Synthesis and anti-cancer, anti-metastatic evaluation of some new fluorinated isocoumarins and 3,4-dihydroisocoumarins

Article abstract of DOI:10.1016/j.jfluchem.2011.11.011

Synthesis of some fluorinated isocoumarins and 3,4-dihydroisocoumarins is reported. Structures of the synthesized compounds were confirmed by spectral and elemental analysis. All the synthesized compounds were evaluated for their antimetastatic activity and anti-cancer activity against breast cell line (MCF-7). Among all the tested compounds 3h [3-(3′,4′-difluorophenyl) isocoumarin] has shown excellent antimetastatic activity, however the growth inhibitory response of the tested compounds [3a-h, 4a-h, 5e-h, 6a-h and 7a, c, d] for MCF-7 human breast carcinoma cell line was moderate at higher concentrations (100-400 μM) and negligible at lower concentrations.

Full text of DOI:10.1016/j.jfluchem.2011.11.011

Journal of Fluorine Chemistry 135 (2012) 240–245
Contents lists available at SciVerse ScienceDirect
Journal of Fluorine Chemistry
journal homepage: www.elsevier.com/locate/fluor
Synthesis and anti-cancer, anti-metastatic evaluation of some new fluorinated
isocoumarins and 3,4-dihydroisocoumarins
Obaid-ur-Rahman Abid a, Muhammad Khalid a, Muhammad T. Hussain b, Muhammad Hanif a,
c
d
Ghulam Qadeer a, Nasim H. Rama a, , Alexander Kornienko , Khalid M. Khan
*
a Department of Chemistry, Quaid-i-Azam University, Islamabad 45320, Pakistan
b Department of Applied Sciences, National Textile University, Faisalabad 37610, Pakistan
c Department of Chemistry, New Mexico Tech, Socorro, NM 87801, Mexico
d H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan
A R T I C L E I N F O
A B S T R A C T
Article history:
Synthesis of some fluorinated isocoumarins and 3,4-dihydroisocoumarins is reported. Structures of the
synthesized compounds were confirmed by spectral and elemental analysis. All the synthesized
compounds were evaluated for their antimetastatic activity and anti-cancer activity against breast cell
line (MCF-7). Among all the tested compounds 3h [3-(30,40-difluorophenyl)isocoumarin] has shown
excellent antimetastatic activity, however the growth inhibitory response of the tested compounds [3a–
h, 4a–h, 5e–h, 6a–h and 7a, c, d] for MCF-7 human breast carcinoma cell line was moderate at higher
Received 10 July 2011
Received in revised form 24 November 2011
Accepted 26 November 2011
Available online 6 December 2011
Keywords:
concentrations (100–400
mM) and negligible at lower concentrations.
Isocoumarins
ß 2011 Elsevier B.V. All rights reserved.
Dihydroisocoumarins
Antimetastatic activity
Anti-cancer activity
MCF-7
Human breast cancer
1. Introduction
on the growth of three human tumor cell lines, MCF-7 (breast
adenocarcinoma), NCI-H460 (non-small cell lung cancer) and SF-
268 (CNS cancer). In a study [9], it was found that bireticulol has
cytotoxic effects against KB (human epidermoid carcinoma, ATCC
CCL-17) and NCI-H187 (human small cell lung cancer, ATCC CRL-
Metastasis is the most deadly aspect of cancer in which it
spreads from its primary site to other places of the body. This
process involves a series of tightly coupled events. These include
the detachment of tumor cells from the primary site, migration of
the cells accompanied by regulated proteolysis of the extracellular
matrix/basement membrane, dissemination of the tumor cells
through the vasculature and finally adhesion and proliferation of
the cells at a secondary site.
Isocoumarins and 3,4-dihydroisocoumarins are a group of
biologically active compounds that have been shown to influence
not only hormone metabolism but also intracellular enzymes [1],
protein synthesis, growth factors, malignant cell proliferation and
angiogenesis [2,3]. These compounds have been successfully
evaluated for antifungal, antibacterial, anti-tumor or cytotoxic
activities [4–6]. Rossi et al. [7] prepared different 3-aryliso-
couamrins which showed cytotoxic activity against human cancer
cell lines in vitro. Queiroz et al. [8] prepared different tricyclic
lactones which showed a high growth inhibitory effect (in vitro)
5804) cell lines with IC50 values of 24.4 and 8.31
m
g mLꢀ1
respectively. In another study [10], an antiangiogenic isocou-
marin, NM-3, has been found to increase the antitumor effects of
radiotherapy without toxicity. NM-3 alters several stages of the
angiogenic process including endothelial cell survival, migration,
and tube formation.
Organic compounds with fluorine moiety are medicinally very
important [11]. Drug-receptor interactions are improved in the
presence of small sized highly electronegative fluorine atom. The
transport of drug is facilitated by the high lipophilicity of
organofluorine compounds [12]. The literature survey revealed
that among the halogenated isocoumarins, tested for anticancer
activities, fluorinated isocoumarins have been rarely screened. So
in continuation of our previous studies [13–21], here we report the
synthesis of some new fluorine containing isocoumarins and their
conversion to the corresponding dihydroisocoumarins in order to
check their antimetastatic activity and anti-cancer activity against
breast cell line (MCF-7).
* Corresponding author. Tel.: +92 51 90642135; fax: +92 51 90642241.
General synthetic scheme is shown as follows (Scheme 1).
E-mail address: nhrama@qau.edu.pk (N.H. Rama).
0022-1139/$ – see front matter ß 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.jfluchem.2011.11.011

O.R. Abid et al. / Journal of Fluorine Chemistry 135 (2012) 240–245
241
O
O
R
OH
R
Cl
R
(iii)
(iv)
(i)
COOH
(ii)
+
O
O
COOH
1(a-h)
2(a-h)
3(a-h)
HO
O
HO
O
O
O
R
(v)
(vi)
Cl
R
R
O
OH
4(a-h)
5(a-h)
6(a-h)
Cl
Cl
F
F
F
Cl
F
(a) R=
(e) R=
(b) R=
(f) R=
(c) R =
(d) R=
(h) R=
F
F
F
F
F
F
F
F
(g) R=
Scheme 1. Synthesis of dihalophenylisocoumarins: (i) SOCl2, 60 8C for 30 min; (ii) reflux 4 h; (iii) 5% KOH/ethanol, reflux 5 h; (iv) Ac2O, reflux 1 h; (v) 1% NaOH/NaBH4,
overnight stirring (vi) Ac2O, reflux 1 h.
2. Chemistry
3.48 ppm and another doublet of doublet was observed at d 5.48–
6.15 ppm for C3-H proton. Thioisocoumarins 7(a–d) were synthe-
sized by thionation of isocoumarins 3(a–d) by refluxing with
Lawesson’s reagent [23] as shown in Scheme 2 and the structures
were confirmed by the appearance of new peaks at 1061–
1109 cmꢀ1 for C55S in IR. The structure elucidations of newly
synthesized compounds were determined by modern spectro-
scopic techniques like IR, 1H NMR and 13C NMR. Further
confirmations of the compounds were carried out by mass
spectrometry and microanalysis. We have already reported
synthesis, anti-oxidant, and anti-analgesic activities of compounds
(3e–h, 4e–h, 5e–h and 6e-h) [20]; however in this paper we want
to report their anti-cancer and anti-metastatic activities.
Synthesis of the target compounds is shown in the general
synthetic Schemes 1 and 2. Condensation of the acid chloride with
homophthalic acid is a useful method for the preparation of 3-
substituated isocoumarins skeleton [22]. A short and efficient
synthesis of 3-(dihalophenyl)isocoumarins 3(a–d) using this
method and conversion of 3(a–d) into corresponding racemic 3-
(dihalophenyl)-3,4-dihydroisocoumarins 6(a–d) were achieved.
Dihalobenzoic acids 1(a–d) were converted into their respective
acid chlorides 2(a–d) by reaction with thionyl chloride. Direct
condensation of acid chlorides 2(a–d) with homophthalic acid at
200 8C afforded 3-dihalophenylisocoumarins 3(a–d) in 63–73%
yield, which were purified by column chromatography. These
isocoumarins 3(a–d) exhibited characteristic 1H-singlet at
d
6.70–
3. Pharmacology
7.20 ppm for C4–H in 1H NMR, while in IR spectra lactonic carbonyl
absorptions were observed at 1716–1728 cmꢀ1. Alkaline hydroly-
sis of isocoumarins 3(a–d) afforded 2-(dihalobenzoylmethyl)ben-
zoic acids 4(a–d). Isocoumarins 3(a–d) were obtained back on
refluxing keto-acids 4(a–d) with acetic anhydride. Sodium
borohydride reduction of keto-acids 4(a–d) afforded the corre-
sponding racemic hydroxy acids 5(a–d), which were cyclodehy-
drated with acetic anhydride to produce 3-dihalophenyl-3,4-
dihydroisocoumarins 6(a–d) which exhibited the carbonyl absorp-
tions at 1712–1731 cmꢀ1 in IR spectra. The typical AB pattern for
C3–H proton and ABX pattern for C4–H protons were observed in
1H NMR spectrum of the compound 6(a-d). Thus, each of the C4–H
3.1. Antimetastatic activity and anticancer activity against breast cell
line MCF-7
3.1.1. Cell culture
MCF-7/AZ is a variant of the human mammary carcinoma cell
family MCF-7 [24]. The cells are maintained on a tissue culture
plastic substrate (Nunc) in a mixture of Dulbecco’s modified Eagle’s
medium (DMEM) and HAMF12 (50/50) (Invitrogen, Carlsbad, CA,
USA) supplemented with 250 IU/mL penicillin, 100
m
g mLꢀ1
streptomycin (Invitrogen) and 10% fetal bovine serum (FBS)
(Invitrogen), at 37 8C in a humidified atmosphere containing
10% CO2.
showed a doublet of doublet at
d
3.01–3.18 ppm and d 3.27–
3.1.2. Assay for cell viability
O
O
R
S
O
R
Thirty-one compounds (3a–h, 4a–h, 5e–h, 6a–h and 7a, c, d)
were evaluated for their anticancer activities against MCF-7 breast
cell line. Cell viability was tested in accordance with Romijn et al.
[25]. Briefly mitochondrial dehydrogenase activities were measured
by an MTT-reagent (Sigma, St. Louis, MO, USA). Cells were seeded in
Lawesson's Reagent
Toluene, Reflux 3h
3(a-d)
7(a-d)
microtiter plates at an initial density of 1.5 ꢁ 104 cells in 200
mL
Scheme 2. Conversion of isocoumarins to thioisocoumarins.
culture medium and treated with increasing concentrations of each

242
O.R. Abid et al. / Journal of Fluorine Chemistry 135 (2012) 240–245
compound. In each experiment, eight wells were used to determine
the mean optical density (OD) referring to cell viability.
4.2. Anti-cancer activity against breast cell line (MCF-7)
Growth inhibitory effect of the tested compounds against
human breast carcinoma cell line MCF-7 is shown in Table 2. It is
clear from the table that the inhibition is moderate at higher
3.1.3. Collagen type I invasion assay
This was performed as described [26]. Briefly, six-well plates
were filled with 1.25 mL neutralized type I collagen (0.09%) (Upstate
Biotechnology, Lake Placid, NY) and incubated for 1 h at 37 8C to
allow gelification. Non-invasive MCF-7/Az cells were pretreated
with ET-18-OMe for 24 h in order to become invasive into collagen
type I [27] and served as control for invasiveness as compared to
untreated MCF-7/Az cells. Single cell suspensions were prepared
with trypsin/EDTA, mixed with different compound solutions, seed
on top of collagen type I gel and cultured at 37 8C for 24 h. The
number of cells penetrating into the gel or remaining at the surface
were counted in 12 field of 0.157 mm2 [27], using an inverted
microscope controlled by a computer programme. This invasion
index expresses the percentage of invading cells over the total
number of cells.
concentrations (100–400
mM) however it is negligible at lower
concentrations (<100
mM).
Table 2
Anticancer activity against breast cell line (MCF-7) of synthesized compounds (3a–
h), (4a–h), (5e–h), (6a–h) and (7a, c–d).
Compd.
Concentration (
mM)
0
2.5
98
25
50
89
100
400
3a
% viability
% std
100
91
83
4
79
5
5
112
3
1
110
5
6
104
6
3b
% viability
% std
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
95
8
84
6
3c
% viability
% std
97
7
96
6
95
5
94
6
89
4
3d
% viability
% std
91
9
80
6
82
11
112
7
80
7
75
10
53
10
48
8
4. Results
3ea
3fa
3ga
3ha
4a
% viability
% std
108
6
118
2
60
6
4.1. Anti-metastatic activity
% viability
% std
104
6
90
2
87
8
77
4
Thirty one compounds (3a–h, 4a–h, 5e–h, 6a–h and 7a, c, d) were
evaluated for their antimetastatic activities. The results for inhibition
of invasion of these cells into collagen, which is an indication of
antimetastatic activity are given in Table 1. All the tested compounds
showed very small antimetastatic activity except the compound 3h
[3-(30,40-difluorophenyl)isocoumarin] which has shown excellent
activity. This activity may be attributed to the presence of two fluoro
groups at metaandparapositionsof thephenylring attached at C-3 in
conjugation with the double bond of isocoumarin nucleus.
% viability
% std
92
5
82
8
80
8
80
4
68
4
% viability
% std
75
7
80
6
82
11
97
8
70
7
62
9
% viability
% std
104
6
100
3
77
4
73
6
4b
% viability
% std
92
5
82
8
80
8
80
4
78
2
4c
% viability
% std
98
6
93
5
90
5
90
6
88
7
4d
% viability
% std
113
6
111
8
99
5
91
4
89
8
4ea
4fa
4ga
4ha
5ea
5fa
5ga
5ha
6a
% viability
% std
104
6
90
2
87
8
77
4
70
8
Table 1
Anti-metastatic activity of the synthesized compounds (3a–h), (4a–h), (5e–h), (6a–
h) and (7a, c–d).
% viability
% std
92
5
82
8
80
8
80
4
75
4
Compd.
Collagen type I invasion
Treatment corresponding
OD80 cells MCF-7/AZ
% viability
% std
98
7
95
5
91
7
90
6
82
8
3a
16
15
16
13
17
16
16
2
2
2
1
3
3
2
2
1
2
2
2
2
2
2
1
2
3
2
2
3
1
2
2
1
1
2
2
1
2
1
1
1
% viability
% std
103
4
98
8
92
9
91
4
89
8
3b
3c
% viability
% std
93
7
93
8
88
9
80
6
79
2
3d
3ea
3fa
3ga
3ha
4a
% viability
% std
91
4
90
8
89
4
86
3
48
5
% viability
% std
89
4
86
9
86
7
75
9
71
8
15
19
18
16
17
16
16
18
18
17
18
18
18
17
17
18
18
16
16
16
15
14
10
19
% viability
% std
100
8
91
3
90
5
85
7
83
8
4b
4c
% viability
% std
105
7
104
5
92
7
80
6
80
4
4d
4ea
4fa
4ga
4ha
5ea
5fa
5ga
5ha
6a
6b
% viability
% std
98
5
92
8
89
6
86
3
76
8
6c
% viability
% std
99
4
98
5
86
7
81
8
79
4
6d
% viability
% std
100
8
97
4
90
5
89
6
83
8
6ea
6fa
6ga
6ha
7a
% viability
% std
96
5
93
6
82
7
83
5
77
9
% viability
% std
93
8
93
4
90
7
86
7
85
7
6b
6c
% viability
% std
75
7
80
6
82
8
83
5
80
9
6d
6ea
6fa
6ga
6ha
7a
% viability
% std
92
5
85
8
80
8
79
4
71
4
% viability
% std
96
5
93
6
82
7
83
5
77
9
7c
% viability
% std
97
6
95
4
90
7
78
8
77
7
7c
7d
7d
% viability
% std
95
6
80
6
82
8
80
5
77
6
+control
a
a
Synthesis of these compounds is reported already [22].
Synthesis of these compounds is reported already [22].

O.R. Abid et al. / Journal of Fluorine Chemistry 135 (2012) 240–245
243
5. Conclusion
4a), 148.2 (C-3), 156.5 (C-1), 162.2 (d, J = 252 Hz, C-40) ppm; MS (EI,
ꢂ
70 eV): m/z (%); =274 (M+ , 85), 246 (93.5), 227 (2.6), 211 (100), 157
A total of thirty one compounds {fifteen new (3a–d, 4a–d, 6a–d,
7a, 7c and 7d) and sixteen already reported (3e–h, 4e–h, 5e–h, 6e–
h) by us [22]} were screened for anti-cancer and anti-metastatic
activities. These compounds have shown MCF-7 human breast
cancer cell inhibition activity at higher concentrations. Antimeta-
static activity of the tested compounds was small except the
compound 3h which has surprisingly shown excellent activity and
it is far better than the control. The compound 3h may be a
potential lead candidate with respect to antimetastatic activity and
for further biological exploration.
(24.3), 145 (1.4), 117 (4.2), 89 (37.4); Elemental Analysis: Found
(Calcd.)%; C: 65.60 (65.01), H: 2.91 (2.85).
6.1.2. 3-(20-Chloro-60-fluorophenyl)isocoumarin (3b)
Yield 69%; m.p. 240–241 8C; IR (KBr)
n
max in cmꢀ1: 1720 (C55O),
1182 (C–F), 1047 (C–Cl); 1H NMR (CDCl3,
d
-values): 6.70 (1H, s, H-
4), 7.12 (1H, dt, J = 8.1, 3.0 Hz, H-40), 7.28–7.43 (2H, m, H-30,50), 7.52
(1H, d, J = 7.8 Hz, H-5), 7.59 (1H, dt, J = 7.8, 1.2 Hz, H-7), 7.78 (1H,
dt, J = 7.5, 1.2 Hz, H-6), 8.37 (1H, dd, J = 7.8, 1.2 Hz, H-8) ppm; 13
C
NMR (CDCl3,
d-values): 102.0 (d, J = 2.4 Hz, C-4), 111.7 (d,
J = 21.9 Hz, C-50), 120.3 (d, J = 21.9 Hz, C-10), 124.2 (C-5), 126.2(d,
J = 3.0 Hz, C-30), 128.9 (C-7), 129.8 (C-8, 8a), 130.3 (d, J = 8.4 Hz, C-
40), 133.1 (d, J = 7.4 Hz, C-3), 133.9 (C-6, 4a), 136.5 (d, J = 8.4 Hz, C-
6. Experimental
All the common chemicals and solvents were of analytical grade
or dry distilled. All the processes involving air or moisture
sensitivity were conducted under the inert atmosphere of dry
nitrogen using oven dried glassware. The solvent evaporation was
performed under reduced pressure using a Bu¨chi rotary evapora-
tor. Melting points were determined on Stuart melting point
apparatus (SMP3) and are uncorrected. The IR spectra were
recorded on Bio-Rad Merlin FTS 3000 MX spectrophotometer. 1H
NMR (300 MHz) and 13C NMR (75 MHz) were recorded on a Bruker
AM-300 as DMSO and CDCl3 solutions using TMS as internal
20), 157.2 (C-1), 161.4 (d, J = 249 Hz, C-60) ppm; MS (EI, 70 eV): m/z
ꢂ
(%); =274 (M+ , 85), 246 (90.7), 227 (3.5), 211 (100), 157 (22.4), 145
(2.5), 117 (3.5), 89 (35.9); Elemental Analysis: Found (Calcd.)%; C:
65.60 (65.20), H: 2.91 (2.89).
6.1.3. 3-(30-Chloro-40-fluorophenyl)isocoumarin (3c)
Yield 73%; m.p. 178–179 8C; IR (KBr)
n
max in cmꢀ1: 1716 (C55O),
1114 (C–F) 1059 (C–Cl); 1H NMR (CDCl3,
d
-values): 6.92 (1H, s, H-
4), 7.22–7.27 (1H, m, H-50), 7.51–7.57 (2H, m, H-20,60), 7.73–7.79
(2H, m, H-5,7), 7.95 (1H, dt, J = 7.5, 1.2 Hz, H-6), 8.32 (1H, dd, J = 7.8,
1.2 Hz, H-8) ppm; 13C NMR (CDCl3,
d-values): 102.0 (C-4), 114.4 (d,
standard. Chemical shifts (
d
) are expressed in units of parts per
million relative to TMS. Electron impact mass spectra (EIMS) were
recorded on a Finnigan MAT-311A Germany. CHN analyses were
performed on a Carlo Erba Strumentazion-Mod-1106 Italy. Thin
layer chromatography was performed on pre-coated silica gel
plates (Kieselgel 60, 254, E. Merck, Germany). Chromatograms
were visualized by, UV at 254 and 365 nm. The synthetic pathway
for the compounds is shown in Schemes 1 and 2.
J = 23.4 Hz, C-50), 126.1 (C-5), 135.1 (C-6), 126.5 (C-7), 127.6 (d,
J = 3.0 Hz C-10), 128.6 (d, J = 8.1 Hz, C-20), 129.2(C-8a), 129.7 (C-8),
133.1 (d, J = 8.1 Hz, C-60), 135.1 (d, J = 23.4 Hz, C-30), 137.0 (C-4a),
151.3 (C-3), 157.2 (C-1), 161.8 (d, J = 250.2 Hz, C-40) ppm; MS (EI,
ꢂ
70 eV): m/z (%); =274 (M+ , 84), 246 (94.0), 227 (2.0), 211 (100), 157
(20.5), 145 (3.5), 117 (5.6), 89 (40.0); Elemental Analysis: Found
(Calcd.)%; C: 65.60 (64.99), H: 2.91 (2.95).
6.1. General synthetic procedure for the isocoumarin derivatives 3(a–
d)
6.1.4. 3-(40-Chloro-20-fluorophenyl)isocoumarin (3d)
Yield 63%; m.p. 137–138 8C; IR (KBr)
n
max in cmꢀ1: 1726(C55O),
1158 (C–F), 1061 (C–Cl); 1H NMR (CDCl3,
d
-values): 7.20 (1H, s, H-
2-Chloro-4-fluorobenzoic acid, 2-chloro-6-fluorobenzoic acid,
3-chloro-4-fluorobenzoic acid and 4-chloro-2-fluorobenzoic acid
1(a–d) (5 g, 28.7 mmol) were converted into their respective acid
chlorides 2(a–d) by reaction with thionyl chloride (4.09 g, 2.49 mL
and 34 mmol) in the presence of a drop of DMF under reflux for
30 min. Completion of the reaction was indicated by the stoppage
of gas evolution. Removal of excess of thionyl chloride was carried
out under reduced pressure to afford dihalobenzoyl chlorides 2(a–
d). Mixtures of homophthalic acid (1.3 g, 7.2 mmol) and dihalo-
benzoyl chlorides 2(a–d) (5.516 g, 28.7 mmol) were heated at
200 8C under reflux for 4 h. The mixtures were dissolved in ethyl
acetate and aqueous solution of sodium carbonate was added in
order to remove the unreacted homophthalic acid. Organic layer
was separated and the aqueous layer was extracted with 2ꢁ 25 mL
ethyl acetate. The combined organic layers were concentrated and
chromatographed on silica gel using pet ether (40–80 8C fraction)
as eluent which gave 3-(dihalophenyl)isocoumarins 3(a–d) as
solids, which were further purified by recrystallization from
methanol.
4), 7.22–7.30 (2H, m, H-30,50), 7.52–7.58 (2H, m, H-5,60), 7.63 (1H,
dt, J = 7.5, 1.2 Hz, H-7), 7.98 (1H, dt, J = 8.4, 1.2 Hz, H-6), 8.33 (1H,
dd, J = 7.5, 1.2 Hz, H-8) ppm; 13C NMR (CDCl3,
d-values): 107.3 (d,
J = 2.4 Hz, C-4), 117.2 (d, J = 21.9 Hz, C-30), 118.8 (d, J = 21.9 Hz, C-
10), 125.1 (d, J = 8.1 Hz, C-60), 125.8 (C-5), 126.5 (C-7), 128.7 (C-8a),
129.1 (d, J = 3.0 Hz, C-50), 129.6 (C-8), 135.0 (C-6), 136.2 (d,
J = 8.1 Hz, C-40), 137.2 (C-4a), 147.2 (d, J = 7.3 Hz, C-3), 157.9 (C-1),
ꢂ
161.6 (d, J = 251 Hz, C-20) ppm; MS (EI, 70 eV): m/z (%); =274 (M+ ,
92), 246 (85.5), 227 (1.2), 211 (100), 157 (32.3), 145 (1.1), 117 (3.2),
89 (57.4); Elemental Analysis: Found (Calcd.)%; C: 65.60 (65.84), H:
2.91 (2.84).
6.2. General synthetic procedure for 2-{2-(dihalophenyl)-20-
oxoethyl}benzoic acids 4(a–d)
Solution of 3-(dihalophenyl)isocoumarins (1.0 g, 3.6 mmol)
4(a–d) in ethanol (25 mL) and potassium hydroxide (5%, 30 mL)
was refluxed for 5 h. After cooling, the reaction mixture was
evaporated under reduced pressure to remove ethanol. Cold water
(20 mL) was then added and the reaction mixture was acidified
with dilute hydrochloric acid and extracted with dichloromethane
(2ꢁ 25 mL). The solvent was rotary evaporated to afford crude
solid which was recrystallized from ethyl acetate to give 2-{2-
(Dihalophenyl)-20-oxoethyl}benzoic acid 4(a–d).
6.1.1. 3-(20-Chloro-40-fluorophenyl)isocoumarin (3a)
Yield 68%; m.p. 163–164 8C; IR (KBr)
n
max in cmꢀ1: 1728 (C55O),
1117 (C–F), 1069 (C–Cl); 1H NMR (CDCl3,
d
-values): 7.20 (1H, s, H-
4), 7.21–7.30 (2H, m, H-30,50), 7.52–7.58 (2H, m, H-5,60), 7.76 (1H,
dt, J = 7.5, 1.2 Hz, H-7), 7.98 (1H, dt, J = 7.5, 1.2 Hz, H-6), 8.33 (1H,
dd, J = 8.1, 1.2 Hz, H-8) ppm; 13C NMR (CDCl3,
d
-values): 103.9 (C-
6.2.1. 2-{2-(200-chloro-400-fluorophenyl)-20-oxoethyl}benzoic acids
4), 114.4 (d, J = 22.5 Hz, C-50), 118.0 (d, J = 22.5 Hz, C-30), 120.6 (C-
5), 128.4 (d, J = 3.9 Hz, C-10), 129.5 (C-7), 129.8 (C-8a), 131.2 (d,
J = 9.0 Hz C-60), 133.4 (C-8), 134.6 (d, J = 9.0 Hz, C-20), 136.8 (C-6,
(4a)
Yield 78%; m.p. 168–169 8C; IR (KBr)
n
max in cmꢀ1: 3115–2680
-values):
(O–H), 1722 (C55O), 1689 (acidic C55O); 1H NMR (CDCl3,
d

244
O.R. Abid et al. / Journal of Fluorine Chemistry 135 (2012) 240–245
4.70 (2H, s, H-10), 7.05–7.12 (1H, m, H-500), 7.23–7.27 (1H, m, H-300),
7.33 (1H, dd, J = 7.5, 1.2 Hz, H-3), 7.42 (1H, dt, J = 7.8, 1.2 Hz, H-5),
7.58 (1H, dt, J = 7.5, 1.5 Hz, H-4), 7.66–7.74 (1H, m, H-600), 8.16 (1H,
dd, J = 7.5, 1.5 Hz, H-6), 11.62 (1H, s, acidic-H) ppm; 13C NMR
(12.1), 91(29.5); Elemental Analysis: Found (Calcd.)%; C: 61.54
(61.13), H: 3.42 (3.57).
6.3. General synthetic procedure for (dl)-3-(dihalophenyl)-3,4-
(CDCl3,
d
-values): 37.2 (C-10), 112.5 (d, J = 22.5 Hz, C-500), 116.7 (d,
dihydroisocoumarins 6(a–d)
J = 22.8 Hz, C-300), 127.6 (C-5), 128.8 (C-6), 129.6 (C-3), 130.5 (C-4),
130.8 (d, J = 9.0 Hz, C-600), 130.9 (C-1), 133.0 (d, J = 3.0 Hz, C-100),
134.2 (d, J = 9.0 Hz, C-200), 138.0 (C-2), 162.7 (d, J = 252 Hz, C-400),
Keto-acids 4(a–d) (60 mg, 0.2 mmol) were stirred over night at
room temperature with sodium borohydride (0.1 g) in sodium
hydroxide (1%, 20 mL). The reaction mixture was chilled and
acidified with dilute hydrochloric acid to yield hydroxy acids 5(a–
d) which were cyclodehydrated by refluxing with acetic anhydride
for 1 h. The reaction mixture was diluted with chilled water
(20 mL) and extracted with dichloromethane. The solvent was
evaporated under reduced pressure to afford (dl)-3-(dihalophe-
nyl)-3,4-dihydroisocoumarins 6(a–d).
168.4 (C-acidic), 201.9 (C-20) ppm; MS (EI, 70 eV): m/z (%); =292
ꢂ
(M+ , 2.2), 274 (92), 246 (100), 211 (74.6), 163 (6), 157 (34.3), 135
(1.4), 129 (26.2), 117 (3.2), 109 (11.2), 93 (10.4), 91(28.8);
Elemental Analysis: Found (Calcd.)%; C: 61.54 (61.03), H: 3.42
(3.40).
6.2.2. 2-{2-(200-Chloro-600-fluorophenyl)-20-oxoethyl}benzoic acid
(4b)
Yield 82%; m.p. 157–158 8C; IR (KBr) nmax in cmꢀ1
:
6.3.1. (dl)-3-(20-Chloro-40-fluorophenyl)-3,4-dihydroisocoumarin
3072–2654 (O–H), 1719 (C55O), 1681 (acidic C55O); 1H NMR
(CDCl3,
-values): 4.71 (2H, s, H-10), 7.06 (1H, dt, J = 8.1, 0.9 Hz,
(6a)
d
Yield 75%; m.p. 136–138 8C. IR (KBr)
n
max in cmꢀ1: 1727 (C55O),
-values): 3.18 (1H, dd,
H-400), 7.23 (1H, d, J = 7.8 Hz, H-300), 7.29–7.38 (2H, m, H-3,500),
7.45 (1H, dt, J = 7.5, 1.2 Hz, H-5), 7.59 (1H, dt, J = 7.5, 1.5 Hz, H-
4), 8.18 (1H, dd, J = 8.1, 1.5 Hz, H-6), 11.77 (1H, s, acidic-H) ppm;
1112 (C–F), 1062 (C–Cl); 1H NMR (CDCl3,
d
J = 10.8, 2.1 Hz, H-4ii), 3.27 (1H, dd, J = 14.1, 7.5 Hz, H-4i), 5.91 (1H,
dd, J = 14.4, 3.9 Hz, H-3), 6.89–6.95(1H, m, H-30), 7.06–7.20 (2H, m,
H-50,60), 7.32 (1H, dd, J = 7.8, 3.0 Hz, H-5), 7.47 (1H, dt, J = 7.8, 1.2 Hz,
H-7), 7.60 (1H, dt, J = 7.5, 1.5 Hz, H-6), 8.19 (1H, d, J = 7.5 Hz, H-
8) ppm; 13C NMR (CDCl3,
d-values): 35.8 (C-4), 70.6 (C-3), 112.5 (d,
J = 22.5 Hz, C-50), 117.5 (d, J = 22.5 Hz, C-30), 124.8 (C-7), 126.8 (C-5),
127.7 (C-8), 128.8 (C-8a), 128.9 (d, J = 9.0 Hz,C-60), 132.1(C-6), 132.8
(d, J = 3.0 Hz, C-10), 133.1 (d, J = 9.0 Hz, C-20), 140.9 (C-4a) 161.7 (d,
J = 252 Hz, C-40), 165.2 (C-1) ppm; MS (EI, 70 eV): m/z (%); =276 (M+ ,
5.60),147(1.20), 129(1.9), 119(10.4),118(100), 90(31.8),89(12.2);
Elemental Analysis: Found (Calcd.)%; C: 65.09(65.00), H:3.62 (3.60).
13C NMR (CDCl3,
d
-values): 37.5 (d, J = 2.2 Hz, C-10), 113.5
(d, J = 21.9 Hz, C-500), 123.4 (d, J = 2.9 Hz, C-300), 124.8 (d,
J = 21.9 Hz, C-100), 127.8 (C-5), 128.6 (C-3), 130.0 (C-6), 130.8
(C-2), 133.9 (C-4), 134.4 (d, J = 8.4 Hz, C-200), 135.2 (d, J = 8.4 Hz,
C-400), 138.5 (C-1), 160.5 (d, J = 249 Hz, C-600), 169.2 (C-acidic-C),
200.7 (d, J = 7.1 Hz, C-20) ppm; MS (EI, 70 eV): m/z (%); =292 (M+ ,
1.7), 274 (89.5), 246 (100), 211 (67.8), 163 (7.2), 157 (39.6), 135
(2.7), 129 (34.2), 117 (5.1), 109 (9.3), 93 (8.9), 91(21.7);
Elemental Analysis: Found (Calcd.)%; C: 61.54 (60.98), H: 3.42
(3.38).
ꢂ
ꢂ
6.3.2. (dl)-3-(20-Chloro-60-fluorophenyl)-3,4-dihydroisocoumarin
6.2.3. 2-{2-(300-chloro-400-fluorophenyl)-20-oxoethyl}benzoic acid
(6b)
(4c)
Yield 82%; m.p. 158–159 8C. IR (KBr)
n
max in cmꢀ1: 1719 (C55O),
-values): 3.01 (1H, dd,
Yield 80%; m.p. 259–260 8C. IR (KBr)
n
max in cmꢀ1: 3095–2634
-values):
1179 (C–F), 1043 (C–Cl); 1H NMR (CDCl3,
d
(O–H), 1728 (C55O), 1686 (acidic C55O); 1H NMR (CDCl3,
d
J = 16.5, 3.3 Hz, H-4ii), 3.48 (1H, dd, J = 15.9, 13.5 Hz, H-4i), 6.15
(1H, dd, J = 12.9, 3.3 Hz, H-3), 7.10 (1H, dt, J = 8.1, 3.0 Hz, H-40),
7.24–7.37 (3H, m, H-30, 5,50), 7.47 (1H, dt, J = 7.5, 1.2 Hz, H-7), 7.61
(1H, dt, J = 7.5, 1.5 Hz, H-6), 8.19 (1H, dd, J = 7.8, 1.2 Hz, H-8) ppm;
4.73 (2H, s, H-10), 7.08–7.19 (1H, m, H-500), 7.33 (1H, d, J = 7.5 Hz, H-
3), 7.41 (1H, dt, J = 7.8, 1.2 Hz, H-5), 7.54 (1H, dt, J = 7.5, 1.5 Hz, H-
4), 7.64–7.72 (1H, m, H-600), 7.78–7.87 (1H, m, H-200), 8.11 (1H, dd,
J = 7.5, 1.2 Hz, H-6) 11.72 (1H, s, acidic-H) ppm; 13C NMR (CDCl3,
d
-
13C NMR (CDCl3,
d-values): 36.0 (d, J = 2.4 Hz, C-4), 61.1 (d,
values): 36.9 (C-10), 115.7 (d, J = 23.4 Hz, C-500), 119.6 (d,
J = 23.4 Hz, C-300), 126.9 (C-5), 127.5 (d, J = 8.1 Hz, C-600), 128.5
(d, J = 8.1 Hz, C-200), 129.7 (C-3), 129.9 (C-6), 130.2 (C-2), 132.8 (d,
J = 3.0 Hz, C-100), 133.8 (C-4), 137.4 (C-1), 163.9 (d, J = 250.2 Hz, C-
J = 7.3 Hz, C-3), 113.0 (d, J = 21.5 Hz, C-50), 124.9 (C-7), 125.0 (d,
J = 3.2 Hz, C-30), 127.2 (C-5), 128.1 (d, J = 21.5 Hz, C-10), 129.2 (C-8),
130.2 (d, J = 8.0 Hz, C-40), 131.9 (C-6), 133.0 (C-8a), 137.3 (d,
J = 8.0 Hz, C-20), 141.2 (C-4a), 160.9 (d, J = 249 Hz, C-60), 164.9 (C-
ꢂ
400), 168.9 (C-acidic), 201.5 (C-20) ppm; MS (EI, 70 eV): m/z (%);
1) ppm; MS (EI, 70ev): m/z (%); =276 (M+ , 7.56), 147 (4.65), 129
ꢂ
=292 (M+ , 2.1), 274 (82), 246 (100), 211 (70.7), 163 (9.1), 157
(4.6), 119 (12.5), 118 (100), 90 (34.0), 89 (10.9); Elemental
Analysis: Found (Calcd.)%; C: 65.09 (65.12), H: 3.62 (3.59).
(47.3), 135 (1.9), 129 (31.7), 117 (2.8), 109 (12.2), 93 (13.1),
91(32.1); Elemental Analysis: Found (Calcd.)%; C: 61.54 (61.45), H:
3.42 (3.43).
6.3.3. (dl)-3-(30-Chloro-40-fluorophenyl)-3,4-dihydroisocoumarin
(6c)
6.2.4. 2-{2-(400-Chloro-200-fluorophenyl)-20-oxoethyl}benzoic acids
(4d)
Yield 77%; m.p. 129–130 8C. IR (KBr) nmax in cmꢀ1: 1717
(C55O), 1110 (C–F), 1057 (C–Cl); 1H NMR (CDCl3,
d-values): 3.11
Yield 82%; m.p. 184–186 8C; IR (KBr) nmax in cmꢀ1: 3185–
(1H, dd, J = 16.5, 3.0 Hz, H-4ii), 3.35 (1H, dd, J = 16.2, 12.0 Hz, H-
4i), 5.48 (1H, dd, J = 12.0, 3.0 Hz, H-3), 6.96 (1H, dd, J = 8.7,
3.0 Hz, H-60), 7.28–7.35 (2H, m, H-5,50), 7.45 (1H, dt, J = 7.8,
1.5 Hz, H-7), 7.51 (1H, d, J = 2.1 Hz, H-20), 7.60 (1H, dt, J = 7.5,
1.2 Hz, H-6), 8.16 (1H, dd, J = 7.8, 1.2 Hz, H-8) ppm; 13C NMR
26540 (O–H), 1734 (C55O), 1690 (acidic C55O); 1H NMR (CDCl3,
d-
values): 4.69 (2H, s, H-10), 7.01 (1H, dd, J = 8.1, 1.8 Hz, H-300), 7.21–
7.30 (2H, m, H-3, 500), 7.43 (1H, dt, J = 7.5, 1.2 Hz, H-5), 7.56 (1H, dt,
J = 7.5, 1.5 Hz, H-4), 7.79–7.88 (1H, m, H-600), 8.14(1H, dd, J = 8.1,
1.5 Hz, H-6), 11.70 (1H, s, acidic-H) ppm; 13C NMR (CDCl3,
d
-
(CDCl3, d-values): 35.7 (C-4), 78.8 (C-3), 116.9 (d, J = 23.7 Hz, C-
values): 38.9 (d, J = 2.3 Hz, C-10), 117.5 (d, J = 21.9 Hz, C-300), 124.5
(d, J = 21.9 Hz, C-100), 125.4 (d, J = 3.0 Hz C-500), 126.8 (C-6), 127.5
(C-4), 129.6 (C-8), 130.5 (C-8a), 131.8 (d, J = 8.1 Hz, C-600), 133.4
(C-4), 137.1 (C-2), 138.4 (d, J = 8.1 Hz, C-400), 158.4 (d, J = 251 Hz,
50), 120.8 (d, J = 3.1 Hz, C-10), 125.2 (C-7), 126.3 (C-5), 127.0 (d,
J = 7.7 Hz, C-60), 128.1 (d, J = 7.7 Hz, C-20), 128.4 (C-8), 128.9 (C-
8a), 133.0 (C-6), 136.9 (d, J = 23.7 Hz, C-30), 140.5 (C-4a) 160.5 (d,
J = 250.2 Hz, C-40), 165.8 (C-1) ppm; MS (EI, 70 eV): m/z (%);
ꢂ
C-200), 167.4 (C-acidic), 198.1 (d, J = 6.9 Hz, C-20) ppm; MS (EI,
=276 (M+ , 8.76), 147 (2.40), 129 (3.5), 119 (9.65), 118 (100), 90
ꢂ
70 eV): m/z (%); =292 (M+ , 1.3), 274 (87.6), 246 (100), 211 (77.6),
(37.0), 89 (14.5); Elemental Analysis: Found (Calcd.)%; C: 65.09
(65.14), H: 3.62 (3.63).
163 (9.2), 157 (44.1), 135 (4.1), 129 (28.3), 117 (4.2), 109 (10.2), 93

O.R. Abid et al. / Journal of Fluorine Chemistry 135 (2012) 240–245
245
6.3.4. (dl)-3-(40-Chloro-20-fluorophenyl)-3,4-dihydroisocoumarin
128.8 (C-7), 130.9 (C-8), 135.5 (C-6), 136.7 (C-4a), 144.3 (C-8a),
(6d)
157.5 (C-3), 162.9 (d, J = 250.2 Hz, C-40), 199.8 (C-1) ppm; MS (EI,
ꢂ
Yield 78%; m.p. 151–153 8C. IR (KBr)
n
max in cmꢀ1: 1731 (C55O),
-values): 3.17 (1H, dd,
70 eV): m/z (%); =290 (M+ , 100), 246 (5.9), 161 (3.5), 157 (18.5),
1117 (C–F), 1080 (C–Cl); 1H NMR (CDCl3,
d
133 (8.63), 129 (25.5), 89 (40.0); Elemental Analysis: Found
(Calcd.)%; C: 61.96 (61.49), H: 2.75 (2.68), S: 11.02 (10.84).
J = 10.5, 4.5 Hz, H-4ii), 3.29 (1H, dd, J = 16.5, 11.4 Hz, H-4i), 5.83
(1H, dd, J = 11.7, 3.6 Hz, H-3), 7.15 (1H, dd, J = 9.9, 1.8 Hz, H-30),
7.23–7.34 (2H, m, H-50,60), 7.46 (1H, dt, J = 6.3, 1.2 Hz, H-7), 7.52–
64 (2H, m, H-5,6), 8.17 (1H, dd, J = 7.8, 1.5 Hz, H-8) ppm; 13C NMR
6.4.4. 3-(40-Chloro-20-fluorophenyl)thioisocoumarin (7d)
Yield 77%; m.p. 190–192 8C. IR (KBr)
max in cmꢀ1: 1171 (C–F),
1109(C55S), 1072 (C–Cl); 1H NMR (CDCl3,
d-values): 7.23–7.28 (1H,
n
(CDCl3,
d
-values): 34.5 (d, J = 2.3 Hz, C-4), 73.7 (d, J = 7.3 Hz, C-3),
116.4 (d, J = 21.9 Hz, C-30), 120.7 (d, J = 21.8 Hz, C-10), 124.7 (d,
J = 2.7 Hz, C-50), 125.8 (C-7), 127.3 (C-5), 128.1 (C-8a), 128.5 (C-8),
132.8 (d, J = 8.1 Hz, C-60), 133.9 (C-6), 135.2 (d, J = 8.1 Hz, C-40),
m, H-30), 7.32 (1H, dd, J = 8.7, 2.1 Hz, H-50), 7.38 (1H, s, H-4), 7.51–
7.58 (2H, m, H-5, 60), 7.77 (1H, dt, J = 7.8, 1.2 Hz, H-7), 8.09 (1H, dt,
J = 7.5, 1.2 Hz, H-6), 8.74 (1H, dd, J = 8.1, 1.2 Hz, H-8) ppm; 13C NMR
139.3 (C-4a), 160.7 (d, J = 251 Hz, C-20), 165.2 (C-1) ppm; MS (EI,
(CDCl3, d
-values): 92.4 (d, J = 2.1 Hz, C-4), 117.1 (d, J = 2.9 Hz, C-50),
ꢂ
70 eV): m/z (%); =276 (M+ , 2.60), 147 (1.30), 129 (3.2), 119 (10.0),
118.3 (d, J = 22.2 Hz, C-30), 125.3 (C-5), 126.9 (d, J = 22.3 Hz, C-10),
129.4 (C-7), 129.9 (d, J = 8.2 Hz, C-60), 131.5 (C-6), 132.7 (C-8),
136.5 (d, J = 8.2 Hz, C-40), 137.8 (C-4a), 140.0 (C-8a), 157.9 (d,
118 (100), 90 (43.1), 89 (18.2); Elemental Analysis: Found
(Calcd.)%; C: 65.09 (64.92), H: 3.62 (3.72).
J = 7.1 Hz, C-3), 161.3 (d, J = 252 Hz, C-20), 199.9 (C-1) ppm; MS (EI,
ꢂ
6.4. General synthetic procedure for 3-
70 eV): m/z (%); =290 (M+ , 100), 246 (4.7), 161 (3.9), 157 (25.5),
(dihalophenyl)thioisocoumarins 7(a–d)
133 (11.7), 129 (20.5), 89 (48.4); Elemental Analysis: Found
(Calcd.)%; C: 61.96 (61.64), H: 2.75 (2.61), S: 11.02 (10.82).
Thioisocoumarins 7(a–d) were synthesized by thionation of
isocoumarins 3(a–d). Thionation was carried out by refluxing
isocoumarins 3(a–d) (0.5 g, 1.82 mmol) with Lawesson’s Reagent
[23] (0.89 g, 2.2 mmol) in dry toluene for four hours. Pure
thioisocoumarins were obtained from reaction mixture by
recrystallization with methanol.
Acknowledgement
The authors gratefully acknowledge the financial assistance
under the project # 20-946/R&D/07/607 from the Higher Educa-
tion Commission Islamabad, Pakistan.
6.4.1. 3-(20-Chloro-40-fluorophenyl)thioisocoumarin (7a)
Yield 82%; m.p. 197–198 8C. IR (KBr)
n
d
max in cmꢀ1: 1107 (C–F),
-values): 7.23–7.27 (1H,
References
1081(C55S), 1057 (C–Cl); 1H NMR (CDCl3,
m, H-50), 7.32 (1H, ddd, J = 8.7, 2.1, 0.6 Hz, H-30), 7.37(1H, s, H-4),
7.50–7.58 (2H, m, H-5,60), 7.76 (1H, dt, J = 7.5, 1.2 Hz, H-7), 8.09
(1H, dt, J = 8.4, 1.5 Hz, H-6), 8.74 (1H, dd, J = 8.1, 1.2 Hz, H-8) ppm;
[1] J.C. Powers, J.L. Asgian, D. Ekici, K.E. James, Chem. Rev. 102 (2002) 4639–4750.
[2] K. Meepagala, G. Sturtz, D. Wedge, J. Agric. Food Chem. 50 (2002) 6989–6992.
[3] O.A. Krupke, A.J. Castle, D.L. Rinker, Mycol. Res. 107 (2003) 1467–1475.
[4] A.C. Whyte, J.B. Gloer, J.A. Scott, D. Malloch, J. Nat. Prod. 59 (1996) 765–769.
[5] Y. Cai, H. Kleiner, D. Johnston, A. Dubowski, S. Botic, W. Ivie, J. DiGiovanni,
Carcinogenesis 18 (1997) 1521–1527.
[6] E.A. Varanda, M.S. Raddi, F. de, L. Dias, M.C. Araujo, S.C. Gibran, C.S. Takahashi, W.
Vilegas, Teratog. Carcinog. Mutagen. 17 (1997) 85–95.
[7] R. Rossi, A. Carpitaa, F. Bellinaa, P. Stabilea, L. Manninab, Tetrahedron 59 (12)
(2003) 2067–2081.
13C NMR (CDCl3, -values): 90.0 (C-4), 114.8 (d, J = 22.5 Hz, C-50),
d
118.2 (d, J = 22.5 Hz, C-30), 125.4 (C-5), 128.5 (d, J = 3.3 Hz, C-10),
129.5 (C-7), 130.0 (d, J = 9.0 Hz, C-60), 130.9 (C-6), 131.4 (C-8),
133.7 (d, J = 9.0 Hz, C-20), 136.8 (C-4a), 140.0 (C-8a), 160.8 (C-3),
164.5 (d, J = 252 Hz, C-40), 200.6 (C-1) ppm; MS (EI, 70 eV): m/z (%);
ꢂ
[8] M.R.P. Queiroz, R.C. Calhelha, L.A. Vale-Silva, E. Pinto, M. Sa˜o-Jose´ Nascimento,
Eur. J. Med. Chem. 44 (2009) 1893–1899.
=290 (M+ , 100), 246 (5.7), 161 (4.7), 157(19.8), 133(9.45),
129(18.9), 89(37.4); Elemental Analysis: Found (Calcd.)%; C:
61.96 (61.58), H: 2.75 (2.59), S: 11.02 (10.96).
[9] C. Boonlarppradab, C. Suriyachadkun, S. Suphothina, P. Tobwor, J. Antibiot. 64
(2011) 267–270.
[10] R.M. Salloum, N.T. Jaskowiak, H.J. Mauceri, S. Seetharam, M.A. Beckett, A.M. Koons,
D.M. Hari, V.K. Gupta, C. Reimer, R. Kalluri, M.C. Posner, S. Hellman, D.W. Kufe, R.R.
Weichselbaum, Cancer Res. 60 (2000) 6958–6963.
6.4.2. 3-(20-Chloro-60-fluorophenyl)thioisocoumarin (7b)
max in cmꢀ1: 1162 (C–F),
[11] (a) R. Filler, Y. Kobayasi, L.M. Yagupolskii (Eds.), Fluorine in Bioorganic Chemistry,
Elsevier, Amsterdam, 1993;
Yield 84%; m.p. 270–271 8C. IR (KBr)
n
1085 (C55S), 1061 (C–Cl); 1H NMR (CDCl3,
d
-values): 6.90 (1H, s, H-
(b) R. Filler, Fluorine Containing Drugs in Organofluorine Chemicals and Their
Industrial Application, Pergamon, New York, 1979 (Chapter 6);
(c) M. Hudlicky, Chemistry of Organic Compounds, EllisHorwood, Chichester, 1992.
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(2005) 41–51.
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29 (2005) 321–325.
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Res. 17 (2003) 207–214.
4), 7.15 (1H, dt, J = 8.1, 3.0 Hz, H-40), 7.32–7.36 (1H, m, H-30), 7.39–
7.46 (1H, m, H-50), 7.51 (1H, dd, J = 7.5, 1.2 Hz, H-5), 7.60 (1H, dt,
J = 8.1, 1.2 Hz, H-7), 7.78 (1H, dt, J = 7.5, 1.2 Hz, H-6), 8.78 (1H, dd,
J = 8.4, 1.2 Hz, H-8) ppm; 13C NMR (CDCl3,
d-values): 90.5 (d,
J = 2.2 Hz, C-4), 113.8 (d, J = 21.7 Hz, C-50), 123.5 (d, J = 21.7 Hz, C-
10), 125.0 (d, J = 3.0 Hz, C-30), 125.8 (C-5), 128.0 (C-7), 129.5 (d,
J = 8.7 Hz, C-40), 130.0 (C-8), 130.7 (C-6), 135.1 (d, J = 8.7 Hz, C-20),
140.1 (C-4a), 149.2 (C-8a), 159.4 (d, J = 7.2 Hz, C-3), 162.6 (d,
J = 249 Hz, C-60), 201.1 (C-1) ppm; MS (EI, 70 eV): m/z (%); =290
[17] M.T. Hussain, N.H. Rama, A. Malik, Indian J. Chem. Sect. B 40B (2001) 372–376.
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9 (1999) 27–32.
ꢂ
(M+ , 100), 246 (3.5), 161 (2.5), 157 (22.4), 133 (10.2), 129 (20.0), 89
(35.9); Elemental Analysis: Found (Calcd.)%; C: 61.96 (61.71), H:
2.75 (2.63), S: 11.02(10.98).
[19] M.T. Hussai, N.H. Rama, A. Malik, Indian J. Heterocycl. Chem. 8 (1999) 273–276.
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Malana, A. Malik, Indian J. Chem. 42B (2003) 611–615.
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200510122353. Priority: CAN 145:188619AN 2006:593756 CAPLUS, p. 24.
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[23] A. Levai, J. Jeko, J. Heterocycl. Chem. 42 (2005) 739.
6.4.3. 3-(30-Chloro-40-fluorophenyl)thioisocoumarin (7c)
Yield 83%; m.p. 203–204 8C. IR (KBr)
n
d
max in cmꢀ1: 1123 (C–F),
1076 (C55S) 1047 (C–Cl); 1H NMR (CDCl3,
-values): 7.08 (1H, s, H-
[24] U. Grafe, L. Radics, J. Antibiot. 39 (1986) 162–163.
4), 7.23–7.29 (1H, m, H-50), 7.48–7.55 (2H, m, H-20,60), 7.72–7.84
[25] J.C. Romijn, C.F. Verkoelen, F.H. Schroeder, Prostate 12 (1988) 99–110.
[26] M.E. Bracke, T. Boterberg, E.A. Bruynell, M.M. Mareel, in: Brooks, U. Schumzcher
(Eds.), Metastasis Methods and Protocols, Hannover press, Totowa, NJ, USA, 1999.
[27] W.F. Steelant, J.L. Goeman, J. Philippe, L.C. Oomem, J. Hilkens, M.A. Krzewinski-
Recchi, G. Huet, J. Van der Eycken, P. Delannoy, E.A. Bruyneel, M.M. Mareel, Int. J.
Cancer 92 (2001) 527–536.
(2H, m, H-5,7), 7.98 (1H, dt, J = 7.8, 1.2 Hz, H-6), 8.71 (1H, dd, J = 8.1,
1.2 Hz, H-8) ppm; 13C NMR (CDCl3,
d-values): 89.8 (C-4), 118.7 (d,
J = 23.6 Hz, C-50), 121.7 (d, J = 23.6 Hz, C-30), 126.0 (C-5), 127.1 (d,
J = 3.0 Hz, C-10), 127.5 (d, J = 8.3 Hz, C-60), 128.0 (d, J = 8.1 Hz, C-20),