Synthesis of 4-aryl-1,2,3-triazolyl appended natural coumarin-related compounds with antiproliferative and radical scavenging activities and intracellular ROS production modification

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Stipaniev, T. Opaak-Bernardi , M. Juki, S. Martinez, L. Glavaš-Obrovac and S. Rai-Mali, New J. Chem.,

2017, DOI: 10.1039/C7NJ01469D.

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ISSN 1144-0546

PAPER

Jason B. Benedict et al.

The role of atropisomers on the photo-reactivity and fatigue of

diarylethene-based metal–organic frameworks

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Synthesis of 4-aryl-1,2,3-triazolyl appended natural coumarin-

related compounds with antiproliferative, radical scavenging

activities and intracellular ROS production modification

Received 00th January 20xx,

Accepted 00th January 20xx

A. Bistrović,^aN. Stipaničev,^aT. Opačak-Bernardi,^bM. Jukić,^bS. Martinez,^cLj. Glavaš-Obrovac*^band

S. Raić-Malić*^a

DOI: 10.1039/x0xx00000x

www.rsc.org/

Novel natural coumarin-based umbelliferone, herniarin and esculetin series linked to hydroxy- and methoxy-substituted

and non-substituted phenyl ring through 1,2,3-triazole spacer were provided by environmentally friendly click chemistry

under microwave irradiation. Antioxidative activity of compounds was evaluated by DPPH-scavenging activity and cyclic

voltammetry assays. While adjacent 6- and 7-hydroxyl groups in coumarins had major contribution on antioxidative power

and decrease of ROS production relative to esculetin, p-methoxy-substituted phenyl moiety had impact on pronounced

and selective antiproliferative activity against chronic leukemia cells in blast crisis (K562) with no significant change in the

ROS generation. 6,7-Dihydroxycoumarin can be considered as a promising scaffold with major contribution to antioxidant

potential and, thereby, further structural modification of 6,7-dihydroxycoumarin linked to aryl-1,2,3-triazole may provide

lead hybrid molecule that may be of interest for potential application to prevent diseases related to the oxidative-stress

imbalance.

imbalance between the production of free radicals and ROS

leads to oxidative stress, which affects various signaling

Introduction

Coumarins (known as 2H-1-benzopyran-2-ones) comprise a

large class of oxygenated heterocyclic secondary metabolites

that are biosynthesized by plants and fruits de novo.^1,2They

are found throughout the plant kingdom, with the richest

sources being the Rutaceae and Umbelliferone.³Most

coumarins can be found as natural components, such as citrus

fruits, vegetables, essential oils, cereals or in food products, as

flavouring additives, bakery and tobacco products, wine and

other alcoholic beverages.⁴Natural and synthetic coumarins

pathways in many diseases like cancer, cardiovascular and

neurodegenerative pathologies.¹³The impact of ROS in cancer

includes gene mutations, structural changes in DNA as well as

abnormal gene expression, modifications of second-messenger

system and blockage of cell–cell communication, depending on

the stage of cancer.^14-16Therefore, antioxidants are very

important for protecting the organisms from oxidative

disorders. Antioxidants are capable of decrease or prevent

oxidation processes through different mechanisms, such as

scavenging free radicals, inhibition of pro-oxidant enzymes or

chelation of transition metal ions.^11,16Both dietary and

enzymatic antioxidants are components of interrelated

systems that interact with each other to control the ROS

production, thereby ensuring adequate defences against

oxidative stress.^17,18Many studies concerning antioxidant

capacity of phenolic compounds of different classes, such as

natural and synthetic coumarins described correlation of their

physico-chemical properties with antioxidant properties and

hydrogen atom transfer (HAT) mechanism by which they are

proposed to act as antioxidants.^19,20Hydroxycoumarins,

produced by many plants, are traditionally used as natural

medicines. Some hydroxycoumarins are powerful chain-

breaking antioxidants that can prevent free radical injury by

scavenging reactive oxygen species.^3,11,21Thus, fraxetin (7,8-

display

a

wide range of biological activities^5-9with the

antioxidant capacity that expresses the ability to deactivate

free radicals, as probably the most important one.^10-12Free

radicals are unstable chemical species, with unpaired

electrons, that are usually highly reactive toward other

species. The action of reactive oxygen species (ROS) results in

alterations and function modulations of key biomolecules. In

healthy organisms, there is a delicate balance between the

production and the removal of free radicals. However, the

^aDepartment of Organic Chemistry, Faculty of Chemical Engineering and

Technology, University of Zagreb, Marulićev trg 20, HR-10000 Zagreb, Croatia. E-

mail: sraic@fkit.hr; Tel.: +385 1 4597 213; Fax: +385 1 4597 250

bDepartment of Medical Chemistry, Biochemistry and Clinical Chemistry, Faculty of

Medicine, Josip Juraj Strossmayer University of Osijek, J. Huttlera 4, HR-31000

Osijek, Croatia

c Department of Electrochemistry, Faculty of Chemical Engineering and Technology,

University of Zagreb, Savska cesta 16, HR-10000 Zagreb, Croatia

† Electronic Supplementary Information (ESI) available: radical scavenging capacity

dihydroxy-6-methylcoumarin)

and

esculetin

(6,7-

dihydroxycoumarin) showed hydroxyl radicals scavenging

activity and inhibited lipid peroxidation in rat brain and liver.^22-

24

(RSC) of compd. after 30 and 60 min, cyclic voltammograms for 16a

18a 18e and 19a

19d, ROS detection of 20d at 10^-4M, ¹H and ¹³C NMR spectra of

novel compounds.

–16e, 17a–17e,

–

Esculetin efficiently attenuated by the oxidative stress

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induced cell damage via its antioxidant protection of cells from molecules. Furthermore, the effect of electron-donating

protein carbonyl and DNA damage induced by H₂O₂.^25,26substituents at both 6 and/or 7 position of coumarin and 4-

Furthermore, 8-aminomethyl substituted esculetin derivatives position of 1,2,3-triazole ring in target 4-aryl-1,2,3-triazole–

improved both the metabolic stability and the antitumor coumarin hybrids on their antioxidative and antiproliferative

activity of esculetin.²⁷Besides, series of hydroxy- and activities was investigated.

polyhydroxy-3-arylcoumarins showed remarkable antioxidant

properties.²⁸In addition, coumarins are known to be potent

lipid peroxidation inhibitors.^29,30It was also found that both

peptide bond and hydroxyl groups in coumarin-3-acylamino

derivatives enhanced their abilities to protect DNA against

radical induced oxidation.³¹On the other hand, among five-

membered heterocycles, compounds with triazole ring have

been reported to show strong antioxidant activity.^32-34

Biological properties of 1,2,3-triazoles may be related to their

specific features, such as active participation in strong

dipole−dipole interacꢀons and hydrogen bonding, that make

1,4-disubstituted 1,2,3-triazoles appropriate isosteres of Z-

amide in which N-3 strongly resembles a carbonyl oxygen.^35,36

The modification on the structure of natural antioxidants and

the conjunction of known antioxidative structures proved to

be efficient ways to increase the antioxidant ability.³⁷

Therefore, coumarin–chalcone and coumarin–dihydropyrazole

hybids exhibited powerful antioxidant effectiveness.^20,38,39

Furthermore, it was found recently that strong

antiproliferative effect of the 7-methylcoumarin–1,2,3-

triazole–2-methylbenzimidazole hybrid was associated with its

inhibition of 5-lipoxygenase activity and perturbation of

sphingolipid signalling pathway.⁴⁰

Results and discussion

Synthesis of coumarin derivatives with 4-phenyl-substituted 1,2,3-

triazole

A series of novel coumarin–1,2,3-triazole hybrids (16a

–16e,

17a 17e 18a–18e, 19a–19d, 20a–20d) was synthesized by the

–

,

copper(I)-catalyzed Huisgen 1,3-dipolar cycloaddition reaction

of azidomethyl coumarin derivatives bearing a hydroxyl or

methoxyl substituent at 6 and/or 7 positions of the coumarin

ring and terminal alkynes (Scheme 1).

4-Azidomethyl coumarin derivatives

prepared by reaction of 4-chloromethyl coumarin derivatives

10) with sodium azide.⁴³Pechmann cyclization catalyzed by

acid was applied for reaction of -ketoester with

corresponding meta- and para-substituted phenol derivatives

and 1,2,3-acetoxybenzene.^27,43Thus, reaction of resorcinol (

),

hydroquinone ( ), 3-methoxyphenol ( ), 4-methoxyphenol (

and 1,2,3-acetoxybenzene with 4-chloroacetoacetate

afforded 4-chloromethyl coumarin precursors 10).

(11–

15

)

were

(6–

a

β

1

2

3

4)

(5)

(

6

–

A

focused library of 23 4-aryl-1,2,3-triazolyl appended coumarin

derivatives was successfully prepared using molecular

hybridization approach based on the coupling of the

pharmacophoric coumarin scaffold and 4-aryl-substituted

1,2,3-triazole moiety.

Taking into consideration the aforementioned and as a

continuation of our research on development of biologically

active molecules,^41,42we herein design a series of natural

coumarin-related compounds, 16a–16e, 17a–17e, 18a–18e,

19a–19d, and 20a–20d, linked to hydroxy- and methoxy-

substituted and non-substituted phenyl ring through 1,2,3-

triazole spacer (Fig. 1) to evaluate their antioxidative and

antiproliferative activities. The 7-hydroxy-, 7-methoxy-, and

6,7-dihydroxy-substituted coumarin skeleton of natural

umbelliferone, herniarin and esculetin, respectively, and 6-

hydroxy- and 6-methoxy-substituted coumarins were applied

for the design of antioxidant molecules. We anticipated that

an inductive effect of nitrogen atoms of 1,2,3-triazole and an

introduction of 4-phenyl-1,2,3-triazole conjugated system

would contribute in radical scavenger ability of novel hybrid

R₂

R₃

H-bond acceptor

R₄

A

N

O

B

C

N

O

H

H-bond donor

R₂, R₃= H, OH, OCH₃

R₄= H, o-OCH, m-OCH₃, p-OCH₃, m-OH

Fig. 1. Structural rational design of 4-aryl-1,2,3-triazolyl–coumarin hybrids (16a–16e,

17a–17e, 18a–18e, 19a–19d, 20a–20d) with indicated structural features that can be

associated with antioxidant activity: the 3,4-double bond in conjugation with carbonyl

group in the A-ring, the presence of hydroxyl, methoxyl groups at the 6 and 7 position

of coumarin (A ring) and 4-phenyl (C ring) and N-3 of the 1,2,3-triazole, which strongly

resembles a carbonyl oxygen (B ring).

Scheme 1. Reagents and conditions: (i) 4-chloroacetoacetate, HClO₄/conc. H₂SO₄, r.t., 6-

8 h; (ii) NaN₃, CH₃CN, reflux, 24 h; (iii) terminal alkyne, Cu(0), 1M CuSO₄, tert-butanol :

H₂O = 1 : 1, DMF, 80 °C, 90 min, MW, 300 W.

2 | J. Name., 2012, 00, 1-3

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Target regioselective 1,4-disubstituted 1,2,3-triazoles (16a

–

can be observed that 4-aryl-1,2,3-triazolyl-substituted

16e, 17a–17e, 18a–18e, 19a–19d, 20a–20d) were prepared by coumarins 20a–20d containing adjacent 6- and 7-hydroxyl

environmentally friendly click chemistry under microwave group exhibited the highest scavenging activities (> 90% after

irradiation in a mixture of water and organic solvent using 30 min) that were comparable to the esculetin and higher than

Cu(I) catalyst, that was generated in situ from copper(II) that of umbelliferone. 4-Phenyl-substituted 1,2,3-triazole

sulfate and metallic copper. Generally, click reaction of 6- and coumarin 20a displayed similar antioxidant potential after 30

7-methoxy- and 7-hydroxy-substituted coumarin azides min relative to the corresponding coumarins

(20b–20d)

proceeded in higher yields ranging from 35% to 98% than containing p-, m- and o-methoxyphenyl-1,2,3-triazole moiety,

reaction of 6-hydroxy- and 6,7-dihydroxycoumarin derivatives but scavenging activity of 20a significantly decreased (65%)

that

coumarin hybrids in the yields of 33-52%. However, derivatives, the presence of one hydroxyl group at 6 or 7

azidomethyl derivatives of 6-hydroxy- and 6,7- position diminished the scavenging activity of tested

dihydroxycoumarins did not afford the target 4-(m- compounds after 30 min of incubation. For 7-hydroxy-

hydroxyphenyl)-1,2,3-triazolyl coumarin derivatives. substituted coumarins, the best scavenging capacity was found

afforded

methoxy-substituted-4-aryl-1,2,3-triazole– after 60 min. In comparison to 6,7-dihydroxycoumarin

Overall, in order to assess the influence of the electron- for 18c (46% at 30 min, 93% at 60 min) and 18e (62% at 30

donating substituents and their position at coumarin and 4- min, 94% at 60 min) bearing m-methoxyphenyl and m-

phenyl ring on both antioxidative and cytostatic activities, hydroxyphenyl substituents at 4-position of 1,2,3-triazole ring,

hydroxyl and methoxyl substituents were introduced at 6- respectively. Furthermore, antioxidant potential of these

and/or 7-position at coumarin core and ortho-, meta- and compounds was somewhat higher than those of their natural

para-position at 4-phenyl-1,2,3-triazole moiety.

analog umbelliferone. Similar substituent effect on scavenging

activity was found for 7-methoxycoumarins showing that m-

position was beneficial for antioxidative effectiveness. Thus, 7-

methoxycoumarin 16c containing m-methoxyphenyl-1,2,3-

triazole fragment and no hydroxyl substituent exhibited

moderate antioxidative effectiveness (56% at 30 min). Notably,

DPPH free radical scavenging activities of 4-aryl-1,2,3-triazolyl-

substituted coumarins

All the synthesized compounds were assessed for their

antioxidant activity by monitoring spectrophotometrically the

ability of the compounds to reduce 1,1-diphenyl-2-

picrylhydrazyl (DPPH), a commonly used stable free radical.

Radical scavenging capacity (RSC), expressed as % of DPPH

all 7-methoxycoumarins 16a–16c and 16e, apart from 16d with

o-methoxyphenyl unit, showed significant enhancement in

their scavenging potencies comparable to reference esculetin

after 60 min. In 6-substituted coumarin series, only 6-

hydroxycoumarins (19a, 73% and 19b, 68%) exhibited marked

scavenging activity at 30 min. Similar enhancement of

antioxidant potential for 16 series after 60 min was also

reduced to DPPH-H, of novel compounds 16a–16e, 17a–17e,

18a–18e, 19a–19d and 20a–20d, and well-known antioxidants

umbelliferone (7-hydroxycoumarin) and esculetin are given in

Table 1.

observed for 7-hydroxy-

substituted 19a 19e

(

18a

, 18c–18e) and 6-hydroxy-

Table 1. DPPH radical scavenging data obtained for compounds 16a–16e, 17a–17e,

18a–18e, 19a–19d, 20a–20d.

(

–

)

coumarin analogs. Overall,

a

comparative assessment of these activities after 30 and 60 min

showed that majority of compounds had radical scavenger

activity higher than 86% after 60 min, while selected coumarin

RSC^a(%)

DPPH % DPPH %

Compd^b

1,2,3-triazole hybrids 20a–20d with the highest antioxidant

30 min

19.4

23.2

56.1

0.0

60 min

100

30 min

7.2

60 min

86.2

93.5

100

capacity exhibited these activities already after 30 min (Table

1, Fig. S1, Supplementary information).

16a

16b

16c

16d

16e

17a

17b

17c

17d

17e

18a

18b

18c

18d

18e

100

61.7

73.2

67.6

32.6

15.8

94.1

92.1

93.5

93.8

81.6

100.0

94.4

23.5

92.8

89.2

48.9

0

19a

19b

100

Electrochemical oxidation potentials of 4-aryl-1,2,3-triazolyl-

substituted coumarins

43.1

13.1

0.1

19c

95.3

94.8

64.6

91.5

94.6

94.7

82.6

100

19d

Cyclic voltammetry (CV) provides a fast and reliable method

for the measurement of the oxidation peak potential of

compounds that is closely related to their antioxidant activity.

Although the electrochemistry of phenol and phenolic

compounds has been intensively studied,^44,45electrochemical

studies describing the mechanistic aspects of coumarin

derivatives are rather limited.⁴⁶The electrochemical oxidation

20a

0

20b

0

4.8

20c

0

15.7

78.5

37.9

92.9

20d

11.1

19.7

46.4

Umbelliferone

Esculetin

^aThe radical scavenging capacity was calculated as follows: [% RSC = 100 -

(A_sample)/A_blank

100], where A_blankis 95% methanol solution; ^b0.2 mM

of 16a–16e, 17a–17e, 18a–18e, 19a–19d, 20a–20d was

x

investigated by CV and their cyclic voltammograms are

presented (Table 2, Fig. 2, Fig. S2-S5, Supplementary

information).

concentration of the tested compounds.

The data clearly indicate that reduction of DPPH by

coumarin–1,2,3-triazole hybrids was time-dependent showing

better antioxidant activities with prolonged time of 60 min. It

Except for 20a, the CV data of evaluated compounds

showed anodic and no cathodic peaks revealing an irreversible

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Table 2. Electrochemical parameters determined from cyclic voltammetric experiments

for compounds 16a–16e, 17a–17e, 18a–18e, 19a–19d, 20a–20d.

1,2,3-triazole–coumarin 20a was the most potent antioxidant.

In consequence, the easiness oxidation rank order was as

follows: 20a

> 20c > 20d > 20b. For the oxidation of the 20a, a

Compd

16a

E_shoulder(V) I_shoulder(μA) E_peak(V) I_peak(μA)

shoulder appeared at less positive potential (0.39 V) before

the main signal at 0.56 V was evidenced. In general, 6- and 7-

hydroxy-monosubstituted coumarin derivatives shifted the

oxidation potentials towards more positive values compared

with 6,7-dihydroxy-substituted coumarins indicating that a

latter compounds are more easily oxidized than the

monosubstituted hydroxycoumarins. It can be observed that

for monosubstituted 7-hydroxy- and 6-hydroxycoumarin

derivatives, 7-position was better suitable deprotonation site

than 6-position. Among 7-substituted coumarins, both 7-

methoxyl and 7-hydroxyl substituents with 4-(m-

hydroxyphenyl)-1,2,3-triazole moiety in 16e and 18e caused

the highest antioxidative capacities. The antioxidative activities

-

0.6957

0.7920

0.7565

0.7676

0.7058

0.7970

0.7362

0.7970

0.7312

0.7159

0.7514

0.7261

0.7869

0.7617

0.7210

0.7464

0.7413

0.7565

0.7210

0.5588

0.7818

-

0.1511

0.0788

0.2803

0.3261

7.417

0.4297

0.1430

0.0821

0.2964

1.781

1.112

2.247

2.192

0.6571

5.699

0.7179

0.5787

2.260

0.4627

40.607

6.117

-

16b

0.4676

0.4980

0.3865

0.0774

0.4980

0.5487

0.5132

0.3815

0.6501

0.6399

0.4017

0.5740

0.5436

0.5284

0.4575

0.4727

0.4626

0.5487

0.3865

0.4119

0.4524

0.3663

0.365

0.0239

0.1110

0.0477

0.5444

0.1837

0.0739

0.0259

0.0460

1.7580

0.5413

0.2797

1.1314

0.1636

3.0615

0.1229

0.1127

0.4922

0.4706

12.454

5.5600

22.309

11.058

49.945

-

16c

16d

16e

17a

17b

17c

17d

17e

18a

18b

18c

18d

for 6-substituted coumarins 17a

substantially lower than those of 7-substituted counterparts

16a and 18a . In the series of 6-substituted coumarins, 6-

hydroxycoumarins 19a exhibited better antioxidative effect

than their 6-methoxycoumarin analogs 17a . It is interesting

–

e

and 19a

–d

were

18e

19a

19b

–

e

–e

19c

–

d

19d

–d

20a

to note that for 7- and 6-methoxy-substituted coumarins 16e

and 17e with m-hydroxyphenyl ring possessed antioxidative

properties showing again that m-hydroxyl group played an

important role in triggering the antioxidative activity. On the

contrary to this, the lack of activity for their analogs 16c and

17c with m-methoxy-substituted phenyl ring was observed.

20b

20c

20d

-

Esculetin

Umbelliferone

0.5954

0.6765

62.851

4.328

-

Biological evaluations

Antiproliferative evaluations

Results of antiproliferative evaluations of 4-aryl-1,2,3-triazolyl-

substituted coumarins 16a–16e, 17a–17e, 18a–18e, 19a–19d,

20a–20d on human cervix adenocarcinoma (HeLa), colon

adenocarcinoma (CaCo-2) and chronic myeloid leukemia in

blast crisis (K562), as well as on the normal Madin Darby

canine kidney (MDCK1) cells are presented in the Table 3.

It can be observed that 1,2,3-triazolyl-substituted 6,7-

dihydroxycoumarins 20a–20d with the most pronounced

antioxidative activity showed also the highest, albeit moderate

cytostatic effects against all treated cells. Compared to other

tumor cell lines, these compounds exhibited better

antiproliferative activity on chronic myeloid leukemia in blast

crisis (K562) with IC₅₀ranging from 17.9 to 28.5 µM. However,

compounds 20a–20d also exhibited similar cytostatic effect

(IC₅₀= 34.3–46 µM) on normal MDCK1 cells. Of the 4-aryl-

Fig. 2. Cyclic voltammograms measured for 20a–20d on carbon paste electrode.

redox reaction at the electrode. For majority of compounds

two anodic hump and peak currents were observed at

and 0.74 V, respectively. The exceptions were m-methoxy- and

o-methoxyphenyl 1,2,3-triazolyl-substituted 6,7-

∼ 0.48 V

dihydroxycoumarins 20c and 20d with one strong oxidation

hump at 0.45 V and 0.37 V, respectively. Besides, 4-phenyl-

1,2,3-triazolyl-substituted 7-methoxycoumarin 16a exhibited

not strong peak at 0.70 V. The presented results showed the

1,2,3-triazolyl-substituted 7-hydroxycoumarin derivatives 18a

–

18e, all compounds were deprived of cytostatic activities

against tested tumor cell lines. Furthermore, 6-methoxy-, 7-

methoxy- and 6-hydroxycoumarin–1,2,3-triazole hybrids

demonstrated the lack of cytotoxic potency. The only

highest oxidation potential of 6,7-dihydroxycoumarins 20a

–

20d indicating their the most pronounced antioxidative

activity, what is in agreement with observed DPPH free radical

scavenging activities for these compounds. Compared to the

natural coumarins, their oxidation potential was higher than

that of umbelliferone and lower relative to esculetin.

Furthermore, in line with results on DPPH method, 4-phenyl-

exceptions were coumarin derivatives 16b

that exerted antiproliferative effect on the growth of the K562

cells. Interestingly, p-methoxyphenyl-1,2,3-triazolyl-

substituted coumarin derivatives 16b 17b and 19b showed to

be the most selective with the highest potency on K562 cells

, 17b, 19a and 19b

,

(

16b: IC₅₀= 8.7 µM, 17b: IC₅₀= 7.5 µM, 19b: IC₅₀= 9.8 µM).

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Table 3. The growth-inhibition effects in vitro of 16a–16e, 17a–17e, 18a–18e, 19a–19d

and 20a–20d on selected tumor cell lines and normal kidney MDCK1 cells.

displayed lower growth inhibition of K562 cells than those of

16b, 17b, 19a and 19b, the former compounds and esculetin

IC₅₀(μM)^a

caused almost complete growth inhibition of K562 cells at

higher applied concentration of 1x10^-4M (Fig. 3).

Compd

ClogP^b

HeLa

>100

90.6 ± 1.0

>100

CaCo-2

>100

K562

>100

MDCK1

54.5 ± 8.1

77.9 ± 16.8

77.2 ± 22.8

>100

Divergences in antiproliferative effects of the investigated

compounds may be explained by their diverse ability to

scavenge radicals and by several other factors, such as

differences in the nature and concentration of ROS production

or their ability to initiate pro-apoptotic signalling pathways.⁴⁷

16a

16b

16c

16d

16e

17a

17b

17c

17d

17e

18a

18b

18c

18d

18e

19a

19b

19c

19d

20a

20b

20c

20d

3.22

3.06

2.91

3.22

3.06

2.91

3.07

2.91

2.77

3.07

2.91

2.77

2.61

8.7 ± 0.9

>100

83.2 ± 30.5

70.7 ± 15.8

48.4 ± 11.7

>100

7.5 ± 0.6

>100

Effects on intracellular ROS production

>100

88.4 ± 24.9

>100

Increased generation of reactive oxygen species (ROS) and an

altered redox status have been observed in cancer cells and,

therefore, this biochemical property of cancer cells can be

used for therapeutic purpose.^48,49With the aim to contribute

to a better understanding of the observed cytotoxic effects of

evaluated coumarin derivatives, ROS (superoxide and hydroxyl

radicals) production of novel structurally related coumarins

>100

63.8 ± 8.1

48.9 ± 15.4

87.3 ± 27.3

84.2 ± 23.2

64.9 ± 12.1

53.3 ± 9.5

67.7 ± 14.8

>100

12.2 ± 1.3

9.8 ± 1.3

16b, 17b, 19b and 20d with strong antiproliferative effects in

K562 cells and radical scavenging capacities was investigated in

treated cells. Measured fluorescence values showed that

compounds 16b and 17b exhibited ROS production level

similar to that of esculetin and lower production of ROS

relative to control untreated cells. However, ROS level in cells

treated with compound 19b was unchanged in comparison to

control cells. Out of the four compounds, selected 4-aryl-1,2,3-

triazolyl-substituted coumarin 20d containing 6,7-dihydroxyl

groups showed a significant shift in the fluorescence (Fig. 4)

compared to both esculetin and control cells.

49.6 ± 9.4 88.1 ± 24.9

>100

81.9 ± 28.3 46.3 ± 4.2 28.5 ± 5.7

59.0 ± 26.2 50.2 ± 5.7 21.8 ± 9.2

41.2 ± 4.9

36.7 ± 2.2

46.0 ± 6.0

34.3 ± 1.5

45.8 ± 2.1

41.6 ± 0.9

44.2 ± 5.2 17.9 ± 5.0

9.7 ± 1.3 24.5 ± 9.3

a

IC₅₀, concentration that inhibited cell’s growth by 50%. Data represents mean

IC₅₀(μM) values ± standard deviation (SD) of three independent experiments.

Exponentially growing cells were treated with compounds during 72 h period.

Cytotoxicity was analysed using MTT survival assay; ^bValues of n-octanol/water

partition coefficients logP were calculated using ChemAxon algorithm

(MarvinView Ver. 6.2.2.).

Namely, total fluorescence value of K562 cells treated with

20d was almost 2-fold lower than that of control cells.

Interestingly, significantly lower fluorescence intensity

Fig. 3. Comparison of growth inhibition of selected compounds and esculetin as a

reference compound on chronic myeloid leukemia in blast crisis (K562) concentration

of 10^-4and 10^-5M.

Besides, these compounds exhibited somewhat enhanced

lipophilicity (ClogP = 2.91–3.07) compared to those of 6,7-

Fig. 4. Intracellular ROS detection in K562 cells. Cells were treated with 10^-5M of tested

substances for 1 h and analyzed by flow cytometry. Fluorescence mean values are

shown in the chart. Histogram overlays show fluorescence patterns for (a) control

(untreated) cells (blue) and esculetin (red), compared to tested compounds (green): (b)

16b, (c) 17b, (d) 19b and (e) 20d. One representative experiment is shown out of three.

Vertical axes: number of events (linear scale); horizontal axes: fluorescence intensity

(log scale).

dihydroxycoumarin derivatives 20a–20d (ClogP = 2.61–2.77).

Cytostatic effects of 16b, 17b and 19b may be correlated with

their radical scavenger capacities (16b: 100%; 17b: 49% and

19b: 100% at 60 min), which were detected by DPPH method.

20d at 10^-5

Although compounds 20a– M concentration

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indicating 4-fold decrease of ROS production was observed selected terminal alkynes. Antioxidant capacity of newly

when K562 cells were treated with compound 20d at prepared compounds was evaluated using DPPH scavenging

concentration of 1 x 10^-4M that caused complete inhibition of activity and cyclic voltammetry assays. It was demonstrated

K562 cells (Fig. S6, Supplementary information).

that 4-aryl-1,2,3-triazolyl 6,7-dihydroxycoumarins 20a–20d

Our results show that compound 20d, besides acting as exhibited the highest antioxidant potency. Moreover, these

strong ROS scavenger, was able to considerably prevent ROS compounds showed unselective cytostatic activities against all

production within the treated cells. On the contrary to this, tested tumor and normal cells. On the contrary, p-

compounds 16b

,

17b and 19b with lower antioxidative methoxyphenyl-1,2,3-triazole subunit in 6- or 7-

17b, and 19b contributed to

activities compared to 20d exhibited also less alteration in ROS monosubstituted coumarins 16b

,

production. This suggests a different mechanism of cytostatic selective inhibitory activity (IC₅₀< 10 µM) against chronic

action for 6- or 7-monosubstituted and 6,7-dihydroxy- leukemia cells in blast crisis (K562 cells). Furthermore,

substituted coumarin series and that strong and selective compound 20d exhibited both considerable radical scavenging

cytostatic effects of 16b

be, along with their radical scavenging activity, ascribed to On the basis of the overall results, we may consider 6,7-

other ROS-independent mechanism. dihydroxycoumarin as promising scaffold that affected

, 17b and 19b against K562 cells may activity and reduction of ROS generation relative to esculetin.

Overall, structure-activity relationship indicated that the antioxidant potential and further structural modification of

type of electron-donating substituents and their position in joined aryl-1,2,3-triazole fragment can lead to improvement in

both coumarin and 1,2,3-triazole scaffolds had impact on their preventing or minimizing the free radicals overproduction that

antioxidative and antiproliferative activities as presented in may be of interest for possible application in oxidative-stress

Fig. 5.

related diseases.

R₂, R₃= OCH₃, OH, H

20a−20d with 6,7-diOH groups led to the highest oxidation

Experimental

R₂

R₃

O

N

O

potential, scavenging activity and unselective

antiproliferative activity

Materials and methods

The order of scavenging activity: 20a−20d with 6,7-diOH >

19a−19d with 6-OH > 16a−16e with 7-OCH₃> 18a−18e

with 7-OH > 17a−17e with 6-OCH₃

All solvents were purified following recommended drying

N

agents and/or distilled over

3 Å molecular sieves. For

m-OH in 16e and 18e increased scavinging and electrochemical activity

o-OCH₃was not beneficial for scavinging activity

monitoring the progress of a reaction and for comparison

purpose, thin layer chromatography (TLC) was performed on

pre-coated Merck silica gel 60F-254 plates using an

appropriate solvent system and the spots were detected under

UV light (254 nm). For column chromatography silica gel

(Fluka, 0.063-0.2 mm) was employed, glass column was slurry-

packed under gravity. Melting points (uncorrected) were

determined with Kofler micro hot-stage (Reichert, Wien). ¹H

and ¹³C NMR spectra were acquired on a Bruker 300 and 600

MHz NMR spectrometer, as well as 300 MHz Agilent

Technologies DD2 NMR spectrometer. All data were recorded

in DMSO-d₆at 298 K. Chemical shifts were referenced to the

R₄

p-OCH₃caused selective effect on K562 cells

R₄= H, p-OCH_3,, o-OCH_3,m-OH

Fig. 5. Insights into structure-activity relationship of a 4-aryl-1,2,3-triazolyl-substituted

coumarin series.

Introduction of adjacent 6- and 7-hydroxyl groups at coumarin

had the crucial effect on scavenging activities in 20a

–20d.

Among the 20a–20d, the highest oxidation potential of 20a

and 20c suggested that unsubstituted- and p-methoxyphenyl-

substituted-phenyl unit at C-4 of 1,2,3-triazole moiety

increased their antioxidative effectiveness. Results on

antiproliferative evaluations on tumor cell lines showed that

residual solvent signal of DMSO at

39.50 ppm for ¹³C. Individual resonances were assigned on the

basis of their chemical shifts, signal intensities, multiplicity of

δ δ

2.50 ppm for ¹H and

compounds 20a–20d with potent antioxidative effect exhibited

resonances and H−H coupling constants. Mass spectra were

unselective cytostatic activities against all tested tumor and

normal cells. While unselective cytotoxicity was concomitant

effect of 6,7-dihydroxyl groups at coumarin ring, p-methoxyl

recorded on an Agilent 6410 instrument equipped with

electrospray interface and triple quadrupole analyzer

(LC/MS/MS). High performance liquid chromatography was

performed on an Agilent 1100 series system with UV detection

(photodiode array detector) using Zorbax C18 reverse-phase

analytical column (2.1 x 30 mm, 3.5 µm). All compounds used

for biological evaluation showed > 95 % purity in this HPLC

system. Microwave-assisted syntheses were performed in a

Milestone start S microwave oven using glass cuvettes at 80 °C

and 800 W under pressure of 1 bar.

substituent at phenyl-1,2,3-triazole in 16b, 17b, and 19b had

impact on selective inhibitory activity against K562 cells.

Conclusions

Novel series of 4-aryl-1,2,3-triazolyl appended natural

coumarin-related derivatives 16a–16e, 17a–17e, 18a–18e,

19a–19d and 20a–20d were successfully synthesized by

convenient, eco-friendly microwave-assisted click reaction of

azidomethyl coumarin derivatives with hydroxyl or methoxyl

substituent at 6 and/or 7 positions of the coumarin ring and

Synthetic procedures

4-(Chloromethyl)-7-hydroxy-2H-chromen-2-one

(

8

),⁴³

27

4-

(chloromethyl)-6,7-dihydroxy-2H-chromen-2-one (10

)

and 4-

6 | J. Name., 2012, 00, 1-3

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ARTICLE

43

)

1

(azidomethyl)-7-hydroxy-2H-chromen-2-one

prepared according to known procedures.

(13

were powder. H NMR (300 MHz, DMSO) δ 7.65 (d, 1H, J = 8.8 Hz,

H5), 7.04 (d, 1H, J = 2.4 Hz, H8), 7.00 (dd, 1H, J = 8.8, 2.5 Hz,

H6), 6.35 (s, 1H, H3), 4.83 (s, 2H, CH₂), 3.87 (s, 3H, OCH₃).¹³C

General procedure for the synthesis of 4-chloromethyl NMR (75 MHz, DMSO) δ 162.63 (C7), 159.94 (C2), 155.11

coumarines (6, 7 and 9). To a mixture of the corresponding (C8a), 150.14 (C4), 125.99 (C5), 112.38 (C6), 110.64 (C4a),

phenol derivate (1 mmol) and ethyl-4-chloroacetate (2 mmol) 110.31 (C3), 101.02 (C8), 55.99 (OCH₃), 49.63 (CH₂). MS (ESI):

perchloric acid (1 mL) was added dropwise. The reaction m/z = 232.1 ([M+H]⁺).

mixture was stirred for 6-8 h at room temperature and

Preparation of 4-(azidomethyl)-6-methoxy-2H-chromen-2-

monitored with TLC. After completion of the reaction, the one (12).⁵³Compound 12 was synthesized according to the

mixture was poured into 50 mL ice-water and the product was general procedure from 4-(chloromethyl)-6-methoxy-2H-

filtered. The crude compound was recrystallized in methanol.

chromen-2-one (7) (600 mg, 2.68 mmol). After purification by

Preparation of 4-(chloromethyl)-7-methoxy-2H-chromen- column chromatography (CH₂Cl₂) compound 12 (725.4 mg,

1

2-one (6) ^50,51

general procedure using 3-methoxyphenol (

mmol). Compound was isolated as light brown solid (1.72 g, = 9.0, 2.9 Hz, H7), 7.19 (d, 1H, J = 2.9 Hz, H5), 6.52 (s, 1H, H3),

.

Compound

6

was synthesized according to the 71%, mp 116–118 °C) was isolated as yellow powder. H NMR

) (1.00 g, 8.05 (300 MHz, DMSO) δ 7.40 (d, 1H, J = 9.0 Hz, H8), 7.26 (dd, 1H, J

1

6

1

95.2%, mp 198–201 °C. H NMR (300 MHz, DMSO-d₆): δ 7.77 4.89 (d, 2H, J = 1.2 Hz, CH₂), 3.83 (s, 3H, OCH₃).¹³C NMR (151

(d, 1H, J = 8.7 Hz, H5), 7.03-7.05 (m, 1H, H6), 7.00 (d, 1H, J = MHz, DMSO) δ 159.61 (C2), 155.57 (C6), 149.71 (C4), 147.43

2.4 Hz, H8), 6.73 (s, 1H, H3), 5.00 (s, 2H, CH₂), 3.87 (s, 3H, (C8a), 119.43 (C8), 117.75 (C7), 117.64 (C4a), 113.85, 107.70,

OCH₃). ¹³C NMR (151 MHz, DMSO-d₆): δ 160.58 (C7), 159.95 55.81 (OCH₃), 49.60 (CH₂). MS (ESI): m/z = 232.1 ([M+H]⁺).

(C2), 155.22 (C8a), 150.82 (C4), 126.34 (C5), 112.30 (C6),

111.96 (C3), 110.41 (C4a), 101.04 (C8), 55.96 (OCH₃), 41.29 one (14). Compound 14 was synthesized according to the

(CH₂). MS (ESI): m/z = 225.0 ([M+H]⁺).

general procedure from 4-(chloromethyl)-6-hydroxy-2H-

Preparation of 4-(chloromethyl)-6-methoxy-2H-chromen- chromen-2-one ( ) (700 mg, 3.33 mmol). After purification by

2-one (7).⁵¹Compound

was synthesized according to the column chromatography (CH₂Cl₂: MeOH = 20: 1) compound 14

general procedure using 4-methoxyphenol (

Preparation of 4-(azidomethyl)-6-hydroxy-2H-chromen-2-

9

7

2

) (2.00 g, 16.11 (450.7 mg, 62%, mp 185–189 °C) was isolated as yellow

mmol). Compound

7

was isolated as light brown solid (613.8 powder. ¹H NMR (300 MHz, DMSO) δ 9.82 (s, 1H, OH), 7.29 (d,

1

mg, 34%, mp 143–146 °C). H NMR (300 MHz, DMSO) δ 7.41 1H, J = 8.9 Hz, H8), 7.08 (dd, 1H, J = 8.9, 2.8 Hz, H7), 7.00 (d,

(d, 1H, J = 9.0 Hz, H8), 7.32 (d, 1H, J = 2.8 Hz, H5), 7.27 (dd, 1H, 1H, J = 2.8 Hz, H5), 6.49 (s, 1H, H3), 4.79 (s, 2H, CH₂).¹³C NMR

J = 9.0, 2.9 Hz, H7), 6.69 (s, 1H, H3), 5.06 (s, 2H, CH₂), 3.84 (s, (151 MHz, DMSO) δ 159.75 (C2), 153.79 (C6), 149.53 (C4),

3H, OCH₃).¹³C NMR (151 MHz, DMSO) δ 159.68 (C2), 155.52 146.41 (C8a), 120.18 (C8), 117.72 (C4a), 117.59 (C7), 113.80

(C6), 150.39 (C4), 147.57 (C8a), 119.41 (C8), 117.82 (C7), (C3), 109.12 (C5), 49.62 (CH₂). MS (ESI): m/z = 218.0 ([M+H]⁺).

117.49 (C4a), 115.68 (C3), 108.26 (C5), 55.83 (OCH₃), 41.33

(CH₂). MS (ESI): m/z = 225.0 ([M+H]⁺).

Preparation

of

4-(azidomethyl)-6,7-dihydroxy-2H-

chromen-2-one (15).⁵⁴Compound 15 was synthesized

Preparation of 4-(chloromethyl)-6-hydroxy-2H-chromen-2- according to the general procedure from 4-(chloromethyl)-6,7-

one (9).⁵²Compound

was synthesized according to the dihydroxy-2H-chromen-2-one (10) (500 mg, 2.21 mmol). After

general procedure from hydroquinone ( ) (2.0 g, 18.16 mmol) purification by column chromatography (CH₂Cl₂: MeOH = 20: 1)

to obtain compound (1.44 g, 38%, mp 219–221 °C) as compound 15 (210.3 mg, 41%, mp > 250 °C) was isolated as

9

4

9

yellowish crystals. ¹H NMR (300 MHz, DMSO) δ 9.84 (s, 1H, light brown powder.¹H NMR (300 MHz, DMSO) δ 10.35 (bs, 1H,

OH), 7.29 (d, 1H, J = 8.9 Hz, H8), 7.15 (d, 1H, J = 2.6 Hz, H5), OH), 9.45 (bs, 1H, OH), 6.98 (s, 1H, H8), 6.78 (s, 1H, H5), 6.25

7.09 (dd, 1H, J = 8.9, 2.7 Hz, H7), 6.65 (s, 1H, H3), 4.97 (s, 2H, (s, 1H, H3), 4.72 (s, 2H, CH₂).¹³C NMR (75 MHz, DMSO) δ

CH₂).¹³C NMR (75 MHz, DMSO) δ 159.83 (C2), 153.74 (C4), 160.35 (C2), 150.54 (C4), 149.87 (C8a), 148.11 (C7), 142.92

150.26 (C6), 146.59 (C8a), 120.24 (C8), 117.67 (C7), 117.57 (C6), 109.56 (C3), 108.97 (C5), 102.85 (C8), 49.77 (CH₂). MS

(C4a), 115.58 (C3), 109.60 (C5), 41.35 (CH₂). MS (ESI): m/z = (ESI): m/z = 234.1 ([M+H]⁺).

211.0 ([M+H]⁺).

General procedure for the synthesis of 4-aryl-1,2,3-triazolyl-

General procedure for the synthesis of 4-azidomethyl substituted coumarins (16a–16e, 17a–17e, 18a–18e, 19a–19d,

coumarins (11, 12, 14 and 15). A mixture of the corresponding 20a–20d. The coumarin azide was dissolved in 0.5 mL DMF and

4-chloromethyl coumarin and sodium azide (2.2 eq) was t-BuOH: H₂O = 1: 1 (2-3 mL). The corresponding terminal

stirred in acetonitrile under reflux overnight. After completion alkyne (1.2 eq), Cu(0) (0.3 eq) and 1M CuSO₄(0.8 eq) were

of the reaction the solvent was removed under reduced added and stirred under microwave irradiation for 1.5 h at 80

pressure and purified by column chromatography.

Preparation of 4-(azidomethyl)-7-methoxy-2H-chromen-2- was purified by column chromatography.

one (11).⁵³Compound 11 was synthesized according to the

Preparation of 7-methoxy-4- (4-phenyl-1H-1,2,3-triazol-1-

general procedure from 4-(chloromethyl)-7-methoxy-2H- yl)methyl -2H-chromen-2-one (16a). Compound 16a was

chromen-2-one ( ) (2.17 g, 9.64 mmol). After purification by synthesized according to the general procedure using 4-

°C and 300 W. The solvent was evaporated and the residue

[

]

6

column chromatography (CH₂Cl₂: MeOH = 50: 1) compound 11 (azidomethyl)-7-methoxy-2H-chromen-2-one (11) (70 mg, 0.30

(718.0 mg, 32%, mp 143–146 °C) was isolated as yellow mmol) and phenylacetylene (0.04 mL, 0.36 mmol). After

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purification by column chromatography (CH₂Cl₂: MeOH = 100: chromatography (CH₂Cl₂: MeOH = 100: 1) compound 16d (33.0

1

1) compound 16a (61.4 mg, 61%, mp 221–224 °C) was isolated mg, 35%, mp 225–228 °C) was isolated as yellow crystals. H

as yellow powder. ¹H NMR (300 MHz, DMSO) δ 8.71 (s, 1H, NMR (300 MHz, DMSO) δ 8.60 (s, 1H, H5'), 8.18 (dd, 1H, J = 7.7,

H5'), 7.86 (d, 2H, J = 7.3 Hz, Ph), 7.77 (d, 1H, J = 8.8 Hz, H5), 1.6 Hz, Ph), 7.84 (d, 1H, J = 8.8 Hz, H5), 7.40–7.32 (m, 1H, Ph),

7.45 (t, 2H, J = 7.5 Hz, Ph), 7.34 (t, 1H, J = 7.3 Hz, Ph), 7.07 (d, 7.15 (d, 1H, J = 8.2 Hz, H8), 7.11–7.00 (m, 3H, H6 and Ph), 6.01

1H, J = 2.3 Hz, H8), 7.01 (dd, 1H, J = 8.8, 2.4 Hz, H6), 5.99 (s, 2H, (s, 2H, CH₂), 5.66 (s, 1H, H3), 3.91 (s, 3H, OCH₃), 3.88 (s, 3H,

CH₂), 5.87 (s, 1H, H3), 3.86 (s, 3H, OCH3).¹³C NMR (151 MHz, OCH₃). ¹³C NMR (75 MHz, DMSO) δ 162.76 (C7), 159.81 (C2),

DMSO) δ 162.73 (C7), 159.80 (C2), 155.12 (C8a), 149.80 (C4), 155.38 (C8a), 155.04 (Ph-q), 150.69 (C4), 142.20 (C4'), 129.16

146.79 (C4'), 130.34 (Ph-q), 128.91 (Ph), 128.07 (Ph), 125.85 (Ph), 126.57 (Ph), 125.90 (C5), 124.99 (C5'), 120.66 (Ph-q),

(C5), 125.23 (Ph), 122.33 (C5'), 112.43 (C6), 111.06 (C3), 118.69, 112.44 (C6), 111.58 (C3), 110.54 (C4a), 110.03 (Ph),

110.54 (C4a), 101.15 (C8), 56.00 (OCH₃), 49.42 (CH₂). MS (ESI): 101.11 (C8), 56.02 (OCH₃), 55.49 (OCH₃), 49.16 (CH₂). MS (ESI):

m/z = 334.1 ([M+H]⁺). Anal. calcd. for C₁₉H₁₅N₃O₃: C, 68.46; H, m/z = 364.1 ([M+H]⁺). Anal. calcd. for C₂₀H₁₇N₃O₄: C, 66.11; H,

4.54; N, 12.61. Found: C, 68.22; H, 4.57; N, 12.73.

Preparation of 7-methoxy-4-{ 4-(4-methoxyphenyl)-1H-

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one (16b). yl

4.72; N, 11.56. Found: C, 66.28; H, 5.01; N, 11.72.

Preparation of 4-{ 4-(3-hydroxyphenyl)-1H-1,2,3-triazol-1-

methyl}-7-methoxy-2H-chromen-2-one (16e). Compound

[

]

Compound 16b was synthesized according to the general 16e was synthesized according to the general procedure using

procedure using 4-(azidomethyl)-7-methoxy-2H-chromen-2- 4-(azidomethyl)-7-methoxy-2H-chromen-2-one (11) (60 mg,

one (11) (70 mg, 0.30 mmol) and 1-ethynyl-4-methoxybenzene 0.23 mmol) and 3-ethynylphenol (0.04 mL, 0.29 mmol). After

(0.05 mL, 0.30 mmol). After purification by column purification by column chromatography (CH₂Cl₂: MeOH = 60: 1)

chromatography (CH₂Cl₂: MeOH = 100: 1) compound 16b compound 16e (61.4 mg, 61%, mp 221–224 °C) was isolated as

(108.1 mg, 98%, mp 235–238 °C) was isolated as white light brown powder.¹H NMR (300 MHz, DMSO) δ 9.54 (s, 1H

1

powder. H NMR (300 MHz, DMSO) δ 8.58 (s, 1H, H5'), 7.86– OH), 8.64 (s, 1H, H5'), 7.77 (d, 1H, J = 8.8 Hz, H5), 7.27–7.21 (m,

7.70 (m, 3H, H5 and Ph), 7.08 (d, 1H, J = 2.4 Hz, H8), 7.05–6.98 2H, Ph), 7.07 (d, 1H, J = 2.4 Hz, H8), 7.05–6.97 (m, 1H, H6),

(m, 3H, H6 and Ph), 5.96 (s, 2H, CH₂), 5.86 (s, 1H, H3), 3.86 (s, 6.97–6.86 (m, 1H, Ph), 6.70–6.77 (m, 1H, Ph), 5.96 (s, 2H, CH₂),

3H, s OCH₃), 3.78 (s, 3H, OCH₃). ¹³C NMR (75 MHz, DMSO) δ 5.85 (s, 1H, H3), 3.86 (s, 3H, OCH₃). ¹³C NMR (75 MHz, DMSO) δ

162.73 (C7), 159.83 (C2), 159.14 (Ph-q), 155.12 (C8a), 149.90 162.73 (C7), 159.82 (C2), 157.76 (Ph-q), 155.12 (C8a), 149.83

(C4), 146.75 (C4'), 126.61 (Ph), 125.86 (C5), 122.92 (Ph-q), (C4), 146.87 (C4'), 131.53 (Ph-q), 129.97 (Ph), 125.86 (C5),

121.35 (C5'), 114.32 (Ph), 112.43 (C6), 111.01 (C3), 110.56 122.30 (C5'), 116.11 (Ph), 115.10 (Ph), 112.43 (C6), 111.93

(C4a), 101.14 (C8), 56.01 (OCH₃), 55.15 (OCH₃), 49.38 (CH₂). MS (Ph), 111.05 (C3), 110.55 (C4a), 101.15 (C8), 56.01 (OCH₃),

(ESI): m/z = 364.1 ([M+H]⁺). Anal. calcd. for C₂₀H₁₇N₃O₄: C, 49.39 (CH₂). MS (ESI): m/z = 350.1 ([M+H]⁺). Anal. calcd. for

66.11; H, 4.72; N, 11.56. Found: C, 66.23; H, 4.70; N, 12.06.

Preparation of 7-methoxy-4-{ 4-(3-methoxyphenyl)-1H- 4.51; N, 12.07.

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one (16c). Preparation of 6-methoxy-4-{

Compound 16c was synthesized according to the general yl methyl}-2H-chromen-2-one (17a). Compound 17a was

C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33; N, 12.03. Found: C, 64.99; H,

[

]

[4-phenyl-1H-1,2,3-triazol-1-

]

procedure using 4-(azidomethyl)-7-methoxy-2H-chromen-2- synthesized according to the general procedure using 4-

one (11) (75 mg, 0.32 mmol) and 1-ethynyl-3-methoxybenzene (azidomethyl)-6-methoxy-2H-chromen-2-one (12) (70 mg, 0.30

(0.05 mL, 0.39 mmol). After purification by column mmol) and phenylacetylene (0.04 mL, 0.36 mmol). After

chromatography (CH₂Cl₂: MeOH = 100: 1) compound 16c purification by column chromatography (CH₂Cl₂: MeOH = 100:

(113.0 mg, 96%, mp 204–207 °C) was isolated as white 1) compound 17a (81.7 mg, 81%, mp 213–215 °C) was isolated

powder. ¹H NMR (300 MHz, DMSO) δ 8.72 (s, 1H, H5'), 7.76 (d, as yellow crystals.¹H NMR (300 MHz, DMSO) δ 8.73 (s, 1H,

1H, J = 8.9 Hz, H5), 7.47–7.40 (m, 2H, Ph), 7.36 (t, 1H, J = 7.8 H5'), 7.87 (d, 2H, J = 7.1 Hz, Ph), 7.53–7.23 (m, 6H, H8, H5, H7

Hz, Ph), 7.07 (d, 1H, J = 2.4 Hz, H8), 7.01 (dd, 1H, J = 8.8, 2.5 Hz, and Ph), 6.11–6.01 (m, 3H, H3 and CH₂), 63.82 (s, 3H, OCH₃).¹³C

H6), 6.95–6.88 (m,1H, Ph), 5.98 (s, 2H, CH₂), 5.89 (s, 1H, H3), NMR (75 MHz, DMSO) δ 159.53 (C2), 155.61 (C6), 149.38 (C4),

3.86 (s, 3H, OCH₃), 3.81 (s, 3H, OCH₃). ¹³C NMR (75 MHz, 147.45 (C8a), 146.88 (C4'), 130.31 (Ph-q), 128.93 (Ph), 128.10

DMSO) δ 162.74 (C7), 159.81 (C2), 159.68 (Ph-q), 155.13 (C8a), (Ph), 125.24 (Ph), 122.38 (C5'), 119.57 (C8), 117.88 (C7),

149.75 (C4), 146.70 (C4'), 131.65 (Ph-q), 130.06 (Ph), 125.85 117.56 (C4a), 114.86 (C3), 107.76 (C5), 55.82 (OCH₃), 49.47

(C5), 122.55 (C5'), 117.56 (Ph), 113.85 (Ph), 112.44 (C6), (CH₂). MS (ESI): m/z = 334.1 ([M+H^]+). Anal. calcd. for

111.14 (C3), 110.53 (C4a), 110.41 (Ph), 101.16 (C8), 56.01 C₁₉H₁₅N₃O₃: C, 68.46; H, 4.54; N, 12.61. Found: C, 68.43; H,

(OCH₃), 55.12 (OCH₃), 49.45 (CH₂). MS (ESI): m/z = 364.1 4.31; N, 12.55.

([M+H]⁺). Anal. calcd. for C₂₀H₁₇N₃O₄: C, 66.11; H, 4.72; N,

11.56. Found: C, 65.94; H, 4.72; N, 11.39.

Preparation of 6-methoxy-4-{

[

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one

4-(4-methoxyphenyl)-1H-

(17b).

]

Preparation of 7-methoxy-4-{

[

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one

4-(2-methoxyphenyl)-1H- Compound 17b was synthesized according to the general

(16d). procedure using 4-(azidomethyl)-6-methoxy-2H-chromen-2-

]

Compound 16d was synthesized according to the general one (12) (70 mg, 0.30 mmol) and 1-ethynyl-4-methoxybenzene

procedure using 4-(azidomethyl)-7-methoxy-2H-chromen-2- (0.04 mL, 0.36 mmol). After purification by column

one (11) (60 mg, 0.23 mmol) and 1-ethynyl-2-methoxybenzene chromatography (CH₂Cl₂: MeOH = 100: 1) compound 17b (86.8

(0.04 mL, 0.29 mmol). After purification by column mg, 79%, mp 237–240 °C) was isolated as yellowish crystals. ¹H

8 | J. Name., 2012, 00, 1-3

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ARTICLE

NMR (300 MHz, DMSO) δ 8.61 (s, 1H, H5'), 7.79 (d, 2H, J = 8.8 8.66 (s, 1H, H5'), 7.42 (d, 1H, J = 8.9 Hz, H8), 7.34–7.19 (m, 5H,

Hz, Ph), 7.42 (d, 1H, J = 9.0 Hz, H8), 7.33 (d, 1H, J = 2.7 Hz, H5), H5, H7 and Ph), 6.78–6.69 (m, 1H, Ph), 6.03–5.99 (m, 3H, H3

7.28 (dd, 1H, J = 9.0, 2.8 Hz, H7), 7.02 (d, 2H, J = 8.8 Hz, Ph), and CH₂), 3.81 (s, 3H, OCH₃). ¹³C NMR (151 MHz, DMSO) δ

6.05 (s, 1H, H3), 6.02 (d, 2H, J = 8.6 Hz, CH₂), 3.82 (s, 3H, OCH₃), 159.53 (C2), 157.77 (Ph-q), 155.60 (C6), 149.37 (C4), 147.45

3.79 (s, 3H, OCH₃). ¹³C NMR (151 MHz, DMSO) δ 159.59 (C2), (C8a), 146.95 (C4'), 131.50 (Ph-q), 129.97 (Ph), 122.33 (C5'),

159.17 (Ph-q), 155.62 (C6), 149.52 (C4), 147.47 (C8a), 146.86 119.54 (C8), 118.47, 117.87 (C7), 117.57 (C4a), 116.11 (Ph),

(C4'), 126.64 (Ph), 122.91 (Ph-q), 121.43 (C5'), 119.59 (C8), 115.12 (Ph), 114.84 (C3), 111.94 (Ph), 107.76 (C5), 55.81

117.92 (C7), 117.60 (C4a), 114.81 (C3), 114.35 (Ph), 107.77 (OCH₃), 49.43 (CH₂). MS (ESI): m/z = 350.1 ([M+H]⁺). Anal.

(C5), 55.84 (OCH₃), 55.18 (OCH₃), 49.46 (CH₂). MS (ESI): m/z = calcd. for C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33; N, 12.03. Found: C,

364.1 ([M+H]⁺). Anal. calcd. for C₂₀H₁₇N₃O₄: C, 66.11; H, 4.72; 65.47; H, 4.44; N, 11.99.

N, 11.56. Found: C, 65.98; H, 4.86; N, 11.43.

Preparation of 6-methoxy-4-{ 4-(3-methoxyphenyl)-1H- yl

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one (17c). synthesized according to the general procedure using 4-

Preparation of 7-hydroxy-4-{

[4-phenyl-1H-1,2,3-triazol-1-

[

]methyl}-2H-chromen-2-one (18a). Compound 18a was

]

Compound 17c was synthesized according to the general (azidomethyl)-7-hydroxy-2H-chromen-2-one (13) (70 mg, 0.30

procedure using 4-(azidomethyl)-6-methoxy-2H-chromen-2- mmol) and phenylacetylene (0.04 mL, 0.30 mmol). After

one (12) (60 mg, 0.26 mmol) and 1-ethynyl-3-methoxybenzene purification by column chromatography (CH₂Cl₂: MeOH = 100:

(0.04 mL, 0.29 mmol). After purification by column 1) compound 18a (59.3 mg, 58%, mp 165–173 °C) was isolated

1

chromatography (CH₂Cl₂: MeOH = 100: 1) compound 17c (72.6 as yellow crystals. H NMR (300 MHz, DMSO) δ 10.70 (bs, 1H,

mg, 77%, mp 200–204 °C) was isolated as light brown OH), 8.70 (s, 1H, H5'), 7.87 (d, 2H, J = 7.1 Hz, Ph), 7.69 (d, 1H, J

powder.¹H NMR (300 MHz, DMSO) δ 8.74 (s, 1H, H5'), 7.47– = 8.7 Hz, H5), 7.46 (t, 2H, J = 7.5 Hz, Ph), 7.35 (t, 1H, J = 7.3 Hz,

7.24 (m, 6H, H8, H5, H7 and Ph), 6.92 (ddd, 1H, J = 8.2, 2.4, 1.2 Ph), 6.84 (dd, 1H, J = 8.7, 2.4 Hz, H6), 6.77 (d, 1H, J = 2.3 Hz,

Hz, Ph), 6.07 (s, 1H, H3), 6.04 (s, 2H, CH₂), 3.81 (m, 6H, OCH₃). H8), 5.95 (s, 2H, CH₂), 5.77 (s, 1H, H3).¹³C NMR (75 MHz,

¹³C NMR (75 MHz, DMSO) δ 159.69 (Ph-q), 159.54 (C2), 155.61 DMSO) δ 161.66 (C7), 159.94 (C2), 155.17 (C8a), 149.99 (C4),

(C6), 149.32 (C4), 147.46 (C8a), 146.80 (C4'), 131.63 (Ph-q), 146.77 (C4'), 130.35 (Ph-q), 128.91 (Ph), 128.07 (Ph), 126.04

130.09 (Ph), 122.59 (C5'), 119.57 (C8), 117.89 (C7), 117.57 (C5), 125.23 (Ph), 122.34 (C5'), 113.21 (C6), 109.97 (C3),

(Ph), 114.95 (C3), 113.89 (Ph), 110.43 (Ph), 107.77 (C5), 55.82 109.40 (C4a), 102.55(C8), 49.40 (CH₂). MS (ESI): m/z = 320.1

(OCH₃), 55.13 (OCH₃), 49.49 (CH₂). MS (ESI): m/z = 364.1 ([M+H]⁺). Anal. calcd. for C₁₈H₁₃N₃O₃: C, 67.71; H, 4.10; N,

([M+H]⁺). Anal. calcd. for C₂₀H₁₇N₃O₄: C, 66.11; H, 4.72; N, 13.16. Found: C, 68.0; H, 4.36; N, 13.09.

11.56. Found: C, 66.17; H, 4.62; N, 11.53.

Preparation of 6-methoxy-4-{ 4-(2-methoxyphenyl)-1H- 1,2,3-triazol-1-yl

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one (17d). Compound 18b was synthesized according to the general

Compound 17d was synthesized according to the general procedure using 4-(azidomethyl)-7-hydroxy-2H-chromen-2-one

procedure using 4-(azidomethyl)-6-methoxy-2H-chromen-2- 13) (70 mg, 0.32 mmol) and 1-ethynyl-4-methoxybenzene

Preparation of 7-hydroxy-4-{

[

4-(4-methoxyphenyl)-1H-

[

]

methyl}-2H-chromen-2-one

(18b).

]

(

one (12) (60 mg, 0.26 mmol) and 1-ethynyl-2-methoxybenzene (0.04 mL, 0.32 mmol). After purification by column

(0.04 mL, 0.29 mmol). After purification by column chromatography (CH₂Cl₂: MeOH = 100: 1) compound 18b (70.0

chromatography (CH₂Cl₂: MeOH = 100: 1) compound 17d (86.8 mg, 63%, mp 110–113 °C) was isolated as white powder. ¹H

mg, 92%, mp 186–189 °C) was isolated as white powder. ¹H NMR (600 MHz, DMSO) δ 10.68 (bs, 1H, OH), 8.57 (s, 1H, H5'),

NMR (300 MHz, DMSO) δ 8.63 (s, 1H, H5'), 8.18 (dd, 1H, J = 7.7, 7.78 (d, 2H, J = 8.8 Hz, Ph), 7.69 (d, 1H, J = 8.8 Hz, H5), 7.01 (d,

1.6 Hz, Ph), 7.47–7.24 (m, 4H, H5 and H6, H7 and Ph), 7.29 (dd, 2H, J = 8.8 Hz, Ph), 6.83 (dd, 1H, J = 8.7, 2.3 Hz, H6), 6.77 (d,

1H, J = 9.0, 2.9 Hz, H7), 7.15 (d, 1H, J = 8.0 Hz, Ph), 7.08 (td, 1H, 1H, J = 2.3 Hz, H8), 5.92 (s, 2H, CH₂), 5.76 (s, 1H, H3), 3.79 (s,

J = 7.6, 0.9 Hz, Ph), 6.06 (s, 2H, CH₂), 5.90 (s, 1H, H3), 3.91 (s, 3H, OCH₃).¹³C NMR (75 MHz, DMSO) δ 161.62 (C7), 159.95

3H, OCH₃), 3.84 (s, 3H, OCH₃). ¹³C NMR (75 MHz, DMSO) δ (C2), 159.13 (Ph-q), 155.16 (C8a), 150.08 (C4), 146.72 (C4'),

159.55 (C2), 155.63 (C6), 155.39 (Ph-q), 150.13 (C4), 147.40 126.60 (Ph), 126.05 (C5), 122.93 (Ph-q), 121.35 (C5'), 114.31

(C8a), 142.25 (C4'), 129.19 (Ph), 126.59 (Ph), 125.05 (C5'), (Ph), 113.19 (C6), 109.94 (C3), 109.43 (C4a), 102.54 (C8), 55.15

120.68 (Ph), 119.65 (C8), 118.67 (Ph-q), 117.88 (C7), 117.57 (OCH₃), 49.36 (CH₂). MS (ESI): m/z = 350.1 ([M+H]⁺). Anal.

(C4a), 114.04 (C3), 111.58 (Ph), 107.75 (C5), 55.85 (OCH₃), calcd. for C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33; N, 12.03. Found: C,

55.48 (OCH₃), 49.25 (CH₂). MS (ESI): m/z = 364.1 ([M+H]⁺). 65.65; H, 4.08; N, 12.19.

Anal. calcd. for C₂₀H₁₇N₃O₄: C, 66.11; H, 4.72; N, 11.56. Found:

C, 66.; H, 4.86; N, 11.43.

Preparation of 7-hydroxy-4-{

[

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one

4-(3-methoxyphenyl)-1H-

(18c).

]

Preparation of 4-{

methyl}-6-methoxy-2H-chromen-2-one (17e). Compound procedure using 4-(azidomethyl)-7-hydroxy-2H-chromen-2-one

17e was synthesized according to the general procedure using 13) (70 mg, 0.32 mmol) and 1-ethynyl-3-methoxybenzene

[4-(3-hydroxyphenyl)-1H-1,2,3-triazol-1- Compound 18c was synthesized according to the general

yl

]

(

4-(azidomethyl)-6-methoxy-2H-chromen-2-one (12) (60 mg, (0.04 mL, 0.32 mmol). After purification by column

0.26 mmol) and 3-ethynylphenol (0.04 mL, 0.29 mmol). After chromatography (CH₂Cl₂: MeOH = 100: 1) compound 18c (81.5

purification by column chromatography (CH₂Cl₂: MeOH = 60: 1) mg, 73%, mp 248–251 °C) was isolated as white powder.¹H

compound 17d (36.5 mg, 40%, mp 235–238 °C) was isolated as NMR (600 MHz, DMSO) δ 10.69 (bs, 1H, OH), 8.71 (s, 1H, H5'),

white powder.¹H NMR (300 MHz, DMSO) δ 9.55 (s, 1H, OH), 7.69 (s, 1H, H5), 7.46–7.42 (m, 2H, Ph), 7.36 (t, 1H, J = 7.9 Hz,

J. Name., 2013, 00, 1-3 | 9

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ARTICLE

Journal Name

Ph), 6.92 (dd, 1H, J = 8.1, 1.9 Hz, H6), 6.83 (d, 1H, J = 6.5 Hz, 149.08 (C4), 146.83 (C4'), 146.46 (C8), 130.34 (Ph-q), 128.93

Ph), 6.77 (s, 1H, H8), 5.94 (s, 2H, CH₂), 5.78 (s, 1H, H3), 3.81 (s, (Ph), 128.09 (Ph), 125.25 (Ph), 122.26 (C5'), 120.38 (C8),

3H, s, OCH₃). ¹³C NMR (151 MHz, DMSO) δ 161.61 (C7), 159.90 117.72 (C7), 114.98 (C3), 109.09 (C5), 49.47 (CH2). MS (ESI):

(C2), 159.65 (Ph-q), 149.87 (C8a), 146.66 (C4'), 131.64 (Ph-q), m/z = 320.1 ([M+H]⁺). Anal. calcd. for C₁₈H₁₃N₃O₃: C, 67.71; H,

130.01 (Ph), 126.00 (C5), 122.51 (C5'), 117.53 (Ph), 113.82 4.10; N, 13.16. Found: C, 67.72; H, 3.91; N, 12.96.

(C3), 113.20 (Ph), 110.41 (Ph), 110.03 (C3), 109.37 (C4a),

102.52 (C8), 55.09 (OCH₃), 49.39 (CH₂). MS (ESI): m/z = 350.1 1,2,3-triazol-1-yl

([M+H]⁺). Anal. calcd. for C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33; N, Compound 19b was synthesized according to the general

12.03. Found: C, 65.25; H, 4.20; N, 12.13. procedure using 4-(azidomethyl)-6-hydroxy-2H-chromen-2-one

Preparation of 7-hydroxy-4-{ 4-(2-methoxyphenyl)-1H- 14) (70 mg, 0.32 mmol) and 1-ethynyl-4-methoxybenzene

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one (18d). (0.05 mL, 0.38 mmol). After purification by column

Preparation of 6-hydroxy-4-{

[

4-(4-methoxyphenyl)-1H-

]

methyl}-2H-chromen-2-one

(19b).

[

(

]

Compound 18d was synthesized according to the general chromatography (CH₂Cl₂: MeOH = 20: 1) compound 19b (41.2

procedure using 4-(azidomethyl)-7-hydroxy-2H-chromen-2-one mg, 37%, mp > 280 °C) was isolated as yellowish powder.¹H

(

13) (70 mg, 0.32 mmol) and 1-ethynyl-2-methoxybenzene NMR (300 MHz, DMSO) δ 9.86 (s, 1H, OH), 8.57 (s, 1H, H5'),

(0.04 mL, 0.32 mmol). After purification by column 7.79 (d, 2H, J = 8.7 Hz, Ph), 7.31 (d, 1H, J = 8.6 Hz, H8), 7.14–

chromatography (CH₂Cl₂: MeOH = 100: 1) compound 18d (55.7 7.05 (m, 2H, H7 and H5), 7.02 (d, 2H, J = 8.8 Hz, Ph), 6.06 (s,

mg, 49%, mp 265–267 °C) was isolated as yellowish powder. ¹H 1H, H3), 5.92 (s, 2H, CH₂), 3.79 (s, 3H, OCH₃).¹³C NMR (151

NMR (600 MHz, DMSO) δ 10.78 (bs, 1H, OH), 8.59 (s, 1H, H5'), MHz, DMSO) δ 159.67 (C2), 159.13 (Ph-q), 153.86 (C6), 149.14

8.18 (dd, 1H, J = 7.7, 1.6 Hz, Ph), 7.75 (d, 1H, J = 8.8 Hz, H5), (C4), 146.74 (C4'), 146.43 (C8a), 126.59 (Ph), 122.92 (Ph-q),

7.40–7.32 (m, 1H, Ph), 7.15 (d, 1H, J = 8.1 Hz, Ph), 7.09–7.06 121.25 (C5'), 120.33 (C8), 117.70 (C4a), 117.68 (C7), 114.92

(m, 1H, Ph), 6.85 (dd, 1H, J = 8.7, 2.3 Hz, H6), 6.77 (d, 1H, J = (C3), 114.31 (Ph), 109.07 (C5), 55.14 (OCH₃), 49.40 (CH₂). MS

2.3 Hz, H8), 5.97 (s, 2H, CH₂), 5.56 (s, 1H, H3), 3.91 (s, 3H, (ESI): m/z = 350.1 ([M+H]⁺). Anal. calcd. for C₁₉H₁₅N₃O₄: C,

OCH₃).¹³C NMR (151 MHz, DMSO) δ 161.87 (C7), 159.94 (C2), 65.32; H, 4.33; N, 12.03. Found: C, 65.55; H, 4.27; N, 11.81.

155.37 (Ph-q), 155.10 (C8a), 150.83 (C4), 142.16 (C4'), 129.12

Preparation of 6-hydroxy-4-{[4-(3-methoxyphenyl)-1H-

(Ph), 126.56 (Ph), 126.01 (C5), 124.96 (C5'), 120.64 (Ph), 1,2,3-triazol-1-yl)methyl)-2H-chromen-2-one (19c). Compound

118.71 (Ph-q), 113.24 (C6), 111.56 (C8), 109.27 (C4a), 108.82 19c was synthesized according to the general procedure using

(C3), 102.51 (Ph), 55.47 (OCH₃), 49.12 (CH₂). MS (ESI): m/z = 4-(azidomethyl)-6-hydroxy-2H-chromen-2-one

350.1 ([M+H]⁺). Anal. calcd. for C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33; 0.28 mmol) and 1-ethynyl-3-methoxybenzene (0.04 mL, 0.30

N, 12.03. Found: C, 65.67; H, 3.97; N, 12.36. mmol). After purification by column chromatography (CH₂Cl₂:

Preparation of 7-hydroxy-4-{ 4-(3-hydroxyphenyl)-1H- MeOH = 20: 1) compound 19c (32.1 mg, 33%, mp 250–253 °C)

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one

(18e). was isolated as light brown powder.¹H NMR (300 MHz, DMSO)

(14) (60 mg,

[

]

Compound 18e was synthesized according to the general δ 9.88 (s, 1H, OH), 8.70 (s, 1H, H5'), 7.37–7.23 (m, 4H, H8 and

procedure using 4-(azidomethyl)-7-hydroxy-2H-chromen-2-one Ph), 7.12–7.05 (m, 2H, H7 and H5), 6.95–6.89 (m, 1H, Ph), 6.08

13) (70 mg, 0.32 mmol) and 3-ethynylphenol (0.03 mL, 0.32 (s, 1H, H3), 5.94 (s, 2H, CH₂), 3.81 (s, 3H, OCH₃). ¹³C NMR (151

(

mmol). After purification by column chromatography (CH₂Cl₂: MHz, DMSO) δ 159.73 (C2), 159.71 (Ph-q), 153.91 (C6), 149.06

MeOH = 10: 1) compound 18e (83.1 mg, 77%, mp 240–243 °C) (C4), 146.76 (C4'), 146.48 (C8a), 131.69 (Ph-q), 130.13 (Ph),

was isolated as light brown powder.¹H NMR (300 MHz, DMSO) 122.51 (C5'), 120.40 (C8), 117.77 (C7), 117.73 (C4a), 117.58

δ 8.63 (s, 1H, H5'), 7.66 (d, 1H, J = 8.8 Hz, H5), 7.32–7.19 (m, (Ph), 115.12 (Ph), 113.90 (C3), 110.43 (Ph), 109.11 (C5), 55.16

3H, Ph), 6.83–6.67 (m, 3H, H6, H8 and Ph), 5.91 (s, 2H, CH₂), (OCH₃), 49.53 (CH₂). MS (ESI): m/z = 350.1 ([M+H]⁺). Anal.

5.68 (s, 1H, H3).¹³C NMR (75 MHz, DMSO) δ 160.12 (C7), calcd. for C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33; N, 12.03. Found: C,

157.79 (C2), 150.12 (C4), 146.84 (C4'), 131.54 (Ph-q), 129.96 65.14; H, 4.12; N, 12.35.

(Ph), 125.87 (C5), 122.31 (C5'), 116.09 (Ph), 115.10 (C6),

111.94 (C8), 102.64 (Ph), 49.38 (CH₂). MS (ESI): m/z = 336.1 1,2,3-triazol-1-yl

([M+H]⁺). Anal. calcd. for C₁₈H₁₃N₃O₄: C, 64.48; H, 3.91; N, Compound 19d was synthesized according to the general

12.53. Found: C, 64.49; H, 3.76; N, 12.29. procedure using 4-(azidomethyl)-6-hydroxy-2H-chromen-2-one

Preparation of 6-hydroxy-4- (4-phenyl-1H-1,2,3-triazol-1- 14) (60 mg, 0.28 mmol) and 1-ethynyl-2-methoxybenzene

yl)methyl -2H-chromen-2-one (19a). Compound 19a was (0.04 mL, 0.30 mmol). After purification by column

Preparation of 6-hydroxy-4-{

[

4-(2-methoxyphenyl)-1H-

]

methyl}-2H-chromen-2-one

(19d).

[

(

]

synthesized according to the general procedure using 4- chromatography (CH₂Cl₂: MeOH = 20: 1) compound 19d (42.6

(azidomethyl)-6-hydroxy-2H-chromen-2-one (14) (70 mg, 0.32 mg, 44%, mp > 280 °C) was isolated as white powder.¹H NMR

mmol) and phenylacetylene (0.04 mL, 0.39 mmol). After (300 MHz, DMSO) δ 9.89 (s, 1H, OH), 8.58 (s, 1H, H5'), 8.18 (dd,

purification by column chromatography (CH₂Cl₂: MeOH = 20: 1) 1H, J = 7.7, 1.6 Hz, Ph), 7.43–7.27 (m, 2H, H8 and Ph), 7.21–

compound 19a (97.5 mg, 95%, mp > 280 °C) was isolated as 7.02 (m, 4H, H7, H5 and Ph), 5.97 (s, 2H, CH₂), 5.83 (s, 1H, H3),

light brown powder.¹H NMR (300 MHz, DMSO) δ 9.87 (s, 1H, 3.91 (s, 3H, OCH₃).¹³C NMR (75 MHz, DMSO) δ 159.67 (C2),

OH), 8.69 (s, 1H, H5'), 7.87 (d, 2H, J = 7.1 Hz, Ph), 7.46 (t, 2H, J 155.39 (Ph-q), 153.89 (C6), 150.07 (C4), 146.35 (C4'), 142.24

= 7.5 Hz, Ph), 7.36 (d, 1H, J = 7.4 Hz, Ph), 7.31 (d, 1H, J = 8.6 Hz, (Ph-q), 129.18 (Ph), 126.59 (Ph), 124.94 (C5'), 120.68 (Ph),

H8), 7.14–7.04 (m, 2H, H7 and H5), 6.07 (s, 1H H3), 5.95 (2H, s, 120.37 (C8), 118.70 (C4a), 117.68 (C7), 113.79 (C3), 111.58

CH2).¹³C NMR (151 MHz, DMSO) δ 159.69 (C2), 153.88 (C6), (Ph), 109.14 (C5), 55.50 (OCH₃), 49.17 (CH₂). MS (ESI): m/z =

10 | J. Name., 2012, 00, 1-3

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350.1 ([M+H]⁺). Anal. calcd. for C₁₉H₁₅N₃O₄: C, 65.32; H, 4.33;

N, 12.03. Found: C, 65.31; H, 4.21; N, 12.43.

Preparation of 6,7-dihydroxy-4-{

1,2,3-triazol-1-yl

[

methyl}-2H-chromen-2-one(20d). Compound

4-(2-methoxyphenyl)-1H-

]

Preparation

of

6,7-dihydroxy-4-[(4-phenyl-1H-1,2,3- 20d was synthesized according to the general procedure using

triazol-1-yl)methyl

]-2H-chromen-2-one (20a). Compound 20a 4-(azidomethyl)-6,7-dihydroxy-2H-chromen-2-one (15) (45 mg,

was synthesized according to the general procedure using 4- 0.19 mmol) and 1-ethynyl-2-methoxybenzen (0.03 mL, 0.23

(azidomethyl)-6,7-dihydroxy-2H-chromen-2-one (15) (42.5 mg, mmol). After purification by column chromatography (CH₂Cl₂:

0.18 mmol) and phenylacetylene (0.02 mL, 0.21 mmol). After MeOH = 20: 1) compound 20d (30.8 mg, 44%, mp > 250 °C)

purification by column chromatography (CH₂Cl₂: MeOH = 20: 1) was isolated as yellow powder.¹H NMR (300 MHz, DMSO) δ

compound 20a (15.6 mg, 26%, mp > 260 °C) was isolated as 10.43 (bs, 1H, OH), 9.47 (bs, 1H, OH), 8.56 (s, 1H, H5'), 8.18

light brown powder.¹H NMR (300 MHz, DMSO) δ 8.67 (s, 1H, (dd, 1H, J = 7.7, 1.7 Hz, Ph), 7.39–7.32 (m, 1H, Ph), 7.16 (s, 1H,

H5'), 7.87 (d, 2H, J = 7.3 Hz, Ph), 7.45 (t, 1H, J = 7.5 Hz, Ph), H8), 7.14–7.04 (m, 2H, Ph), 6.80 (s, 1H, H5), 5.91 (s, 2H, CH₂),

7.33 (d, 1H, J = 7.2 Hz, Ph), 7.09 (s, 1H, H8), 6.78 (s, 1H, H5), 5.55 (s, 1H, H3), 3.91 (s, 3H, OCH₃). ¹³C NMR (151 MHz, DMSO)

5.88 (s, 2H, CH₂), 5.77 (s, 1H, H3).¹³C NMR (75 MHz, DMSO) δ δ 160.26 Ph-q), 155.38 (C2), 150.73 (C7), 150.53 (C6), 148.07

160.40 (C2), 150.81 (C4), 149.72 (C8a), 148.26 (C7), 146.86 (C4), 143.02 (C4'), 142.17 (Ph-q), 129.14 (Ph), 126.57 (Ph),

(C6), 143.11 (C4'), 130.41 (Ph-q), 129.02 (Ph), 128.18 (Ph), 124.91 (C5'), 120.66 (Ph), 118.72 (Ph-q), 111.57 (Ph), 111.17

125.32 (Ph), 122.34 (C5'), 110.31 (C3), 109.01 (C4a), 108.95 (C3), 108.94 (C5), 108.90 (C4a), 102.87 (C8), 55.48 (OCH₃),

(C5), 102.98 (C8), 49.67 (CH₂). MS (ESI): m/z = 336.1 ([M+H]⁺). 49.26 (CH₂). MS (ESI): m/z = 366.1 ([M+H]⁺). Anal. calcd. for

Anal. calcd. for C₁₈H₁₃N₃O₄: C, 64.48; H, 3.91; N, 12.53. Found: C₁₉H₁₅N₃O₅: C, 62.46; H, 4.14; N, 11.50. Found: C, 62.73; H,

C, 64.52; H, 3.79; N, 12.24.

3.85; N, 11.38.

Preparation of 6,7-dihydroxy-4-{

[

4-(4-methoxyphenyl)-1H-

1,2,3-triazol-1-yl]methyl}-2H-chromen-2-one (20b). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay

Compound 20b was synthesized according to the general

procedure using 4-(azidomethyl)-6,7-dihydroxy-2H-chromen-2-

one (15) (35 mg, 0.15 mmol) and 1-ethynyl-4-methoxybenzen

(0.02 mL, 0.18 mmol). After purification by column

chromatography (CH₂Cl₂: MeOH = 20: 1) compound 20b (26.9

mg, 49%, mp > 260 °C) was isolated as yellow powder.¹H NMR

(300 MHz, DMSO) δ 8.55 (s, 1H, H5'), 7.79 (2H, d, J = 8.8 Hz,

Ph), 7.09 (s, 1H, H8), 7.01 (d, 2H, J = 8.8 Hz, Ph), 6.76 (s, 1H,

H5), 5.84 (s, 2H, CH₂), 5.74 (s, 1H, H3), 3.79 (s, 3H, OCH₃).¹³C

NMR (151 MHz, DMSO) δ 160.29 (Ph-q), 159.10 (C2), 151.01

(C4), 149.65 (C8a), 148.26 (C7), 146.68 (C6), 143.07 (C4'),

126.58 (Ph), 122.95 (Ph-q), 121.23 (C5'), 114.29 (Ph), 110.01

(C3), 108.76 (C4a), 108.72 (C5), 102.82 (C8), 55.13 (OCH₃),

49.52 (CH₂). MS (ESI): m/z = 366.1 ([M+H]⁺). Anal. calcd. for

C₁₉H₁₅N₃O₅: C, 62.46; H, 4.14; N, 11.50. Found: C, 62.51; H,

4.10; N, 11.48.

The antiradical activity of the tested compounds was

determined according to the described procedure (Sharma &

Bhat, 2009). Briefly, a 0.1 mM DPPH radical solution in 95%

methanol was prepared and 0.5 mL of this solution was mixed

with 0.5 mL of compounds 16a–16e, 17a–17e, 18a–18e, 19a–

19d, 20a–20d and reference compounds umbelliferon (7-

hydroxycoumarin) and esculetin (6,7-dihydroxycoumarin) in

0.2 mM concentrations. After 30 or 60 minutes of incubation

in the dark and at room temperature, absorbance (A) was

measured at 518 nm using a UV/VIS spectrometer (Lambda 25

UV/VIS, PerkinElmer, USA). The percentage of the radical

scavenging capacity (RSC) was calculated based on the

following equation:

RSC (%) = 100 - (A_samplex 100/A_blank),

where A_blankis an absorbance values (at 518 nm) of 1 mL of

95% methanol. Results were obtained from three replicate

assays.

Preparation of 6,7-dihydroxy-4-{

[

1,2,3-triazol-1-yl methyl}-2H-chromen-2-one

4-(3-methoxyphenyl)-1H-

(20c).

]

Compound 20c was synthesized according to the general

procedure using 4-(azidomethyl)-6,7-dihydroxy-2H-chromen-2-

one (15) (45 mg, 0.19 mmol) and 1-ethynyl-3-methoxybenzen

(0.03 mL, 0.23 mmol). After purification by column

chromatography (CH₂Cl₂: MeOH = 20:1) compound 20c (37.1

mg, 53%, mp > 260 °C) was isolated as yellow powder.¹H NMR

(300 MHz, DMSO) δ 10.41 (s, 1H, OH), 9.46 (s, 1H, OH), 8.68 (s,

1H, H5'), 7.48–7.41 (m, 2H, Ph), 7.36 (t, 1H, J = 7.8 Hz, Ph), 7.09

(s, 1H, H8), 6.91 (dd, 1H, J = 8.1, 1.4 Hz, Ph), 6.79 (s, 1H, H5),

5.87 (s, 2H, CH₂), 5.80 (s, 1H, H3), 3.80 (s, 3H, OCH₃). ¹³C NMR

(75 MHz, DMSO) δ 160.28 (Ph-q), 159.68 (C2), 150.74 (C4),

149.52 (C8a), 148.20 (C7), 146.68 (C6), 143.02 (C4'), 131.70

(Ph-q), 130.06 (Ph), 122.44 (C5'), 117.56 (Ph), 113.85 (Ph),

110.41 (Ph), 109.41 (C3), 108.93 (C4a), 108.89 (C5),

102.91(C8), 55.13 (OCH₃), 49.63 (CH₂). MS (ESI): m/z = 366.1

([M+H]⁺). Anal. calcd. for C₁₉H₁₅N₃O₅: C, 62.46; H, 4.14; N,

11.50. Found: C, 62.22; H, 4.42; N, 11.87.

Cyclic voltammetry

Carbon paste electrode (CPE, ALS-Japan) of diameter equal to

6 mm was filled with carbon paste (Metrohm) using a spatula.

A fresh layer of carbon paste was used at the electrode surface

for every measurement. After filling, the electrode was

prepared by rubbing the exposed carbon paste surface on a

wet filter paper and then by polishing on a glass prior to

sample attachment. Approximately 300 mg of each compound

was spread on a fine paper sheet, forming a thin layer of finely

distributed particles. The particles were then attached to the

surface of the CPE by gently rubbing the layer. Cyclic

voltammetry measurements were carried out in

a

conventional three electrode electrochemical cell consisting of

a carbon paste electrode, a saturated calomel electrode (SCE),

and a platinum wire as the working, the reference, and the

counter electrodes, respectively. PalmSense potentiostat has

been used for measuring cyclic voltammograms. The

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electrolyte was phosphate buffer of pH 7.4 prepared from

KH₂PO₄and NaOH of analytical grade. All the measurements

were carried out at room temperature of 25 °C ± 1 °C.

Voltammetric cycles were measured, at a scan rate of 100mV

s⁻¹, in the potential range between 0 and +0.8 V v.s. SCE,

starting from the cathodic potential limit. Three voltammetric

cycles were performed for each sample, yielding the

reproducibility of the calculated parameters within ±10%.

Acknowledgements

Financial support for this study was provided by Croatian

Science Foundation project No. IP-2013-11-5596 and VIF-2016-

MEFOS-25 project.

Notes and references

1

2

3

I. Svinyarov and M. G. Bogdanov, Eur. J. Med. Chem., 2014,

78, 198–206.

Biological assays

F. Borges, F. Roleira, N. Milhazes, E. Uriarte and L. Santana,

Cytotoxicity evaluation. Cytotoxic effects on the normal and

tumor cells’ growth were determined using the colorimetric

methyltetrazolium (MTT) assay. Experiments were carried out

on three tumor human cell lines (HeLa, CaCo-2, and K562) and

on one canine cell line (MDCK1) as normal cells. The adherent

cells, MDCK1, HeLa, CaCO-2, K562 were seeded in 96 micro-

well plates at a concentration of 2 × 10⁴cells/mL and allowed

Front. Med. Chem., 2009, 4, 23–85.

F. G. Medina, J. G. Marrero, M. Macías-Alonso, M. C.

González, I. Córdova-Guerrero, A. G. Teissier García and S.

Osegueda-Robles, Nat. Prod. Rep., 2015, 32, 1472–1507.

4

5

6

N. Z. Ballin and A. T. Sørensen, Food Control, 2014, 38,

198–203.

J. Grover and S. M. Jachak, RSC Adv., 2015, 5, 38892–

to attach overnight in the CO₂incubator (IGO 150 CELLlife^TM

JOUAN, Thermo Fisher Scientific Waltham, MA, USA). After 72

,

38905.

,

X. W. Ye, Y. C. Zheng, Y. C. Duan, M. M. Wang, B. Yu, J. L.

h of exposure to tested compounds, medium was replaced

with 5 mg/mL MTT solution and the resulting formazane

Ren, J. L. Ma, E. Zhang and H. M. Liu, Med. Chem.

Commun., 2014, 5, 650–654.

crystals were dissolved in DMSO. Leukemia cells at

a

7

8

9

J. Dandriyal, R. Singla, M. Kumar and V. Jaitak, Eur. J. Med.

Chem., 2016, 119, 141–168.

concentration of 1 x 10⁵cells/mL were plated onto 96 micro-

well plates and after 72 h of incubation, 5 mg/mL MTT solution

was added to each well and incubated 4 hours in CO₂

incubator. To each well, 10% SDS with 0.01 mol/L HCl was

added to dissolve water-insoluble MTT-formazane crystals

overnight. Elisa micro plate reader (iMark, BIO RAD, Hercules,

CA, USA) was used for measurement of absorbance at 595 nm.

All experiments were performed at least three times in

triplicates. The percentage of cell growth (PG) was calculated

using the following equation:

S. Emami and S. Dadashpour, Eur. J. Med. Chem., 2015,

102, 611–630.

E. Calcio Gaudino, S. Tagliapietra, K. Martina, G. Palmisano

and G. Cravotto, RSC Adv., 2016, 6, 46394–46405.

10 A. Torres de Pinedo, P. Peñalver and J. C. Morales, Food

Chem., 2007, 103, 55–61.

11 M.J. Matos, F. Pérez-Cruz, S. Vazquez-Rodriguez, E.

Uriarte, L. Santana, F. Borges and C. Olea-Azar, Bioorg.

Med. Chem., 2013, 21, 3900–3906.

PG = (A _compound– A _background/A _control– A _background) x 100

12 K. Venkata Sairam, B. M. Gurupadayya, B. I. Vishwanathan,

where A_backgroundat the adherent cells is absorbance of MTT

solution and DMSO; A_backgroundat the suspension cells is the

absorbance of the medium without cells, but containing MTT

and 10% SDS with 0.01 mol/L HCl; and A_controlis the

absorbance of cell suspension grown without tested

compounds. Calculation of IC₅₀values curves and QC analysis is

performed by using Excel tools and GraphPadPrism software

(La Jolla, CA), v. 5.03. Briefly, individual concentration effect

curves are generated by plotting the logarithm of the

concentration of tested compounds.

R. S. Chandana and D. K. Nageshab, RSC Adv., 2016, 6,

98816–98828.

13 Y. K. Tyagi, A. Kumar, H. G. Raj, P. Vohra, G. Gupta, R.

Kumari, P. Kumar and R. K. Gupta, Eur. J. Med. Chem.,

2005, 40, 413–420.

14 E. Geromichalou, N. Sayyad, E. Kyriakou, A.V.

Chatzikonstantinou, E. Giannopoulou, N. Vrbjar, H.P.

Kalofonos, H. Stamatis and A.G. Tzakos, Eur. J. Med.

Chem., 2015, 96, 47–57.

15 S. Reuter, S. C. Gupta, M. M. Chaturvedi and B. B.

Aggarwal, Free Radical Biol. Med., 2010, 49, 1603–1616.

16 N. Hamdi, M. C. Puerta and P. Valerga, Eur. J. Med. Chem.,

2008, 43, 2541–2548.

Measurement of intracellular ROS by flow cytometry.

Intracellular ROS in K562 cells were detected by Fluorimetric

Intracellular ROS Kit (Sigma-Aldrich, St. Louis, MO, USA)

according to maunfacturer's instruction. Briefly, to induce ROS

17 M. Lodovici, S. Menichetti, C. Viglianisi, S. Caldini and E.

Giuliani, Bioorg. Med. Chem. Lett., 2006, 16, 1957–1960.

18 K. E. Heim, A. R. Tagliaferro and D. J. Bobilya, J. Nutr.

Biochem., 2002, 13, 572–584.

1 x 10⁶K562 cells were treated for 60 min with 1 x 10^-5

esculetin as a standard compound and with 1 x 10^-5

M

compounds 16b, 17b, 19b and 20d. After treatment, were

19 M. Leopoldini, N. Russo and M. Toscano, Food Chem.,

2011, 125, 288–306.

added 1 µL of 500 x ROS detection reagent stock solution were

added per 1 mL of cells and incubated in the CO₂incubator for

30 min. The fluorescence intensity was measured by flow

cytometry (FacsCanto II, BD Biosciences, USA) and analyzed

using Flowlogic software (Inivai Technologies, USA).

20 G. Mazzone, N. Malaj, A. Galano, N. Russo and M. Toscano,

RSC Adv., 2015, 5, 565–575.

12 | J. Name., 2012, 00, 1-3

Please do not adjust margins

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Page 13 of 14

Journal Name

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DOI: 10.1039/C7NJ01469D

ARTICLE

21 T. Symeonidis, M. Chamilos, D. J. Hadjipavlou-Litina, M. 42 S. Maračić, T. G. Kraljević, H. Č. Paljetak, M. Perić, M.

Kallitsakis and K.E. Litinas, Bioorg. Med. Chem. Lett.,

2009, 19, 1139–1142.

Matijašić, D. Verbanac, M. Cetina and S. Raić-Malić, Bioorg.

Med. Chem., 2015, 23, 7448–7463.

22 M. F. Molina-Jiménez, M.I . Sánchez-Reus, D. Andres, M. 43 Y. C. Duan, Y. C. Ma, E. Zhang, X. J. Shi, M.M. Wang, X. W.

Cascales and J. Benedi, Brain Res., 2004, 1009, 9–16. Ye and H.M. Liu, Eur. J. Med. Chem., 2013, 62, 11–9.

23 A. R. Proteggente, A. S. Pannala, G. Paganga, L. V. Buren, E. 44 J. L. Boudenne, O. Cerclie and P. Bianco, J. Electrochem.

Wagner, S. Wiseman, F. V. D. Put, C. Dacombe and C.A.

Rice-Evans, Free Radical Res., 2002, 36, 217–233.

24 T. Kaneko, N. Baba and M. Matsuo, Chem.-Biol.

Interact., 2003, 142, 239–254.

Soc., 1998, 1145, 2763–2768.

45 V. Pardo-Jiménez, C. Barrientos, J. A. Squella, P. A.

Navarrete-Encina, and L. J. Nuñez-Vergara, J. Electrochem.

Soc., 2011, 158, 166–172.

25 R. Zhang, Acta Pharmacol. Sin., 2008, 29, 1319–1326.

26 A. D. Kim, X. Han, M. J. Piao, M. S. R. K. M. Hewage, C.L.

Hyun, S.J. Cho and J.W. Hyun, Environ. Toxicol.

Pharmacol., 2015, 39, 982–989.

46 V. Pardo-Jiménez, C. Barrientos, K. Pérez-Cruz, P. A.

Navarrete-Encina, C. Olea-Azar, L. J. Nuéez-Vergara and J.

A. Squella, Electrochim. Acta, 2014, 125, 457–464.

47 A. Bast and G. R. M. M. Haenen, Environ. Toxicol.

Pharmacol., 2002, 11, 251–258.

27 P. Wang, Y. L. Xia, Y. Yu, J. X. Lu, L. W. Zou, L. Feng, G. Ge

and L. Yang, RSC Adv., 2015,

5, 53477–53483.

48 M. Y. Liou and P. Storz, Free Radical Res., 2010, 44

,

479–

28 M. Roussaki, K. Zelianaios, E. Kavetsou, S. Hamilakis, D.

476.

Hadjipavlou-Litina, C. Kontogiorgis, T. Liargkova and A. 49 M. Marvibaigi, N. Amini, E. Supriyanto, F. A. A. Majid, S. K.

Detsi, Bioorg. Med. Chem., 2014, 22, 6586–6594.

Jaganathan, S. Jamil, J. H. Almaki and R. Nasiri, PloS One,

2016, 11, e0158942.

29 D. Ribeiro, M. Freitas, S.M. Tomé, A.M.S. Silva, G. Porto, E.

J. Cabrita, M. M. B. Marques and E. Fernandes, Eur. J. Med. 50 A. M. Piloto, A. S. Fonseca, S. P. Costa and M. S. T.

Chem., 2014, 72, 137–145. Gonçalves, Tetrahedron, 2006, 62, 9258–9267.

30 A. Balabani, D.J. Hadjipavlou-Litina, K. E. Litinas, M. 51 N. H. Nam, Y. Kim, Y. J. You, D. H. Hong, H. M. Kim and B. Z.

Mainou, C.C. Tsironi and A. Vronteli, Eur. J. Med.

Chem., 2011, 46, 5894–5901.

Ahn, Bioorg. Med. Chem. Lett., 2002, 12, 2345–2348.

52 M. S. Frasinyuk, S. P. Bondarenko and V. P. Khilya, Chem.

Heterocyc. Comp., 2009, 45, 290–296.

31 Y. Yang, Q.W. Liu, Y. Shi, Z.G. Song, Y.H. Jin and Z.Q. Liu,

Eur. J. Med. Chem., 2014, 84, 1–7.

53 R. A. Kusanur, M. V. Kulkarni, G. M. Kulkarni, S. K. Nayak, T.

N. Guru Row, K. Ganesan and C. M. Sun, J. Het.

Chem., 2010, 47, 91–97.

32 I. Khan, S. Ali, S. Hameed, N.H. Rama, M.T. Hussain, A.

Wadood, R. Uddin, Z. Ul-Haq, A. Khan, S. Ali and M.I.

Choudhary, Eur. J. Med. Chem., 2010, 45, 5200–5207.

33 P. Valentina, K. Ilango, M. Deepthi, P. Harusha, G. Pavani,

K.L. Sindhura and C. Keerthanan, J. Pharm. Sci. Res.,

54 X. Liu, D. G. Smith and K. A. Jolliffe, Chem. Comm., 2016,

52, 8463–8466.

2009,

34 H. Yüksek, S. Kolaylı, M. Küçük, M. O. Yüksek, U. Ocak, E.

Şahinbaş, E. Sivrikaya and M. Ocak, Ind. J. Chem., 2006, 45

1, 74–77.

,

715–718.

35 S. Raic-Malic and A. Mescic, Curr. Med. Chem., 2015, 22

,

1462–1499.

36 V. K. Tiwari, B. B. Mishra, K. B. Mishra, N. Mishra, A. C.

Singh and X. Chen, Chem. Rev., 2016, 116, 3086–3240.

37 K. Inami, Y. Iizuka, M. Furukawa, I. Nakanishi, K. Ohkubo, K.

Fukuhara, S. Fukuzumi and M. Mochizuki, Bioorg. Med.

Chem., 2012, 20, 4049–4055.

38 F. Pérez-Cruz, S. Vazquez-Rodriguez, M. J. Matos, A.

Herrera-Morales,

F.A.

Villamena,

A.,

Das,

B.

Gopalakrishnan, C. Olea-Azar, L. Santana, E. Uriarte, J. Med.

Chem., 2013, 56, 6136–6145.

39 H. Wei, J. Ruan and X. Zhang, RSC Adv., 2016, 6, 10846–10860.

40 T. Gazivoda Kraljević, A. Harej, M. Sedić, S. Kraljević Pavelić, V.

Stepanić, D. Drenjančević, J. Talapko and S. Raić-Malić, Eur. J.

Med. Chem., 2016, 124, 794–808.

41 T. Gregorić, M. Sedić, P. Grbčić, A. T. Paravić, S. Kraljević

Pavelić, M. Cetina, R. Vianello and S. Raić-Malić, Eur. J.

Med. Chem., 2017, 125, 1247–1267.

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Graphical abstract

Diverse natural coumarin-based compounds linked to aryl via 1,2,3-triazole spacer with antiproliferative activity on K562 cells,

radical scavenging activity and decrease of ROS production were provided.

14 | J. Name., 2012, 00, 1-3

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Article Doi

Synthesis of 4-aryl-1,2,3-triazolyl appended natural coumarin-related compounds with antiproliferative and radical scavenging activities and intracellular ROS production modification

DOI: 10.1039/c7nj01469d

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