Novel quinolinone–pyrazoline hybrids: synthesis and evaluation of antioxidant and lipoxygenase inhibitory activity

Molecular Diversity

https://doi.org/10.1007/s11030-020-10045-x

ORIGINAL ARTICLE

Novel quinolinone–pyrazoline hybrids: synthesis and evaluation

of antioxidant and lipoxygenase inhibitory activity

1

2

Ioanna Kostopoulou · Antonia Diassakou · Eleni Kavetsou · Eftichia Kritsi · Panagiotis Zoumpoulakis ·

3

1

Eleni Pontiki · Dimitra Hadjipavlou‑Litina · Anastasia Detsi

Received: 3 December 2019 / Accepted: 29 January 2020

©

Springer Nature Switzerland AG 2020

Abstract

The present project deals with the investigation of structure–activity relationship of several quinolinone–chalcone and quin-

olinone–pyrazoline hybrids, in an eꢀort to discover promising antioxidant and anti-inꢁammatory agents. In order to accom-

plish this goal, four bioactive hybrid quinolinone–chalcone compounds (8a–8d) were synthesized via an aldol condensation

reaction, which were then chemically modiꢂed, forming ꢂfteen new pyrazoline analogues (9a–9o). All the synthesized

analogues were in vitro evaluated in terms of their antioxidant and soybean lipoxygenase (LOX) inhibitory activity. Among

all the pyrazoline derivatives, compounds 9b and 9m were found to possess the best combined activity, whereas 9b ana-

logue exhibited the most potent LOX inhibitory activity, with IC value 10 μM. The in silico docking results revealed that

5

0

the synthetic pyrazoline analogue 9b showed high AutoDock Vina score (−10.3 kcal/mol), while all the tested derivatives

presented allosteric interactions with the enzyme.

Graphic Abstract

R

3

NH

HN

2

HCl

R

1,

R

2

= OH, OCH

3

OHC

R

1

2

OH

N

O

N

OH

O

R = H, OCH CH

OH

O

3

3,

R

3

R

1

2

R

1

2

OCH

2

Ar, CN, Cl

R

Potent lipoxygenase inhibitors

IC50 (9b) = 10 µM

Aldol condensation

N

Y

R

Chemical modif ication

N

Y

R

N

Y

8a-8d

9a-9o

IC50 (9j) = 15 µM

Y= H, CH

3

Quinolinone-Chalcone Hybrids

Novel Pyrazoline Hybrids

Keywords Quinolinones · Chalcones · Pyrazolines · Lipoxygenase · LOX inhibition · Antioxidant activity

Introduction

Electronic supplementary material The online version of this

article (https://doi.org/10.1007/s11030-020-10045-x) contains

supplementary material, which is available to authorized users.

Inꢁammation is part of the body’s defense mechanism, and it

can be beneꢂcial as a healing process, by which the immune

system recognizes and removes harmful stimuli. Acute

inꢁammation is induced by tissue damage due to trauma,

microbial invasion or noxious compounds. Chronic inꢁam-

mation is a persistent phenomenon that can last from months

to years and plays a central role in some of the most chal-

lenging diseases of our time, including neurodegenerative

and heart diseases, rheumatoid arthritis, diabetes, asthma,

and even cancer.

*

Anastasia Detsi

adetsi@chemeng.ntua.gr

1

Laboratory of Organic Chemistry, School of Chemical

Engineering, National Technical University of Athens,

Zografou Campus, 15780 Athens, Greece

2

3

Institute of Chemical Biology, National Hellenic Research

Foundation, 48, Vas. Constantinou Avenue, 11635 Athens,

Greece

Laboratory of Pharmaceutical Chemistry, School

of Pharmacy, Faculty of Health Sciences, Aristotle

University of Thessaloniki, 54124 Thessaloníki, Greece

Lipoxygenases (LOXs) are a heterogeneous family of

structurally related non-heme iron-containing enzymes that

Vol.:(0

1

123456789)

3

Molecular Diversity

catalyze the insertion of molecular oxygen into polyunsatu-

rated fatty acids, such as arachidonic acid or linoleic acid.

They are classiꢂed as 5-, 8-, 12, and 15- LOXs according to

their selectivity to oxygenate fatty acids in a speciꢂc posi-

tion [1–4].

that have shown potential activity in many biological tests,

and they are widely used in the development of new hybrid

compounds.

Quinolinones are heterocyclic nitrogen compounds which

are mainly found as alkaloids in a variety of natural prod-

ucts, as well as synthetic analogues (Fig. 1) [24, 25]. Fur-

thermore, compounds containing the heterocyclic system

of 4-substituted-2-quinolinone as a building block exhibit

a wide variety of biological and pharmacological proper-

ties and are useful intermediates in the synthesis of a large

number of bioactive molecules [26–28].

Over the years, several strategies have been developed in

order to block the arachidonic acid pathway and diꢀerent

compounds have been identiꢂed as 5-LOX inhibitors [5–8].

Up to now, Zileuton is the only approved potent and selec-

tive 5-LOX inhibitor for the treatment of asthma [9, 10].

However, several 5-LOX inhibitors are now under clinical

investigation for the treatment of cardiovascular diseases and

vascular inꢁammation, atherosclerosis, asthma and for knee

osteoarthritis [11–13]. Furthermore, recent studies examine

the relationship between arachidonic acid cascade and car-

cinogenesis, revealing novel targets for the treatment of can-

cer [14–16]. It has been demonstrated that 5-LOX plays an

important role in regulating cellular proliferation and there

are 5-LOX inhibitor pharmacophores which can also cause

cell death in prostate cancer cells [2].

Chalcones are natural products which have been recog-

nized as the precursors to the biosynthesis of ꢁavonoids and

isoꢁavonoids [29]. They are α,β-unsaturated carbonyl com-

pounds, which are characterized by the presence of a three

carbon bridge with a double bond (Fig. 1) [30]. Chalcones

and their derivatives consist “privileged structures” and they

show many interesting biological properties, such as anti-

inꢁammatory [20, 31, 32], anti-parasitic [33], antimicrobial

[34], antibacterial [32, 35], antioxidant [31, 36], cytotoxic

[37], anticancer [31, 38] and so on.

Chronic inꢁammation and oxidative stress are two com-

monly associated conditions involved in the pathophysiol-

ogy of cancers, diabetes, cardiovascular and pulmonary dis-

eases, and others. Oxidative stress occurs when the balance

between antioxidants and reactive oxygen species (ROS)

is disrupted because of either depletion of antioxidants or

accumulation of ROS.

Following the above trend, the aim of the present research

is the synthesis of bioactive hybrid quinolinone–chalcone

compounds, and the chemical modiꢂcation of the above

system, in order to form new pyrazoline analogues (Fig. 1).

Pyrazolines (Fig. 1) are well-known and important ꢂve-

membered heterocyclic compounds, possessing two adjacent

nitrogen atoms in the ring and only one endocyclic double

bond. [39]. Depending on the position of the double bond,

there are three possible partially reduced forms of pyrazoline

structure, which are namely 1-pyrazoline, 2-pyrazoline or

3-pyrazoline which can exist in equilibrium with each other.

Among them, 2-pyrazoline is more stable than the rest of

the reduced forms, while it seems to be the most frequently

studied form of pyrazoline [40].

The main feature of an antioxidant is its ability to bind

free radicals, which are produced during the inꢁammation

process by phagocytic leukocytes that invade the tissue.

Moreover, ROS are involved in the biosynthesis of prosta-

glandins and in the cyclooxygenase (COX) and LOX medi-

ated conversion of arachidonic acid into proinꢁammatory

intermediates [17–19]. Therefore, the development of novel

drugs that combine anti-inꢁammatory and antioxidant activ-

ity could be beneꢂcial for the treatment of several diseases

Pyrazoline derivatives have been reported to exhibit a

wide range of pharmacological activities such as antimi-

crobial [41–43], anticancer [44, 45], anti-inꢁammatory

[46], antioxidant [47, 48], antidepressant [49] and so on.

Out of all these biological activities of pyrazolines and

their derivatives, the anti-inꢁammatory activity seems to

concentrate the interest of researchers, while there is a

[

20].

Hybrid molecules combine the structural frame of two

or more different pharmacophores which have already

been exploited in drug development, and they are designed

in order to provide novel drugs with enhanced activity

[

21–23]. Quinolinones and chalcones are two scaffolds

Fig. 1 General structure of the

quinolinone (I), chalcone (II)

and pyrazoline (III) molecular

scaꢀolds

OH

O

3

2

N

X

4

A

B

NH

N

Y

O

5

1

(III)

(I)

(II)

1

3

Molecular Diversity

Scheme 1 Synthesis of

3

-acetyl-4-hydroxy-2-quin-

olinone (3). Reagents and con-

ditions: (i) (CH CO) O, 130 °C,

3

2

h (ii) CH COCH COOEt,

3 2

t-BuOK, t-BuOH, rt (iii) aq

Na CO /NaOH, rt

2

3

Scheme 2 Synthesis of

3

2

-acetyl-4-hydroxy-1-methyl-

-quinolinone (6). Reagents and

conditions: (i) toluene, 110 °C,

2

h (ii) NaOCH CH , 77 °C, 2 h

2 3

plethora of information available in the literature which is

growing steadily over the years [50–56].

Table 1 New quinolinone–

Code

Y

R₁

R₂

chalcone hybrids

8

a

b

c

H

^C^H3

^O^C^H3

OH

^O^C^H3

OH

In 2019, Stefanes and coworkers synthesized a series

of new pyrazoline derivatives, which were then evalu-

ated for their anti-leukemic activity, revealing two of the

compounds as promising candidates against acute leuke-

mia [57]. Mumtaz and his group focused on the current

need of new potential agents over the treatment of neu-

rodegenerative disorders, such as Parkinson’s disease or

Alzheimer’s disease, synthesizing several derivatives of

thioureas and pyrazolines, which were then evaluated via

two assays. Results showed one of the pyrazoline deriva-

tives as the most potent acetylcholinesterase inhibitor, with

d

^C^H3

Results and discussion

Chemistry

The desired starting quinolinones diꢀer to the substituent

Y attached to the nitrogen of the heterocyclic ring, and

they were synthesized through diꢀerent synthetic routes.

IC = 123 ± 51 nM [58].

5

0

Furthermore, the evaluation of the anti-inꢁammatory

activity of pyrazolines still stands out in the literature,

since several corresponding studies were published in

3

-Acetyl-4-hydroxy-2-quinolinone (3) was synthe-

sized using our previously developed methodology which

includes C-acylation of ethyl acetoacetate by 2-methyl-

2

019. Cai et al. [59] synthesized a number of novel steroi-

dal derivatives bearing pyrazoline structure, among others,

identifying one of them as the most potent anti-inꢁamma-

3

,1-benzoxazin-4-one (1) in a basic environment, followed

by a cyclization reaction of the C-acylation product 2 in

tory agent, with an IC value of 0.86 μM on NO produc-

5

0

aqueous solution of Na CO and NaOH (Scheme 1) [33,

2

3

tion in LPS induced RAW 264.7 cells. Earlier the same

year, Sethiya and his research group synthesized some new

pyrazoline derivatives, using ultrasonic irradiation, which

demonstrated remarkable anti-inꢁammatory activity [60].

Chandel et al. [61] synthesized coumarin-based pyrazo-

lines and evaluated them in terms of their in vitro and

in vivo anti-inꢁammatory activity. Experimental results

revealed one of the tested compounds as highly active anti-

inꢁammatory agent and suggested it to be used as lead

compound for the development of eꢀective inhibitors.

With regard to the synthesis of pyrazolines, literature

oꢀers a wide variety of methods depending on the reactiv-

ity of molecules and the need of the chemist [62]. How-

ever, the most popular method is the one of Fischer and

Knoevenagel, i.e., the reaction of α,β-unsaturated ketones

with phenyl hydrazine in acetic acid under reꢁuxing con-

ditions [63, 64].

6

5].

In order to investigate the role of the substituent Y, we

synthesized quinolinone 6, which bears a methyl group at

this position. The synthesis was accomplished using our

previous methodology, via an acylation reaction of sec-

ondary amine 4, followed by a cyclization reaction of the

acylation product 5, in basic environment (Scheme 2) [33].

The desired four quinolinone–chalcone hybrids

(

Table 1) were ꢂnally synthesized according to Scheme 3,

using compounds 3 and 6 as starting materials in an aldol

condensation reaction with two diꢀerent benzaldehydes.

It is a methodology that has already been developed in a

previous work of our laboratory, [33].

Proceeding with the synthesis and in order to further

investigate the structure–activity relationship of the ꢂnal

compounds, we attempted a structural modiꢂcation, which

1

3

Molecular Diversity

Scheme 3 Synthesis of

OH

O

CHO

R₂

OH

O

quinolinone-chalcone hybrid

compounds and of new pyra-

zoline analogues. Reagents

and conditions: (i) Piperidine,

6

9

2'

4

12

(i)

5

3

1'

3' ^R1

7

8

11

13

+

2

4'

N

Y

6'

R₁

10 N₁

O

R₂

5'

7

8 °C, 5–24 h (ii) CH COOH/

3

Y

EtOH, reꢁux, 120 °C

8a-8d

3

6

. Y= H

. Y= CH

7

a. R =R = OCH

1 2

3

7b. R = OCH R = OH

3,

1 2

NH₂

HN

HCl

N

(

ii)

R₃

^R3

2

1

OH

N

H

6'

X

2''

4'

5

^R1

5'

3'

3

4

1''

7'

3

''

H_B

B

2

'

^HA

8'

10'

6''

4''

R₂

N₁_'

Y

O

9'

5''

9a-9o

was carried out to the α,β-unsaturated carbonyl system

The ꢂrst one is a fast, simple, cost-eꢀective and widely

of compounds 8, leading to pyrazoline derivatives 9

used method, where the DPPH stable free radical reacts

directly with the antioxidant and is decolorized. The scav-

enging eꢀect of the synthesized compounds on the DPPH

radical was evaluated according to the methods of Hadjipav-

lou et al. [66, 67].

(

Table 2).

Various conditions were investigated in order to synthe-

size the target pyrazoline derivatives. After optimization

of the reaction conditions, we ꢂnally synthesized 15 new

molecules by reꢁuxing the corresponding quinolinone–chal-

cone compound (8) with a variety of hydrazine derivatives

in glacial acetic acid (Scheme 3).

AAPH-induced linoleic acid oxidation has been devel-

oped as a quick and reliable method for measuring the anti-

oxidant activity. The thermal free radical producer (AAPH)

generates free radicals in the solution which cause the oxi-

dation of linoleic acid, and the method is a measure of how

effective antioxidants protect against lipid peroxidation

in vitro. Oxidation of exogenous linoleic acid by AAPH is

followed by UV spectrophotometry in a highly diluted sam-

ple [68].

The structures of all the new pyrazolines were elucidated

1

13

1

by H-NMR, C-NMR and HR-MS. In the H-NMR of the

compounds, all the three protons H , H and H attached to

A

B

X

the C and C carbon atoms of the pyrazoline ring, respec-

4

5

tively, were found to give an ABX spin system. The methyl-

ene protons of C resonated as a pair of doublet of doublet

4

peaks at regions 3.43–3.66 ppm (C -H ) and 4.14–4.33 ppm

Furthermore, for the evaluation of their anti-inꢁamma-

tory activity, all new molecules were tested as inhibitors of

soybean LOX, which is a plant enzyme with satisfactory

homology with the human 5-LOX, and the results can be

considered as an indication of the anti-inꢁammatory activ-

ity of new analogues. The results of this study are presented

in Table 3.

4

A

(

C -H ). The methine proton of C also appeared as a dou-

4 B 5

blet of doublet peak at the region 4.96–5.48 ppm, due to its

vicinal coupling with the two magnetically non-equivalent

protons of the position C of the pyrazoline ring. Among the

4

three signals of H , H and H protons, the most deshielded

A

B

X

one is attributed to the methine of C , due to its close prox-

5

imity to the nitrogen of the pyrazoline ring and to ring B of

the chalcone moiety.

Evaluation of antioxidant and soybean LOX inhibi-

tory activity

Biology

The results of the DPPH radical scavenging activity of the

majority of the tested analogues showed good interaction

with the DPPH radical at 100 μM concentration. In general,

the interaction is altered in relation to the diꢀerent substitu-

ents and more speciꢂcally in relation to the presence or not

The new derivatives were tested for their antioxidant activity

in vitro based on their capacity to scavenge the stable free

radical DPPH, as well as their ability to inhibit lipid peroxi-

dation of linoleic acid induced by AAPH radical.

1

3

Molecular Diversity

Table 2 New pyrazoline

analogues

1

3

Molecular Diversity

Table 2 (continued)

of labile protons, such as phenolic hydroxyl groups, which

react directly with the free radical.

by quinolinone–chalcones 8b and 8d, which diꢀer only in

the substituent at the heterocyclic N, indicate that the most

important structural feature for both activities is the presence

of the phenolic OH. Moreover, substitution of the phenolic

Among the four synthesized quinolinone–chalcones, it

seems that the presence of a phenolic hydroxyl group in

compounds 8b and 8d leads to good DPPH radical scav-

engers (81% and 78% after 20-min incubation at a concen-

tration of 0.1 mM, respectively) and LOX inhibitors (IC

OH with a OCH group (compounds 8a and 8c) leads to

3

complete loss of DPPH and LOX inhibitory activity. On the

other hand, the lipid peroxidation ability of compounds 8b

5

0

5

6.0 μM and 50.0 μM, respectively). The similar values of

and 8d is low, but the replacement of the OH by a OCH

3

DPPH scavenging and LOX inhibition ability presented

group (compounds 8a and 8c) leads to signiꢂcant inhibitors

1

3

Molecular Diversity

Table 3 In vitro biological

evaluation of all the synthesized

analogues

CODE Interaction with the free radical

DPPH (%)

Inibition of lipid peroxidation of

linoleic acid induced by AAPH radical bean lipoxygenase

%) IC (μΜ)

.1 mM

Inhibition of soy-

(

5

0

.1 mM/20 min 0.1 mM/60 min

0

8

9

a

b

c

d

a

b

c

d

e

f

g

h

i

j

k

l

–

81.0

–

84.0

–

100.0

23.0

100.0

31.0

74.7

82.9

93.3

83.8

88.2

71.4

25.0

92.3

100.0

77.0

87.2

97.0

79.0

59.0

59.1

–

(19.2% at 0.1 mM)

56.0

(13.0% at 0.1 mM)

50.0

(27.3% at 0.1 mM)

10.0

(43.8% at 0.1 mM)

52.0

63.0

74.0

65.0

67.5

78.0

27.3

85.3

86.2

95.0

76.0

63.0

99.0

60.0

87.0

74.0

77.0

83.0

85.0

67.0

74.5

82.0

12.7

86.2

98.9

95.3

84.0

71.0

44.0

71.0

91.0

8.0

79.0

87.0

100.0

73.0

85.5

97.0

–

(16% at 0.1 mM)

15.0

57.0

57.5

39.0

50.0

90.0

0.5

m

n

o

NDGA 88.0

Trolox

–

88.0

–

of lipid peroxidation (100% inhibition at a concentration of

.1 mM for both compounds) but inactive DPPH scavengers

quinolinone–chalcone 8a: 8a is inactive, whereas 9b and 9c

are among the most active DPPH scavengers (85.3% and

86.2%, respectively, at a concentration of 0.1 mM and after

20 min incubation) of the series. Pyrazoline 9a is not a good

DPPH scavenger; however, it is not completely inactive as

8a. The same trend is followed in the case of the very good

DPPH scavengers pyrazolines 9g and 9i (99% and 87% inhi-

bition, respectively), which are derived from the inactive

quinolinone–chalcone 8c.

0

and LOX inhibitors. From the above observations, it can be

postulated that in the case of quinolinone–chalcones, LOX

inhibition is related to the antioxidant activity as measured

by the DPPH scavenging ability and can probably be cor-

related with their H atom transfer capacity.

An important observation emerging from the evalua-

tion of the antioxidant activity of all the new analogues is

related to the 4-hydroxyl group of the quinolinone moiety.

This proton creates a strong hydrogen bond with the adjacent

carbonyl group, and for this reason it cannot react with the

free DPPH radical, so it does not participate in the enhance-

ment of the antioxidant activity of quinolinone–chalcones.

In the case of the pyrazoline analogues produced from

the good radical scavengers 8b and 8d, namely compounds

9d–9f and 9j–9o, respectively, the contribution of the OH

group at position 4 of the quinolinone heterocyclic ring in

the DPPH scavenging ability does not seem to be as sig-

niꢂcant as the one of the phenolic OH. Thus, the above

pyrazolines were found to possess analogous or even less

antioxidant activity compared to their precursors 8b and 8d.

As far as the ability of the tested pyrazolines to inhibit

lipid peroxidation of linoleic acid induced by a thermal free

radical producer (AAPH), ꢂve pyrazolines, namely 9c, 9e,

9h, 9i and 9m, were found to be potent inhibitors showing

activity equal or higher than the reference compound Trolox.

Insertion of the pyrazoline moiety at quinolinone–chalcone

8b (with very low lipid peroxidation ability, 23% at 0.1 mM)

resulted in compounds 9d–9f which had remarkably higher

activity. The same is true for pyrazolines arising from

1

The stability of this proton is easily proven by the H NMR

analysis, where it appears through a singlet peak at very

low ꢂelds of the spectrum, 17–18 ppm. In the spectra of

pyrazoline analogues this peak is shifted to 13–14 ppm,

indicating that the proton in this case creates a weaker

hydrogen bond with the nitrogen of the pyrazoline ring.

This is a plausible explanation to account for the observa-

tion that the majority of the tested pyrazolines show a better

DPPH radical scavenger capacity, compared to their qui-

nolinone–chalcone precursors. As a striking example, one

can compare the DPPH scavenging ability of pyrazolines 9a,

9

b and 9c, which were prepared from the same precursor,

1

3

Molecular Diversity

quinolinone–chalcone 8d: the parent compound had very

low activity (31% at 0.1 mM), whereas the corresponding

pyrazolines 9j–9o showed moderate to high activity (>59%).

However, pyrazolines 9a–9c and 9g–9i which were derived

from 8a and 8c, respectively, showed analogous or lower

activity with the parent quinolinone–chalcones.

Among all the pyrazoline analogues tested, the most

potent LOX inhibitor is compound 9b (IC =10 μΜ) which

50

possesses a p-methoxy-phenyl substituent attached to the

nitrogen of the pyrazoline moiety. In addition, pyrazoline 9b

exhibits good antioxidant activity in both DPPH and lipid

peroxidation assays; thus, it can be considered as a prom-

ising lead compound for further development. Regarding

structural modiꢂcations on 9b, replacement of the p-meth-

oxy group by a p-cyano (9a) or a p-benzyloxy (9c) group or

insertion of a methyl group on the heterocyclic NH of the

quinolinone ring (compound 9i) resulted in loss of activity.

However, the insertion of a methyl group on the nitrogen

of the heterocyclic ring of the quinolinone moiety seems to

enhance LOX inhibitory activity in the case of pyrazolines

that share the p-cyano-phenyl or the p-benzyloxy-phenyl

substituent as a common structural feature: N–H pyrazolines

Fig. 2 Docking orientation of 9b (depicted in turquoise) bound to

soybean LOX

therefore, it can serve as the starting point for the develop-

ment of potential agents with combined anti-inꢁammatory

and antioxidant activity.

9

a, 9d and 9e (IC =27.3% at 0,1 mM, 52.0 and 63.0 μΜ,

Conclusions

5

0

respectively) are less potent than their N-CH analogues 9g,

3

9

j and 9m (IC =65.0, 15.0 and 39.0 μΜ, respectively).

In conclusion, this work reports the synthesis of four qui-

nolinone–chalcones and ꢂfteen novel structurally modiꢂed

pyrazoline analogues, which to our knowledge have not

been reported in the literature. In order to classify the new

molecules as promising antioxidant and anti-inꢁammatory

agents, we studied how the ꢂnal biological eꢀect is aꢀected

in relation to the diꢀerent substituents at several positions

of the framework, like the nitrogen heteroatom of the qui-

nolinone ring, the aromatic ring B of the chalcone moiety,

as well as the phenyl group attached to the pyrazoline ring.

The evaluation of all the synthesized compounds showed

that pyrazolines 9b and 9m possess the best combined activ-

ity (IC 10.0 and 39.0 μΜ for LOX inhibition, 86% and

5

0

The comparison of pyrazoline analogues possessing two

methoxy groups at the aromatic ring B (compounds 9a, 9c,

9

g, 9h, 9i) with their analogues which bear one methoxy and

one hydroxyl group at ring B (compounds 9d, 9e, 9j, 9k, 9l)

shows that as a general trend, the presence of the hydroxyl

group dramatically increases the ability of the compounds to

inhibit LOX. In fact, compound 9j is the second best lipoxy-

genase inhibitor in this series.

Molecular modeling of the synthesized derivatives

in soybean LOX

5

0

All the synthesized derivatives have been subjected to in sil-

ico docking. The docking results provided useful interpreta-

tion of the experimental study. Figure 2 depicts the preferred

docking pose for the most potent derivative 9b in soybean

LOX (PDB code: 3PZW) presenting a high AutoDock Vina

score (− 10.3 kcal/mol). From the docking results, it can

be concluded that the novel synthesized derivatives pre-

sent allosteric interactions with the enzyme. It seems that

100% for DPPH scavenging ability and 83% and 79% in lipid

peroxidation inhibition, respectively). The in silico dock-

ing results revealed that the pyrazoline analogue 9b showed

high AutoDock Vina score (−10.3 kcal/mol), while all the

tested derivatives presented allosteric interactions with the

enzyme.

9

b accommodates to an extensively hydrophobic cavity

Experimental

Chemistry

with possible hydrophobic interactions (π–π stacking). It is

possible that 9b extends into the hydrophobic domain and

prevents access of substrates to the active site and hence pre-

vents lipoxygenation. It is well known that LOX inhibitory

activity is based on a carbon-centered radical on a lipid chain

and most LOX inhibitors are antioxidants or free radical

scavengers [69]. Pyrazoline 9b is also a potent antioxidant;

General methods

The reagents and solvents used for synthesis and analy-

sis were commercially available and used without further

1

3

Molecular Diversity

purification. NMR spectra were recorded on a Varian

COCH = CH-Ar), 7.00 (d, 1H, J = 8.4 Hz, Ar–H), 7.93 (d,

1H, J = 15.6 Hz, COCH = CH-Ar), 7.67 (t, 1H, J = 7.2 Hz,

Ar–H), 7.29 (m, 3H, Ar–H), 7.22 (t, 1H, J=7.8 Hz, Ar–H),

6

00 MHz spectrometer at the National Hellenic Research

Foundation. The HR-MS spectrum was obtained using a

UHPLC-MSn Orbitrap Velos-Thermo mass spectrometer at

the National Hellenic Research Foundation. Melting points

were determined on a Gallenkamp MFB-595 melting point

apparatus and are uncorrected.

6.90 (d, 1H, J = 7.8 Hz, Ar–H), 3.85 (s, 3H, 2OCH );

3

(E)‑3‑(3‑(3,4‑dimethoxyphenyl)acryloyl)‑4‑hydroxy‑1‑meth‑

ylquinolin‑2(1H)‑one (8c)

Synthesis

This compound was prepared following the general pro-

General procedure (A) for the synthesis of quinolinone–

chalcone analogues (8a–8d)

cedure A. 3-acetyl-4-hydroxy-2-(1-CH )-quinolinone (6)

3

(500 mg, 2.30 mmol) and 3,4-dimethoxy benzaldehyde

(

7a) (382.2 mg, 2.30 mmol) were dissolved in 15 ml EtOH,

Equimolar amounts of 3-acetyl-4-hydroxy-2-(1H)-quin-

olinone (3, 6) and the appropriate benzaldehyde (7) are dis-

solved in absolute ethanol (EtOH), and a catalytic amount

of piperidine is added. The reaction mixture is stirred for

and 18 drops of piperidine were added. The mixture was

stirred overnight at 78 °C. The solid produced upon acidi-

ꢂcation was triturated from methanol/dichloromethane to

aꢀord chalcone 8c as orange powder. Yield: 357 mg (71%);

1

5

–24 h at 78 °C with a reꢁux condenser, while the reaction

m.p. > 250 °C H NMR (CDCl , 600 MHz): δ ppm 18.20

3

is monitored by thin-layer chromatography (TLC). After the

reaction is complete, the mixture is cooled in an ice bath

and acidiꢂed with aq. HCl 10% v/v. Finally, the product is

obtained from the acidiꢂed aqueous solution in solid form.

(s, 1H, OH), 8.58 (d, 1H, J = 15.6 Hz, COCH = CH-Ar),

8.25 (d, 1H, J = 7.8 Hz, Ar–H), 7.96 (d, 1H, J = 15.6 Hz,

COCH = CH-Ar), 7.68 (t, 1H, J = 7.8 Hz, Ar–H), 7.32 (m,

2H, Ar–H), 7.26 (m, 2H, Ar–H), 6.89 (d, 1H, J = 8.4 Hz,

Ar–H), 3.97 (s, 3H, 2OCH ), 3.93 (s, 3H, OCH ), 3.67 (s,

3

(E)‑3‑(3‑(3,4‑dimethoxyphenyl)acryloyl)‑4‑hydroxyquino‑

3H, NCH );

3

lin‑2(1H)‑one (8a)

This compound was prepared following the general pro-

(E)‑4‑hydroxy‑3‑(3‑(4‑hydroxy‑3‑methoxyphenyl)

acryloyl)‑1‑methylquinolin‑2(1H)‑one (8d)

cedure A. 3-acetyl-4-hydroxy-2-(1H)-quinolinone (3)

(

1000 mg, 4,92 mmol) and 3,4-dimethoxy benzaldehyde

7a) (810 mg, 4.92 mmol) were dissolved in 20 ml EtOH

This compound was prepared following the general pro-

and 40 drops of piperidine were added. The mixture was

cedure A. 3-acetyl-4-hydroxy-2-(1-CH )-quinolinone (6)

3

stirred at 78 °C for 5 h. The solid (chalcone 8a) was obtained

(600 mg, 2.76 mmol) and 4-hydroxy-3-methoxy benzalde-

hyde (7b) (419.9 mg, 2.76 mmol) were dissolved in 18 ml

EtOH, and 22 drops of piperidine were added. The mixture

was stirred overnight at 78 °C. The solid produced upon

acidiꢂcation was triturated from methanol/dichlorometh-

upon acidiꢂcation as yellow powder. Yield: 680 mg (68%);

1

m.p.>250 °C; H NMR (DMSO-d , 600 MHz): δ ppm 18.32

6

(

s, 1H, OH), 11.44 (s, 1H, NH), 8.54 (d, 1H, J = 15 Hz,

COCH = CHAr), 8.01 (d, 1H, J = 4.2 Hz, Ar–H), 7.94 (d,

H, J = 15.6 Hz, COCH = CHAr), 7.67 (t, 1H, J = 6.0 Hz,

Ar–H), 7.37 (d, 1H, J=8.4 Hz, Ar–H), 7.27 (m, 3H, Ar–H),

.08 (d, 1H, J=8.4 Hz, Ar–H), 3.83 (s, 6H, 2OCH ).

1

ane to aꢀord chalcone 8d as orange powder. Yield: 286 mg

1

(48%); m.p. > 250 °C; H NMR (DMSO-d , 600 MHz): δ

6

7

ppm 18.25 (s, 1H, OH), 9.90 (s, 1H, Ar-OH), 8.44 (d,

3

1

H, J=15.6 Hz, COCH=CH-Ar), 8.09 (d, 1H, J=7.8 Hz,

(E)‑4‑hydroxy‑3‑(3‑(4‑hydroxy‑3‑methoxyphenyl)acryloyl)

Ar–H), 7.89 (d, 1H, J = 15.6 Hz, COCH = CH-Ar), 7.77

(t, 1H, J = 7.8 Hz, Ar–H), 7.50 (d, 1H, J = 8.4 Hz, Ar–H),

quinolin‑2(1H)‑one (8b)

7

.27 (m, 3H, Ar–H), 6.88 (d, 1H, J = 7.8 Hz, Ar–H), 3.85

1

3

This compound was prepared following the general pro-

(s, 3H, OCH ), 3.55 (s, 3H, NCH ); C NMR (DMSO-

3 3

cedure A. 3-acetyl-4-hydroxy-2-(1H)-quinolinone (3)

d , 600 MHz): δ ppm 193.0 (C, C-11), 175.7 (C, C-4),

6

(

500 mg, 2.46 mmol) and 4-hydroxy-3-methoxy benza-

160.7 (C, C-2), 150.4 (C, C-3′, C-4′), 148.0 (CH, C-12),

146.1 (C, C-5), 141.4 (C, C-1′), 135.4 (CH, C-8), 126.3

(CH, C-13), 125.3 (CH, C-2′), 123.7 (CH, C-5′), 122.1

(CH, C-6′), 121.1 (CH, C-6), 116.0 (CH, C-7), 115.2 (CH,

ldehyde (7b) (374.3 mg, 2.46 mmol) were dissolved in

0 ml EtOH, and 20 drops of piperidine were added. The

1

mixture was stirred at 78 °C for 5 h. The solid (chalcone

8

b) was obtained upon acidiꢂcation as orange powder.

C-9), 112.3 (C, C-10), 104.9 (C, C-3), 55.7 (CH , Ar–O-

3

1

Yield: 290 mg (58%); m.p. > 250 °C; H NMR (DMSO-

CH ), 29.1 (CH , N-CH ); HR-MS m/z (neg): 350.10314

3

d , 600 MHz): δ ppm 18.46 (s, 1H, OH), 11.42 (s, 1H,

C H NO (calcd. 351.1107).

6

20 17 5

NH), 9.90 (s, 1H, Ar-OH), 8.51 (d, 1H, J = 15.6 Hz,

1

3

Molecular Diversity

General procedure (B) for the synthesis of pyrazoline ana‑

logues (9a–9o)

ethanol. Quinolinone-chalcone 8a (120 mg, 0.34 mmol) was

dissolved in 1.9 ml of acetic acid, and it was added dropwise

to the ethanolic solution of hydrazine. The resulting solu-

tion was stirred at 120 °C for 24 h. The solid produced upon

ꢂltration was triturated from methanol/dichloromethane to

The appropriate hydrazine derivative (2.5 eq.) is dissolved

in absolute ethanol. The appropriate quinolinone–chalcone

8

(1 eq.) is dissolved in acetic acid, and it is added dropwise

aꢀord the pure pyrazoline 9b as yellow powder. Yield: 82 mg

1

to the ethanolic solution of hydrazine. The resulting mixture

is stirred at 120 °C for 24 h. The reaction is monitored by

TLC. After completion of the reaction, the mixture is poured

in ice and a yellow or orange precipitate is formed. The solid

is ꢂltered and washed with ice water. If no solid precipitates,

the mixture is extracted with dichloromethane, the organic

extracts are dried with Na SO and the solvent is evaporated

(68.3%); m.p. 230 °C; H NMR (DMSO-d , 600 MHz): δ

6

ppm 13.54 (s, 1H, OH), 11.54 (s, 1H, NH), 7.96 (d, 1H,

J=7.8 Hz, Ar–H), 7.55 (t, 1H, J=7.8 Hz, Ar–H), 7.29 (d,

1H, J=8.7 Hz, Ar–H), 7.23 (t, 1H, J=7.8 Hz, Ar–H), 6.99

(s, 1H, Ar–H), 6.86 (m, 6H, Ar–H), 5.16 (dd, 1H, J=11.4,

9.0 Hz, C -H ), 4.19 (dd,1H, J=18.6, 12.0 Hz, C -H ), 3.72

5

X

4

B

(br, 6H, 2OCH ), 3.67 (s, 3H, OCH ), 3.44 (dd, 1H, J=18.0,

2

4

3

1

3

under reduced pressure to give the pyrazoline analogue as

a solid product. Final compounds are further puriꢂed by

trituration with methanol and dichloromethane.

8.4 Hz, C -H ); C NMR (DMSO-d , 600 MHz): δ ppm

4 A 6

162.4 (C, C-4′), 161.3 (C, C-3), 153.4 (C, C-2′), 150.6 (C,

C-4′″), 149.1 (C, C-3″), 148.1 (C, C-4″), 138.4 (C, C-10′),

1

38.2 (C, C-1′″), 134.3 (CH, C-8′), 123.3 (C, C-1″), 123.1

4

‑(5‑(3,4‑Dimethoxyphenyl)‑3‑(4‑hydroxy‑2‑oxo‑1,2‑dihy‑

droquinolin‑3‑yl)‑4,5‑dihydro‑1H‑pyrazol‑1‑yl)benzonitrile

9a)

(CH, C-6′), 122.1 (C, C-7′), 121.6 (CH, C-6″), 118.2 (CH,

C-3′″, C-5′″), 115.4 (CH, C-9′), 115.2 (CH, C-5″), 114.8

(CH, C-2′″, C-6′″), 114.4 (C, C-5′), 112.2 (CH, C-2″),

(

1

01.4 (C, C-3′), 63.2 (CH, C-5), 55.6 (CH , Ar–O-CH ),

3 3

This compound was prepared following the general proce-

dure B. 4-cyano phenyl-hydrazine hydrochloride (89.1 mg,

55.4 (CH , Ar–O-CH ), 55.1 (CH , Ar–O-CH ), 46.6 (CH ,

3 3 3 3 2

C-4); HR-MS m/z (pos): 471.17808 C H N O (calcd.

2

7

25

3

5

0

.53 mmol) was dissolved in 0.67 ml of absolute ethanol.

471.1794).

Quinolinone-chalcone 8a (73 mg, 0.21 mmol) was dissolved

in 1.3 ml of acetic acid and it was added dropwise to the

ethanolic solution hydrazine. The resulting solution was

stirred at 120 °C for 24 h. The solid produced upon ꢂltra-

tion was triturated from methanol/dichloromethane to aꢀord

3‑(1‑(4‑(Benzyloxy)phenyl)‑5‑(3,4‑dimethoxyphenyl)‑4,

5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxyquinolin‑2(1H)‑one

(9c)

the pure pyrazoline 9a as yellow powder. Yield: 35.8 mg

This compound was prepared following the general pro-

cedure B. 4-benzyloxy phenyl hydrazine hydrochloride

(214.1 mg, 0.85 mmol) was dissolved in 0.9 ml of absolute

ethanol. Quinolinone-chalcone 8a (120 mg, 0.34 mmol) was

dissolved in 1.9 ml of acetic acid, and it was added drop-

wise to the ethanolic solution of hydrazine. The resulting

solution was stirred at 120 °C for 24 h. The solid produced

upon ꢂltration was triturated from methanol/dichlorometh-

1

(

49%); m.p. >250 °C; H NMR (DMSO-d , 600 MHz,): δ

6

ppm 13.04 (s, 1H, OH), 11.61 (s, 1H, NH), 7.98 (br, 1H,

Ar–H), 7.62 (m, 3H, Ar–H), 7.27 (m, 2H, Ar–H), 6.94 (m,

4

H, Ar–H), 6.71 (br, 1H, Ar–H), 5.48 (br, 1H, C -H ), 4.21

5 X

(

br, 1H, C -H ), 3.72 (br, 6H, 2OCH ), 3.59 (br, 1H, C -H );

4 B 3 4 A

1

3

C NMR (DMSO-d , 600 MHz): δ ppm 163.7 (C, C-4′),

6

1

61.7 (C, C-3), 154.2 (C, C-2′), 149.7 (C, C-3″), 148.7 (C,

C-4″), 146.1 (C, C-1′″), 139.0 (C, C-10′), 133.9 (CH, C-3′″,

C-5′″), 133.8 (CH, C-8′), 132.8 (C, C-1″), 123.9 (CH, C-6′),

ane to afford the pure pyrazoline 9c as yellow powder.

1

Yield: 73.2 mg (61%); m.p. 245 °C; H NMR (DMSO-d ,

6

1

22.4 (CH, C-7′), 120.2 (C, C-6″), 117.7 (C, Ar–C ≡ N),

15.7 (CH, C-9′), 114.3 (C, C-5′) 113.1 (CH, C-2′″, C-6′″),

12.6 (CH, C-5″), 110.0 (C, C-2″), 101.5 (CH, C-4′″), 100.0

600 MHz): δ ppm 13.53 (s, 1H, OH), 11.55 (s, 1H, NH),

7.96 (d, 1H, J = 7.2 Hz, Ar–H), 7.55 (t, 1H, J = 7.2 Hz,

Ar–H), 7.38 (m, 4H, Ar–H), 7.30 (m, 2H, Ar–H), 7.23 (t,

1H, J = 7.2 Hz, Ar–H), 6.98 (s, 1H, Ar–H), 6.90 (m, 5H,

Ar–H), 6.81 (d, 1H, J=7.2 Hz, Ar–H), 5.17 (br, 1H, C -H ),

(

C, C-3′), 60.9 (CH, C-5), 56.0 (CH , Ar–O-CH , Ar–O-

3 3

CH ), 55.9 (CH , Ar–O-CH , Ar–O-CH ), 47.2 (CH ,

3

2

5

X

C-4); HR-MS m/z (neg): 465.15554 C H N O (calcd.

5.0 (s, 2H, Ar-CH -O-Ar), 4.19 (dd, 1H, J=18.6, 12.0 Hz,

2

7

22

4

2

4

66.1641).

C -H ), 3.72 (s, 6H, OCH ), 3.44 (dd, 1H, J=18.0, 8.4 Hz,

4

B

3

1

3

C -H ); C NMR (DMSO-d , 600 MHz): δ ppm 162.4 (C,

4

A

6

3‑(5‑(3,4‑Dimethoxyphenyl)‑1‑(4‑methoxyphenyl)‑4,5‑di‑

C-4′), 161.3 (C, C-3), 152.4 (C, C-2′), 150.6 (C, C-4′″),

hydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxyquinolin‑2(1H)‑one (9b)

149.1 (C, C-3″), 148.1 (C, C-4″), 138.5 (C, C-10′), 138.2

(

C, C-1″″), 137.3 (C, C-1′″), 134.3 (CH, C-8′), 131.8 (C,

This compound was prepared following the general pro-

cedure B. 4-methoxy phenyl hydrazine hydrochloride

C-1″), 128.3 (CH, C-3″″, C-5″″), 127.7 (CH, C-4″″), 127.6

(CH, C-2″″, C-6″″), 123.2 (CH, C-6′), 121.8 (CH, C-7′),

118.0 (CH, C-6″), 115.6(CH, C-2′″, C-6′″), 115.1 (CH,

(

149.1 mg, 0.85 mmol) was dissolved in 0.94 ml of absolute

1

3

Molecular Diversity

C-9′), 114.8 (CH, C-3′″, C-5′″), 114.0 (CH, C-5″), 112.0

600 MHz): δ ppm 13.54 (s, 1H, OH), 11.54 (s, 1H, NH),

8.95 (s, 1H, Ar-OH), 7.95 (d, 1H, J = 7.8 Hz, Ar–H), 7.55

(t, 1H, J = 7.8 Hz, Ar–H), 7.41 (m, 2H, Ar–H), 7.37 (t,

2H, J = 7.2 Hz, Ar–H), 7.30 (m, 2H, Ar–H), 7.23 (t, 1H,

J = 7.2 Hz, Ar–H), 6.93 (m, 3H, Ar–H), 6.88 (m, 2H,

Ar–H), 6.71 (m, 2H, Ar–H), 5.11 (br, 1H, C -H ), 5.00

(

C, C-5′), 109.8 (CH, C-2″), 101.4 (C, C-3′), 69.5 (CH ,

2

Ar–O-CH -Ar), 63.3(CH, C-5), 55.4 (CH , Ar–O-CH ,

2

3

Ar–O-CH ), 30.7 (CH , C-4); HR-MS m/z (pos): 548.21717

3

2

C H N O (calcd. 547.2107).

3

29

3

5

X

4‑(3‑(4‑Hydroxy‑2‑oxo‑1,2‑dihydroquino‑

(s, 2H, Ar-CH -O-Ar), 4.17 (dd, 1H, J = 18.6, 12.0 Hz,

2

lin‑3‑yl)‑5‑(4‑hydroxy‑3‑methoxyphenyl)‑4,5‑dihy‑

dro‑1H‑pyrazol‑1‑yl)benzonitrile (9d)

C -H ), 3.73 (s, 3H, OCH ), 3.43 (dd, 1H, J = 18.6,

4

B

3

1

3

8.4 Hz, C -H ); C NMR (DMSO-d , 600 MHz): δ ppm

4 A 6

1

62.3 (C, C-4′), 161.3 (C, C-3), 152.4 (C, C-2′), 150.6

This compound was prepared following the general proce-

dure B. 4-cyano phenyl hydrazine hydrochloride (150.8 mg,

(C, C-4′″), 147.9 (C, C-3″), 145.9 (C, C-4″), 138.6 (C,

C-10′), 138.2 (C, C-1″″), 137.3 (C, C-1′″), 132.8 (CH,

C-8′), 131.8 (C, C-1″), 128.3 (CH, C-3″″, C-5″″), 127.7

(CH, C-4″″), 127.6 (CH, C-2″″, C-6″″), 123.1 (CH, C-6′),

121.8 (CH, C-7′), 118.4 (CH, C-6″), 115.6 (CH, C-2′″,

C-6′″), 115.1 (CH, C-9′), 114.8 (CH, C-3′″, C-5′″), 114.0

(CH, C-5″), 112.9 (C, C-5′), 110.2 (CH, C-2″), 101.4 (C,

C-3′), 69.5 (CH , Ar–O-CH -Ar), 63.4 (CH, C-5), 55.6

0

.89 mmol) was dissolved in 1 ml of absolute ethanol. Qui-

nolinone-chalcone 8b (120 mg, 0.36 mmol) was dissolved

in 2 ml of acetic acid, and it was added dropwise to the

ethanolic solution of hydrazine. The resulting solution was

stirred at 120 °C for 24 h. The solid produced upon ꢂltra-

tion was triturated from methanol/dichloromethane to aꢀord

2

the pure pyrazoline 9d as orange powder. Yield: 62 mg

(CH , Ar–O-CH ,), 46.6 (CH , C-4); HR-MS m/z (pos):

3 3 2

1

(

51.7%); m.p. > 250 °C; H NMR (DMSO-d , 600 MHz):

533.19400 C H N O (calcd. 533.1951).

32 27 3 5

6

δ ppm 13.08 (s, 1H, OH), 11.61 (s, 1H, NH), 9.01 (s, 1H,

Ar-OH), 7.98 (d, 1H, J = 8.4 Hz, Ar–H), 7.62 (d, 2H,

J=9.0 Hz, Ar–H), 7.58 (t, 1H, J=8.4 Hz, Ar–H), 7.29 (d,

3‑(1‑(4‑Chlorophenyl)‑5‑(4‑hydroxy‑3‑methoxyphenyl)‑4,

5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxyquinolin‑2(1H)‑one

(9f)

1

H, J=8.4 Hz, Ar–H), 7.25 (t, 1H, J=7.8 Hz, Ar–H), 6.98

(

d, 2H, J=9.0 Hz, Ar–H), 6.90 (d, 1H, J=1.2 Hz, Ar–H),

.71 (d, 1H, J=8.4 Hz, Ar–H), 6.59 (dd, 1H, J=7.8, 1.2 Hz,

Ar–H), 5.42 (dd, 1H, J = 11.4, 5.4 Hz, C -H ), 4.19 (dd,

6

This compound was prepared following the general

procedure B. 4-chloro-phenyl hydrazine hydrochloride

(100.9 mg, 0.56 mmol) was dissolved in 0.6 ml of absolute

ethanol. Quinolinone-chalcone 8b (76 mg, 0.2 mmol) was

dissolved in 1.2 ml of acetic acid and was added dropwise

to the ethanolic solution of hydrazine. The resulting solu-

tion was stirred at 120 °C for 24 h. The solid produced

upon ꢂltration was triturated from methanol/dichlorometh-

ane to aꢀord the pure pyrazoline 9f as orange powder.

5

X

1

H, J = 18.6, 12.0 Hz, C -H ), 3.73 (s, 3H, OCH ), 3.58

4 B 3

1

3

(

dd, 1H, J = 19.2,6.0 Hz, C -H ); C NMR (DMSO-d ,

4

A

6

00 MHz): δ ppm 163.5 (C, C-4′), 161.6 (C, C-3), 154.2 (C,

C-2′), 148.5 (C, C-3″), 146.5 (C, C-4″), 146.0 (C, C-1′″),

1

38.9 (C, C-10′), 133.9 (CH, C-3′″, C-5′″), 132.8 (C, C-1″),

32.3 (CH, C-8′), 123.8 (CH, C-6′), 122.4 (CH, C-7′), 120.3

1

(

CH, C-6″), 118.1 (C, Ar–C≡N), 116.3 (CH, C-9′), 115.7

CH, C-5″), 114.2 (C, C-5′), 113.1 (CH, C-2′″, C-6′″), 110.4

C, C-2″), 101.5 (C, C-4′″), 99.9 (C, C-3′), 61.0 (CH, C-5),

1

Yield: 32 mg (42.1%); m.p. > 250 °C; H NMR (DMSO-

5

6.1 (CH , Ar–O-CH ,), 47.2 (CH , C-4); HR-MS m/z (neg):

d , 600 MHz): δ ppm 13.29 (s, 1H, OH), 11.58 (s, 1H,

3

2

6

4

51.13988 C H N O (calcd. 452.1485).

NH), 9.0 (s, 1H, Ar-OH), 7.96 (d, 1H, J = 7.8 Hz, Ar–H),

2

6

20

4

7

.56 (t, 1H, J = 7.2 Hz, Ar–H), 7.25 (m, 4H, Ar–H), 6.91

3‑(1‑(4‑(Benzyloxy)phenyl)‑5‑(4‑hydroxy‑3‑methoxyp

(m, 3H, Ar–H), 6.71 (d, 1H, J = 8.4 Hz, Ar–H), 6.64 (dd,

1H, J = 7.8, 0.6 Hz, Ar–H), 5.26 (dd, 1H, J = 12.0, 7.2 Hz,

C -H ), 4.18 (dd, 1H, J = 18.6, 12.0 Hz, C -H ), 3.73 (s,

henyl)‑4,5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxyquino‑

lin‑2(1H)‑one (9e)

5

X

4

B

1

3

H, OCH ), 3.51 (dd, 1H, J = 19.2, 7.2 Hz, C -H );

C

3

4

A

This compound was prepared following the general pro-

cedure B. 4-benzyloxy phenyl hydrazine hydrochloride

NMR (DMSO-d , 600 MHz): δ ppm 162.6 (C, C-4′),

6

161.3 (C, C-3), 151.7 (C, C-2′), 148.0 (C, C-3″), 146.0

(C, C-4″), 142.4 (C, C-1′″), 138.3 (C, C-10′), 132.4 (CH,

C-8′), 132.1 (C, C-4′″), 130.1 (C, C-1″), 128.9 (CH, C-3′″,

C-5′″), 123.2 (CH, C-6′), 122.9 (CH, C-7′), 121.9 (CH,

C-6″), 119.6 (CH, C-2′″, C-6′″), 118.0 (CH, C-9′), 115.2

(CH, C-5″), 113.9 (C, C-5′), 111.6 (CH, C-2″), 101.3 (C,

C-3′), 61.8 (CH, C-5), 55.6 (CH , Ar–O-CH ,), 46.8 (CH ,

(

223 mg, 0.89 mmol) was dissolved in 1 ml of absolute

ethanol. Quinolinone-chalcone 8b (120 mg, 0.36 mmol)

was dissolved in 2 ml of acetic acid and it was added drop-

wise to the ethanolic solution of hydrazine. The resulting

solution was stirred at 120 °C for 24 h. The solid produced

upon ꢂltration was triturated from methanol/dichlorometh-

3

2

ane to aꢀord the pure pyrazoline 9e as orange powder.

C-4); HR-MS m/z (neg): 460.10629 C H ClN O (calcd.

25 20 3 4

1

Yield: 68 mg (56.7%); m.p.>250 °C; H NMR (DMSO-d ,

461.1142).

6

1

3

Molecular Diversity

4

‑(5‑(3,4‑Dimethoxyphenyl)‑3‑(4‑hydroxy‑1‑me‑

149.5 (C, C-2′), 149.2 (C, C-3″), 148.7 (C, C-4″), 143.0 (C,

C-1′″), 139.0 (C, C-10′), 137.7 (C, C-3′″), 137.5 (C, C-4′″),

136.7 (CH, C-5′″), 131.3 (C, C-1″), 130.1 CH, C-8′), 126.4

thyl‑2‑oxo‑1,2‑dihydroquinolin‑3‑yl)‑4,5‑dihydro‑1H‑pyra‑

zol‑1‑yl)benzonitrile (9g)

(

CH, C-6′), 124.6 (CH, C-7′), 122.8 (CH, C-6′″), 121.8

This compound was prepared following the general proce-

dure B. 4-cyano phenyl hydrazine hydrochloride (116.1 mg,

(CH, C-2′″), 116.6 (CH, C-6″), 114.0 (CH, C-9′), 112.1 (C,

C-5′), 111.0 (CH, C-5″), 108.2 (CH, C-2″), 100.8 (C, C-3′),

70.7 (CH, C-5), 56.0 (CH , C-4), 55.9 (CH , Ar–O-CH ),

0

.68 mmol) was dissolved in 0.9 ml of absolute ethanol. Qui-

2

3

nolinone–chalcone 8c (100 mg, 0.27 mmol) was dissolved

in 1.7 ml of acetic acid and it was added dropwise to the

ethanolic solution of hydrazine. The resulting solution was

stirred at 120 °C for 24 h. The solid produced upon ꢂltration

was triturated from methanol/dichloromethane to aꢀord the

55.9 (CH , Ar–O-CH ), 29.4 (CH , N-CH ), 20.0 (CH ,

3 3 3 3 3

Ar-CH ), 19.6 (CH , Ar-CH ); HR-MS m/z (neg): 482.20754

3

C H N O (calcd. 483.2158).

2

9

29

3

4

3‑(5‑(3,4‑Dimethoxyphenyl)‑1‑(4‑methoxyphenyl)‑4,

5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxy‑1‑methylquino‑

lin‑2(1H)‑one (9i)

pure pyrazoline 9g as orange powder. Yield: 73 mg (73%);

1

m.p.>250 °C; H NMR (DMSO-d , 600 MHz): δ ppm 13.07

6

(

s, 1H, OH), 8.07 (br, 1H, Ar–H), 7.69 (br, 1H, Ar–H), 7.62

d, 2H, J = 7.8 Hz, Ar–H), 7.50 (br, 1H, Ar–H), 7.33 (br,

This compound was prepared following the general proce-

dure B. 4-methoxy phenyl hydrazine hydrochloride (239 mg,

1.37 mmol) was dissolved in 1.7 ml of absolute ethanol. Qui-

nolinone-chalcone 8c (200 mg, 0.55 mmol) was dissolved

in 3.5 ml of acetic acid, and it was added dropwise to the

ethanolic solution of hydrazine. The resulting solution was

stirred at 120 °C for 24 h. The solid produced upon ꢂltration

was triturated from methanol/dichloromethane to aꢀord the

1

H, Ar–H), 6.96 (m, 3H, Ar–H), 6.88 (d, 1H, J = 8.4 Hz,

Ar–H), 6.71 (d, 1H, J = 8.4 Hz, Ar–H), 5.46 (br, 1H,

C -H ), 4.21 (dd, 1H, J=18.6, 12.6 Hz, C -H ), 3.71 (br,

5

X

4

B

1

3

6

H, 2OCH ), 3.59 (br, 1H, C -H ), 3.57 (s, 3H, NCH ); C

3 4 A 3

NMR (DMSO-d , 600 MHz): δ ppm 162.2 (C, C-3), 160.8

6

(

C, C-4′), 154.4 (C, C-2′), 149.6 (C, C-3″), 148.7 (C, C-4″),

1

46.0 (C, C-1′″), 139.8 (C, C-10′), 134.0 (CH, C-3′″, C-5′″),

33.8 (C, C-1″), 133.3 (CH, C-8′), 124.3 (CH, C-6′), 122.6

1

pure pyrazoline 9i as orange powder. Yield: 152 mg (76%);

1

(

CH, C-7′), 117.8 (C, Ar–C≡N), 115.4 (CH, C-6″), 115.0

CH, C-9′), 113.1 (CH, C-2′″, C-6′″), 112.6 (CH, C-5″),

m.p. >250 °C; H NMR (CDCl , 600 MHz): δ ppm 13.74

3

(s, 1H, OH), 8.17 (dd, 1H, J = 7.8, 0.6 Hz, Ar–H), 7.58

(t, 1H, J = 8.4 Hz, Ar–H), 7.283 (m, 2H, Ar–H), 6.92 (m,

2H, Ar–H), 6.87 (m, 2H, Ar–H), 6.81 (m, 3H, Ar–H), 4.96

(dd, 1H, J = 12.0, 9.6 Hz, C -H ), 4.33 (dd, 1H, J = 18.6,

1

10.0 (C, C-5′), 101.4 (C, C-4′″), 100.1 (CH, C-2″), 99.5 (C,

C-3′), 60.9 (CH, C-5), 55.9 (CH , Ar–O-CH ), 55.9 (CH ,

3

Ar–O-CH ), 47.4 (CH , C-4), 29.5 (CH , N-CH ); HR-MS

3

2

3

5

X

m/z (neg): 479.17102 C H N O (calcd. 480.1798).

12.0 Hz, C -H ), 3.86 (s, 3H, OCH ), 3.83 (s, 3H, OCH ),

2

8

24

4

4 B 3 3

3

.74 (s, 3H, OCH ), 3.64 (s, 3H, NCH ), 3.60 (br, 1H,

3 3

1

3

5

‑(5‑(3,4‑Dimethoxyphenyl)‑1‑(3,4‑dimethylphenyl)‑4,

‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxy‑1‑methylquino‑

C -H ); C NMR (CDCl , 600 MHz): δ ppm 162.6 (C,

4 A 3

C-3), 161.9 (C, C-4′), 154.1 (C, C-2′), 151.4 (C, C-4′″),

149.6 (C, C-3″), 149.5 (C, C-4″), 148.5 (C, C-10′), 139.4

lin‑2(1H)‑one (9h)

(

C, C-1′″), 139.4 (C, C-1″), 134.8 (CH, C-8′), 124.7 (CH,

This compound was prepared following the general proce-

dure B. 1-(3,4-dimethylphenyl) hydrazine hydrochloride

C-6′), 124.6 (CH, C-7′), 121.9 (CH, C-6″), 118.8 (CH, C-9′),

116.1 (C, C-5′), 115.6 (CH, C-5″), 114.8 (CH, C-2′″, C-6′″),

111.7 (CH, C-3′″, C-5′″), 109.3 (CH, C-2″), 101.7 (C, C-3′),

65.3 (CH, C-5), 56.0 (CH , Ar–O-CH ), 55.8 (CH , Ar–O-

(

118.2 mg, 0.68 mmol) was dissolved in 0.9 ml of absolute

ethanol. Quinolinone-chalcone 8c (100 mg, 0.27 mmol) was

dissolved in 1.7 ml of acetic acid, and it was added dropwise

to the ethanolic solution of hydrazine. The resulting solu-

tion was stirred at 120 °C for 24 h. The solid produced upon

ꢂltration was triturated from methanol/dichloromethane

3

CH ), 55.6 (CH , Ar–O-CH ), 47.4 (CH , C-4), 29.1 (CH ,

3

2

3

N-CH ); HR-MS m/z (neg): 484.18742 C H N O (calcd.

3

28 27

3

5

485.1951).

to aꢀord the pure pyrazoline 9h as orange powder. Yield:

4‑(3‑(4‑Hydroxy‑1‑methyl‑2‑oxo‑1,2‑dihydroquino‑

lin‑3‑yl)‑5‑(4‑hydroxy‑3‑methoxyphenyl)‑4,5‑dihy‑

dro‑1H‑pyrazol‑1‑yl)benzonitrile (9j)

1

3

4 mg (34%); m.p.>250 °C; H NMR (CDCl , 600 MHz):

3

δ ppm 13.74 (br, 1H, OH), 8.17 (d, 1H, J=7.8 Hz, Ar–H),

.56 (t, 1H, J = 7.8 Hz, Ar–H), 7.25 (m, 2H, Ar–H), 6.94

d, 1H, J = 7.8 Hz, Ar–H), 6.81 (m, 4H, Ar–H), 6.64 (d,

H, J = 7.8 Hz, Ar–H), 5.01 (dd, 1H, J = 12.0, 9.0 Hz,

7

(

This compound was prepared following the general proce-

dure B. 4-cyano phenyl hydrazine hydrochloride (120.7 mg,

0.71 mmol) was dissolved in 0.9 ml of absolute ethanol.

Quinolinone-chalcone 8d (100 mg, 0.28 mmol) was dis-

solved in 1.8 ml of acetic acid, and it was added dropwise to

the ethanolic solution of hydrazine. The resulting solution

1

C -H ), 4.29 (dd, 1H, J=18.6, 12.6 Hz, C -H ), 3.82 (br,

5

X

4

B

6

2

H, 2OCH ), 3.76 (br, 1H, C -H ), 3.61 (s, 3H, NCH ),

3 4 A 3

1

3

.18 (s, 3H, Ar-CH ), 2.145 (s, 3H, Ar-CH ); C NMR

3

(

CDCl , 600 MHz): δ ppm 162.1 (C, C-3), 160.8 (C, C-4′),

3

1

3

Molecular Diversity

was stirred at 120 °C for 24 h. The solid produced upon

ꢂltration was triturated from methanol/dichloromethane to

(CH, C-2″), 101.6 (C, C-3′), 62.7 (CH, C-5), 56.0 (CH ,

3

Ar–O-CH ), 47.1 (CH , C-4), 29.4 (CH , N-CH ), 20.3

3

2

3

aꢀord the pure pyrazoline 9j as orange powder. Yield: 82 mg

(CH , Ar-CH ), 18.9 (CH , Ar-CH ); HR-MS m/z (neg):

3 3 3 3

1

(

82%); m.p. > 250 °C; H NMR (DMSO-d , 600 MHz): δ

468.19315 C H N O (calcd. 469.2002).

28 27 3 4

6

ppm 13.06 (s, 1H, OH), 9.01 (s, 1H, Ar-OH), 8.05 (d, 1H,

J=7.2 Hz, Ar–H), 7.67 (t, 1H, J=6.6 Hz, Ar–H), 7.60 (d,

4‑Hydroxy‑3‑(5‑(4‑hydroxy‑3‑methoxyphenyl)‑1‑(4‑meth

oxyphenyl)‑4,5‑dihydro‑1H‑pyrazol‑3‑yl)‑1‑methylquino‑

lin‑2(1H)‑one (9l)

2

H, J=7.8 Hz, Ar–H), 7.48 (d, 1H, J=7.8 Hz, Ar–H), 7.31

(

t, 1H, J=7.2 Hz, Ar–H), 6.95 (d, 2H, J=7.8 Hz, Ar–H),

.90 (br, 1H, Ar–H), 6.71 (d, 1H, J =7.8 Hz, Ar–H), 6.60

d, 1H, J= 7.2 Hz, Ar–H), 5.38 (dd, 1H, J= 10.8, 4.2 Hz,

6

(

This compound was prepared following the general pro-

cedure B. 4-methoxy phenyl hydrazine hydrochloride

(124.3 mg, 0.71 mmol) was dissolved in 0.9 ml of absolute

ethanol. Quinolinone-chalcone 8d (100 mg, 0.28 mmol) was

dissolved in 1.8 ml of acetic acid, and it was added dropwise

to the ethanolic solution of hydrazine. The resulting solu-

tion was stirred at 120 °C for 24 h. The solid produced upon

ꢂltration was triturated from methanol/dichloromethane to

C -H ), 4.18 (dd, 1H, J= 18.6, 12.6 Hz, C -H ), 3.73 (s,

5

X

4

B

3

H, OCH ), 3.57 (dd, 1H, J=19.8, 3.6 Hz, C -H ), 3.52 (s,

3 4 A

1

3

H, NCH ); C NMR (DMSO-d , 600 MHz): δ ppm 162.2

3

6

(

C, C-3), 160.8 (C, C-4′), 154.4 (C, C-2′), 148.5 (C, C-3″),

1

46.5 (C, C-4″), 146.0 (C, C-1′″), 139.8 (C, C-10′), 133.9

(

CH, C-3′″, C-5′″), 133.1 (C, C-1″), 132.3 (CH, C-8′), 124.3

CH, C-6′), 122.5 (CH, C-7′), 120.2 (CH, C-5″), 118.2 (C,

(

Ar–C≡N), 116.3 (CH, C-9′), 115.2 (CH, C-6″), 115.0 (C,

aꢀord the pure pyrazoline 9l as orange powder. Yield: 48 mg

1

C-5′), 113.1 (CH, C-2′″, C-6′″), 110.4 (CH, C-2″), 101.3 (C,

(48%); m.p. > 250 °C; H NMR (DMSO-d , 600 MHz): δ

6

C-4′″), 100.0 (C, C-3′), 61.0 (CH, C-5), 56.1 (CH , Ar–O-

ppm 13.57 (s, 1H, OH), 8.95 (s, 1H, Ar-OH), 8.04 (d, 1H,

J = 7.2 Hz, Ar–H), 7.66 (t, 1H, J = 7.2 Hz, Ar–H), 7.48

(d, 1H, J=8.4 Hz, Ar–H), 7.31 (t, 1H, J=7.8 Hz, Ar–H),

3

CH ), 47.4 (CH , C-4), 29.4 (CH , N-CH ); HR-MS m/z

3

2

3

(

neg): 465.15552 C H N O (calcd. 466.1641).

27 22 4 4

6

.94 (br, 1H, Ar–H), 6.86 (m, 4H, Ar–H), 6.71 (m, 2H,

3‑(1‑(3,4‑Dimethylphenyl)‑5‑(4‑hydroxy‑3‑methoxypheny

Ar–H), 5.08 (dd, 1H, J = 11.4, 9.0 Hz, C -H ), 4.16 (dd,

5 X

l)‑4,5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxy‑1‑methylquino‑

lin‑2(1H)‑one (9k)

1H, J = 18.6, 12.0 Hz, C -H ), 3.73 (s, 3H, OCH ), 3.67

4 B 3

(s, 3H, OCH ), 3.54 (s, 3H, NCH ), 3.44 (dd, 1H, J=18.6,

3 3

1

3

8

.4 Hz, C -H ); C NMR (DMSO-d , 600 MHz): δ ppm

4 A 6

This compound was prepared following the general proce-

dure B. 1-(3,4-dimethylphenyl) hydrazine hydrochloride

161.5 (C, C-3), 160.9 (C, C-4′), 153.8 (C, C-2′), 151.2 (C,

C-4′″), 148.4 (C, C-3″), 146.3 (C, C-4″), 139.4 (C, C-10′),

138.8 (C, C-1′″), 133.2 (CH, C-8′), 132.6 (C, C-1″), 124.1

(CH, C-6′), 122.4 (CH, C-7′), 118.9 (CH, C-6″), 116.2 (CH,

C-9′), 115.4 (CH, C-2′″, C-6′″), 115.3 (C, C-5′), 115.2 (CH,

C-5″), 115.0 (CH, C-3′″, C-5′″), 110.7 (CH, C-2″), 101.6 (C,

C-3′), 63.9 (CH, C-5), 56.0 (CH , Ar–O-CH ), 55.7 (CH ,

(

122.85 mg, 0.71 mmol) was dissolved in 0.9 ml of absolute

ethanol. Quinolinone-chalcone 8d (100 mg, 0.28 mmol) was

dissolved in 1.8 ml of acetic acid and it was added dropwise

to the ethanolic solution of hydrazine. The resulting solu-

tion was stirred at 120 °C for 24 h. The solid produced upon

ꢂltration was triturated from methanol/dichloromethane to

3

Ar–O-CH ), 47.2 (CH , C-4), 29.4 (CH , N-CH ); HR-MS

3

2

3

aꢀord the pure pyrazoline 9k as orange powder. Yield: 67 mg

m/z (pos): 472.18606 C H N O (calcd. 471.1794).

27 25 3 5

1

(

67%); m.p. > 250 °C; H NMR (DMSO-d , 600 MHz): δ

6

ppm 13.55 (s, 1H, OH), 8.95 (s, 1H, Ar-OH), 8.04 (d, 1H,

3‑(1‑(4‑(Benzyloxy)phenyl)‑5‑(4‑hydroxy‑3‑methoxypheny

l)‑4,5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxy‑1‑methylquino‑

lin‑2(1H)‑one (9m)

J=7.2 Hz, Ar–H), 7.65 (t, 1H, J=7.2 Hz, Ar–H), 7.46 (d,

1

H, J=8.4 Hz, Ar–H), 7.30 (t, 1H, J=7.2 Hz, Ar–H), 6.95

(

d, 1H, J = 7.8 Hz, Ar–H), 6.91 (br, 1H, Ar–H), 6.75 (br,

H, Ar–H), 6.70 (d, 1H, J = 7.8 Hz, Ar–H), 6.65 (d, 1H,

1

This compound was prepared following the general procedure

B. 4-benzyloxy phenyl hydrazine hydrochloride (178.4 mg,

0.71 mmol) was dissolved in 0.9 ml of absolute ethanol.

Quinolinone-chalcone 8d (100 mg, 0.28 mmol) was dis-

solved in 1.8 ml of acetic acid, and it was added dropwise to

the ethanol solution of hydrazine. The resulting solution was

stirred at 120 °C for 24 h. The solid produced upon ꢂltration

was triturated from methanol/dichloromethane to aꢀord the

J = 7.8 Hz, Ar–H), 6.60 (d, 1H, J = 7.2 Hz, Ar–H), 5.14

(

dd, 1H, J = 10.8, 7.8 Hz, C -H ), 4.14 (dd, 1H, J = 18.0,

5 X

1

3

2.0 Hz, C -H ), 3.72 (s, 3H, OCH ), 3.52 (s, 3H, NCH ),

4 B 3 3

.46 (dd, 1H, J=18.6, 7.8 Hz, C -H ), 2.11 (br, 6H, 2Ar-

4

A

1

3

CH ); C NMR (DMSO-d , 600 MHz): δ ppm 161.4 (C,

3

6

C-3), 160.8 (C, C-4′), 151.0 (C, C-2′), 148.3 (C, C-3″),

46.2 (C, C-4″), 142.3 (C, C-1′″), 139.4 (C, C-10′), 137.1

C, C-3′″), 133.5 (C, C-4′″), 132.6 (CH, C-5′″), 130.4 (CH,

1

(

pure pyrazoline 9 m as orange powder. Yield: 38 mg (38%);

1

C-8′), 127.7 (C, C-1″), 124.1 (CH, C-6′), 122.4 (CH, C-7′),

m.p.>250 °C; H NMR (DMSO-d , 600 MHz): δ ppm 13.51

6

1

18.6 (CH, C-6′″), 116.2 (CH, C-6″), 115.2 (CH, C-2′″),

15.1 (CH, C-9′), 115.0 (CH, C-5″), 111.1 (C, C-5′), 110.5

(s, 1H, OH), 8.93 (br, 1H, Ar-OH), 8.03 (br, 1H, Ar–H), 7.64

(br, 1H, Ar–H), 7.37 (m, 7H, Ar–H), 6.89 (m, 5H, Ar–H), 6.69

1

3

Molecular Diversity

(

br, 2H, Ar–H), 5.09 (br, 1H, C -H ), 4.99 (s, 2H, Ar-CH -

hydrazine were dissolved. The resulting solution was stirred

at 118 °C for 4 h. The reaction was monitored by thin-layer

chromatography (TLC). After completion of the reaction,

2 ml of aqueous solution of absolute ethanol 50% was added

to the mixture, and an orange precipitate was formed. The

solid was ꢂltered, washed with ice water and further tritu-

5

X

2

O-Ar), 4.17 (br, 1H, C -H ), 3.72 (s, 3H, OCH ), 3.55 (s,

4

B

3

1

3

6

H, NCH ), 3.45 (br, 1H, C -H ); C NMR (DMSO-d ,

3 4 A 6

00 MHz): δ ppm 161.0 (C, C-3), 160.4 (C, C-4′), 152.4 (C,

C-2′), 150.8 (C, C-4′″), 147.9 (C, C-3″), 145.9 (C, C-4″), 138.9

(

C, C-10′), 138.5 (C, C-1′″), 137.3 (C, C-1″″), 132.8 (CH,

C-8′), 132.2 (C, C-1″), 128.4 (CH, C-3″″, C-5″), 127.7 (CH,

C-4″″), 127.6 (CH, C-2″″, C-6″″), 123.6 (CH, C-6′), 122.0

rated from methanol and dichloromethane. Yield: 17 mg

1

(34%); m.p. > 250 °C; H NMR (DMSO-d , 600 MHz): δ

6

(

CH, C-7′), 118.4 (CH, C-6″), 115.7 (CH, C-9′), 115.6 (CH,

ppm 13.48 (s, 1H, OH), 11.55 (s, 1H, NH), 8.95 (s, 1H,

Ar-OH), 7.96 (d, 1H, J = 7.8 Hz, Ar–H), 7.55 (t, 1H,

J=7.2 Hz, Ar–H), 7.29 (d, 1H, J=7.8 Hz, Ar–H), 7.23 (m,

3H, Ar–H), 6.92 (m, 3H, Ar–H), 6.80 (t, 1H, J = 7.2 Hz,

Ar–H), 6.72 (d, 1H, J = 8.4 Hz, Ar–H), 6.67 (d, 1H,

J=7.8 Hz, Ar–H), 5.24 (dd, 1H, J=11.4, 7.2 Hz, C -H ),

C-2′″, C-6′″), 114.8 (CH, C-5″), 114.8 (CH, C-3′″, C-5′″),

14.7 (C, C-5′), 110.2 (CH, C-2″), 101.1 (C, C-3′), 69.5 (CH ,

1

2

Ar–O-CH -Ar), 63.4 (CH, C-5), 55.6 (CH , Ar–O-CH ), 46.8

2

3

(CH , C-4), 28.9 (CH , N-CH ); HR-MS m/z (pos): 548.21901

2 3 3

C H N O (calcd. 547.2107).

33

29

3

5

X

4

.18 (dd, 1H, J=18.6, 12.0 Hz, C -H ), 3.73 (s, 3H, OCH ),

4 B 3

1

3

4

‑(1‑(4‑Chlorophenyl)‑5‑(4‑hydroxy‑3‑methoxyphenyl)‑

,5‑dihydro‑1H‑pyrazol‑3‑yl)‑4‑hydroxy‑1‑methylquino‑

3.49 (dd, 1H, J=19.2, 7.8 Hz, C -H ); C NMR (DMSO-

4 A

d , 600 MHz): δ ppm 163.0 (C, C-4′), 161.7 (C, C-3), 151.5

6

lin‑2(1H)‑one (9n)

(C, C-2′), 148.4 (C, C-3″), 146.3 (C, C-4″), 144.2 (C, C-1′″),

1

38.7 (C, C-10′), 133.3 (CH, C-8′), 132.3 (C, C-1″), 129.5

This compound was prepared following the general proce-

dure B. 4-chloro phenyl hydrazine hydrochloride (255.2 mg,

(CH, C-3′″, C-5′″), 123.6 (CH, C-6′), 122.3 (CH, C-4′″),

119.8 (CH, C-7′), 118.5 (CH, C-6″), 116.2 (CH, C-9′), 115.6

(CH, C-5″), 114.4 (C, C-5′), 113.4 (CH, C-2′″, C-6′″), 110.5

1

.43 mmol) was dissolved in 1.80 ml of absolute ethanol.

Quinolinone-chalcone 8d (200 mg, 0.57 mmol) was dissolved

in 3.60 ml of acetic acid, and it was added dropwise to the

ethanolic solution of hydrazine. The resulting solution was

stirred at 120 °C for 24 h. The solid produced upon ꢂltra-

tion was triturated from methanol/dichloromethane to aꢀord

(CH, C-2″), 101.8 (C, C-3′), 62.4 (CH, C-5), 56.1 (CH ,

3

Ar–O-CH ), 47.1 (CH , C-4); HR-MS m/z (neg): 426.14538

3

2

C H N O (calcd. 427.1532).

2

5

21

3

4

Biological evaluation

the pure pyrazoline 9n as yellow powder. Yield: 180 mg

1

(

90%); m.p. >250 °C; H NMR (DMSO-d , 600 MHz): δ

Determination of the reducing activity of the stable radical

2,2‑Diphenyl‑1‑picrylhydrazyl (DPPH)

6

ppm 13.26 (s, 1H, OH), 8.99 (s, 1H, Ar-OH), 8.00 (d, 1H,

J=7.8 Hz, Ar–H), 7.64 (t, 1H, J=7.2 Hz, Ar–H), 7.43 (d,

1

H, J=8.4 Hz, Ar–H), 7.28 (t, 1H, J=7.2 Hz, Ar–H), 7.24

To an ethanolic solution of DPPH (100 μM) in absolute etha-

nol, an equal volume of the compounds dissolved in DMSO

was added (100 μM). The mixture was shaken vigorously

and allowed to stand for 20 or 60 min; absorbance at 517 nm

was determined spectrophotometrically, and the percentage

of activity was calculated. All tests were undertaken on three

replicates, and the results presented in Table 3 were aver-

aged and compared with the appropriate standard nordihy-

droguaiaretic acid (NDGA) [66].

(

d, 2H, J=8.4 Hz, Ar–H), 6.89 (br, 1H, Ar–H), 6.87 (d, 2H,

J=8.4 Hz, Ar–H), 6.71 (d, 1H, J=7.8 Hz, Ar–H), 6.63 (d,

1

4

3

H, J=8.4 Hz, Ar–H), 5.18 (dd, 1H, J=11.4, 7.2 Hz, C -H ),

5 X

.15 (dd, 1H, J=18.6, 12.0 Hz, C -H ), 3.72 (s, 3H, OCH ),

4

B

3

13

.49 (s, 3H, NCH ), 3.47 (br, 1H, C -H ); C NMR (DMSO-

3

4

A

d , 600 MHz): δ ppm 161.3 (C, C-3), 160.4 (C, C-4′), 151.9

6

(

C, C-2′), 148.0 (C, C-3″), 146.0 (C, C-4″), 142.3 (C, C-1′″),

1

39.1 (C, C-10′), 132.4 (CH, C-8′), 128.9 (CH, C-3′″, C-5′″),

23.7 (CH, C-6′), 122.9 (CH, C-7′), 122.1 (CH, C-6″), 118.0

1

(

C, C-4′″), 115.8 (CH, C-1″), 114.8 (C, C-9′), 114.7 (C, C-5″),

Inhibition of linoleic acid lipid peroxidation

1

14.4 (CH, C-2′″, C-6′″), 110.0 (C, C-5′), 101.0 (CH, C-2″),

9.1 (C, C-3′), 61.8 (CH, C-5), 55.6 (CH , Ar–O-CH ), 47.0

9

Production of conjugated diene hydroperoxide by oxidation

of linoleic acid in an aqueous dispersion is monitored at

234 nm. 2,2′-Azobis(2-amidinopropane) dihydrochloride

3

(CH , C-4), 29.0 (CH , N-CH ); HR-MS m/z (pos): 476.13721

2 3 3

C H ClN O (calcd. 475.1299).

26

22

3

4

(

AAPH) is used as a free radical initiator. Ten microliters

Experimental procedure for the synthesis

of 4‑hydroxy‑3‑(5‑(4‑hydroxy‑3‑methoxyphenyl)‑1‑phe‑

nyl‑4,5‑dihydro‑1H‑pyrazol‑3‑yl)quinolin‑2(1H)‑one (9o)

of the 16 mM linoleic acid sodium salt solution was added

to the UV cuvette containing 0.93 ml of 0.05 M phosphate

buꢀer, pH 7.4 prethermostated at 37 °C. The oxidation reac-

tion was initiated at 37 °C under air by the addition of 50 µl

of 40 mM AAPH solution. Oxidation was carried out in

the presence of the synthesized compounds (10 µl, from a

In 4.5 ml of acetic acid, 0.15 mmol (50 mg) of quin-

olinone–chalcone 8b and 0.30 mmol (32.4 mg) of phenyl

1

3

Molecular Diversity

stock solution of 10 mM in DMSO). In the assay without

antioxidant, lipid oxidation was measured in the presence of

the same level of DMSO. The rate of oxidation at 37 °C was

monitored by recording the increase in absorption at 234 nm

caused by conjugated diene hydroperoxides and compared

with the appropriate standard trolox [66].

2. Wisastra R, Dekker FJ (2014) Inꢁammation, cancer and oxidative

lipoxygenase activity are intimately linked. Cancers 6(3):1500–

1

521. https://doi.org/10.3390/cancers6031500

3

4

5

.

Yanes DA, Mosser-Goldfarb JL (2018) Emerging therapies for

atopic dermatitis: the prostaglandin/leukotriene pathway. J Am

Acad Dermatol 78(3 Suppl 1):S71–S75. https://doi.org/10.1016/j.

jaad.2017.12.021

Radmark O, Werz O, Steinhilber D, Samuelsson B (2007)

5

-Lipoxygenase: regulation of expression and enzyme activity.

Soybean LOX inhibition study in vitro

Trends Biochem Sci 32(7):332–341. https://doi.org/10.1016/j.

tibs.2007.06.002

Dahlén SE (2006) Treatment of asthma with antileukotrienes: ꢂrst

line or last resort therapy? Eur J Pharmacol 533(1–3):40–56. https

The tested compounds dissolved in DMSO were incubated at

room temperature with sodium linoleate (0.1 ml) and 0.2 ml

:

//doi.org/10.1016/j.ejphar.2005.12.070

−

4

of enzyme solution (1/9×10 w/v in saline) in Tris buꢀer

pH 9. The conversion of sodium linoleate to 13-hydroper-

oxylinoleic acid at 234 nm was recorded and compared with

the appropriate standard inhibitor NDGA [66].

6. Lötzer K, Funk CD, Habenicht AJ (2005) The 5-lipoxygenase

pathway in arterial wall biology and atherosclerosis. Biochim

Biophys Acta 1736(1):30–37. https://doi.org/10.1016/j.bbali

p.2005.07.001

7

.

Fürstenberger G, Krieg P, Müller-Decker K, Habenicht AJ (2006)

What are cyclooxygenases and lipoxygenases doing in the driver’s

seat of carcinogenesis? Int J Cancer 119(10):2247–2254. https://

doi.org/10.1002/ijc.22153

Molecular docking simulations for soybean LOX

8

9

.

Pontiki E, Hadjipavlou-Litina D (2013) Targeting Lipoxygenases

(LOs): drug design and discovery. Curr Enzym Inhib 9(2):89–105.

https://doi.org/10.2174/1573408011309020003

Molecular docking studies of soybean LOX were performed

selecting 3PZW from the Protein Data Bank (PDB) and

using UCSF Chimera for the visualization of the protein

Lu P, Schrag ML, Slaughter DE, Raab CE, Shou M, Rodrigues

AD (2003) Mechanism-based inhibition of human liver microso-

mal cytochrome P450 1A2 by zileuton, a 5-lipoxygenase inhibitor.

Drug Metab Dispos 31(11):1352–1360. https://doi.org/10.1124/

dmd.31.11.1352

(

PDB code: 3PZW) [70]. Water molecules were removed,

missing residues were added with Modeller [71], hydrogen

atoms and AMBER99SB-ILDN charges were added while

the iron charge was set to +2.0, with no restraint applied to

the iron atom and the ligands.

1

0. Zaman K, Hanigan MH, Smith A, Vaughan J, Macdonald T, Jones

DR, Hunt JF, Gaston B (2006) Endogenous S-nitrosoglutathione

modiꢂes 5-lipoxygenase expression in airway epithelial cells. Am

J Respir Cell Mol Biol 34(4):387–393. https://doi.org/10.1165/

rcmb.2005-0336RC

OpenBabel was used to generate and minimize Ligand

3

D coordinates [72] using the MMFF94 force ꢂeld [73],

1. Gaztanaga J, Farkouh M, Rudd JH, Brotz TM, Rosenbaum D,

Mani V, Kerwin TC, Taub R, Tardif JC, Tawakol A, Fayad ZA

while ACPYPE (AnteChamber PYthon Parser interfacE)

74] was used to generate ligand topologies and parame-

[

(

2015) A phase 2 randomized, double-blind, placebo-controlled

ters using Antechamber [75]. Energy minimizations were

achieved using the AMBER99SB-ILDN force ꢂeld [76] with

GROMACS 4.6.5 [77] as the molecular dynamics simulation

toolkit. Docking studies were carried out with AutoDock

Vina (1.1.2) [78] using a grid box of size 100 Å, 70 Å, 70 Å

in X, Y, Z dimensions for soybean LOX. UCSF Chimera was

used for the generation of docking input ꢂles and analysis of

the obtained docking results. Docking was carried out with

an exhaustiveness value of 10 and a maximum output of 20

docking modes soybean LOX.

study of the eꢀect of VIA-2291, a 5-lipoxygenase inhibitor, on

vascular inꢁammation in patients after an acute coronary syn-

drome. Atherosclerosis 240(1):53–60. https://doi.org/10.1016/j.

atherosclerosis.2015.02.027

1

2. Matsumoto S, Ibrahim R, Grégoire JC, L’Allier PL, Pressacco J,

Tardif JC, Budoꢀ MJ (2017) Eꢀect of treatment with 5-lipoxyge-

nase inhibitor VIA-2291 (atreleuton) on coronary plaque progres-

sion: a serial CT angiography study. Clin Cardiol 40(4):210–215.

https://doi.org/10.1002/clc.22646

3. Bruno F, Spaziano G, Liparulo A, Roviezzo F, Nabavi SM, Sureda

A, Filosa R, D’Agostino B (2018) Recent advances in the search

for novel 5-lipoxygenase inhibitors for the treatment of asthma.

Eur J Med Chem 153:65–72. https://doi.org/10.1016/j.ejmec

h.2017.10.020

Acknowledgements I.K. gratefully acknowledges ꢂnancial support

4. Tavolari S, Bonafè M, Marini M, Ferreri C, Bartolini G, Brighenti

E, Manara S, Tomasi V, Laufer S, Guarnieri T (2008) Licofelone,

a dual COX/5-LOX inhibitor, induces apoptosis in HCA-7 colon

cancer cells through the mitochondrial pathway independently

from its ability to aꢀect the arachidonic acid cascade. Carcino-

genesis 29(2):371–380. https://doi.org/10.1093/carcin/bgm265

5. Merchant N, Bhaskar LVKS, Momin S, Sujatha P, Reddy ABM,

Nagaraju GP (2018) 5-Lipoxygenase: its involvement in gastroin-

testinal malignancies. Crit Rev Oncol Hematol 127:50–55. https

://doi.org/10.1016/j.critrevonc.2018.05.012

from the Eugenides Foundation (scholarship for postgraduate studies).

Compliance with ethical standards

Conflict of interest The authors have declared no conꢁict of interest.

1

References

6. Kim W, Son B, Lee S, Do H, Youn B (2018) Targeting the

enzymes involved in arachidonic acid metabolism to improve

radiotherapy. Cancer Metastasis Rev 37(2–3):213–225. https://

doi.org/10.1007/s10555-018-9742-0

1

.

Brash AR (1999) Lipoxygenases: occurrence, functions, catalysis,

and acquisition of substrate. J Biol Chem 274(34):23679–23682.

https://doi.org/10.1074/jbc.274.34.23679

1

3

Molecular Diversity

1

7. Azad MB, Chen Y, Gibson SB (2009) Regulation of autophagy by

reactive oxygen species (ROS): implications for cancer progres-

sion and treatment. Antioxid Redox Signal 11(4):777–790. https

32. Bandgar BP, Patil SA, Korbad BL, Nile SH, Khobragade CN

(2010) Synthesis and biological evaluation of beta-chloro vinyl

chalcones as inhibitors of TNF-alpha and IL-6 with antimicro-

bial activity. Eur J Med Chem 45(6):2629–2633. https://doi.

org/10.1016/j.ejmech.2010.01.050

:

//doi.org/10.1089/ars.2008.2270

8. Weber V, Rubat C, Duroux E, Lartigue C, Madesclaire M, Coudert

P (2005) New 3- and 4-hydroxyfuranones as anti-oxidants and

anti-inꢁammatory agents. Bioorg Med Chem 13(14):4552–4564.

https://doi.org/10.1016/j.bmc.2005.04.055

33. Roussaki M, Hall B, Lima SC, Cordeiro da Silva A, Wilkinson S,

Detsi A (2013) Synthesis and anti-parasitic activity of a novel qui-

nolinone–chalcone series. Bioorg Med Chem Lett 23(23):6436–

6441. https://doi.org/10.1016/j.bmcl.2013.09.047

9. Pontiki E, Hadjipavlou-Litina D, Patsilinakos A, Tran TM, Mar-

son CM (2015) Pteridine-2,4-diamine derivatives as radical

scavengers and inhibitors of lipoxygenase that can possess anti-

inꢁammatory properties. Future Med Chem 7(14):1937–1951.

https://doi.org/10.4155/fmc.15.104

34. Siddiqui ZN, Musthafa TNM, Ahmad A, Khan AU (2011) Ther-

mal solvent-free synthesis of novel pyrazolyl chalcones and pyra-

zolines as potential antimicrobial agents. Bioorg Med Chem Lett

21(10):2860–2865

2

0. Detsi A, Majdalani M, Kontogiorgis CA, Hadjipavlou-Litina D,

Kefalas P (2009) Natural and synthetic 2′-hydroxy-chalcones

and aurones: synthesis, characterization and evaluation of

the antioxidant and soybean lipoxygenase inhibitory activity.

Bioorg Med Chem 17(23):8073–8085. https://doi.org/10.1016/j.

bmc.2009.10.002

35. Dave SS, Rahatgaonkar AM (2011) Syntheses and anti-microbial

evaluation of new quinoline scaꢀold derived pyrimidine deriva-

tives. Arab J Chem Res 9:S451–S456. https://doi.org/10.1016/j.

arabjc.2011.06.009

36. Sonawane V, Hiwarale D, Sonawane B, Kamble S, Sonawane K,

Bhosale R (2018) Synthesis, characterization and biological eval-

uation of novel imidazo thiazole chalcones as antioxidant agent.

Int J Pharm Sci Res 10(5):2439–2446. https://doi.org/10.13040/

IJPSR.0975-8232

2

1. Bansal Y, Silakari O (2014) Multifunctional compounds: smart

molecules for multifactorial diseases. Eur J Med Chem 76:31–42.

https://doi.org/10.1016/j.ejmech.2014.01.060

2. Fortin S, Bérubé G (2013) Advances in the development of hybrid

anticancer drugs. Expert Opin Drug Discov 8(8):1029–1047. https

37. Ramalho SD, Bernades A, Demetriusa G, Perez CN, Vieirab PC,

Santos CY, Silva JA, Moraes MO, Mousinhod KC (2013) Syn-

thetic chalcone derivatives as inhibitors of cathepsins K and B,

and their cytotoxic evaluation. Chem Biodivers 10:1999–2006.

https://doi.org/10.1002/cbdv.201200344

:

//doi.org/10.1517/17460441.2013.798296

3. Rana A, Alex JM, Chauhan M, Joshi G, Kumar R (2015) A review

on pharmacophoric designs of antiproliferative agents. Med Chem

Res 24(3):903–920. https://doi.org/10.1007/s00044-014-1196-5

4. Nakashima K, Oyama M, Ito T, Akao Y, Witono JR, Darnaedi D,

Tanaka T, Murata J, Iinuma M (2012) Novel quinolinone alkaloids

bearing a lignoid moiety and related constituents in the leaves of

Melicope denhamii . Tetrahedron 68(10):2421–2428. https://doi.

org/10.1016/j.tet.2012.01.007

38. Champelovier P, Chauchet X, Puch FH, Vergnaud S, Garrel C,

Laporte F, Boutonnat J, Boumendjel A (2013) Cellular and molec-

ular mechanisms activating the cell death processes by chalcones:

critical structural eꢀects. Toxicol In Vitro 27(8):2305–2315. https

://doi.org/10.1016/j.tiv.2013.09.021

39. Yusuf M, Jain P (2014) Synthetic and biological studies of pyrazo-

596. https://doi.org/10.1016/j.arabjc.2011.09.013

2

5. Gao H, Zhang L, Zhu T, Gu Q, Li D (2012) Unusual pyrrolyl

4

-quinolinone alkaloids from the marine-derived fungus Penicil-

lium sp. ghq208. Chem Pharm Bull 60(11):1458–1460. https://

doi.org/10.1248/cpb.c12-00487

40. Singh P, Singh J, Pant GJ, Rawat MSM (2018) 2-Pyrazolines

as biologically active and fluorescent agents, an overview.

Anticancer Agents Med Chem 18(10):1366–1385. https://doi.

org/10.2174/1871520618666180313153407

6. Detsi A, Bouloumbasi D, Prousis KC, Koufaki M, Athanasellis

G, Melagraki G, Afantitis A, Igglessi-Markopoulou O, Konto-

giorgis C, Hadjipavlou-Litina DJ (2007) Design and synthesis

of novel quinolinone-3-aminoamides and their alpha-lipoic acid

adducts as antioxidant and antiiꢁammatory agents. J Med Chem

41. Almahdi MM, Saeed AEM, Metwally NH (2019) Synthesis and

antimicrobial activity of some new pyrazoline derivatives bear-

ing sulfanilamido moiety. Eur J Chem 10(1):30–36. https://doi.

org/10.5155/eurjchem.10.1.30-36.1791

5

0(10):2450–2458. https://doi.org/10.1021/jm061173n

2

7. Angeleska S, Kefalas P, Detsi A (2013) Crude peroxidase from

onion solid waste as a tool for organic synthesis. Part III: syn-

thesis of tetracyclic heterocycles (coumestans and benzofuro-

quinolinones). Tetrahedron Lett 54(19):2325–2328. https://doi.

org/10.1016/j.tetlet.2013.02.081

42. Abdel-Wahab BF, Abdel-Latif E, Mohamed HA, Awad GE

(2012) Design and synthesis of new 4-pyrazolin-3-yl-1,2,3-tri-

azoles and 1,2,3-triazol-4-yl-pyrazolin-1-ylthiazoles as potential

antimicrobial agents. Eur J Med Chem 52:263–268. https://doi.

org/10.1016/j.ejmech.2012.03.023

8. Jönsson S, Andersson G, Fex T, Fristedt T, Hedlund G, Jansson

K, Abramo L, Fritzson I, Pekarski O, Runström A, Sandin H,

Thuvesson I, Björk A (2004) Synthesis and biological evaluation

of new 1,2-dihydro-4-hydroxy-2-oxo-3-quinolinecarboxamides

for treatment of autoimmune disorders: structure-activity rela-

tionship. J Med Chem 47(8):2075–2088. https://doi.org/10.1021/

jm031044w

43. Azarifar D, Shaebanzadeh M (2002) Synthesis and characteriza-

tion of new 3,5-dinaphthyl substituted 2-pyrazolines and study of

their antimicrobial activity. Molecules 7(12):885–895. https://doi.

org/10.3390/71200885

44. Rathore P, Yaseen S, Ovais S, Bashir R, Yaseen R, Hameed AD,

Samim M, Gupta R, Hussain F, Javed K (2014) Synthesis and

evaluation of some new pyrazoline substituted benzenesulfonylu-

reas as potential antiproliferative agents. Bioorg Med Chem Lett

24(7):1685–1691. https://doi.org/10.1016/j.bmcl.2014.02.059

45. Wang H, Zheng J, Xu W, Chen C, Wei D, Ni W, Pan Y (2017) A

new series of cytotoxic pyrazoline derivatives as potential antican-

cer agents that induce cell cycle arrest and apoptosis. Molecules

22(10):1635. https://doi.org/10.3390/molecules22101635

46. Khalil NA, Ahmed EM, El-Nassan HB, Ahmed OK, Al-Abd

AM (2012) Synthesis and biological evaluation of novel pyra-

zoline derivatives as anti-inꢁammatory and antioxidant agents.

Arch Pharm Res 35(6):995–1002. https://doi.org/10.1007/s1227

2-012-0606-9

2

3

9. Rahman MA (2011) Chalcone: a valuable insight into the recent

advances and potential pharmacological activities. J Chem Sci

2

9:1–16

0. Chetana BP, Mahajan SK, Suvarna AK (2009) Chalcone: a

versatile molecule. J Pharm Sci Res 1(3):11–22. https://doi.

org/10.1002/chin.201025251

1. Bandgar BP, Gawande SS, Bodade RG, Totre JV, Khobragade

CN (2010) Synthesis and biological evaluation of simple meth-

oxylated chalcones as anticancer, anti-inꢁammatory and anti-

oxidant agents. Bioorg Med Chem 18(3):1364–1370. https://doi.

org/10.1016/j.bmc.2009.11.066

1

3

Molecular Diversity

4

7. Shringare SN, Chavan HV, Bhale PS, Dongare SB, Mule YB, Patil

SB, Bandgar BP (2018) Synthesis and pharmacological evaluation

of combretastatin-A4 analogs of pyrazoline and pyridine deriva-

tives as anticancer, anti-inꢁammatory and antioxidant agents.

Med Chem Res 27(4):1226–1237. https://doi.org/10.1007/s0004

irradiation. Int J Pharm Sci Res 10:3290–3294. https://doi.

org/10.13040/IJPSR.0975-8232.10(7).3290-94

61. Chandel P, Kumar A, Singla N, Kumar A, Singh G, Gill RK

(2019) Rationally synthesized coumarin based pyrazolines ame-

liorate carrageenan induced inꢁammation through COX-2/pro-

inꢁammatory cytokine inhibition. Medchemcomm 10(3):421–

430. https://doi.org/10.1039/c8md00457a

4

-018-2142-8

4

5

8. Jeong TS, Kim KS, Kim JR, Cho KH, Lee S, Lee WS (2004)

Novel 3,5-diaryl pyrazolines and pyrazole as low-density lipo-

protein (LDL) oxidation inhibitor. Bioorg Med Chem Lett

62. Sharma S, Kaur S, Bansal T, Gaba J (2014) Review on synthesis

of bioactive pyrazoline derivatives. Chem Sci Trans 3(3):861–

875. https://doi.org/10.7598/cst2014.796

1

4(11):2719–2723. https://doi.org/10.1016/j.bmcl.2004.03.072

9. Kaplancikli ZA, Ozdemir A, Turan-Zitouni G, Altintop MD,

Can OD (2010) New pyrazoline derivatives and their antide-

pressant activity. Eur J Med Chem 45(9):4383–4387. https://doi.

org/10.1016/j.ejmech.2010.06.011

63. Weber FG, Brosehe K, Seedorf Ch, Itinow A (1969)

2

1,3,5-Triaryl-Δ -pyrazoline. Monatsh Chem 100(6):1924–1927.

https://doi.org/10.1007/BF01151742

64. Bhatnagar I, George MV (1968) Oxidation with metal oxides—

II: oxidation of chalcone phenylhydrazones, pyrazolines,

o-aminobenzylidine anils and o-hydroxy benzylidine anils with

manganese dioxide. Tetrahedron 24(3):1293–1298. https://doi.

org/10.1016/0040-4020(68)88080-9

0. Bandgar BP, Adsul LK, Chavan HV, Jalde SS, Shringare SN,

Shaikh R, Meshram RJ, Gacche RN, Masand V (2012) Synthe-

sis, biological evaluation, and docking studies of 3-(substituted)-

aryl-5-(9-methyl-3-carbazole)-1H-2-pyrazolines as potent anti-

inꢁammatory and antioxidant agents. Bioorg Med Chem Lett

65. Detsi A, Bardakos V, Markopoulos J, Igglessi-Markopoulou O

(1996) Reactions of 2-methyl-3,1-benzoxazin-4-one with active

methylene compounds: a new route to 3-substituted 4-hydrox-

yquinolin-2(1H)-ones. J Chem Soc Perkin Trans 1(24):2909–

2913. https://doi.org/10.1039/P19960002909

2

2(18):5839–5844. https://doi.org/10.1016/j.bmcl.2012.07.080

5

1. Kharbanda C, Alam MS, Hamid H, Javed K, Bano S, Dhulap A,

Ali Y, Nazreen S, Haider S (2014) Synthesis and evaluation of

pyrazolines bearing benzothiazole as anti-inꢁammatory agents.

Bioorg Med Chem 22(21):5804–5812. https://doi.org/10.1016/j.

bmc.2014.09.028

66. Hadjipavlou D, Garnelis T, Athanassopoulos CM, Papaioannou

D (2009) Kukoamine a analogs with lipoxygenase inhibitory

activity. J Enzyme Inhib Med Chem 24(5):1188–1193. https://

doi.org/10.1080/14756360902779193

2. Viveka S, Dinesha Shama P, Nagaraja GK, Ballav S, Kerkar S

(

2015) Design and synthesis of some new pyrazolyl-pyrazolines

as potential anti-inꢁammatory, analgesic and antibacterial agents.

Eur J Med Chem 101(28):442–451. https://doi.org/10.1016/j.

ejmech.2015.07.002

67. Kontogiorgis C, Hadjipavlou-Litina D (2003) Biological evalu-

ation of several coumarin derivatives designed as possible anti-

inꢁammatory/antioxidant agents. J Enzyme Inhib Med Chem

18(1):63–69. https://doi.org/10.1080/1475636031000069291

68. Roussaki M, Kontogiorgis CA, Hadjipavlou-Litina D, Hami-

lakis S, Detsi A (2010) A novel synthesis of 3-aryl coumarins

and evaluation of their antioxidant and lipoxygenase inhibitory

activity. Bioorg Med Chem Lett 20(13):3889–3892. https://doi.

org/10.1016/j.bmcl.2010.05.022

3. He J, Ma L, Wei Z, Zhu J, Peng F, Shao M, Lei L, He L, Tang

M, He L, Wu Y, Chen L (2015) Synthesis and biological evalua-

tion of novel pyrazoline derivatives as potent anti-inꢁammatory

agents. Bioorg Med Chem Lett 25(11):2429–2433. https://doi.

org/10.1016/j.bmcl.2015.03.087

5

4. Eid NM, George RF (2018) Facile synthesis of some pyrazo-

line-based compounds with promising anti-inꢁammatory activ-

ity. Future Med Chem 10(2):183–199. https://doi.org/10.4155/

fmc-2017-0144

69. Denisov ET, Afanas’ev IB (2005) Oxidation and antioxidants

in organic chemistry and biology. CRC Press, Boca Raton

70. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt

DM, Meng EC, Ferrin TE (2004) UCSF Chimera–a visualiza-

tion system for exploratory research and analysis. J Comput

Chem 25(13):1605–1612. https://doi.org/10.1002/jcc.20084

71. Fiser A, Sali A (2003) Modeller: generation and reꢂnement of

homology-based protein structure models. Methods Enzymol

374:461–491. https://doi.org/10.1016/S0076-6879(03)74020-8

72. O’Boyle NM, Banck M, James CA, Morley C, Vandermeersch

T, Hutchison GR (2011) Open Babel: an open chemical toolbox.

J Cheminform 3:33. https://doi.org/10.1186/1758-2946-3-33

73. Halgren TA (1996) Merck molecular force ꢂeld. I. Basis, form,

scope, parameterization, and performance of MMFF94. J Com-

put Chem 17(5–6):490–515. https://doi.org/10.1002/(sici)1096-

987x(199604)17:5/6%3c490::aid-jcc1%3e3.0.co;2-p

5. Turkan F, Cetin A, Taslimi P, Karaman HS, Gulçin İ (2019)

Synthesis, characterization, molecular docking and biological

activities of novel pyrazoline derivatives. Arch Pharm (Weinheim)

3

52(6):e1800359. https://doi.org/10.1002/ardp.201800359

6. Ozmen Ozgun D, Gul HI, Yamali C, Sakagami H, Gulcin I,

Sukuroglu M, Supuran CT (2019) Synthesis and bioactivities of

pyrazoline benzensulfonamides as carbonic anhydrase and ace-

tylcholinesterase inhibitors with low cytotoxicity. Bioorg Chem

8

4:511–517. https://doi.org/10.1016/j.bioorg.2018.12.028

5

7. Stefanes NM, Toigo J, Maioral MF, Jacques AV, Chiaradia-

Delatorre LD, Perondi DM, Ribeiro AAB, Bigolin Á, Pirath

IMS, Duarte BF, Nunes RJ, Santos-Silva MC (2019) Synthesis

of novel pyrazoline derivatives and the evaluation of death mecha-

nisms involved in their antileukemic activity. Bioorg Med Chem

74. Sousa de Silva AW, Vranken WF (2012) ACPYPE—AnteCham-

ber PYthon Parser interface. BMC Res Notes 5:367. https://doi.

org/10.1186/1756-0500-5-367

2

7(2):375–382. https://doi.org/10.1016/j.bmc.2018.12.012

8. Mumtaz A, Majeed A, Zaib S, Ur Rahman S, Hameed S, Saeed

A, Raꢂque H, Mughal E, Maalik A, Hussain I, Iqbal J (2019)

Investigation of potent inhibitors of cholinesterase based on thio-

urea and pyrazoline derivatives: synthesis, inhibition assay and

molecular modeling studies. Bioorg Chem 90:103036. https://doi.

org/10.1016/j.bioorg.2019.103036

75. Wang J, Wang W, Kollman PA, Case DA (2006) Automatic

atom type and bond type perception in molecular mechanical

calculations. J Mol Graph Model 25(2):247–260. https://doi.

org/10.1016/j.jmgm.2005.12.005

76. Lindorꢀ-Larsen K, Piana S, Palmo K, Maragakis P, Klepeis

JL, Dror RO, Shaw DE (2010) Improved side-chain torsion

potentials for the Amber ꢀ99SB protein force ꢂeld. Proteins

78(8):1950–1958. https://doi.org/10.1002/prot.22711

5

6

9. Cai X, Zhao S, Cai Zheng J, Zhu Z, Wei D, Zheng Z, Zhu H, Chen

Y (2019) Synthesis and evaluation of novel D-ring substituted ste-

roidal pyrazolines as potential anti-inꢁammatory agents. Steroids

1

46:70–78. https://doi.org/10.1016/j.steroids.2019.03.012

77. Hess B, Kutzner C, van der Spoel D, Lindhal E (2008)

GROMACS 4: algorithms for highly eꢃcient, load-balanced,

0. Sethiya JP, Bhavsar SP, Shahare HV (2019) Synthesis of poten-

tial anti-inꢁammatory pyrazoline derivatives under ultrasonic

1

3

Molecular Diversity

and scalable molecular simulation. J Chem Theory Comput

Publisher’s Note Springer Nature remains neutral with regard to

4

(3):435–447. https://doi.org/10.1021/ct700301q

jurisdictional claims in published maps and institutional aꢃliations.

7

8. Trott O, Olson AJ (2010) AutoDock Vina: improving the speed

and accuracy of docking with a new scoring function, eꢃcient

optimization, and multithreading. J Comput Chem 31(2):455–461.

https://doi.org/10.1002/jcc.21334

1

3

Article Doi

Novel quinolinone–pyrazoline hybrids: synthesis and evaluation of antioxidant and lipoxygenase inhibitory activity

DOI: 10.1007/s11030-020-10045-x

Source and publish data:

Authors:

Article abstract of DOI:10.1007/s11030-020-10045-x

Full text of DOI:10.1007/s11030-020-10045-x

Products guided by the article

R&D Labs maybe for 26138-64-7

Relevant to this article

Hot Product