Synthesis, complexation, and biological activity studies of 4-aminomethyl-7,8-dihydroxy coumarines and their crown ether derivatives

Article abstract of DOI:10.1002/jhet.435

(Chemical Equation Presented) The article focuses synthesis of novel 4-aminomethyl-7,8-dihydroxy coumarins and their crown ether derivatives. The purified novel coumarins and their crown ether derivatives were identified by ¹H NMR, ¹³C NMR, mass spectrometry and elemental analysis. The 1:1 binding constants of Ca²⁺, Mg²⁺, Fe²⁺, Zn²⁺, Ni²⁺, Cd²⁺, Co²⁺, and Mn ²⁺ ions at 25°C ± 0.1 with the 4-aminomethyl-7,8-dihydroxy coumarins and their crown ether derivatives estimated using extraction procedure with Inductively Coupled Plasma-Atomic Emission Spectroscopy in dichloromethane/water membrane systems. Synthesized compounds were investigated for complexation and biological activity properties. Best results in biological activity studies were observed for antioxidant activity.

Full text of DOI:10.1002/jhet.435

September 2010
Synthesis, Complexation, and Biological Activity Studies of
4-Aminomethyl-7,8-dihydroxy Coumarines and Their
Crown Ether Derivatives
1127
a
a
b
c
_
˘
Arzu Gu¨mu¨ꢀs, ꢀSeref Karadeniz, Halil Ibrahim Ugraꢀs, * Mustafa Bulut,
a
Umit C¸ akır, and Ahmet Ceyhan Goren
d
¨
¨
aDepartment of Chemistry, Faculty of Arts and Sciences, Balikesir University,
10160 Balikesir, Turkey
bDepartment of Chemistry, Faculty of Arts and Sciences, Giresun University,
28049 Giresun, Turkey
cDepartment of Chemistry, Faculty of Arts and Sciences, Marmara University,
34722 Istanbul, Turkey
dTubitak—UME, Chemical Group Laboratory, PO Box 54, 41470 Gebze, Turkey
*E-mail: halil.ibrahim.ugras@giresun.edu.tr
Received October 14, 2009
DOI 10.1002/jhet.435
Published online 16 July 2010 in Wiley Online Library (wileyonlinelibrary.com).
The article focuses synthesis of novel 4-aminomethyl-7,8-dihydroxy coumarins and their crown ether
1
derivatives. The purified novel coumarins and their crown ether derivatives were identified by H NMR,
13C NMR, mass spectrometry and elemental analysis. The 1:1 binding constants of Ca2þ, Mg2þ, Fe2þ
,
Zn2þ, Ni2þ, Cd2þ, Co2þ, and Mn2þ ions at 25ꢀC 6 0.1 with the 4-aminomethyl-7,8-dihydroxy coumar-
ins and their crown ether derivatives estimated using extraction procedure with Inductively Coupled
Plasma-Atomic Emission Spectroscopy in dichloromethane/water membrane systems. Synthesized com-
pounds were investigated for complexation and biological activity properties. Best results in biological
activity studies were observed for antioxidant activity.
J. Heterocyclic Chem., 47, 1127 (2010).
INTRODUCTION
activities, and anticoagulant effect on blood and diuretic
properties. Extensive work has been done on the synthe-
sis [8] of these classes of compounds. Methylene halide
that is attached at the 4th position is a very highly reac-
tive group in coumarins. Some coumarine compounds
obtained from reaction of coumarin-4-methylene halide
group with various amino compounds have been investi-
gated for complexation [9], biological [10], photopysical
[11,12], and spectroscopic [13–15] properties.
Coumarins are members of the class of compounds
called benzopyrones and display a variety of pharmaco-
logical properties [1] depending on their substitution
pattern. They are also known to possess antifungal and
antibacterial properties [2,3], natural [4], and synthetic
origin [5]. The diverse biological activity of chromenone
derivatives as anticoagulants and antithrombotics is well
known [6].
Macrocyclic molecules have attracted much attention
because of their potential use in a variety of chemical
processes, complexation ability, selective complexing
3-Alkyl and 4-alkylcoumarins are well known [7] for
their anthelmintic, hypnotic, insecticidal, antifungal
C
V
2010 HeteroCorporation

¨
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A. Gu¨mu¨ꢀs, Sꢀ. Karadeniz, H. I. Ugraꢀs, M. Bulut, U. C¸ akır, and A. C. Goren
Vol 47
agents for metal ions, and photoinduced electron transfer
since its discovery by Pedersen [16,17]. The complexa-
tion selectivity of crown ethers has often been explained
in terms of the size-fit concept that the crown ether
forms a more stable complex with the cation which is
more suitable in size for the crown ether cavity. From
many investigations of crown ether and cation interac-
tions it was determined that crown ethers can form vari-
able complexes with cations that are too large to fit into
the macrocyclic cavity [18–23]. This led to the synthesis
of new crown ethers derivatives which are attachment of
functional side arm.
Kex
þ LðorgÞ þ mAðaqÞ ꢀ ðMLAmÞðorgÞ
þm
M
;
(1:1)
ðaqÞ
½MLAmꢁðorgÞ
Kex
¼
;
(1:2)
(1:3)
m
½MþmꢁðaqÞ½LꢁðorgÞ½AꢁðaqÞ
KD;L ¼ ½LꢁðorgÞ=½LꢁðLÞ
:
The complexation of metal cations Ca2þ, Mg2þ, Fe2þ
,
Zn2þ, Ni2þ, Cd2þ, Co2þ, and Mn2þ have been examined
with the synthesized ligands and extracting solvent.
Association constants based on 1:1 stoichiometry of var-
ious metal complexes of these ligands were calculated
from eqs. (1.1)–(1.3) (see Table 1) as the interactions of
the ligands with different metal ions in dichloromethane
solvent media. Assuming that formation of 1:1 com-
plexes of the metal cations have been extracted at the
natural pH of the aqueous metal salt solutions. The
results are interesting, namely, the binding order of
hydroxycoumarin compounds observed for only 3,7,8,
In ion-pair extractive separation of metal cations
using a neutral chelation reagent and a counter anion,
selection of the chelation reagent is one of the most im-
portant factors to realize preferable separation. Espe-
cially, investigation of effect of steric structure around
electron donor atoms in the reagent on the separation
ability is very important for the development of novel
reagents having high separation performance. The deter-
mination of complexation constants of organic ligands-
metal ion complexes in water can be examined with dif-
ferent methods by following the extraction of metals to
the varied organic solvents with organic ligands [24–29].
The solvent extraction of metal cations which contains
macrocyclic ligands is preferred to use for its easy deter-
mination by spectrophotometric methods [23,30,31].
We report in this study the synthesis, complexation,
and biological activity studies of novel 4-aminomethyl-
7,8-dihydroxy coumarins and their crown ether
derivatives.
and 9 as (Ni2þ > Co2þ > Mn2þ > Fe2þ > Ca2þ
Zn2þ >Cd2þ > Mg2þ ), (Cd2þ > Ca2þ > Co2þ > Fe2þ
> Ni2þ > Zn2þ > Mn2þ > Mg2þ), (Co2þ > Ni2þ
Cd2þ > Mn2þ > Fe2þ > Ca2þ > Zn2þ > Mg2þ),
(Co2þ > Cd2þ > Fe2þ > Ca2þ > Zn2þ >>> Ni2þ
Mn2þ ¼ Mg2þ), respectively. It is found that the values
of stability constants of coumarine crown derivatives
were nearly in the same range for most examined metal
cations except for Cd2þ and Co2þ cations. Cd2þ is
found to be the best extracted metal cation for the cou-
marine crowns. It followed by Co2þ for all synthesized
coumarine crown derivatives. One of the most important
findings was the increase in Mg2þ selectivity for only 8
and 11 among the all examined ligands. However, the
five hydroxycoumarins (1,2,4,5, and 6) and two couma-
rin crown compounds (10 and 13) did not show any
complexation towards metal cations.
>
>
¼
RESULTS AND DISCUSSION
Synthesis of compounds. 4-chloromethyl 7,8-dihy-
droxy coumarin compound (2) was synthesized by reac-
tion of 1,2,3-trihydroxybenzene with ethyl-4-chloro-
acetoacetate. The 4-aminomethyl coumarins (2–9) were
synthesized at room temperature by stirring and then
refluxing the reaction mixture of amino compound and
triethylamine with 4-chloromethyl coumarin in dry ace-
tone. (Scheme 1). Next stage, the substituted 4-amino-
methyl coumarin crown ether derivatives (10–17) were
synthesized by refluxing 4-aminomethyl coumarins, tet-
raethyleneglycole ditosylate and Na2CO3 in CH3CN for
3–4 days.
The synthesized compounds were tested for antibacte-
rial (Table 2), antituberculosis (Table 2), antifungal (Ta-
ble 2), and antioxidant (Table 3) activity. High values
were obtained in antimicrobial activity studies especially
for some hydroxycoumarin compounds. Best results in
biological activity studies were observed for antioxidant
activity. In comparison with the control substance,
extremely high values have been obtained for 2,3,4,6,
and 8 ligands.
Complexation studies. Kex is extraction equilibrium
constant; [Mþm] and [MLAm] are the concentrations of
metal cation in aqueous phase and organic phase,
respectively. KD,L denotes a distribution constant of
ligand between organic solvent and water [25,32,33].
Extraction values, complexation, and distribution con-
stants of synthesized ligands collected in Table 1.
EXPERIMENTAL
General. The starting chemicals were purchased from
Aldrich or Merck unless otherwise cited. CaCl2, MgCl2,
ZnCl2, CoCl2, FeSO4, MnCl2, NiCl2, and CdCl2 were analyti-
cal grade reagents from Fluka dried over P2O5 for 48 h at 0.1
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

September 2010
Synthesis, Complexation, and Biological Activity Studies of
4-Aminomethyl-7,8-dihydroxy Coumarines and Their Crown Ether Derivatives
1129
Scheme 1. Synthesized compounds.
torr. The CH2Cl2 used was of analytical reagent grade. FT-IR
spectra have taken as a KBr pellet with a Perkin Elmer Spec-
trum spectrometer, model BX-II, High resolution EI mass
spectra have been obtained with Agilent 1100 LC/MSD, NMR
spectra have been obtained with a Bruker-Specrospin Avan-
ceDPX-400 Ultra-Shield 1H: 400 MHz 13C: 100 MHz. CPX
and TMS was the initial standard. All melting points reported
are uncorrected. The concentrations of metal ions in the aque-
ous phases have been determined spectroscopically: ICP-AES
(Inductively Coupled Plasma-Atomic Emission Spectroscopy
(ICP-AES): Perkin Elmer Optima 3100 XL).
1H NMR (400 MHz, Acetone/TMS) d(ppm): 7.5 (s), 7.2 (d),
6.9 (d), 6.6 (s), 6.4 (s), 4.9 (s).
General synthesis of 4-aminomethyl 7,8-dihydroxy
coumarins. A mixture of 4-chloromethyl-7,8-dihydroxy cou-
marin (0.010 mol), amino compound (0.010 mol) and acetone
(300 mL) was stirred to room temparature under N2 for 24 h.
Triethyl amine (0.010 mol) added and stirred for 1 h and then
the mixture was refluxed for 3 h. After the removal of solution
by evaporation, the resulting mixture was triturated with water,
and the precipitates were collected by filtration. The crude
product was dried under vacuum.
Organic
synthesis. 4-Chloromethyl-7,8-dihydroxy-
7,8-Dihydroxy-4-piperidine-1-ylmethyl-2H-chromen-2-
one(2). Yield (brown); 71%. mp; 65–68ꢀC. 1H NMR (400
MHz, Acetone/TMS) d(ppm): 7.73 (s), 7.28 (dd), 6.85 (dd),
6.38 (s), 6.33 (s), 4.72 (s), 2.65 (m), 1.7 (m), 1.36 (m).
4-{(Bis-(2-hydroxy-ethyl)-amino)-methyl}-7,8-dihydroxy-
2H-chromen-2-one (3). Yield (darkgreen); 67%. mp; 155–
159ꢀC. 1H NMR (400 MHz, Acetone/TMS) d(ppm): 7.78 (s),
2H-chromen-2-one(1). A mixture of pyrogallol (0.03 mol),
ethyl 4-chloroacetoacetate (0.06 mol) and HClO4 (10 mL,
70%) was heated to 90ꢀC for 4 h. The resulting mixture was
cooled, diluted with water and the precipitates were collected
by filtration. The dried crude product was purified by recry-
stallisation from ethanol. Yield (brown):48%. mp; 105–107ꢀC.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

¨
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¨
1130
A. Gu¨mu¨ꢀs, Sꢀ. Karadeniz, H. I. Ugraꢀs, M. Bulut, U. C¸ akır, and A. C. Goren
Vol 47
Table 1
D,L, % Ext, and Log Kex values for extraction of synthesized compounds in CH2Cl2 with Ca2þ, Mg2þ, Fe2þ, Zn2þ, Ni2þ, Cd2þ, Co2þ, and Mn2þ
ions at 25ꢀC 6 0.1.a
K
Cations
Ligand
Value
Zn2þ
Cd2þ
Ni2þ
Ca2þ
Mn2þ
Co2þ
Fe2þ
Mg2þ
1
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
2
3
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
0.26
20.49
8.41
–
0.14
12.30
8.06
–
2.65
72.57
10.34
–
0.68
40.52
9.08
–
1.19
54.37
9.55
–
1.78
64.09
9.94
–
0.77
43.46
9.18
–
–
–
–
–
4
% Ext
Log Kex
KD,L
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
5
–
–
–
–
–
–
–
–
% Ext
Log Kex
KD,L
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
6
–
–
–
–
–
–
–
–
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
7
0.63
38.69
9.02
0.70
41.02
9.10
11.22
85.21
5.76
–
1.98
66.39
10.04
1.34
57.23
9.66
1.88
65.24
9.99
–
1.41
58.56
9.71
2.33
69.93
10.21
–
1.86
65.02
9.98
0.86
46.13
9.27
1.41
58.48
9.71
–
0.31
23.86
8.53
0.95
48.72
9.35
–
1.80
64.35
9.95
3.63
78.40
10.69
3.01
75.08
10.48
–
1.64
62.10
9.85
0.88
46.91
9.29
1.50
59.99
9.77
–
–
–
8
0.09
8.23
7.82
–
9
–
–
–
–
–
–
10
11
12
13
14
15
16
17
–
–
–
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
KD,L
% Ext
Log Kex
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
1.27
55.96
9.61
1.03
50.84
9.43
–
2.49
71.31
10.28
2.51
71.49
10.28
–
2.19
68.61
10.14
1.35
57.41
9.67
–
0.48
32.23
8.81
0.85
45.95
9.26
–
0.90
47.27
9.30
0.56
35.97
8.93
–
2.46
71.14
10.27
1.95
66.09
10.02
–
1.04
50.99
9.43
1.10
52.28
9.48
–
0.04
4.12
7.47
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
1.72
63.23
9.90
1.32
56.92
9.65
1.46
59.40
9.74
1.68
62.69
9.88
2.73
73.22
10.38
3.12
75.71
10.52
3.23
76.33
10.56
2.72
73.13
10.37
1.71
63.03
9.89
1.55
60.82
9.80
1.88
65.29
9.99
1.64
62.18
9.86
1.06
51.56
9.45
1.24
55.30
9.59
0.63
38.59
9.02
0.86
46.26
9.27
0.67
40.21
9.07
0.72
41.71
9.12
0.66
39.71
9.05
0.97
49.32
9.37
2.28
69.52
10.19
1.80
64.26
9.94
2.49
71.31
10.28
2.32
69.86
10.20
0.95
48.79
9.36
1.72
63.22
9.90
0.85
45.92
9.26
0.91
47.76
9.32
–
–
–
–
–
–
–
–
–
–
–
a Corr. coefficient 0.999.
7.24 (d), 6.9 (d), 6.71 (s), 6.38 (s), 4.89 (s), 3.15 (m), 2.82
(br).
Bis-(4-piperazinyl methyl-7,8-dihydroxy)-2H-chromen-2-
MHz, Acetone/TMS) d(ppm): 7.38 (d), 6.93 (d), 6.86 (d), 6.41
(s), 6.3 (s), 4.91 (s), 3.84 (t).
4-((N,N-diphenylamino)-methyl)-7,8-dihydroxy-2H-chromen-
2-one(5). Yield (brown); 77%. mp; 60–62ꢀC. 1H NMR (400
1
one(4). Yield (darkgreen); 63%. mp; 82–84ꢀC. H NMR (400
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

September 2010
Synthesis, Complexation, and Biological Activity Studies of
4-Aminomethyl-7,8-dihydroxy Coumarines and Their Crown Ether Derivatives
1131
Table 2
Antibacterial, antituberculosis, antifungal, and antimicrobial activities results of synthesized compounds.
Inhibiton zone (mm)
Ligands
M. tuberculosisa M. simiae M. kansasii M. terrae M. szulgai E. coli S. aureus C. albicans M. smegmatis
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
NT
NT
NA
NA
NT
NT
NA
NA
NT
NA
NT
NT
NT
NT
NA
NT
NA
NT
NT
NT
NA
NA
NT
NT
NA
NA
NT
NA
NT
NA
NT
NT
NA
NT
NA
NT
NT
NT
9
NA
NT
NT
6
NT
NT
NA
NA
NT
NT
NA
3
NT
NA
NT
NA
NT
NT
NA
NT
NA
NT
NT
NT
9
NA
7
NA
NA
7
NA
NA
NA
6
NA
NA
7
5
6
NA
NA
7
NA
NA
NA
NA
NT
NA
NA
NA
NA
NA
NA
NA
NA
NA
NT
7
NT
NT
5
NA
NA
7
NA
NA
7
6
NA
NT
NA
NT
NA
NT
NT
NA
NT
NA
NT
5
NT
9
5
NA
7
6
7
6
NA
8
NA
13
NA
NA
12
NA
7
NA
8
NT
NA
NT
NT
NA
NT
NA
NT
NA
NA
NA
NA
6
NA
NA
NT
NA
NA
6
NA
6
17
Rifampicin
NA
NT
35
NA: not active, NT: not tested.
a MIC values (lg/mL).
MHz, Acetone/TMS) d(ppm): 7.41 (s), 7.22 (dd), 7.11 (d),
6.91 (d), 6.84 (dd), 6.38 (s), 4.88 (s).
4-(N,N-diethylamino)methyl-7,8-dihydroxy-2H-chromen-2-
one(6). Yield (yellow); 82%. mp; 79–82ꢀC. 1H NMR (400
MHz, Acetone/TMS) d(ppm): 7.56 (s), 7.25 (d), 6.93 (d), 6.65
(s), 6.4 (s), 4.91 (s), 3.1 (m), 2.12 (t).
42, 21.9. EI-MS (m/z). Mþ :434(8), 351(100), 219(47). Ele-
mental Analysis: Anal. Calcd. for C, 63.73; H, 7.21; N, 3.23;
O, 25.84; Found: C, 63.69; H, 7.18; N, 3.28; O, 25.85.
20-{(Bis-(2-hydroxy-ethyl)-amino)-methyl}-2,5,8,11,14,17-
hexaoxa-tricyclo(13.8.0.016,21)tricosa-1(2),15,19,21-tetraen-18-
1
on (11). Yield (yellow); 19%, mp; 217–220ꢀC. H NMR (400
1-(7,8-Dihydroxy-2-oxo-coumarin-4-ylmethyl)-piperidine-3-
carboxylic acid ethyl ester (7). Yield (brown); 71%. mp; 101–
103ꢀC. 1H NMR (400 MHz, acetone/TMS) d(ppm): 7.65 (s).
7.03 (s), 6.84 (s), 6.65 (s), 6.33 (s), 4.53 (s), 3.4 (m), 2.52 (m),
1.68 (m), 1.50 (t).
MHz, CDCl3/TMS) d(ppm): 7.4 (d), 7.08 (d), 6.79 (s), 4.72
(s), 3.35 (m), 2.47 (t), 4.2 (m), 3.6 (m), 2.64 (t). 13C NMR
(100 MHz) d(ppm): 166.78, 153.43, 144.4, 132, 131.2, 129,
4-(4-(4-Fluoro-phenyl)-piperazin-1-ylmethyl)-7,8-dihydroxy-
1
Table 3
2H-chromen-2-one (8). Yield (brown); 70%. mp; 94–96ꢀC. H
Antioxidant activity results.
NMR (400 MHz, Acetone/TMS) d(ppm): 7.36 (d), 6.99 (m),
6.85 (d), 6.32 (s), 3.73 (s), 3.18 (m), 2.72 (m).
Compounds
TEAC CUPRAC
7,8-dihydroxy-4-(morpholinomethyl)-2H-chromen-2-one(9).
Yield (yellow); 78%. mp; 199–202ꢀC. 1H NMR (400 MHz,
CDCl3/TMS) d(ppm): 7.8 (d), 7.2 (d), 6.7 (s), 4.7 (s), 3.2 (t),
2.4 (t).
General synthesis of 4-aminomethyl 7,8-coumarin crown
ethers. A mixture of 4-alkylamino substitued 7,8-dihydroxy
coumarin (0.005 mol), tetraethylenglycole ditosylate (0.005
mol), Na2CO3 (0.01 mol) and 700 mL CH3CN was refluxed
for 3–4 days under N2. After the removal of solution by evap-
oration, the residue was extracted with CHCl3, the organic
layer washed with water and dried on MgSO4. After the evap-
oration of CHCl3, the residue was purified by column chroma-
tography on silicagel (chloroform–methanol).
20-Piperidine-1-ylmethyl-2,5,8,11,14,17-hexaoxa-tricyclo(13.
8.0.016,21)tricosa-1(23),15,19,21-tetraen-18-on (10). Yield (yel-
low solid); 33%, mp; >300ꢀC. 1H NMR (400 MHz, CDCl3/
TMS) d(ppm): 7.7 (d), 7.15 (d), 6.3 (s), 4.78 (s), 4.03 (t), 3.93
(t), 3.6 (m),1.87 (m), 1.69 (m). 13C NMR (100 MHz) d(ppm):
158, 149, 143.5, 139.34, 128.64, 125.95, 122, 112.88, 103,
71.88, 71.00, 70.9, 70.48, 70.24, 68.6, 69.1, 65, 60.22, 60.02,
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
NA
1.28
2.40
3.64
NA
2.26
NA
0.89
NA
NA
NA
NA
NA
NA
NA
NA
NA
17
Askorbik acid
Tocopherol
0.93/0.96a
0.97/1.01a
a Values in literature.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

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1132
A. Gu¨mu¨ꢀs, Sꢀ. Karadeniz, H. I. Ugraꢀs, M. Bulut, U. C¸ akır, and A. C. Goren
Vol 47
110, 108.65, 102.73, 71.65, 71.19, 70.88, 70.32, 69.73, 68.69,
68.16, 67.66, 53, 49.8, 39.65. EI-MS (m/z). Mþ: 453(7),
391(57), 358(100), 351(75). Elemental Analysis: Anal. Calcd.
for C, 58.27; H, 6.89; N, 3.09; O, 31.75; Found: C, 58.20; H,
6.99; N, 3.00; O, 31.81.
TMS) d(ppm): 7.5 (d), 7.1 (d), 7.0 (s), 4.5 (s), 3.5 (m), 2.4 (t). 13
C
NMR (100 MHz) d(ppm): 161, 150.22, 146.26, 137.98, 133,
128.59, 110, 108.9, 103, 71.84, 71.12, 70.95, 70.42, 69.75, 69.4,
69, 68.78, 67.66, 56.63, 44.3. EI-MS (m/z). Mþ: 436(100). Elemen-
tal Analysis: Anal. Calcd. for C, 60.68; H, 6.71; N, 3.22; O, 29.39;
Found: C, 60.61; H, 6.75; N, 3.27; O, 29.37.
16,160-(piperazin-1,4-diilbis(methylen))bis(5,6,8,9,11,12-hexa-
hidro-2H-(1,4,7,10,13) pentaoxa cyclopentadeca (2,3-h)chro-
men-18(3H)-one) (12). In this procedure tetraethylenglycole
ditosylate and Na2CO3 was used twofold. Yield (yellow);
19%, mp; 285–290ꢀC. 1H NMR (400 MHz, CDCl3/TMS)
d(ppm): 7.75 (d), 7.11 (d), 6.4 (s), 4.4 (s), 3.9 (m), 3.7 (m),
2.6 (m). 13C NMR (100 MHz) d(ppm): 163.55, 142, 140.4,
138.55, 130.23, 128.6, 126, 113.45, 101.11, 70.77, 70.44,
70.29, 69.88, 69.08, 68.73, 68.11, 67.24, 53, 43, 39.87. EI-MS
(m/z). Mþ: 783(100), 611.5(74). Elemental Analysis: Anal.
Calcd. for C, 61.37; H, 6.44; N, 3.58; O, 28.61; Found: C,
61.27; H, 6.53; N, 3.58; O, 28.62.
EXTRACTION PROCEDURE
The extraction measurements were done in 100 mL
glass thermostated cell compartment with a mechanical
stirrer where a solution 10 mL (1 ꢂ 10ꢃ5 M) of an
aqueous salt and ligand in CH2Cl2 organic solvent in
appropriate concentration were placed and stirred for
120 min at 25 6 0.1ꢀC and subsequently allowed to
stand for 60 min to complete the phase separation. The
optimum concentrations of the ligands were determined
by extracting the alkali salts with 10 mL aliquot of vari-
ous concentrations of the ligands (1 ꢂ 10ꢃ5 M).
20-((Diphenylamino)-methyl)-2,5,8,11,14,17-hexaoxa-tri-
cyclo(13.8.0.016,21)tricosa-1(23),15,19,21-tetraen-18-on
(13). Yield (yellow); 29%, mp; 270–275ꢀC. 1H NMR (400
MHz, CDCl3/TMS) d(ppm): 7.45 (d), 7.09 (d), 6.9 (m), 6.2
(s), 4.65 (s), 4.1 (m), 3.32 (m). 13C NMR (100 MHz) d(ppm):
161, 147.65, 145.23, 142.95, 127, 123.2, 118.2, 116.56,
110.43, 109.33, 107.46, 107.33, 101, 71.39, 71.28, 71.05, 70.8,
70.46, 68.77, 68.39, 68.04, 36.55. EI-MS (m/z). Mþ: 517(17),
439(21), 416(48), 384(100), 300(52). Elemental Analysis:
Anal. Calcd. for C, 69.62; H, 6.04; N, 2.71; O, 21.64; Found:
C, 69.56; H, 6.01; N, 2.78; O, 21.65.
After extraction, the metal concentrations in the aqueous
phase were determined using ICP-AES. Each value was the
average of three subsequent measurements. Complexation
and distribution constants summarized in Table 1.
BIOLOGICAL ASSESSMENTS
20-Diethylaminomethyl-2,5,8,11,14,17-hexaoxa-tricyclo (13.8.
0.016,21) tricosa-1(23),15,19,21-tetraen-18-on (14). Yield (yel-
low); 23%, mp; 278–280. 1H NMR (400 MHz, CDCl3/TMS)
d(ppm): 7.49 (d), 7.1 (d), 6.87 (s), 6.3 (s), 4.7 (s), 4.1 (m), 3.5
(t), 2.48 (q), 2.26 (t). 13C NMR (100 MHz) d(ppm): 161,
145.4, 142.2, 138.55, 132.22, 128.61, 125, 101, 107, 71.37,
71.06, 70.65, 70.29, 70, 69.64, 68.91, 68.21, 56, 41.21, 36.55.
EI-MS (m/z). Mþ: 422.5(100), 407(32), 253(45), 219(47). Ele-
mental Analysis: Anal. Calcd. for C, 62.69; H, 7.41; N, 3.32;
O, 26.57; Found: C, 62.75; H, 7.43; N, 3.35; O, 26.47.
1-(18-Oxo-2,5,8,11,14,17-hexaoxa-tricyclo(13.8.0.016,21)tricosa-
1(23),15,19,21-tetraen-20-ylmethyl)-piperidine-3-carboxylic acid
Antibacterial and antifungal activities studies; Disc diffu-
sion method was used [34]. The compounds were tested
against standard bacterial strains; E. coli, S. aureus, M.
smegmatis, M. tuberculosis, M. simiae, M. kansasii, M. ter-
rae, M. szulgai, and a fungi C. albicans. Disc diffusion
method was applied for the determination of antimicrobial
activities of the samples. Compounds were dissolved in
dichloromethane (CH2Cl2) and then filter-sterilized using a
0.20-lm membrane filter. A suspension of the tested micro-
organism (0.1 mL of 108 cells/mL) was spread over the sur-
face of agar plates (MHA and SDA). Filter papers having a
diameter of 6 mm, soaked with 10 lL of samples and 10 lg
compound in solution were placed on the inoculated agar
plates. Before incubation all petri dishes were kept in the re-
frigerator (4ꢀC) for 2 h. Then they were incubated at 37ꢀC
for 24 h for bacteria and at 30ꢀC for 48 h for the yeasts. The
diameters of the inhibition zones were measured in milli-
meters. The biological activity results of synthesized com-
pounds are displayed in Table 2.
1
ethyl ester (15). Yield (brown); 35%, mp; >310. H NMR (400
MHz, CDCl3/TMS) d(ppm): 7.75 (d), 7.13 (d), 6.27 (s), 4.67
(s), 4.1 (m), 3.6 (m), 2.32 (m), 1.25 (m). 13C NMR (100 MHz)
d(ppm): 172, 161, 145, 142.65, 140.11, 128.65, 125.94, 113,
112, 109, 71.85, 71.22, 70.75, 70.50, 70.32, 69.86, 68.56,
68.24, 59.60, 58, 57.7, 56.5, 55.29, 40, 36.35. EI-MS (m/z).
Mþ: 506(17), 489(52), 475(17), 316(45), 302(100). Elemental
Analysis: Anal. Calcd. for C, 61.77; H, 6.98; N, 2.77; O,
28.48; Found: C, 61.70; H, 7.02; N, 2.71; O, 28.57.
20-(4-(4-Fluoro-phenyl)-piperazin-1-ylmethyl)-2,5,8,11,14,17-
hexaoxa-tricyclo (13.8.0.016,21
) tricosa-1(23),15,19,21-tetraen-
18-on (16). Yield (brown); 37%, mp; 276–280ꢀC. 1H NMR
(400 MHz, CDCl3/TMS): 7.47 (d), 7.1 (d), 7.0 (m), 4.7 (s), 3.7
(m), 3.6 (m), 2.5 (m). 13C NMR (100 MHz): 167, 140, 139, 133,
131.55, 130.46, 128.9, 128.34, 126.65, 124.98, 116, 115.89,
111.76, 70.79, 70.68, 70.43, 7þ0.12, 69.69, 69.17, 68.49, 68.04,
54, 50.88, 39. EI-MS (m/z). M : 529(100). Elemental Analysis:
Anal. Calcd. for C, 63.62; H, 6.29; F, 3.59; N, 5.30; O, 21.19;
Found: C, 63.57; H, 6.27; F, 3.61; N, 5.33; O, 21.22.
ANTIOXIDANT ACTIVITY STUDIES
Antioxidant activity studies were measured by using
the cuprac method in the literature [35]. Antioxidant ac-
tivity studies results are showed in Table 3.
Acknowledgment. The authors are grateful to the Research
Fund of the TUBITAK for their support with the TBAG-
105T214.
16-(morpholinomethyl)-5,6,8,9,11,12-hexahidro-2H-(1,4,7,10,13)
pentaoxacyclopentadeca (2,3-H) chromen-18(3H)-on (17). Yield
(yellow); 27%, mp; 255–260ꢀC. 1H NMR (400 MHz, CDCl3/
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

September 2010
Synthesis, Complexation, and Biological Activity Studies of
4-Aminomethyl-7,8-dihydroxy Coumarines and Their Crown Ether Derivatives
1133
[15] Sui, G.; Kele, P.; Orbulescu, J.; Huo, Q.; Leblanc, R. M.
Lett Pept Sci 2002, 8, 47.
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet