Investigation on the substitution effects of the flavonoids as potent anticancer agents: A structure-activity relationships study

MEDICINAL

CHEMISTRY

RESEARCH

Med Chem Res (2012) 21:1833–1849

DOI 10.1007/s00044-011-9701-6

ORIGINAL RESEARCH

Investigation on the substitution effects of the ﬂavonoids as potent

anticancer agents: a structure–activity relationships study

•

Xiao-Bing Wang Wei Liu Lei Yang

•

Qing-Long Guo Ling-Yi Kong

Received: 29 January 2011 / Accepted: 9 June 2011 / Published online: 25 June 2011

Ó Springer Science+Business Media, LLC 2011

Abstract Three series of ﬂavonoid analogues substituted

with different aminomethyl substitutions at C-6, C-7, and

C-8 were designed and synthesized for the structure–

activity relationship studies as potent anticancer agents.

The prepared analogues were evaluated for their in vitro

inhibitory activity against the growth of the hepatic cancer

cell lines HepG2 and SMMC-7721. Structure–activity

relationships indicated that not only the compounds with

amino methyl groups were more active than those without

the groups in the same series but also the compounds

substituted by aminomethyl groups at position C-8 were

more active than those at positions C-6 and C-7.

antiallergic, antiinﬂammatory, antimicrobial, anticancer

effects (Ververidis et al., 2007; Spencer, 2008; Cushnie

and Lamb, 2005; Lotito and Frei, 2006). Noteworthily,

ﬂavonoids bearing alkyl substituents at C-8 were fre-

quently found with interesting pharmacological properties

(Adinarayana and Ramachandraiah, 1987; Barron and Ibrahim,

1996). Some good examples (Scheme 1) are kushenol C

(1) with potent antitumor activity both in vitro and in vivo

(Sun et al., 2007), phellamurin (2) with DNA cleavage

activity (Ma et al., 2005), icariin (3), the main active

component of Epimedium koreanum, with myriads of

activities (Makarova et al., 2007; He et al., 1995; Liu et al.,

2005), puerarin (4) with preventive and treating effects on

arrhythmia (Chai et al., 1985), and the synthetic ﬂavonoids,

ﬂavone-8-acetate acid (5) having strong antitumor activity

(Atassi et al., 1985; Naik et al., 1988). Of these com-

pounds, ﬂavopiridol (6), a semisynthetic ﬂavonoid that

inhibits tumor growth both in vitro and in vivo, is the ﬁrst

potent cyclin-dependent kinase (CDK) inhibitor entering

clinical trials. It inhibits the activity of several CDKs,

speciﬁcally CDK1, CDK2, and CDK4, inducing a G₁- or

G₂-phase cell cycle arrest (De Azevedo et al., 1996;

Losiewicz et al., 1994; Carlson et al., 1996; Kaur et al.,

1992; Czech et al., 1995; Parker et al., 1998). Its success

stimulates ensuing researches on the synthesis of related

ﬂavonoid analogues. Its SAR studies have shown that

N-methyl-piperidine attached to the skeleton with carbon–

carbon bond formation was the key pharmacophore. Some

ideas that the substitution at position C-8 may be the key

pharmacophore to their bioactivity and the bioactivity may

increase progressively with the substitution changed from

position C-6 to C-7 and ﬁnally to C-8 were achieved based

on the above investigations. Bearing this in mind, we

focused our attention on the substitution effects of the

ﬂavonoids as potent anticancer agents and designed three

Keywords Flavonoid ꢀ Drug design ꢀ Cytotoxicity ꢀ

Structure–activity relationship

Introduction

Flavonoids are a large class of natural compounds with a

C6–C3–C6 skeleton substituted by various groups. They

have been receiving great interests because of a wide

spectrum of biological activities exhibited, such as

X.-B. Wang ꢀ L. Yang ꢀ L.-Y. Kong (&)

Department of Natural Medicinal Chemistry,

China Pharmaceutical University, 24 Tong Jia Xiang,

210009 Nanjing, People’s Republic of China

e-mail: cpu_lykong@126.com

W. Liu ꢀ Q.-L. Guo

Department of Physiology, Jiangsu Key Laboratory

of Carcinogenesis and Intervention, China Pharmaceutical

University, 24 Tong Jia Xiang, 210009 Nanjing,

People’s Republic of China

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Med Chem Res (2012) 21:1833–1849

series of ﬂavonoids analogues. Compounds substituted by

various groups containing nitrogen at position C-6 (series

A, Table 1), C-7 (series B, Table 2), and C-8 (series C,

Table 3) were designed. We reported herein the synthesis,

biological evaluation, and structure–activity relationship of

these ﬂavonoid analogues as anticancer agents.

Results and discussion

Chemistry

Mannich reaction, a directive tactics to make an amino

alkylation of an acidic proton placed next to a carbonyl

Table 1 The target compounds

in series A

Compound

R

H

X

Compound

R

X

4 -NO₂

4 -Cl

17a

17n

H

4 -Cl

2 -Cl

H

4 -Cl

17b

17c

17d

17e

17f

17o

17p

17q

17r

17s

4 -NH₂

4 -NO₂

4 -Cl

2 -Cl

17g

17h

17t

17u

4 -NO₂

H

17i

17j

17k

17v

17w

17x

4 -NH₂

1-3B

4 -NO₂

4 -Cl

1-6B

17l

17y

17m

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1835

Table 2 The target compounds

in series B

Compound

R

H

X

Compound

R

X

18a

4 -NO₂

18j

4 -NO₂

2 -Cl

4 -Cl

18b

18c

18d

18e

18f

H

4 -Cl

2 -Cl

18k

18l

H

18m

18n

18o

18p

18q

4 -NH₂

4 -NO₂

18g

18h

18i

4 -NO₂

18r

4 -Cl

functional group, was used as the normal preparation

method of these kinds of compounds. For example, series

of C-8 nitrogen-containing ﬂavonoid analogues had been

designed and synthesized through Mannich reaction

(Zhang et al., 2008; Liu et al., 2007). In this study, three

series of compounds with the substitution positions varied

among C-6, C-7, and C-8 were designed to investigate the

substitution effects of these ﬂavonoids on their activities.

However, Mannich reaction is not suitable for the direct

synthesis of these compounds. So the oriented synthesis

routes were outlined as shown in Schemes 1 and 2. The

synthetic procedures of the target compounds can be

divided into two parts: (i) the synthesis of the methylﬂav-

ones, 17a–17d, 18a–18d, and 19a–19d, and (ii) the amino

methylﬂavones, 17e–17y, 18e–18r, and 19e–19w

(Schemes 2, 3, and 4).

esteriﬁed with acetic anhydride to yield p-tolyl acetate 21a.

The Fries rearrangement of 21a led to a high yield of

2-hydroxy-5-methylacetophenone 22a, which was con-

densed with different substituted benzoyl chlorides through

the Baker–Venkataraman method to produce b-diketone

23a–23d. The classic Baker–Venkataraman transformation

involves two steps: 2-Hydroxy-5-methyl- acetophenone

and benzoyl chloride are ﬁrst converted into a benzoyl ester

in pyridine, and then treated with a base, such as KOH,

affording b-diketone through Claisen rearrangement. In

this study, the two Baker–Venkataraman transformation

steps were reduced to one by using K₂CO₃in acetone, and

then the obtained b-diketone 23a–23d was treated with

NaOAc/HOAc to give desired methylﬂavones 17a–17d as

the important intermediates. The bromomethylﬂavones

24a–24c were prepared through free-radical reaction of

methylﬂavones 17a–17c with N-bromosuccinimide (NBS)

in CCl₄using benzoyl peroxide (BPO) as an inducer. The

Compounds 17a–17y in series A were prepared using

p-cresol as the starting material. Compound 20a was ﬁrst

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Med Chem Res (2012) 21:1833–1849

Table 3 The target compounds

in series C and their CDK/cyclin

B inhibitory activity

CDK/ Cyclin B

Compound

R

H

X

Y

(IC₅₀, μM)

19a

4′-NO₂

H

ND^*

19b

19c

19d

19e

H

4′-NO₂

4′-Cl

Cl

ND

2′-Cl

4′-NH₂

19f

19g

19h

19i

4′-NO₂

2′-Cl

H

2.21

8.13

ND

6.22

ND

4.84

ND

2.14

2.06

ND

2.17

H

Cl

19j

19k

19l

19m

19n

19o

19p

19q

19r

19s

19t

2′-Cl

19u

19v

19w

2′-Cl

ND the CDK/cyclin B inhibitory

activity was not detected in the

assay

4′-Cl

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Med Chem Res (2012) 21:1833–1849

1837

Scheme 1 Some bioactivity ﬂavonoids

Scheme 2 Synthesis of the

target compounds in A

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Med Chem Res (2012) 21:1833–1849

Scheme 3 Synthesis of the

target compounds in B

CDK1/cyclin B inhibitory activity

target molecules 17f–17y were prepared by reaction of

24a–24c with optional amines, diethylamine, piperidine,

piperazine, N-methylpiperazine, morphine, dimethylamine,

imidazole, triethylamine, and so on. Compounds 17a and

17f–17i were reduced by sodium dithionite to give the cor-

responding aminoﬂavones 17e and 17w–17y, respectively.

Similar to the protocols for the synthesis of the com-

pounds in series A, compounds 18a–18r in series B were

synthesized using m-cresol as the starting material. In the

preparation of the compounds in series C, 8-aminometh-

ylﬂavones, an initial attempt to use o-cresol as the starting

material was made (Scheme 5). The result was disap-

pointing as the Fries rearrangement of 21c majorly pro-

duced the para product 22c⁰. Although the ortho product

22c was the key intermediate for the synthesis of the

8-alkylated ﬂavones, as a result, only four compounds

19f–19h were obtained using this way. To enhance the

yield of ortho products, 4-chloro-2-methylphenol (20d)

was used as the starting material in terms of the charac-

teristic of the Fries rearrangement, which was exclusively

converted to its ortho products. In this manner, 8-substituted

ﬂavone derivatives 19i–19w were also successfully

synthesized.

Compounds 19a–19w in series C were assayed for their

CDK1/cyclin B inhibitory activity with Z⁰-LYTE^TMkinase

assay kits (Table 3). Some compounds, especially those

containing a piperazine moiety, e.g., 19g (IC₅₀= 8.13 lM),

19k (IC₅₀= 6.22 lM), 19m (IC₅₀= 4.84 lM), exhibited

potent CDK1/cyclin B inhibitory activity. However, their

activities were still many times weaker than that of ﬂavo-

piridol (Kim et al., 2000), which might be because of the

lack of the hydrogen-bonding donors, e.g., hydroxyl groups.

Cytotoxicity

The anticancer activities of all the synthesized compounds

were evaluated by MTT method using hepatic cancer cell

lines HepG2 and SMMC-7721 (Table 4). Almost all the

ﬂavones with amino methyl groups, such as 17e–17y,

18e–18r, and 19e–19w, were more active than those without

amino methyl groups, such as 17a–17d, 18a–18d, and

19a–19d, which suggested the key role of the N-containing

substituents for activities. Potential bioactivities were found

for 11 out of 50 tests assays in series A (ca. 22%), 13 of 36

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Med Chem Res (2012) 21:1833–1849

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Scheme 4 Synthesis of the

target compounds in C

Scheme 5 Synthetic routes of the key methylﬂavones in series C

(ca. 36%) in series B, and 31 of 46 (ca. 67%) in series C in

two cell lines. Compounds with substitution at position C-8

were more active than those at positions C-7 and C-6.

Compounds 17h, 18h, 19g, and 19k, for example, have the

same substituent group, 4-methylpiperazinylmethyl, at ring

A of the ﬂavone nucleus but with different substitution

positions. The 8-substituted derivatives 19g and 19k

exhibited potent cytotoxic activity against hepatic cancer

cell line SMMC-7721 with IC₅₀at 1.19 and 5.21 lM,

respectively, as compared with those for 18h (7-substituted)

and 17h (6-substituted) at 10.7 and 19.0 lM, respectively.

The bioactivity of all the target compounds increased when

the substituent positions varied from C-6 to C-7 and C-8,

indicating the essential role of the substituent position for the

biological activity of these ﬂavonoids as anticancer agents.

Furthermore, almost all the target compounds containing

piperidine, piperazine, and N-methylpiperazine groups

exhibited cytotoxic activity to HepG2 and SMMC-7721

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Table 4 Cytotoxicity of the present compounds against two tumor cells

Compound

MTT/IC₅₀(lM)

Compound

MTT/IC₅₀(lM)

Compound

MTT/IC₅₀(lM)

HepG2

SMMC-7721

HepG2

SMMC-7721

HepG2

SMMC-7721

17a

17b

17c

17d

17e

17f

ND

6.23

ND

21.3

ND

14

ND

15.1

ND

19

17w

17x

17y

18a

18b

18c

18d

18e

18f

ND

2.64

ND

6.83

28.3

ND

40.5

ND

4.53

10.7

3.64

14.2

ND

2.58

ND

3.68

ND

10.7

ND

2.03

ND

12.9

ND

1.91

28.2

ND

19b

19c

19d

19e

19f

ND

11.9

28.8

2.94

ND

19g

19h

19i

1.19

ND

17g

17h

17i

1.19

8.91

1.76

ND

2.99

1.09

5.21

ND

16.2

ND

1.51

ND

3.20

19j

17j

18g

18h

18i

19k

19l

17k

17l

19m

19n

19o

19p

19q

19r

19s

19t

0.243

6.19

2.94

7.94

10.7

ND

1.1

17m

17n

17o

17p

17q

17r

17s

18j

56.53

1.2

18k

18l

ND

2.69

ND

61

0.81

3.4

18m

18n

18o

18p

18q

18r

19a

3.1

6.81

18.7

1.87

3.27

6.33

13.2

2.69

1.14

1.07

15.3

17t

19u

19v

19w

17u

17v

Control^a

ND

6.11

ND the bioactivity of the compound was not detected

a

Gambogic acid was chosen as the control in the MTT assay

cells. For example, the following IC₅₀values have been

obtained for HepG2 and SMMC-7721 cells, respectively

using the following compounds: 17q with a piperazine 2.69

and 1.51 lM, 18j with a piperazine 4.53 and 2.03 lM,

18o with a piperazine 2.58 and 1.91 lM, 19g with an

N-methylpiperazine as 2.94 and 1.19 lM, 19j with a

piperidine 8.91 and 1.09 lM, 19k with an N-methylpipera-

zine 1.76 and 5.21 lM, and 19m with a piperazine as 0.243

and 1.1 lM. The results combined suggested that 8-ami-

nomethyl ﬂavonoids with free hydroxyl groups have the

potential to be developed as anticancer agents in the future.

that compounds with substitutions at position C-8 were

more active than those at positions C-7 and C-6, and in the

same series, compounds with the amino methyl groups

were more active than those without such functionalities.

Experimental

General

NMR spectra were recorded on Bruker AC 300 and 500

spectrometer using TMS as internal standard. ESI–MS was

detected with HP-1100 Series LC/MSD ion trap mass

spectrometer. The IR (KBr) spectra were obtained on a

Nicolet Impact 410 spectrometer. Flash chromatography

was performed with 230–400 mesh silica gel. TLC was

carried out using commercially available precoated silica

gel 60 F₂₅₄(Qingdao Marine Chemical Co. Ltd).

Conclusions

In conclusion, three series of ﬂavonoid analogues were

designed and synthesized. The CDK1/cyclin B inhibitory

activity of the compounds in series C was measured using

Z⁰-LYTE^TMkinase assay kits, and some compounds were

found to show inhibitory activity. The cytotoxic activity of

all the synthesized compounds was assayed using MTT

method. Structure–activity relationships studies indicated

Unless otherwise stated, chemicals and solvents were of

reagent grade and used as obtained from commercial

sources without further puriﬁcation. All the reactions were

monitored by TLC and MS.

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Med Chem Res (2012) 21:1833–1849

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General procedure for the synthesis of the methylﬂavones

(d, 1H, J = 8.6 Hz, H-7), 7.69 (d, 1H, J = 8.6 Hz, H-8),

7.60 (m, 3H, H-3⁰, H-4⁰, H-5⁰, H-6⁰), 6.61 (s, 1H, H-3), and

2.46 (s, 3H, CH₃); ESI–MS m/z 271 [M ? H]^?.

One drop of concentrated sulfuric acid was added to a

mixture of optionally substituted phenol (0.10 mol) and

acetic anhydride (10 g, 0.1 mol). After the mixture was

stirred for h, water was added to the solution. The

solution was extracted with EtOAc, washed with water and

brine, dried over Na₂SO₄, then the solvent was evaporated

under reduce pressure to obtain phenyl acetates 21a–21d as

an oil. Optional phenyl acetates 21a–21d (1 mol) were

mixed with anhydrous AlCl₃(2 mol) at 120*180°C for

2 h, respectively, then cooled down to room temperature.

The solution of ice water and HCl with 1:1 (V:V) was

added to the mixture slowly, o-hydroxyacetophenone

derivatives 22a–22d, the most important intermediate for

preparing the designed ﬂavone, were synthesized. A solu-

tion of the o-hydroxyacetophenone derivatives 22a–22d

(20 mmol) in acetone (60 ml) was added with appropri-

ately substituted benzoyl chloride (20 mmol) using dry

K₂CO₃(12 g) as the base under reﬂuxing for 12 h. The

mixture was ﬁltered and the precipitate was decomposed

with 10% glacial acetic acid to give yellow powder as

b-diketone 23a–23l. The resulted b-diketone 23a–23l was

treated with NaOAc/HOAc (8 g/40 ml) under reﬂux for

12 h and cooled to room temperature. Water was added to

the mixture, then ﬁltered, and washed with hot water,

saturated NaHCO₃solution, and cold water successively to

give the methylﬂavones 17a–17d, 18a–18d, and 19a–19d.

6-Methyl-2-phenyl-4H-chromen-4-one (17d)

Yield 78%; IR (KBr) t_max3,451, 1,644, 1,615, 1,568,

1,483, 1,451, 1,360, 1,223, 1,139, 1,041, 902, 822,

771 cm^-^{1 1}H NMR (CDCl₃, 300 MHz) d 7.91 (s, 1H,

;

H-5), 7.89 (m, 2H, H-7, H-8), 7.49 (m, 5H, H-2⁰, H-3⁰,

H-4⁰, H-5⁰, H-6⁰), 6.79 (s, 1H, H-3), and 2.44 (s, 3H, CH₃);

ESI–MS m/z 237 [M ? H]^?.

7-Methyl-2-(4-nitrophenyl)-4H-chromen-4-one (18a)

Yield 72%; IR (KBr) t_max3,450, 3,073, 1,632, 1,568,

1,518, 1,424, 1,376, 1,346, 1,046, 852, 834, 692 cm^-1; H

1

NMR (CDCl₃, 300 MHz) d 8.38 (d, 2H, J = 8.1 Hz, H-2⁰,

H-6⁰), 8.11 (d, 2H, J = 8.1 Hz, H-3⁰, H-5⁰), 7.42 (s, 1H,

H-5), 7.28 (m, 2H, H-6, H-8), 6.88 (s, 1H, H-3), and 2.55

(s, 3H, CH₃); ESI–MS m/z 282 [M ? H]^?.

2-(4-Chlorophenyl)-7-methyl-4H-chromen-4-one (18b)

Yield 76%; IR (KBr) t_max3,449, 3,069, 1,639, 1,571, 1,491,

1,411, 1,370, 1,277, 1,233, 1,161, 1,093, 1,046, 1,012, 907,

1

826 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.07 (d, 2H,

J = 8.4 Hz, H-2⁰, H-6⁰), 7.91 (d, 1H, J = 8.5 Hz, H-5), 7.62

(d, 2H, J = 8.4 Hz, H-3⁰, H-5⁰), 7.55 (s, 1H, H-8), 7.30 (d,

1H, J = 8.5 Hz, H-6), 7.00 (s, 1H, H-3), and 2.47 (s, 3H,

CH₃); ESI–MS m/z 271 [M ? H]^?.

6-Methyl-2-(4-nitrophenyl)-4H-chromen-4-one (17a)

Yield 74.9%; IR (KBr) t_max3,442, 1,640, 1,618, 1,573,

1

1,522, 1,483, 1,347, 1,042, 851, 824, 691 cm^-1; H NMR

2-(2-Chlorophenyl)-7-methyl-4H-chromen-4-one (18c)

Yield 69%; IR (KBr) t_max3,442, 1,643, 1,612, 1,418, 1,365,

(CDCl₃, 300 MHz) d 8.38 (d, 2H, J = 8.1 Hz, H-3⁰, H-5⁰),

8.11 (d, 2H, J = 8.1 Hz, H-2⁰, H-6⁰), 8.03 (d, 1H,

J = 2.1 Hz, H-5), 7.55 (dd, 1H, J = 8.6, 2.1 Hz, H-7),

7.50 (d, 1H, J = 8.6 Hz, H-8), 6.89 (s, 1H, H-3), and 2.49

(s, 3H, CH₃); ESI–MS m/z 282 [M ? H]^?.

1

1,230, 1,140, 1,039, 911, 868, 823, 752 cm^-1; H NMR

(DMSO-d₆, 300 MHz) d 7.98 (d, 1H, J = 8.1 Hz, H-5), 7.80

(m, 1H, H-6⁰), 7.61 (m, 3H, H-3⁰, H-4⁰, H-5⁰), 7.50 (s, 1H,

H-8), 7.37 (d, 1H, J = 8.1 Hz, H-6), 6.59 (s, 1H, H-3), and

2.54 (s, 3H, CH₃); ESI–MS m/z 271 [M ? H]^?.

2-(4-Chlorophenyl)-6-methyl-4H-chromen-4-one (17b)

Yield 82% IR (KBr) t_max3,444, 3,075, 1,641, 1,620, 1,574,

7-Methyl-2-phenyl-4H-chromen-4-one (18d)

1,486, 1,432, 1,364, 1,090, 1,039, 1,009, 902, 820 cm^-1

;

¹H NMR (DMSO-d₆, 500 MHz) d 8.09 (d, 2H, J = 8.5 Hz,

H-2⁰, H-6⁰), 7.82 (s, 1H, H-5), 7.65 (m, 2H, H-7, H-8), 7.63

(d, 2H, J = 8.5 Hz, H-3⁰, H-5⁰), 7.01 (s, 1H, H-3), and 2.43

(s, 3H, CH₃); ESI–MS m/z 271 [M ? H]^?.

Yield 73%; ¹H NMR (DMSO-d₆, 300 MHz) d 8.12 (1H, s,

H-8), 7.95(d, 1H, J = 7.8 Hz, H-5), 7.61(br, 5H, H-2⁰,

H-3⁰, H-4⁰, H-5⁰, H-6⁰), 7.34(d, 1H, J = 7.8 Hz, H-6), 7.02

(s, 1H, H-3), and 2.46 (s, 3H, CH₃); ESI–MS m/z 237

[M ? H]^?.

2-(2-Chlorophenyl)-6-methyl-4H-chromen-4-one (17c)

8-Methyl-2-(4-nitrophenyl)-4H-chromen-4-one (19a)

Yield 80%; IR (KBr) t_max3,444, 3,070, 1,658, 1,620,

1,485, 1,440, 1,340, 1,224, 1,072, 1,025, 813, 761 cm^-1

;

Yield 78%; IR (KBr) t_max3,442, 3,076, 1,662, 1,523,

¹H NMR (DMSO-d₆, 500 MHz) d 7.89 (s, 1H, H-5), 7.82

1,484, 1,416, 1,347, 1,328, 1,037, 850, 758 cm^-1; ¹H NMR

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Med Chem Res (2012) 21:1833–1849

(DMSO-d₆, 300 MHz) d 8.40 (br, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 7.90 (d, 1H, J = 2.4 Hz, H-5), 7.74 (d, 1H,

J = 2.4 Hz, H-7), 7.43(m, 1H, H-6), 7.26 (s, 1H, H-3), and

2.62 (s, 3H, –CH₃); ESI–MS m/z 282 [M ? H]^?.

ﬁltered and recrystallized to give the target molecules 17f–

17v, 18f–18r, and 19f–19w, respectively.

6-((Diethylamino)methyl)-2-(4-nitrophenyl)-4H-chromen-

4-one (17f)

6-Chloro-8-methyl-2-(4-nitrophenyl)-4H-chromen-4-one

(19b)

Yield 42.3%; IR (KBr) t_max3,450, 2,968, 2,796, 1,639,

1

1,573, 1,522, 1,481, 1,347, 1,042, 851, 822, 690 cm^-1; H

Yield 78%; IR (KBr) t_max3,443, 3,071, 1,667, 1,582,

NMR (CDCl₃, 300 MHz) d 8.39 (d, 2H, J = 8.2 Hz, H-3⁰,

H-5⁰), 8.12 (d, 2H, J = 8.2 Hz, H-2⁰, H-6⁰), 7.83 (d, 1H,

J = 8.1 Hz, H-8), 7.57 (d, 1H, J = 8.1 Hz, H-7), 7.27 (s,

1H, H-5), 6.91 (s, 1H, H-3), 3.67 (s, 2H, Ar–CH₂–), 2.54

(q, 4H, J = 6.9 Hz, N(CH₂CH₃)₂), and 1.06 (t, 6H,

J = 6.9 Hz, NCH₂(CH₃)₂); ESI–MS m/z 353 [M ? H]^?.

1

1,523, 1,465, 1,347, 1,300, 1,041, 882, 864, 852 cm^-1; H

NMR (DMSO-d₆, 300 MHz) d 8.41 (d, 2H, J = 9.0 Hz,

H-3⁰, H-5⁰), 8.02 (d, 2H, J = 9.0 Hz, H-2⁰, H-6⁰), 7.98 (d,

1H, J = 2.1 Hz, H-5), 7.48 (d, 1H, J = 2.1 Hz, H-7), 6.83

(s, 1H, H-3), and 2.15 (s, 3H, CH₃); ESI–MS m/z 316

[M ? H]^?.

2-(4-Nitrophenyl)-6-(piperidin-1-ylmethyl)-4H-chromen-4-

one (17g)

6-Chloro-2-(4-chlorophenyl)-8-methyl-4H-chromen-4-one

(19c)

Yield 37.6%; IR (KBr) t_max3,443, 1,640, 1,569, 1,518,

1

Yield 76%; IR (KBr) t_max3,444, 3,069, 1,636, 1,594,

1,492, 1,465, 1,415, 1,361, 1,291, 1,095, 1,042, 880,

1,483, 1,414, 1,346, 1,038, 851, 757, 693 cm^-1; H NMR

(DMSO-d₆, 500 MHz) d 8.39 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 7.96 (s, 1H, H-8), 7.79 (s, 1H, H-5), 7.23 (s, 1H, H-3),

3.56 (s, 2H, Ar–CH₂–), 2.36 (m, 4H, N(CH₂CH₂)₂CH₂),

1.69 (m, 2H, N(CH₂CH₂)₂CH₂), and 1.50 (m, 4H,

N(CH₂CH₂)₂CH₂); ESI–MS m/z 365 [M ? H]^?.

836 cm^-^{1 1}H NMR (CDCl₃, 300 MHz) d 8.03 (s, 1H,

;

H-5), 7.85 (d, 2H, J = 8.4 Hz, H-3⁰, H-5⁰), 7.52(d, 2H,

J = 8.4 Hz, H-2⁰, H-6⁰), 7.27 (s, 1H, H-7), 6.81 (s, 1H,

H-3), and 2.59 (s, 3H, CH₃); ESI–MS m/z 305 [M ? H]^?.

6-((4-Methylpiperazin-1-yl)methyl)-2-(4-nitrophenyl)-4H-

chromen-4-one (17h)

6-Chloro-2-(2-chlorophenyl)-8-methyl-4H-chromen-4-one

(19d)

Yield 34.6%; IR (KBr) t_max3,453, 2,929, 1,639, 1,521, 1,482,

1,452, 1,349, 1,133, 1,041, 904, 853 cm^-1; ¹H NMR (DMSO-

d₆, 500 MHz) d 8.39 (m, 4H, H-2⁰, H-3⁰, H-5⁰, H-6⁰), 7.96 (s,

1H, H-8), 7.79(s, 2H, H-5, H-7), 7.23(s, 1H, H-3), 3.59(s, 2H,

Ar–CH₂–), 2.38 (m, 8H, N(CH₂CH₂)₂NCH₃), and 2.16 (s, 3H,

NCH₃); ESI–MS m/z 380 [M ? H]^?.

Yield 76%; IR (KBr) t_max3,442, 1,637, 1,595, 1,581,

1,493, 1,465, 1,405, 1,361, 1,291, 1,163, 1,095, 1,041,

1

1,011, 880, 837 cm^-1; H NMR (CD₃COCD₃, 300 MHz)

d 7.89 (m, 3H, H-3⁰, H-5, H-6⁰), 7.65 (m, 3H, H-4⁰, H-5⁰,

H-7), 6.64 (s, 1H, H-3), and 2.54 (s, 3H, CH₃); ESI–MS m/z

305 [M ? H]^?.

6-(Morpholinomethyl)-2-(4-nitrophenyl)-4H-chromen-

4-one (17i)

General procedure for the synthesis

of the aminomethylﬂavones

Yield 45.0%; IR (KBr) t_max3,423, 1,639, 1,618, 1,520,

To a solution of methylﬂavones 17a–17c, 18a, 18b,

and 19a–19d (5 mmol) in CCl₄(150 ml) were added

N-bromosuccinimide (5 mmol) and a catalytic amount of

benzyl peroxide, and the mixture was reﬂuxed for 12 h.

The mixture was hot-ﬁltered and evaporated to dryness

to give 24a–24i, respectively, and they were used with-

out further puriﬁcation. Bromomethylﬂavones 24a–24j

(1 mmol) were individually mixed with different amine

(10 mmol) in CH₂Cl₂at room temperature with stirring for

h, and the solvent was evaporated to dryness. HCl

solution was added to adjust the pH 3*4 and ﬁltered. Then,

the pH of the solution was adjusted to 10 with ammonia

water, and precipitation appeared. The precipitation was

1,452, 1,346, 1,120, 1,084, 904, 866 cm^-^{1 1}H NMR

;

(DMSO-d₆, 500 MHz) d 8.39 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 8.30 (s, 1H, H-5), 8.12 (d, 1H, J = 8.5 Hz, H-8),

7.94 (d, 1H, J = 8.5 Hz, H-7), 7.30 (s, 1H, H-3), 4.52 (s,

2H, Ar–CH₂–), 3.95 (m, 2H, N(CH₂CH₂)₂O), 3.75 (m, 2H,

N(CH₂CH₂)₂O), 3.26 (m, 2H, N(CH₂CH₂)₂O), and 3.13

(m, 2H, N(CH₂CH₂)₂O); ESI–MS m/z 367 [M ? H]^?.

2-(4-Aminophenyl)-6-((dimethylamino)methyl)-4H-

chromen-4-one (17j)

Yield 27.1%; IR (KBr) t_max3,451, 1,641, 1,520, 1,482,

1

1,448, 1,349, 1,124, 1,037, 901, 852, 694 cm^-1; H NMR

123

Med Chem Res (2012) 21:1833–1849

1843

(DMSO-d₆, 500 MHz) d 8.40 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 8.11 (s, 1H, H-5), 7.93 (d, 1H, J = 8.6 Hz, H-8),

7.85 (d, 1H, J = 8.6 Hz, H-7), 7.26 (s, 1H, H-3), 3.95 (s,

2H, Ar–CH₂–), and 2.44 (s, 6H, N(CH₃)₂); ESI–MS m/z

325 [M ? H]^?.

2-(4-Chlorophenyl)-6-((4-methylpiperazin-1-yl)methyl)-

4H-chromen-4-one (17o)

Yield 52%; IR (KBr) t_max3,443, 2,930, 2,797, 1,638,

1,619, 1,575, 1,492, 1,452, 1,407, 1,361, 1,278, 1,091,

1,040, 1,011, 903, 832 cm^-1

;

¹H NMR (DMSO-d₆,

500 MHz) d 8.13 (d, 2H, J = 8.5 Hz, H-2⁰, H-6⁰), 7.94 (s,

1H, H-5), 7.66 (s, 2H, H-7, H-8), 7.61 (d, 2H, J = 8.5 Hz,

H-3⁰, H-5⁰), 7.07 (s, 1H, H-3), 3.59 (s, 2H, Ar–CH₂–), 2.42

(br, 8H, N(CH₂CH₂)₂NCH₃), and 2.18 (s, 3H, NCH₃); ESI–

MS m/z 369 [M ? H]^?.

6-((1H-imidazol-1-yl)methyl)-2-(4-nitrophenyl)-4H-

chromen-4-one (17k)

Yield 30.5%; IR (KBr) t_max3,443, 1,639, 1,572, 1,516,

1,483, 1,444, 1,347, 1,184, 1,040, 912, 852, 823,

693 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.39 (m, 4H,

H-2⁰, H-3⁰, H-5⁰, H-6⁰), 7.94 (s, 1H, H-2⁰⁰), 7.86 (s, 1H,

H-5), 7.78 (m, 2H, H-7, H-8), 7.59 (s, 1H, H-5⁰⁰), 7.24 (s,

1H, H-4⁰⁰), 6.69 (s, 1H, H-3), and 5.38 (s, 2H, Ar–CH₂–);

ESI–MS m/z 348 [M ? H]^?.

2-(4-Chlorophenyl)-6-(morpholinomethyl)-4H-chromen-

4-one (17p)

Yield 56%; IR (KBr) t_max3,443, 2,852, 2,803, 1,638,

1,575, 1,494, 1,454, 1,409, 1,131, 1,112, 1,091, 1,010, 856,

1

833 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.14 (d, 2H,

N,N-Diethyl-N-((2-(4-nitrophenyl)-4-oxo-4H-chromen-

6-yl)methyl)ethanaminium (17l)

J = 8.5 Hz, H-2⁰, H-6⁰), 7.97 (s, 1H, H-5), 7.77 (m, 2H,

H-7, H-8), 7.66 (d, 2H, J = 8.5 Hz, H-3⁰, H-5⁰), 7.08 (s,

1H, H-3), 3.59 (s, 6H, Ar–CH₂–, N(CH₂CH₂)₂O), and 2.39

(br, 4H, N(CH₂CH₂)₂O); ESI–MS m/z 356 [M ? H]^?.

Yield 51.2%; IR (KBr) t_max3,442, 1,639, 1,572, 1,521,

1,488, 1,448, 1,411, 1,347, 1,198, 1,037, 910, 852,

694 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.41 (m, 4H,

H-2⁰, H-3⁰, H-5⁰, H-6⁰), 8.26 (s, 1H, H-5), 7.98 (m, 2H, H-7,

H-8), 7.32 (s, 1H, H-3), 4.70 (s, 2H, Ar–CH₂–), 3.23

(q, 6H, J = 7.0 Hz, N(CH₂CH₃)₃), and 1.35 (t, 9H,

J = 7.0 Hz, N(CH₂CH₃)₃); ESI–MS m/z 381 [M ? H]^?.

2-(4-Chlorophenyl)-6-(piperazin-1-ylmethyl)-4H-chromen-

4-one (17q)

Yield 51%; IR (KBr) t_max3,439, 2,923, 1,645, 1,550,

1,494, 1,454, 1,408, 1,362, 1,113, 1,091, 1,038, 1,010,

1

756 cm^-1; H NMR (DMSO-d₆, 300 MHz) d 8.13 (d, 2H,

2-(4-Chlorophenyl)-6-((diethylamino)methyl)-4H-

chromen-4-one (17m)

J = 8.1 Hz, H-2⁰, H-6⁰), 7.93 (s, 1H, H-5), 7.75 (m, 2H,

H-7, H-8), 7.65 (d, 2H, J = 8.1 Hz, H-3⁰, H-5⁰), 7.08 (s,

1H, H-3), 3.55 (s, 2H, Ar–CH₂–), 2.70 (br, 4H,

N(CH₂CH₂)₂NH), 2.48(s, 1H, NH), and 2.32(br, 4H,

N(CH₂CH₂)₂NH); ESI–MS m/z 355 [M ? H]^?.

Yield 46%; IR (KBr) t_max3,444, 2,967, 1,642, 1,621,

1,576, 1,495, 1,448, 1,408, 1,360, 1,095, 1,037, 910,

1

827 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.12 (d, 2H,

J = 8.5 Hz, H-2⁰, H-6⁰), 7.97 (s, 1H, H-5), 7.78 (d, 1H,

J = 9.0 Hz, H-7), 7.73 (d, 1H, J = 9.0 Hz, H-8), 7.65 (d,

2H, J = 8.5 Hz, H-3⁰, H-5⁰), 7.06 (s, 1H, H-3), 3.65 (s, 2H,

Ar–CH₂–), 2.50 (q, 4H, J = 7.0 Hz, N(CH₂CH₃)₂), and

0.99 (t, 6H, J = 7.0 Hz, N(CH₂CH₃)₂); ESI–MS m/z 342

[M ? H]^?.

2-(4-Chlorophenyl)-6-((dimethylamino)methyl)-4H-

chromen-4-one (17r)

Yield 39%; IR (KBr) t_max3,441, 2,969, 2,770, 1,642,

1,622, 1,577, 1,495, 1,455, 1,408, 1,364, 1,134, 1,095,

1

1,029, 1,012, 903, 840, 820 cm^-1; H NMR (DMSO-d₆,

500 MHz) d 8.13 (d, 2H, J = 8.5 Hz, H-2⁰, H-6⁰), 7.94 (s,

1H, H-5), 7.74 (m, 2H, H-7, H-8), 7.65 (d, 2H, J = 8.5 Hz,

H-3⁰, H-5⁰), 7.06 (s, 1H, H-3), 3.52 (s, 2H, Ar–CH₂–), and

2.18 (s, 6H, N(CH₃)₂); ESI–MS m/z 314 [M ? H]^?.

2-(4-Chlorophenyl)-6-(piperidin-1-ylmethyl)-4H-

chromen-4-one (17n)

Yield 48%; IR (KBr) t_max3,431, 2,935, 1,643, 1,620,

1,594, 1,494, 1,453, 1,408, 1,366, 1,094, 1,038, 1,011, 906,

6-((1H-imidazol-1-yl)methyl)-2-(4-chlorophenyl)-4H-

chromen-4-one (17s)

1

829 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.15 (d, 2H,

J = 8.5 Hz, H-2⁰, H-6⁰), 8.05 (s, 1H, H-5), 7.87 (m, 2H,

H-7, H-8), 7.68 (d, 2H, J = 8.5 Hz, H-3⁰, H-5⁰), 7.12 (s,

1H, H-3), 3.32 (s, 2H, Ar–CH₂–), 2.50 (m, 4H,

N(CH₂CH₂)₂CH₂), and 1.73 (br, 6H, N(CH₂CH₂)₂CH₂);

ESI–MS m/z 354 [M ? H]^?.

Yield 47%; IR (KBr) t_max3,433, 1,640, 1,620, 1,578,

1,493, 1,452, 1,408, 1,369, 1,285, 1,232, 1,093, 1,038, 911,

1

827 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.12 (d, 2H,

J = 8.5 Hz, H-2⁰, H-6⁰), 7.91 (s, 1H, H-2⁰⁰), 7.84 (s, 1H,

123

1844

Med Chem Res (2012) 21:1833–1849

H-5), 7.79 (d, 1H, J = 8.5 Hz, H-5⁰⁰), 7.73 (d, 1H,

J = 8.5 Hz, H-4⁰⁰), 7.64 (d, 2H, J = 8.5 Hz, H-3⁰, H-5⁰),

7.26 (s, 1H, H-8), 7.07 (s, 1H, H-7), 6.95(s, 1H, H-3), and

5.36 (s, 2H, Ar–CH₂–); ESI–MS m/z 337 [M ? H]^?.

2-(4-Nitrophenyl)-7-(piperidin-1-ylmethyl)-4H-chromen-4-

one (18g)

Yield 34.8%; IR (KBr) t_max3,425, 2,937, 1,635, 1,521,

1

1,438, 1,346, 1,106, 1,043, 910, 851, 756, 692 cm^-1; H

NMR (DMSO-d₆, 500 MHz) d 8.39 (m, 4H, H-2⁰, H-3⁰,

H-5⁰, H-6⁰), 8.01 (d, 1H, J = 9.0 Hz, H-5), 7.77 (s, 1H,

H-8), 7.48 (d, 1H, J = 9.0 Hz, H-6), 7.22 (s, 1H, H-3),

3.68 (s, 2H, Ar–CH₂–), 2.46 (m, 4H, N(CH₂CH₂)₂CH₂),

1.56 (m, 4H, N(CH₂CH₂)₂CH₂), and 1.43 (m, 2H,

N(CH₂CH₂)₂CH₂); ESI–MS m/z 365 [M ? H]^?.

N-((2-(4-chlorophenyl)-4-oxo-4H-chromen-6-yl)methyl)-

N,N-diethylethanaminium (17t)

Yield 52%; IR (KBr) t_max3,439, 2,997, 1,642, 1,618,

1,577, 1,495, 1,449, 1,408, 1,360, 1,095, 1,036, 908,

829 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.20 (br, 1H,

H-5), 8.15 (d, 2H, J = 8.5 Hz, H-2⁰, H-6⁰), 7.92 (m, 2H,

H-7, H-8), 7.65 (d, 2H, J = 8.5 Hz, H-3⁰, H-5⁰), 7.15

(s, 1H, H-3), 4.65 (s, 2H, Ar–CH₂–), 3.19 (q, 6H,

J = 7.0 Hz, N(CH₂CH₃)₃), and 1.31 (t, 9H, J = 7.0 Hz,

N(CH₂CH₃)₃); ESI–MS m/z 370 [M]^?.

7-((4-Methylpiperazin-1-yl)methyl)-2-(4-nitrophenyl)-4H-

chromen-4-one (18h)

Yield 36.3%; IR (KBr) t_max3,443, 2,938, 2,803, 1,635,

1,519, 1,437, 1,347, 1,160, 1,135, 1,045, 909, 850, 812,

755, 691 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.39 (m,

4H, H-2⁰, H-3⁰, H-5⁰, H-6⁰), 8.02 (d, 1H, J = 9.2 Hz, H-5),

7.75 (s, 1H, H-8), 7.47 (d, 1H, J = 9.2 Hz, H-6), 7.23 (s,

1H, H-3), 3.67 (s, 2H, Ar–CH₂–), 2.50 (brs, 8H,

N(CH₂CH₂)₂NCH₃), and 2.30 (s, 3H, NCH₃); ESI–MS m/z

380 [M ? H]^?.

N-((2-(2-chlorophenyl)-4-oxo-4H-chromen-6-yl)methyl)-

N,N-diethylethanaminium (17u)

Yield 51.2%; IR (KBr) t_max3,442, 1,639, 1,572, 1,521,

1,488, 1,448, 1,411, 1,347, 1,198, 1,037, 910, 852,

694 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.41 (m, 4H,

H-2⁰, H-3⁰, H-5⁰, H-6⁰), 8.26 (s, 1H, H-5), 7.98 (m, 2H, H-7,

H-8), 7.32 (s, 1H, H-3), 4.70 (s, 2H, Ar–CH₂–), 3.23

(q, 6H, J = 7.0 Hz, N(CH₂CH₃)₃), and 1.35 (t, 9H,

J = 7.0 Hz, N(CH₂CH₃)₃); ESI–MS m/z 381 [M ? H]^?.

7-(Morpholinomethyl)-2-(4-nitrophenyl)-4H-chromen-4-

one (18i)

Yield 43.1%; IR (KBr) t_max3,451, 2,964, 1,634, 1,521,

1,436, 1,346, 1,113, 1,008, 910, 850 cm^-^{1 1}H NMR

;

(DMSO-d₆, 500 MHz) d 8.40 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 8.03 (d, 1H, J = 9.1 Hz, H-5), 7.77 (s, 1H, H-8),

7.49 (d, 1H, J = 9.1 Hz, H-6), 7.23 (s, 1H, H-3), 3.66 (s,

2H, –CH₂–), 3.61 (t, 4H, J = 4.5 Hz, N(CH₂CH₂)₂O), and

2.43 (t, 4H, J = 4.5 Hz, N(CH₂CH₂)₂O); ESI–MS m/z 367

[M ? H]^?.

6-((Diethylamino)methyl)-2-phenyl-4H-chromen-4-one

(17v)

Yield 51.2%; IR (KBr) t_max3,442, 2,974, 2,793, 1,647,

1,621, 1,483, 1,454, 1,364, 1,225, 1,198, 1,043, 901, 831,

771, 687 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.11 (m,

2H, H-2⁰, H-6⁰), 7.99 (s, 1H, H-5), 7.76 (m, 2H, H-7, H-8),

7.60 (br, 3H, H-3⁰, H-4⁰, H-5⁰), 7.03 (s, 1H, H-3), 3.66

(s, 2H, Ar–CH₂–), 2.49 (q, 4H, J = 6.5 Hz, N(CH₂CH₃)₂,

and 1.00 (t, 6H, J = 6.5 Hz, N(CH₂CH₃)₂); ESI–MS m/z

308 [M ? H]^?.

2-(4-Nitrophenyl)-7-(piperazin-1-ylmethyl)-4H-chromen-

4-one (18j)

Yield 30.7%; IR (KBr) t_max3,443, 2,807, 1,634, 1,521,

1,436, 1,346, 1,139, 1,008, 910, 850 cm^-^{1 1}H NMR

;

(DMSO-d₆, 300 MHz) d 8.40 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 8.03 (d, 1H, J = 8.1 Hz, H-5), 7.77 (d, 1H,

J = 8.1 Hz, H-6), 7.48 (s, 1H, H-8), 7.23 (s, 1H, H-3), 3.65

(s, 2H, Ar–CH₂–), 2.83 (m, 4H, N(CH₂CH₂)₂NH)), and

2.43 (m, 4H, N(CH₂CH₂)₂NH)); ESI–MS m/z 366

[M ? H]^?.

7-((Diethylamino)methyl)-2-(4-nitrophenyl)-4H-chromen-

4-one (18f)

Yield 43.2%; IR (KBr) t_max3,443, 2,971, 2,797, 1,636,

1,523, 1,437, 1,346, 1,115, 1,042, 910, 851, 757,

692 cm^-1; ¹H NMR (DMSO-d₆, 500 MHz) d 8.37 (m, 4H,

H-2⁰, H-3⁰, H-5⁰, H-6⁰), 8.00 (d, 1H, J = 8.0 Hz, H-5), 7.74

(s, 1H, H-8), 7.47 (d, 1H, J = 8.0 Hz, H-6), 7.20 (s, 1H,

H-3), 3.70 (s, 2H, Ar–CH₂–), 2.56 (q, 4H, J = 6.9 Hz,

N(CH₂CH₃)₂), and 1.01 (t, 6H, J = 6.9 Hz, N(CH₂CH₃)₂);

ESI–MS m/z 353 [M ? H]^?.

2-(4-Chlorophenyl)-7-((diethylamino)methyl)-4H-

chromen-4-one (18k)

Yield 41.2%; IR (KBr) t_max3,444, 2,970, 2,803, 1,638,

1,572, 1,492, 1,439, 1,371, 1,094, 1,042, 909, 826 cm^-1

;

123

Med Chem Res (2012) 21:1833–1849

1845

¹H NMR (DMSO-d₆, 500 MHz) d 8.16 (d, 2H, J = 7.8 Hz,

H-2⁰, H-6⁰), 8.00 (d, 1H, J = 9.0 Hz, H-5), 7.73 (s, 1H,

H-8), 7.66 (d, 2H, J = 7.8 Hz, H-3⁰, H-5⁰), 7.47 (d, 1H,

J = 9.0 Hz, H-6), 7.06 (s, 1H, H-3), 3.71 (s, 2H, Ar–CH₂–),

2.54 (q, 4H, J = 6.9 Hz, N(CH₂CH₃)₂), and 1.02 (t, 6H,

J = 6.9 Hz, N(CH₂CH₃)₂); ESI–MS m/z 342 [M ? H]^?.

H-6), 7.07 (s, 1H, H-3), 3.61 (s, 2H, Ar–CH₂–), 2.74 (m,

4H, CH₂N(CH₂CH₂)₂NH)), and 2.36 (m, 4H,

CH₂N(CH₂CH₂)₂NH)); ESI–MS m/z 355 [M ? H]^?.

2-(4-Chlorophenyl)-7-((dimethylamino)methyl)-4H-

chromen-4-one (18p)

2-(4-chlorophenyl)-7-(piperidin-1-ylmethyl)-4H-chromen-

4-one (18l)

Yield 38%; IR (KBr) t_max3,443, 2,976, 1,637, 1,572,

1,492, 1,442, 1,373, 1,096, 1,032, 908, 823 cm^-1; ¹H NMR

(DMSO-d₆, 500 MHz) d 8.15 (d, 2H, J = 9.2 Hz, H-2⁰,

H-6⁰), 8.00 (d, 1H, J = 9.0 Hz, H-5), 7.73 (s, 1H, H-8),

7.65 (d, 2H, J = 9.2 Hz, H-3⁰, H-5⁰), 7.46 (d, 1H,

J = 9.0 Hz, H-6), 7.07 (s, 1H, H-3), 3.63 (s, 2H, Ar–CH₂),

and 2.25 (s, 6H, N(CH₃)₂); ESI–MS m/z 314 [M ? H]^?.

Yield 43%; IR (KBr) t_max3,442, 2,933, 2,794, 1,642,

1,570, 1,492, 1,437, 1,345, 1,106, 1,042, 909, 831 cm^-1

;

¹H NMR (DMSO-d₆, 500 MHz) d 8.15 (d, 2H, J = 7.8 Hz,

H-2⁰, H-6⁰), 7.99 (d, 1H, J = 9.0 Hz, H-5), 7.70 (s, 1H,

H-8), 7.65(d, 2H, J = 7.8 Hz, H-3⁰, H-5⁰), 7.44 (d, 1H,

J = 9.0 Hz, H-6), 7.06 (s, 1H, H-3), 3.60 (s, 2H, Ar–CH₂–),

2.38 (br, 4H, N(CH₂CH₂)₂CH₂), 1.52(br, 4H,

N(CH₂CH₂)₂CH₂), and 1.41 (br, 2H, N(CH₂CH₂)₂CH₂);

ESI–MS m/z 354 [M ? H]^?.

7-((1H-imidazol-1-yl)methyl)-2-(4-chlorophenyl)-4H-

chromen-4-one (18q)

Yield 39%; IR (KBr) t_max3,441, 1,629, 1,571, 1,490,

1,437, 1,410, 1,372, 1,096, 1,011, 909, 831 cm^-1; ¹H NMR

(DMSO-d₆, 500 MHz) d 8.09 (d, 2H, J ,= 8.6 Hz, H-2⁰,

H-6⁰), 8.00 (d, 1H, J = 9.0 Hz, H-5), 7.87 (s, 1H, H-8),

7.64 (d, 2H, J = 8.6 Hz, H-3⁰, H-5⁰), 7.62 (s, 1H, H-2⁰⁰),

7.57 (d, 1H, J = 9.0 Hz, H-6), 7.36 (d, 1H, J = 5.5 Hz,

H-4⁰⁰), 7.06 (s, 1H, H-3), 7.00 (d, 1H, J = 5.5 Hz, H-5⁰⁰),

and 5.42 (s, 2H, Ar–CH₂–); ESI–MS m/z 337 [M ? H]^?.

2-(4-Chlorophenyl)-7-((4-methylpiperazin-1-yl)methyl)-

4H-chromen-4-one (18m)

Yield 46%; IR (KBr) t_max3,428, 2,790, 1,641, 1,569,

1,491, 1,436, 1,409, 1,370, 1,283, 1,093, 1,042, 1,011, 907,

1

830 cm^-1; H NMR (DMSO-d₆, 300 MHz) d 8.14 (d, 2H,

J = 6.9 Hz, H-2⁰, H-6⁰), 8.00 (d, 1H, J = 7.8 Hz, H-5),

7.69 (s, 1H, H-8), 7.65 (d, 2H, J = 6.9 Hz, H-3⁰, H-5⁰),

7.43 (d, 1H, J = 7.8 Hz, H-6), 7.06 (s, 1H, H-3), 3.63 (s,

2H, Ar–CH₂–), 2.65 (br, 4H, N(CH₂CH₂)₂NCH₃), 2.44 (br,

4H, N(CH₂CH₂)₂NCH₃), and 2.21 (s, 3H, NCH₃); ESI–MS

m/z 369 [M ? H]^?.

N-((2-(4-chlorophenyl)-4-oxo-4H-chromen-7-yl)methyl)-

N,N-diethylethan-amiium (18r)

Yield 46%; IR (KBr) t_max3,444, 2,985, 1,639, 1,562,

1,492, 1,440, 1,411, 1,369, 1,311, 1,095, 1,012, 909,

2-(4-Chlorophenyl)-7-(morpholinomethyl)-4H-chromen-4-

one (18n)

1

834 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.15 (d, 2H,

J = 8.5 Hz, H-2⁰, H-6⁰), 8.10 (d, 1H, J = 9.2 Hz, H-5),

8.03 (d, 1H, J = 1.8 Hz, H-8), 7.66 (d, 2H, J = 8.5 Hz,

H-3⁰, H-5⁰), 7.62 (dd, 1H, J = 9.2, 1.8 Hz, H-6), 7.13 (s,

1H, H-3), 4.67 (s, 2H, Ar–CH₂–), 3.26 (q, 6H, J = 7.2 Hz,

N(CH₂CH₃)₃, and 1.33 (t, 9H, J = 7.2 Hz, N(CH₂CH₃)₃;

ESI–MS m/z 370 [M]^?.

Yield 52%; IR (KBr) t_max3,444, 2,805, 1,641, 1,570,

1,492, 1,437, 1,409, 1,371, 1,351, 1,112, 1,005, 907, 865,

1

810 cm^-1; H NMR (DMSO-d₆, 500 MHz) d 8.13 (d, 2H,

J = 9.1 Hz, H-2⁰, H-6⁰), 7.99 (d, 1H, J = 9.0 Hz, H-5),

7.71 (s, 1H, H-8), 7.64 (d, 2H, J = 9.1 Hz, H-3⁰, H-5⁰),

7.45 (d, 1H, J = 9.0 Hz, H-6), 7.04 (s, 1H, H-3), 3.64 (s,

2H, Ar–CH₂–), 3.62 (t, 4H, N(CH₂CH₂)O), and 2.51(t, 4H,

J = 4.4 Hz, N(CH₂CH₂)₂O); ESI–MS m/z 356 [M ? H]^?.

8-((Diethylamino)methyl)-2-(4-nitrophenyl)-4H-chromen-

4-one (19f)

2-(4-Chlorophenyl)-7-(piperazin-1-ylmethyl)-4H-chromen-

4-one (18o)

Yield 43.2%; IR (KBr) t_max3,445, 2,970, 1,655, 1,522,

1

1,417, 1,347, 1,117, 1,066, 851, 758, 696 cm^-1; H NMR

(DMSO-d₆, 300 MHz) d 8.41 (br, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 7.98 (d, 1H, J = 8.1 Hz, H-5), 7.91 (d, 1H,

J = 8.1 Hz, H-7), 7.51(d, 1H, J = 8.1 Hz, H-6), 7.25 (s,

1H, H-3), 3.98 (s, 2H, Ar–CH₂–), 2.60 (q, 4H, J = 6.8 Hz,

N(CH₂CH₃)₂), and 1.03 (t, 6H, J = 6.8 Hz, N(CH₂CH₃)₂);

ESI–MS m/z 353 [M ? H]^?.

Yield 36.7%; IR (KBr) t_max3,443, 2,927, 1,640, 1,569,

1,436, 1,410, 1,371, 1,112, 1,094, 908, 829 cm^-1; ¹H NMR

(DMSO-d₆, 300 MHz) d 8.15 (d, 2H, J = 8.4 Hz, H-2⁰,

H-6⁰), 8.00 (d, 1H, J = 8.1 Hz, H-5), 7.72 (s, 1H, H-8),

7.66 (d, 1H, J = 8.4, H-3⁰, H-5⁰), 7.45 (d, 1H, J = 8.1 Hz,

123

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Med Chem Res (2012) 21:1833–1849

8-((4-Methylpiperazin-1-yl)methyl)-2-(4-nitrophenyl)-4H-

chromen-4-one (19g)

N(CH₂CH₂)₂NCH₃), 2.39 (br, 4H, N(CH₂CH₂)₂NCH₃),

and 2.15 (br, 3H, N(CH₂CH₂)₂NCH₃); ESI–MS m/z 414

[M ? H]^?.

Yield 45%; IR (KBr) t_max3,439, 2,923, 1,645, 1,481,

1,460, 1,416, 1,376, 1,281, 1,145, 1,036, 1,009, 847,

756 cm^-1; ¹H NMR (DMSO-d₆, 300 MHz) d 8.20 (br, 2H,

H-3⁰, H-5⁰), 7.95 (br, 1H, H-5), 7.88 (m, 2H, H-2⁰, H-6⁰),

7.55 (m, 1H, H-6), 6.73 (s, 1H, H-3), 3.69 (s, 2H, Ar–CH₂–),

2.34 (br, 4H, N(CH₂CH₂)₂NCH₃), 2.05 (br, 4H,

N(CH₂CH₂)₂NCH₃), and 1.01 (s, 3H, NCH₃); ESI–MS m/z

380 [M ? H]^?.

6-Chloro-8-(morpholinomethyl)-2-(4-nitrophenyl)-4H-

chromen-4-one (19l)

Yield 62%; IR (KBr) t_max3,442, 2,854, 1,649, 1,581, 1,525,

1,451, 1,347, 1,115, 1,010, 850 cm^-1; ¹H NMR (DMSO-d₆,

300 MHz) d 8.42 (m, 4H, H-2⁰, H-3⁰, H-5⁰, H-6⁰), 7.94 (s, 1H,

H-5), 7.91 (d, 1H, H-7), 7.34 (s, 1H, H-3), 3.91(s, 2H,

Ar–CH₂–), 3.61(t, 4H, J = 4.2 Hz, N(CH₂CH₂)₂O), and

2.51 (t, 4H, J = 4.2 Hz, N(CH₂CH₂)₂O); ESI–MS m/z 401

[M ? H]^?.

8-(Morpholinomethyl)-2-(4-nitrophenyl)-4H-chromen-4-

one (19h)

Yield 56%; IR (KBr) t_max3,444, 2,855, 1,640, 1,582,

6-Chloro-2-(4-nitrophenyl)-8-(piperazin-1-ylmethyl)-4H-

chromen-4-one (19m)

1,520, 1,377, 1,346, 1,116, 1,009, 853 cm^-^{1 1}H NMR

;

(DMSO-d₆, 300 MHz) d 8.42 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 8.01 (d, 1H, J = 7.8 Hz, H-5), 7.85 (d, 1H,

J = 7.8 Hz, H-7), 7.50 (t, 1H, J = 7.8 Hz, H-6), 3.90 (s,

2H, Ar–CH₂), 3.60 (t, 4H, J = 4.2 Hz, N(CH₂CH₂)₂O),

and 2.50 (t, 4H, J = 4.2 Hz, N(CH₂CH₂)₂O); ESI–MS m/z

367 [M ? H]^?.

Yield 46%;IR (KBr) t_max3,435, 1,651, 1,523, 1,447, 1,405,

1

1,347, 1,163, 1,116, 1,089, 851 cm^-1; H NMR (DMSO-

d₆, 300 MHz) d 8.41 (m, 4H, H-2⁰, H-3⁰, H-5⁰, H-6⁰), 7.92

(br, 2H, H-5, H-7), 7.32 (s, 1H, H-3), 3.93(s, 2H, Ar–CH₂),

2.63(s, 4H, N(CH₂CH₂)₂NH), 2.50 (t, 4H, J = 4.2 Hz,

N(CH₂CH₂)₂NH), and 2.27(s, 1H, NH); ESI–MS m/z 400

[M ? H]^?.

6-Chloro-8-((diethylamino)methyl)-2-(4-nitrophenyl)-4H-

chromen-4-one (19i)

6-Chloro-8-((dimethylamino)methyl)-2-(4-nitrophenyl)-

4H-chromen-4-one (19n)

Yield 43.2%; IR (KBr) t_max3,444, 2,970, 1,651, 1,523,

1,450, 1,346, 1,117, 849 cm^-^{1 1}H NMR (DMSO-d₆,

;

300 MHz) d 8.41 (br, 4H, H-2⁰, H-3⁰, H-5⁰, H-6⁰), 7.90 (br,

2H, H-5, H-7), 7.29 (s, 1H, H-3), 3.97 (s, 2H, Ar–CH₂–),

2.60 (q, 4H, J = 6.9 Hz, N(CH₂CH₃)₂), and 1.03 (t, 6H,

J = 6.9 Hz, N(CH₂CH₃)₂); ESI–MS m/z 387 [M ? H]^?.

Yield 40%; IR (KBr) t_max3,442, 2,943, 2,821, 1,649,

1,580, 1,523, 1,451, 1,346, 1,037, 849 cm^-^{1 1}H NMR

;

(DMSO-d₆, 300 MHz) d 8.39 (m, 4H, H-2⁰, H-3⁰, H-5⁰,

H-6⁰), 7.89 (s, 1H, H-5), 7.85 (s, 1H, H-7), 7.29 (s, 1H,

H-3), 3.82 (s, 2H, Ar–CH₂), and 2.29 (s, 6H, N(CH₃)₂);

ESI–MS m/z 359 [M ? H]^?.

6-Chloro-2-(4-nitrophenyl)-8-(piperidin-1-ylmethyl)-4H-

chromen-4-one (19j)

8-((1H-imidazol-1-yl)methyl)-6-chloro-2-(4-nitrophenyl)-

4H-chromen-4-one (19o)

Yield 38%; IR (KBr) t_max3,442, 2,936, 1,649, 1,524,

1

1,450, 1,346, 1,299, 11,107, 849 cm^-1; H NMR (DMSO-

d₆, 300 MHz) d 8.41 (br, 4H, H-2⁰, H-3⁰, H-5⁰, H-6⁰), 7.92

(s, 1H, H-5), 7.87 (s, 1H, H-7), 7.32 (s, 1H, H-3), 3.87 (s,

2H, Ar–CH₂), 2.51 (br, 4H, N(CH₂CH₂)₂CH₂), 1.55 (br,

4H, N(CH₂CH₂)₂CH₂), and 1.43 (br, 2H, N(CH₂CH₂)₂

CH₂); ESI–MS m/z 399 [M ? H]^?.

Yield 49%; IR (KBr) t_max3,431, 1,649, 1,580, 1,523,

1,460, 1,347, 1,081, 851 cm^-^{1 1}H NMR (DMSO-d₆,

;

300 MHz) d 8.38 (m, 4H, H,-2⁰, H-3⁰, H-5⁰, H-6⁰), 8.08 (m,

1H, H-2⁰⁰), 7.96 (s, 1H, H-5), 7.56 (s, 1H, H-5⁰⁰), 7.33

(br, 2H, H-4⁰⁰, H-7), 7.02 (s, 1H, H-3), and 5.72 (s, 2H,

Ar–CH₂–); ESI–MS m/z 382 [M ? H]^?.

6-Chloro-8-((4-methylpiperazin-1-yl)methyl)-2-(4-

nitrophenyl)-4H-chromen-4-one (19k)

6-Chloro-2-(2-chlorophenyl)-8-((diethylamino)methyl)-

4H-chromen-4-one (19p)

Yield 47%; IR (KBr) t_max3,424, 2,939, 2,803, 1,645,

1

1,579, 1,524, 1,454, 1,347, 1,298, 1,142, 849 cm^-1; H

Yield 49%; IR (KBr) t_max3,444, 2,972, 1,667, 1,649,

1,595, 1,455, 1,363, 1,194, 1,120, 1,071, 1,033, 857,

778 cm^-1; ¹H NMR (DMSO-d₆, 300 MHz) d 7.88 (m, 3H,

H-3⁰, H-5, H-6⁰), 7.64 (m, 3H, H-4⁰, H-5⁰, H-7), 6.74 (s, 1H,

NMR (DMSO-d₆, 300 MHz) d 8.40 (br, 4H, H-2⁰, H-3⁰,

H-5⁰, H-6⁰), 7.91 (s, 1H, H-5), 7.86 (s, 1H, H-7), 7.31 (s,

1H, H-3), 3.88 (s, 2H, Ar–CH₂–), 2.51 (br, 4H,

123

Med Chem Res (2012) 21:1833–1849

1847

1

H-3), 3.82 (s, 2H, Ar–CH₂–), 2.54 (q, 4H, J = 6.9 Hz,

N(CH₂CH₃)₂), and 0.95 (t, 6H, J = 6.9 Hz, N(CH₂CH₃)₂);

ESI–MS m/z 376 [M ? H]^?.

1,073, 1,028, 857, 770, 715 cm^-1; H NMR (DMSO-d₆,

300 MHz) d 7.93 (s, 1H, H-5), 7.86 (m, 2H, H-3⁰, H-6⁰),

7.65 (m, 3H, H-4⁰, H-5⁰, H-7), 6.76 (s, 1H, H-3), 3.72 (s,

2H, Ar–CH₂), and 2.23 (s, 6H, N(CH₃)₂); ESI–MS m/z 348

[M ? H]^?.

6-Chloro-2-(2-chlorophenyl)-8-(piperidin-1-ylmethyl)-4H-

chromen-4-one (19q)

8-((1H-imidazol-1-yl)methyl)-6-chloro-2-(2-chlorophenyl)-

4H-chromen-4-one (19v)

Yield 35%; IR (KBr) t_max3,444, 2,937, 1,644, 1,593, 1,449,

1,430, 1,352, 1,271, 1,030, 855, 796 cm^-^{1 1}H NMR

;

(DMSO-d₆, 300 MHz) d 7.94 (s, 1H, H-5), 7.90 (m, 2H, H-3⁰,

H-6⁰), 7.62 (m, 3H, H-4⁰, H-5⁰, H-7), 6.76 (s, 1H, H-3), 3.74

(s, 2H, Ar–CH₂–), 2.42 (br, 4H, N(CH₂CH₂)₂CH₂), 1.50 (br,

4H, N(CH₂CH₂)₂CH₂), and 1.39 (br, 2H, N(CH₂CH₂)₂CH₂);

ESI–MS m/z 388 [M ? H]^?.

Yield 41%; IR (KBr) t_max3,419, 1,649, 1,593, 1,462,

1,439, 1,361, 1,345, 1,286, 1,232, 1,158, 1,111, 1,088,

1,026, 898, 856, 784, 760 cm^-^{1 1}H NMR (DMSO-d₆,

;

300 MHz) d 7.99 (s, 1H, H-5⁰⁰), 7.83 (s, 1H, H-5), 7.68 (m,

4H, H-2⁰, H-3⁰, H-4⁰, H-5⁰), 7.49 (s, 1H, H-5⁰⁰), 7.20 (s, 1H,

H-7), 6.94 (s, 1H, H-4⁰⁰), 6.80 (s, 1H, H-3), and 5.54 (s, 2H,

Ar–CH₂–); ESI–MS m/z 348 [M ? H]^?.

6-Chloro-2-(2-chlorophenyl)-8-((4-methylpiperazin-1-

yl)methyl)-4H-chromen-4-one (19r)

6-Chloro-2-(4-chlorophenyl)-8-(morpholinomethyl)-4H-

chromen-4-one (19w)

Yield 48%; IR (KBr) t_max3,448, 2,939, 2,802, 1,710, 1,637,

1,581, 1,462, 1,355, 1,280, 1,161, 1,142, 1,012, 853,

1

762 cm^-1; H NMR (DMSO-d₆, 300 MHz) d 7.93 (s, 1H,

Yield 62%; IR (KBr) t_max3,445, 1,650, 1,595, 1,493,

H-5), 7.86 (m, 2H, H-3⁰, H-6⁰), 7.64 (m, 3H, H-4⁰, H-5⁰, H-7),

6.76 (s, 1H, H-3), 3.76 (s, 2H, Ar–CH₂–), 2.46 (br, 4H,

N(CH₂CH₂)₂NCH₃), 2.36 (br, 4H, N(CH₂CH₂)₂NCH₃), and

2.17 (s, 3H, NCH₃); ESI–MS m/z 403 [M ? H]^?.

1,451, 1,357, 1,117, 1,093, 1,012, 827 cm^-^{1 1}H NMR

;

(DMSO-d₆, 300 MHz) d 8.16 (d, 2H, J = 8.7 Hz, H-3⁰,

H-5⁰), 7.90 (d, 1H, J = 2.4 Hz, H-5), 7.85 (d, 1H,

J = 2.4 Hz, H-7), 7.68 (d, 2H, J = 8.7 Hz, H-2⁰, H-6⁰),

7.15 (s, 1H, H-3), 3.88 (s, 2H, Ar–CH₂–), 3.61 (t, 4H,

J = 4.5 Hz, N(CH₂CH₂)₂O), and 2.51 (t, 4H, J = 4.5 Hz,

N(CH₂CH₂)₂O); ESI–MS m/z 390 [M ? H]^?.

6-Chloro-2-(2-chlorophenyl)-8-(morpholinomethyl)-

4H-chromen-4-one (19s)

Yield 61%; IR (KBr) t_max3,444, 2,968, 2,817, 1,646,

1,592, 1,450, 1,354, 1,271, 1,138, 1,116, 1,014, 854,

General procedure for the reduction of the nitro group

1

767 cm^-1; H NMR (DMSO-d₆, 300 MHz) d 7.94 (s, 1H,

Compounds 17a, 17f, 17g, 17i, 18a, and 19b (5 mmo1)

were individually suspended in 100 ml of a solution of

ethanol in water [V(EtOH):V(H₂O) = 2:1] and heated

under reﬂux. Sodium dithionite (6 mmoI) was gradually

added to the ﬂask until the compound was dissolved

completely to give a yellow solution. An excess of

approximately 0.5 g of sodium dithionite was then added to

the solution, and the solution was reﬂuxed for another 1 h.

Ethanol was distilled at a reduced pressure, and the solu-

tion was poured to 80 ml of concentrated HCl and heated

further until the smell of SO₂disappeared. The solution

was ﬁltered hot and cooled, then neutralized with ammonia

to give 17e, 17w, 17x, 17y, 18e, and 19e, respectively.

H-5), 7.88 (m, 2H, H-3⁰, H-6⁰), 7.64 (m, 3H, H-4⁰, H-5⁰,

H-7), 6.76 (s, 1H, H-3), 3.77 (s, 2H, Ar–CH₂), 3.59 (t, 4H,

J = 4.5 Hz, N(CH₂CH₂)₂O), and 2.44 (t, 4H, J = 4.5 Hz,

N(CH₂CH₂)₂O); ESI–MS m/z 390 [M ? H]^?.

6-Chloro-2-(2-chlorophenyl)-8-(piperazin-1-ylmethyl)-

4H-chromen-4-one (19t)

Yield 52%; IR (KBr) t_max3,442, 2,937, 2,808, 1,648,

1,593, 1,450, 1,354, 1,294, 1,197, 1,131, 1,072, 854,

1

762 cm^-1; H NMR (DMSO-d₆, 300 MHz) d 7.94 (s, 1H,

H-5), 7.87 (m, 2H, H-3⁰, H-6⁰), 7.65 (m, 3H, H-4⁰, H-5⁰,

H-7), 6.76 (s, 1H, H-3), 3.75 (s, 2H, Ar–CH₂–), 2.78 (br,

4H, N(CH₂CH₂)₂NH), 2.43 (br, 4H, N(CH₂CH₂)₂NH), and

2.10 (s, 1H, NH); ESI–MS m/z 389 [M ? H]^?.

2-(4-Aminophenyl)-6-methyl-4H-chromen-4-one (17e)

Yield 89%; IR (KBr) t_max3,435, 3,355, 3,235, 1,633,

1,604, 1,558, 1,516, 1,484, 1,448, 1,367, 1,338, 1,255,

6-Chloro-2-(2-chlorophenyl)-8-((dimethylamino)methyl)-

4H-chromen-4-one (19u)

1

1,187, 1,035, 902, 826, 812 cm^-1; H NMR (DMSO-d₆,

300 MHz) d 6.93 (br, 3H, H-5, H-2⁰, H-6⁰), 6.74 (br, 2H,

H-7, H-8), 5.83 (br, 3H, H-3, H-3⁰, H-5⁰), 5.15 (s, 2H,

NH₂), and 2.50 (s, 3H, CH₃); ESI–MS m/z 252 [M ? H]^?.

Yield 51%; IR (KBr) t_max3,448, 3,065, 2,930, 2,826,

1,640, 1,508, 1,452, 1,396, 1,353, 1,266, 1,199, 1,086,

123

1848

Med Chem Res (2012) 21:1833–1849

(d, 2H, J = 8.1 Hz, H-3⁰, H-5⁰), 6.75 (s, 1H, H-3), 6.07 (s,

2H, NH₂), and 2.49 (s, 3H, CH₃); ESI–MS m/z 286

[M ? H]^?.

2-(4-Aminophenyl)-6-((diethylamino)methyl)-4H-chromen-

4-one (17w)

Yield 81%; IR (KBr) t_max3,443, 2,968, 1,630, 1,617,

1

1,563, 1,515, 1,483, 1,452, 1,365, 1,186, 833 cm^-1; H

CDK1/cyclin B activity assays

NMR (DMSO-d₆, 300 MHz) d 7.92 (s, 1H, H-5), 7.78 (d,

2H, J = 7.7 Hz, H-2⁰, H-6⁰), 7.70 (d, 1H, J = 7.4 Hz,

H-7), 7.64 (d, 1H, J = 7.4 Hz, H-8), 6.68 (d, 2H,

J = 7.4 Hz, H-3⁰, H-5⁰), 5.99 (s, 2H, NH₂), 2.50 (q, 4H,

J = 6.9 Hz, N(CH₂CH₃)₂), and 1.06 (t, 6H, J = 6.9 Hz,

N(CH₂CH₃)₂); ESI–MS m/z 323 [M ? H]^?.

The CDK1/cyclin B (Invitrogen Corporation, CA, USA)

enzyme activity was measured by a ﬂuorescence kinetic

assay using the Z⁰-LYTE^TMkinase assay kit (Invitrogen

Corporation, CA, USA) according to the protocol. The

assay was carried out at room temperature (25°C) in a

Corning^Ò384-well assay plate. The ﬁnal assay volume was

20 ll, including 2.5 ll 49 test compound, 5 ll 29 peptide

substrate/CDK1/cyclin B mixture, 2.5 ll 49 ATP solution,

5 ll development reagent solution, and 5 ll stop reagent.

The ﬁnal concentration of the assay constituents was

250 ng/ml CDK1/cyclin B, 2-lM peptide substrate, and

25-lM ATP. Continuous kinetic monitoring of enzyme

activity was performed on Thermo Scientiﬁc Varioskan

Flash with k_ex= 400, k_em= 445, and k_em= 520 nm.

2-(4-Aminophenyl)-6-(piperidin-1-ylmethyl)-4H-chromen-

4-one (17x)

Yield 80%; IR (KBr) t_max3,455, 2,934, 1,633, 1,565,

1,514, 1,453, 1,365, 1,187, 1,038, 834 cm^-^{1 1}H NMR

;

(DMSO-d₆, 500 MHz) d 8.41 (s, 1H, H-5), 8.18 (m, 2H,

H-7, H-8), 7.80 (d, 2H, J = 8.54 Hz, H-2⁰, H-6⁰), 6.75 (s,

1H, H-3), 6.70 (d, 2H, J = 8.54 Hz, H-3⁰, H-5⁰), 6.10 (s,

2H, NH₂), 2.50 (br, 4H, N(CH₂CH₂)₂CH₂), and 1.64 (br,

6H, N(CH₂CH₂)₂CH₂); ESI–MS m/z 335 [M ? H]^?.

Cytotoxicity

2-(4-Aminophenyl)-6-(morpholinomethyl)-4H-chromen-4-

one (17y)

The cytotoxic activities of the compounds were determined

using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium

bromide (MTT) reduction assay. Cells HepG2 and SMMC-

7721, growing at the logistic stage, were seeded in 100 ll

Dulbecco’s Modiﬁed Eagle’s Medium (DMEM) supple-

mented with 10% FBS in each well of 96-well microculture

plates and incubated for 24 h at 37°C in a CO₂incubator.

Compounds 17a–17y, 18a–18r, and 19a–19w and gambogic

acid as control were diluted to the desired concentrations in

the culture medium. After 48 h of incubation, media were

removed, 10-ll MTT (5 mg/ml) was added to each well, and

the plates were further incubated for 4 h. The supernatant

from each well was carefully removed, formazon crystals

were dissolved in 100 ll of DMSO, and absorption at

570-nm wavelength were recorded, respectively.

Yield 45.0%; IR (KBr) t_max3,455, 1,633, 1,571, 1,452,

1,366, 1,188, 1,111, 862 cm^-^{1 1}H NMR (DMSO-d₆,

;

500 MHz) d 7.91 (s, 1H, H-5), 7.78 (d, 2H, J = 8.5 Hz,

H-2⁰, H-6⁰), 7.70 (d, 1H, J = 8.5 Hz, H-7), 7.66 (d, 1H,

J = 8.5 Hz, H-8), 6.71 (s, 1H, H-3), 6.68 (d, 2H,

J = 8.5 Hz, H-3⁰, H-5⁰), 3.58 (m, 4H, NCH₂CH₂O), 3.35

(m, 2H, Ar–CH₂–), and 2.38 (m, 4H, N(CH₂CH₂)₂O); ESI–

MS m/z 337 [M ? H]^?.

2-(4-Aminophenyl)-7-methyl-4H-chromen-4-one (18e)

Yield 86%; IR (KBr) t_max3,443, 1,631, 1,518, 1,496, 1,425,

1,375, 1,346, 1,251, 1,187, 1,161, 852, 820 cm^-1; ¹H NMR

(DMSO-d₆, 300 MHz) d 7.87 (d, 1H, J = 7.8 Hz, H-5), 7.76

(d, 2H, J = 11.4 Hz, H-2⁰, H-6⁰), 7.51 (s, 1H, H-8), 7.25 (d,

1H, J = 7.8 Hz, H-6), 6.68 (d, 2H, J = 11.4 Hz, H-3⁰, H-5⁰),

6.66 (s, 1H, H-3), 6.00 (s, 2H, NH₂), and 2.45 (s, 3H, CH₃);

ESI–MS m/z 525 [2 M ? Na]^?.

Acknowledgments This work was supported by the Fundamental

Research Funds for the Central Universities of People’s Republic of

China (Program No: JKQ2009020).

2-(4-Aminophenyl)-6-chloro-8-methyl-4H-chromen-

4-one (19e)

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Investigation on the substitution effects of the flavonoids as potent anticancer agents: A structure-activity relationships study

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