5), 8.00 (d, J =
13
9.2 Hz, 1H, H3), 8.07 (s, 1H, H6), 8.29 (d, J = 5.6 Hz, 1H, H1); C NMR (DMSO-d6, 125
MHz): 21.4, 39.8, 41.8, 47.2, 55.2, 61.6, 79.1, 99.1, 105.4, 111.1, 112.2, 116.4, 121.0, 123.1,
124.2, 125.4, 125.8, 126.0, 126.4, 128.6, 135.8, 139.8, 143.2, 146.8, 149.7, 150.2, 151.2,
154.9, 160.4, 167.8; HRMS: Calcd for C32H30ClN7O3 [M]+595.2099 and [M+2] 597.2099,
found 595.2094 and 597.2110; Anal. Calcd (%) for: C, 64.48; H, 5.07; N, 16.45, found: C,
64.55; H, 5.21; N, 16.38.
3.1.22 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-5-(4-methoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9f): Yield 83%;
1
Light Yellow Solid; m.p. 190-191 °C; H NMR (CDCl3, 400 MHz): δ 2.37 (s, 3H, -CH3),
2.96 (dd, J = 4.0, 17.6 Hz, 1H, H7), 3.46 (dd, J = 11.6, 17.6 Hz, 1H, H8), 3.71 (s, 3H, -OCH3),
3.73 (s, 3H, -OCH3), 3.78-3.81 (m, 2H, -CH2), 4.68-4.71 (m, 2H, -CH2), 5.16 (s, 2H, -OCH2),
5.50 (dd, J = 4.0, 11.6 Hz, H9), 6.61 (d, J = 5.6 Hz, 1H, H2), 6.77 (d, J = 8.8 Hz, 2H, ArH),
7.03 (d, J = 8.0 Hz, 1H, ArH), 7.42 (d, J = 9.2 Hz, 1H, H4), 7.49-7.53 (m, 4H, ArH), 7.95-
13
7.98 (m, 2H, H3+H5), 8.06 (s, 1H, H6), 8.22 (d, J = 5.6 Hz, 1H, H1); C NMR (DMSO-d6,
125 MHz): 21.6, 41.2, 42.3, 48.2, 55.2, 55.5, 62.5, 79.4, 99.1, 105.4, 111.0, 112.3, 115.9,
119.8, 123.3, 124.3, 125.4, 126.5, 126.8, 128.4, 135.1, 139.7, 142.9, 147.2, 149.6, 150.1,
151.4, 153.0, 157.5, 160.3, 168.0; HRMS: Calcd for C33H32ClN7O4 [M]+625.2204 and [M+2]
627.2204, found 625.2216 and 627.2189; Anal. Calcd (%) for: C, 63.30; H, 5.15; N, 15.66,
found: C, 63.20; H, 5.23; N, 15.63.
3.1.23 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
3-methoxy-phenyl)-5-(3,4-dimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9g): Yield
79%; Light yellow Solid; m.p. 197-198 °C; 1H NMR (DMSO-d6, 300 MHz): δ 2.31 (s, 3H, -
CH3), 3.09 (dd, J = 4.0, 17.6 Hz, 1H, H7), 3.42 (dd, J = 11.6, 17.6 Hz, 1H, H8), 3.82-3.86 (m,
11H, -CH2+3×-OCH3), 4.70-4.72 (m, 2H, -CH2), 5.24 (s, 2H, -OCH2), 5.46 (dd, J = 4.0, 11.6
Hz, H9), 6.64 (d, J = 5.6 Hz, 1H, H2), 7.01 (d, J = 7.8 Hz, 1H, ArH), 7.15 (d, J = 8.4 Hz, 1H,
ArH), 7.40 (d, J = 9.0 Hz, 1H, H4), 7.50-7.54 (m, 3H, ArH), 7.76 (s, 1H, ArH), 8.02 (s, 1H,
13
H5), 8.09 (d, J = 9.2 Hz, 1H, H3), 8.22 (s, 1H, H6), 8.41 (d, J = 5.6 Hz, 1H, H1); C NMR
(DMSO-d6, 125 MHz): 21.4, 40.5, 42.6, 48.0, 55.3, 55.6, 55.7, 62.3, 79.6, 99.0, 105.4, 110.9,
111.8, 112.3, 120.4, 123.1, 123.6, 124.5, 125.1, 125.3, 125.7, 128.0, 134.9, 139.6, 143.2,
147.3, 149.0, 149.9, 151.2, 151.6, 155.5, 158.8, 161.4, 168.4; HRMS: Calcd for
C34H34ClN7O5 [M]+655.2310 and [M+2] 657.2310, found 655.2317 and 657.2305; Anal.
Calcd (%) for: C, 62.24; H, 5.22; N, 14.94, found: C, 62.16; H, 5.39; N, 14.90.
13

ACCEPTED MANUSCRIPT
3.1.24 1-[3-(4-{1-[2-(7-Chloro-quinolin-4-ylamino)-ethyl]-1H-[1,2,3]triazol-4-ylmethoxy}-3-
methoxy-phenyl)-5-(3,4,5-trimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9h): Yield
90%; Light yellow Solid; m.p. 184-185 °C; 1H NMR (DMSO-d6, 400 MHz): δ 2.36 (s, 3H, -
CH3), 2.89 (dd, J = 4.0, 17.6 Hz, 1H, H7), 3.40 (dd, J = 11.6, 17.6 Hz, 1H, H8), 3.74-3.77 (m,
14H, -CH2+4×-OCH3), 4.83-4.85 (m, 2H, -CH2), 5.25 (s, 2H, -OCH2), 5.58 (dd, J = 4.0, 11.6
Hz, H9), 6.44 (s, 2H, ArH), 6.54 (d, J = 5.6 Hz, 1H, H2), 7.00 (d, J = 8.0 Hz, 1H, ArH), 7.44
(d, J = 9.2 Hz, 1H, H4), 7.46-7.48 (m, 2H, ArH), 7.89 (s, 1H, H5), 7.96 (d, J = 9.2 Hz, 1H,
13
H3), 8.01 (s, 1H, H6), 8.25 (d, J = 5.6 Hz, 1H, H1); C NMR (DMSO-d6, 125 MHz): 21.6,
41.1, 42.4, 48.8, 55.3, 55.5, 56.0, 60.7, 62.7, 79.6, 98.8, 106.2, 111.0, 112.4, 116.4, 120.5,
122.9, 124.2, 125.6, 126.4, 126.8, 128.0, 134.7, 139.7, 142.6, 146.8, 150.3, 151.4, 154.5,
156.3, 159.2, 161.6, 168.3; HRMS: Calcd for C35H36ClN7O6 [M]+685.2416 and [M+2]
687.2416, found 685.2411 and 687.2408; Anal. Calcd (%) for: C, 61.27; H, 5.29; N, 13.99,
found: C, 61.39; H, 5.21; N, 13.86.
3.1.25 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-phenyl-4,5-dihydro-pyrazol-1-yl]-ethanone (9i): Yield 79%; Light yellow Solid;
m.p. 204-205 °C; 1H NMR (DMSO-d6, 500 MHz): δ 2.27-2.30 (m, 2H, -CH2), 2.40 (s, 3H, -
CH3), 2.95 (dd, J = 4.0, 17.5 Hz, 1H, H7), 3.51 (dd, J = 11.5, 17.5 Hz, 1H, H8), 3.81-3.83 (m,
2H, -CH2), 4.72-4.74 (m, 2H, -CH2), 5.22 (s, 2H, -OCH2), 5.59 (dd, J = 4.5, 11.5 Hz, 1H, H9),
6.41 (d, J = 5.5 Hz, 1H, H2), 7.13 (d, J = 8.5 Hz, 2H, ArH), 7.37 (d, J = 9.0 Hz, 1H, H4),
7.55-7.59 (m, 5H, ArH), 7.88 (s, 1H, H5), 7.92 (d, J = 9.2 Hz, 1H, H3), 8.11 (d, J = 8.5 Hz,
13
2H, ArH), 8.23(s, 1H, H6), 8.40 (d, J = 5.5 Hz, 1H, H1); C NMR (DMSO-d6, 125MHz):
22.0, 28.6, 40.4, 42.7, 48.3, 62.5, 78.9, 98.9, 108.2, 115.4, 117.7, 121.4, 124.2,125.1, 125.9,
126.3, 126.7, 128.5, 129.0, 134.8, 139.0, 142.1, 147.6, 150.3, 151.7, 154.8,160.4, 168.5;
HRMS: Calcd for C32H30ClN7O2 [M]+579.2149 and [M+2] 581.2149, found 579.2155 and
581.2141; Anal. Calcd (%) for: C, 66.26; H, 5.21; N, 16.90, found: C, 66.41; H, 5.26; N,
17.01.
3.1.26 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-(4-methoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9j):Yield 93%; Light
1
Yellow Solid; m.p. 177-178 °C; H NMR (CDCl3, 500 MHz): δ 2.26-2.28 (m, 2H, -CH2),
2.40 (s, 3H, -CH3), 2.91 (dd, J = 4.0, 17.5 Hz, 1H, H7), 3.48 (dd, J = 11.5, 17.5 Hz, 1H, H8),
3.71 (s, 3H, -OCH3), 3.76-3.79 (m, 2H, -CH2), 4.78-4.80 (m, 2H, -CH2), 5.19 (s, 2H, -OCH2),
5.56 (dd, J = 4.5, 11.5 Hz, 1H, H9), 6.45 (d, J = 5.5 Hz, 1H, H2), 7.05 (d, J = 8.5 Hz, 2H,
ArH), 7.31 (d, J = 9.0 Hz, 1H, H4), 7.41 (d, J = 8.5 Hz, 2H, ArH), 7.73 (d, J = 8.5 Hz, 2H,
14

ACCEPTED MANUSCRIPT
ArH), 7.92 (s, 1H, H5), 8.00 (s, 1H, -NH- exchangeable with D2O), 8.09 (d, J = 9.0 Hz, 1H,
13
H3), 8.19 (d, J = 8.5 Hz, 2H, ArH), 8.30 (s, 1H, H6), 8.41 (d, J = 5.5 Hz, 1H, H1); C NMR
(DMSO-d6, 125MHz): 21.5, 28.2, 40.0, 41.6, 48.6, 55.4, 61.9, 78.7, 99.0, 108.5, 115.4, 117.7,
121.0, 123.8, 124.2, 125.4, 126.3, 126.8, 128.5, 134.1, 139.0, 142.9, 147.6, 150.1,
151.5,154.3, 157.0, 160.2, 168.7; HRMS: Calcd for C33H32ClN7O3 [M]+609.2255 and [M+2]
611.2255, found 609.2247 and 611.2252; Anal. Calcd (%) for: C, 64.96; H, 5.29; N, 16.07,
found: C, 65.07; H, 5.21; N, 16.20.
3.1.27 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-(3,4-dimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9k): Yield 76%;
Light yellow Solid; m.p. 210-211 °C; 1H NMR (DMSO-d6, 500 MHz): δ 2.29-2.32 (m, 2H, -
CH2), 2.44 (s, 3H, -CH3), 2.94 (dd, J = 4.0, 17.5 Hz, 1H, H7), 3.33-3.35 (m, 2H, -CH2), 3.59
(dd, J = 11.5, 18.0 Hz, 1H, H8), 3.70 (s, 6H, 2×-OCH3), 4.47-4.50 (m, 2H, -CH2), 5.17 (s, 2H,
-O-CH2), 5.69 (dd, J = 4.0, 11.5 Hz, H9), 6.30-6.32 (m, 2H, ArH), 6.35 (d, J= 2.0 Hz, 1H,
ArH), 6.63 (d, J = 4.5 Hz, 1H, H2), 6.86 (d, J = 8.5 Hz, 1H, H4), 6.96 (d, J = 8.5 Hz, 2H,
ArH), 7.63 (d, J = 8.5 Hz, 2H, ArH), 7.73 (s, 1H, H5), 7.86 (s, 2H, H6), 7.91 (d, J = 8.5 Hz,
13
1H, H3), 8.38 (d, J = 4.5 Hz, 1H, H1); C NMR (DMSO-d6, 125 MHz): 22.0, 28.3, 40.0,
41.6, 48.0, 55.3, 55.4, 61.8, 65.8, 98.8, 104.0, 114.8, 117.1, 121.4, 122.5, 123.5, 124.7, 125.5,
126.2, 126.8, 128.2, 135.5, 139.2, 143.7, 147.4, 150.2, 150.6, 154.8, 156.9, 159.7, 160.2,
168.7; HRMS: Calcd for C34H34ClN7O4 [M]+639.2361 and [M+2] 641.2361, found 639.2370
and 641.2375; Anal. Calcd (%) for: C, 63.79; H, 5.35; N, 15.32, found: C, 63.87; H, 5.46; N,
15.25.
3.1.28 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
phenyl)-5-(3,4,5-trimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9l): Yield 86%;
1
Light yellow Solid; m.p. 187-188 °C; H NMR (CDCl3, 500 MHz): δ 2.24-2.27 (m, 2H, -
CH2), 2.42 (s, 3H, -CH3), 2.99 (dd, J = 4.0, 17.5 Hz, 1H, H7), 3.47 (dd, J = 11.5, 18.0 Hz, 1H,
H8), 3.72 (s, 3H, -OCH3), 3.78 (s, 6H, 2×-OCH3), 3.86-3.89 (m, 2H, -CH2), 4.79-4.81 (m, 2H,
-CH2), 5.18 (s, 2H, -OCH2), 5.66 (dd, J = 4.0, 11.5 Hz, H9), 6.38 (s, 2H, ArH), 6.48 (d, J =
6.0 Hz, 1H, H2), 7.05 (d, J = 9.0 Hz, 2H, ArH), 7.30 (dd, J = 2.0, 8.5 Hz, 1H, H4), 7.74 (s,
1H, H5), 7.90 (d, J = 8.5 Hz, 2H, H3), 8.11 (d, J = 9.0 Hz, 2H, ArH), 8.22 (s, 1H, H6), 8.32 (d,
J = 5.5 Hz, 1H, H1); 13C NMR (DMSO-d6, 125 MHz): 21.6, 28.0, 40.3, 42.6, 48.1, 56.1, 56.4,
61.4, 62.2, 79.0, 98.9, 107.1, 115.5, 117.4, 121.3, 124.4, 125.3, 125.7, 126.2, 135.0, 139.4,
143.6, 146.9, 150.0, 151.6, 152.8, 154.5, 157.6, 159.4, 160.3, 167.7; HRMS: Calcd for
15

ACCEPTED MANUSCRIPT
C35H36ClN7O5 [M]+669.2466 and [M+2] 671.2466, found 669.2472 and 671.2453; Anal.
Calcd (%) for: C, 62.73; H, 5.41; N, 14.63, found: C, 62.68; H, 5.47; N, 14.51.
3.1.29 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
3-methoxy-phenyl)-5-phenyl-4,5-dihydro-pyrazol-1-yl]-ethanone (9m): Yield 79%; Light
yellow Solid; m.p. 201-202 °C; 1H NMR (DMSO-d6, 400 MHz): δ 2.38-2.41 (m, 2H, -CH2),
2.44 (s, 3H, -CH3), 3.00 (dd, J = 4.0, 17.6 Hz, 1H, H7), 3.47 (dd, J = 11.6, 17.6 Hz, 1H, H8),
3.72 (s, 3H, -OCH3), 3.83-3.85 (m, 2H, -CH2), 4.78-4.81 (m, 2H, -CH2), 5.20 (s, 2H, -OCH2),
5.55 (dd, J = 4.0, 11.6 Hz, H9), 6.59 (d, J = 5.6 Hz, 1H, H2), 6.89-6.93 (m, 2H, ArH), 7.06 (d,
J = 8.0 Hz, 1H, ArH), 7.41 (d, J = 9.2 Hz, 1H, H4), 7.52-7.56 (m, 5H, ArH), 7.90 (s, 1H, H5),
7.96 (d, J = 9.2 Hz, 1H, H3), 8.11 (s, 1H, H6), 8.29 (d, J = 5.6 Hz, 1H, H1); 13C NMR
(DMSO-d6, 125 MHz): 21.6, 28.5, 40.1, 42.0, 48.4, 55.3, 62.0, 79.5, 99.2, 105.6, 111.3,
112.5, 117.2, 120.7, 123.1, 124.0, 125.4, 125.7, 126.1, 126.5, 128.8, 135.7, 140.2, 142.9,
147.4, 149.6, 150.3, 151.3, 154.5, 160.9, 167.6; HRMS: Calcd for C33H32ClN7O3
[M]+609.2255 and [M+2] 611.2255, found 609.2264 and 611.2249; Anal. Calcd (%) for: C,
64.96; H, 5.29; N, 16.07, found: C, 64.90; H, 5.21; N, 16.17.
3.1.30 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
3-methoxy-phenyl)-5-(4-methoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9n): Yield
1
92%; Light Yellow Solid; m.p. 195-196 °C; H NMR (CDCl3, 400 MHz): δ 2.21-2.24 (m,
2H, -CH2-), 2.32 (s, 3H, -CH3), 2.88 (dd, J = 4.0, 17.6 Hz, 1H, H7), 3.39 (dd, J = 11.6, 18.0
Hz, 1H, H8), 3.70 (s, 3H, -OCH3), 3.75 (s, 3H, -OCH3), 3.81-3.84 (m, 2H, -CH2), 4.72-4.75
(m, 2H, -CH2), 5.15 (s, 2H, -OCH2), 5.44 (dd, J = 4.0, 11.6 Hz, H9), 6.63 (d, J = 5.6 Hz, 1H,
H2), 7.04-7.07 (m, 3H, ArH), 7.35 (d, J = 9.2 Hz, 1H, H4), 7.52-7.57 (m, 4H, ArH), 7.80 (s,
1H, H5), 8.15 (d, J = 9.2 Hz, 1H, H3), 8.08 (s, 1H, H6), 8.34 (d, J = 5.6 Hz, 1H, H1); 13C NMR
(DMSO-d6, 125 MHz): 21.6, 27.8, 40.0, 41.9, 48.3, 55.1, 55.4, 62.5, 79.1, 98.6, 106.2, 111.2,
112.4, 116.4, 120.2, 123.5, 124.1, 125.3, 126.4, 126.7, 128.0, 135.5, 139.9, 143.2, 147.4,
149.4, 150.0, 151.6, 152.5, 157.7, 160.4, 167.7; HRMS: Calcd for C34H34ClN7O4
[M]+639.2361 and [M+2] 641.2361, found 639.2367 and 641.2373; Anal. Calcd (%) for: C,
63.79; H, 5.35; N, 15.32, found: C, 63.92; H, 5.45; N, 15.27.
3.1.31 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
3-methoxy-phenyl)-5-(3,4-dimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone (9o): Yield
89%; Light yellow Solid; m.p. 192-193 °C; 1H NMR (DMSO-d6, 400 MHz): δ 2.26-2.29 (m,
2H, -CH2), 2.45 (s, 3H, -CH3), 2.99 (dd, J = 4.0, 17.2 Hz, 1H, H7), 3.40 (dd, J = 11.6, 17.2
16

ACCEPTED MANUSCRIPT
Hz, 1H, H8), 3.72-3.75 (m, 11H, 3×-OCH3+-OCH2), 4.77-4.81 (m, 2H, -CH2), 5.17 (s, 2H, -
OCH2), 5.52 (dd, J = 4.0, 11.6 Hz, H9), 6.49 (d, J = 8.0 Hz, 1H, ArH), 6.57 (d, J = 5.6 Hz,
1H, H2), 7.01 (d, J = 8.4 Hz, 1H, ArH), 7.26 (d, J = 8.0 Hz, 1H, ArH), 7.33 (d, J = 9.0 Hz,
1H, H4), 7.52-7.56 (m, 3H, ArH), 7.87 (s, 1H, H5), 8.06 (d, J = 9.2 Hz, 1H, H3), 8.22 (s, 1H,
13
H6), 8.36 (d, J = 5.6 Hz, 1H, H1); C NMR (DMSO-d6, 125 MHz): 21.7, 27.8, 39.5, 42.1,
47.9, 55.2, 55.4, 55.7, 62.1, 79.2, 99.1, 104.6, 111.2, 111.9, 112.5, 118.9, 122.4, 123.2, 124.0,
125.1, 125.4, 125.4, 128.3, 135.0, 139.2, 143.1, 146.6, 149.3, 150.7, 151.4, 152.1, 156.4,
159.2, 162.3, 167.0; HRMS: Calcd for C35H36ClN7O5 [M]+ 669.2466 and [M+2] 671.2466,
found 669.2460 and 671.2479; Anal. Calcd (%) for: C, 62.73; H, 5.41; N, 14.63, found: C,
62.80; H, 5.49; N, 14.87.
3.1.32 1-[3-(4-{1-[3-(7-Chloro-quinolin-4-ylamino)-propyl]-1H-[1,2,3]triazol-4-ylmethoxy}-
3-methoxy-phenyl)-5-(3,4,5-trimethoxy-phenyl)-4,5-dihydro-pyrazol-1-yl]-ethanone
(9p):
1
Yield 86%; Light yellow Solid; m.p. 159-160 °C; H NMR (DMSO-d6, 400 MHz): δ 2.27-
2.30 (m, 2H, -CH2), 2.43 (s, 3H, -CH3), 2.86 (dd, J = 4.0, 18.0 Hz, 1H, H7), 3.36 (dd, J =
11.6, 18.0 Hz, 1H, H8), 3.71-3.76 (m, 14H, 4×-OCH3+ -CH2), 4.65-4.67 (m, 2H, -CH2), 5.19
(s, 2H, -OCH2), 5.48 (dd, J = 4.0, 11.6 Hz, H9), 6.48 (d, J = 5.6 Hz, 1H, H2), 6.53 (s, 2H,
ArH), 7.09 (d, J = 8.0 Hz, 1H, ArH), 7.32 (d, J = 9.2 Hz, 1H, H4), 7.49-7.53 (m, 2H, ArH),
7.93-7.96 (m, 2H, H3+H5), 8.12 (s, 1H, H6), 8.30 (d, J = 5.6 Hz, 1H, H1); 13C NMR (DMSO-
d6, 100 MHz): 22.0, 28.4, 39.4, 42.3, 48.5, 55.0, 55.5, 55.8, 60.4, 61.9, 79.2, 98.8, 105.6,
111.1, 112.2, 116.8, 119.5, 123.5, 124.0, 125.2, 126.1, 126.7, 128.3, 135.2, 140.4, 142.5,
146.7, 149.7, 150.9, 154.6, 156.5, 159.0, 161.2, 167.9; HRMS: Calcd for C36H38ClN7O6
[M]+699.2572 and [M+2] 701.2572, found 699.2561 and 701.2579; Anal. Calcd (%) for: C,
61.75; H, 5.47; N, 14.00, found: C, 61.64; H, 5.33; N, 14.10.
3.2 Biological evaluation:
3.2.1 Methods for assessment of antimalarial activity of test compounds:
The W2 strain of P. falciparum was cultured at 37 °C in human red blood cells at 2%
hematocrit in medium RPMI-1640 supplemented with 0.5% Albumax®, 100 uM
hypoxanthine, 2 µM L-glutamine and 25 µM HEPES pH 7.4 under atmosphere of 3% O2, 5%
CO2 balance nitrogen. The parasites were synchronized with 5% D-sorbitol at ring stage [30]
and incubated with different concentrations of compounds for 48 h. The compounds were
added from DMSO stocks; the maximum concentration of DMSO used was 0.1 %. Controls
without inhibitors included 0.1% DMSO. After 48 h when control cultures had progressed to
new rings, the culture were fixed for 48 h by adding equal volume of 2% formaldehyde in
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PBS, pH 7.4, at room temperature. Fixed parasites were then transferred to 0.1% Triton X-
100 in PBS containing 100 µM ammonium chloride and 1 nM YOYO-1 dye (Molecular
Probes). Parasitemia was determined from dot plots (forward scatter vs. fluorescence)
acquired on a FACSort flow cytometer using Cell Quest software (Beckton Dickinson). IC50
values for growth inhibition were determined from plots of percent control parasitemia over
inhibitor concentration using the Prism 3.0 program, (GraphPad Software), with data from
duplicate experiments fitted by non linear regression [31].
3.2.2 In vitro analysis of cytotoxicity on HeLa cells
HeLa cells were cultured in 60mm × 15mm tissue culture dishes containing 5 mL of
Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with penicillin and
streptomycin. Compounds were dissolved in DMSO to 100 µM concentrations. Once cell
cultures reached 70% confluency, 5 µL of compound was added to the DMEM in the tissue
culture dish for a final concentration of 100 µM. Cells were incubated for 24 h in a 37 °C
CO2 incubator. After 24 h incubation, the media was removed from the HeLa cells and the
cells were then washed with 5 mL of 1X PBS. The cells were then cleaved off of the bottom
of the plate via 5-minute incubation with 0.5 mL of 0.25% trypsin. Cells were re-suspended
in 1 mL of 1X PBS and transferred to a microcentrifuge tube. 100 µL of trypan blue solution
were added to the re-suspended cells and allowed to incubate at room temperature for
approximately 10 minutes. Viable and dead cells were visualized and counted with a
hemacytometer. IC50 values were determined using GraphPad PRISM.
Acknowledgment:
Financial assistance from SERB (Science and Engineering Research Board) under Start-Up
Research grant Scheme No. YSS/2015/000879/CS (RR) is gratefully acknowledged.
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Captions:
Table 1. Antimalarial activity of synthesized conjugates
Table 2. Cytotoxicity and Selective index of potent conjugates
Figure 1. General structure of lead compound and target hybrid compounds
Scheme 1: Synthesis of (2-Azido-ethyl/propyl)-(7-chloro-quinolin-4-yl)-amine 3
Scheme 2: Synthesis of precursors acetylenic chalcones 6 and their corresponding
pyrazolines 7
Scheme 3: Synthesis of 4-aminoquinoline-chalcone conjugates 8 and 4-aminoquinoline-
pyrazoline conjugates 9
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Table 1: Antimalarial activities of synthesized compounds
Compound
R
R1
n
W2 (CQ-R)a
IC50(nM)±Std. deviation
172.1± 3.4
8a
8b
8c
8d
8e
8f
H
H
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
2
2
2
2
2
2
H
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
114.4± 2.6
H
140.9± 6.6
H
175.3± 28.1
354.8± 0.6
OCH3
OCH3
OCH3
OCH3
H
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
1909.0± 56.6
830.1± 5.1
8g
8h
8i
355.4± 105.9
>10000
8j
H
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
>10000
8k
8l
H
>10000
H
5275.5± 188
6960.0± 425.7
>10000
8m
8n
8o
8p
9a
9b
9c
9d
9e
9f
OCH3
OCH3
OCH3
OCH3
H
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
337.1± 30.8
831.4± 119.6
237.4± 122.5
130.7± 18.3
151.7± 22.9
425.8± 0.9
H
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
H
H
OCH3
OCH3
OCH3
OCH3
H
285.5± 34.4
58.7± 22.7
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
9g
9h
9i
53.7± 25.0
139.0± 9.9
3003.5± 68.6
498.0± 49.2
503.5± 119.1
2424.0± 125.9
77.0± 49.4
9j
H
4-OCH3
3,4-(OCH3)2
3,4,5-(OCH3)3
H
9k
9l
H
H
9m
9n
OCH3
OCH3
4-OCH3
1571.5± 72.8
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9o
OCH3
OCH3
3,4-(OCH3)2
2
2
1684.0± 411.5
1741± 79.2
150± 20.0
9p
3,4,5-(OCH3)3
CQ
ART
32.1± 17.0
aCQ-R: Chloroquine-resistant strain
Table 2: Cytotoxicity and Selectivity index of potent conjugates
Compound
Cytotoxicitya
(µM)
35.6
P. falciparum (CQ-R)b
SIc
IC50 (nM)
114.4
140.9
130.7
58.7
8b
311
90
223
558
795
598
8c
9b
9f
12.7
29.2
32.8
9g
42.7
53.7
9m
46.1
77.0
Doxorubicin
8.30
a Cytotoxicity against HeLa cell line
b CQ-R: Chloroquine resistant strain
c SI: Selective index is ratio of IC50 value of HeLa cell line to that of W2 resistant strain
Figure 1. General structure of target hybrid compounds
O
CH3
N
N3
O
HN
N
N
n
R
O
R
O
R1
A
B
R1
A
B
Cl
Molecular
Hybridization
O
R
R
O
N
N
N
N
N
CH3
N
O
A
O
N
N
A
HN
N
n
R1
HN
N
B
n
B
R1
Cl
Cl
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Scheme 1: Synthesis of (2-Azido-ethyl/propyl)-(7-chloro-quinolin-4-yl)-amine 3
N3
OH
Cl
HN
HN
n
n
(a)
(b)
(c)
Cl
N
Cl
N
Cl
N
1
3
2
n = 1,2
Reagents and conditions: (a) Ethanol/propanolamine, Et3N, 120 °C, 12 h; (b) MsCl, Et3N, dry CHCl3, 0 °C-rt, 2-
3 h; (c) NaN3, DMF, 120 °C, 12 h
Scheme 2: Synthesis of precursors acetylenic chalcones 6 and their corresponding
pyrazolines 7
O
O
O
O
H
R
O
R1
R
R
O
CH3
CH3
R1
Br
(a)
(b)
HO
6
4
5
(c)
H3C
O
N N
R
O
R1
7
R = H, -OCH3
R1 = H, 4-OCH3, 3,4-(OCH3)2, 3,4,5-(OCH3)3
Reagents and conditions: (a) K2CO3, DMF, rt, 2 h.; (b) KOH, EtOH, 30min.-2 h; (c) Hydrazine hydrate, Acetic
acid, reflux, 2-3 h
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Scheme 3: Synthesis of 4-aminoquinoline-chalcone conjugates 8 and 4-aminoquinoline-
pyrazoline conjugates 9
Reagents and conditions: (a) CuSO4.5H2O, Sodium ascorbate, EtOH:H2O, rt, 7-8 h
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•
Synthesis and antiplasmodial evaluation of 4-aminoquinoline-chalcone/-pyrazoline
conjugates
•
•
Conjugates have comparable antiplasmodial activities with that of Chloroquine
Most potent and non-cytotoxic conjugate exhibited an IC50 of 53.7 nM