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1189
dried in vacuo to afford (2), yield 47%. M.p. 150–151 °C IR (cm−1):
3266 m, ν(NH); 3070 m 2983 m, ν(CH3); 1615 s, νasym(COO); 1394 s,
yield 95%. M.p. 191–192 °C. IR (cm−1): 3251 m, ν(NH); 3068 m 2986 m,
ν(CH3); 1618 s, νasym(COO); 1390 s, νsym(COO); 430 ms, ν(Cd-OH2O);
368 m, 304 m, ν(Cd-Ooco). 1H NMR (DMSO): 10.33 (s, NH); 8.02 (dd,
H–C(3), JH(3)–H(4) =7.4); 6.74 (t, H–C(4), JH(4)–H(5) =7.4); 7.21 (t,
H–C(5)), JH(5)–H(45) =7.4);), 6.15 (d, H–C(6), JH(6)–H(5) =8.1); 7.32
(d, H4′), JH(4′)–H(5′) =8.2); 7.48 (d, H5′), JH(5′)–H(4′) =8.2); 2.38 (s,
3′ Me); 13C NMR: 173.9 (COOH); 146.1(C(1)); 131.9 (C(3)); 116.7
(C(4)); 135.5 (C(5)); 112.2 (C(6)); 133.3 (C(1′)); 132.4 (C(2′));
130.2 (C(3′)); 128.6 (C(4′)); 128.0 (C(5′)); 136.3 (C(6′)); 20.2 (3′ Me);
νsym(COO); 438 ms, ν(Mn-OH2O); 366, 277 ms, ν(Mn-Ooco). UV–vis:
λ, nm (ε/logε) in DMF:415sh (1.51), 364 (2.37), 321 (4.10), 294
(4.08); in CHCl3: 493sh (1.63), 381 (2.46), 337 (3.84), 283 (3.85).
μ
eff =5.66 MB. Mass spectrum, MS (atmospheric pressure chemical
ionization, APCI, m/z): 664 [2+H3O]+, 296 [1+H]+. Anal. calc. for
C28H20Cl4MnN2O4 (645.2 g mol–1): C, 52.1; H, 3.1; N, 4.3; Mn, 8.5
found: C, 52.3, H, 3.2; N, 4.2; Mn, 8.8%. The complex is soluble in
DMSO, DMF, MeOH, Benzene, toluene, CH2Cl2, and CHCl3.
Mass spectrum, MS (electrospray ionization, ESI, m/z): 632 [5-2Cl− +
] ,
764 [5+Na]+, 741[5+2H]+. Anal. calc. for C28H24CdCl4N2O6
(738.67.6 g mol–1): C, 45.5; H, 3.3; N, 3.8; Zn, 15.2 found: C, 45.6, H,
3.4; N, 3.9; Zn, 15.3%. The complex is soluble in DMSO and DMF and
slightly soluble in CH2Cl2, CHCl3. Recrystallization of 5 from DMSO
solution gives single crystals of [Cd(meclo)2(DMSO)3] (5a) suitable for
X-ray structure determination.
2.2.3. Synthesis of [Cu(meclo)2(H2O)2]; (3)
Copper(II) acetate monohydrate ([Cu(CH3COO)2(H2O)]2)
(0.0250 g, 0.125 mM) was dissolved in methanol (2 mL) and this
solution was added to a solution of meclofenamic acid (0.0740 g,
0.25 mM) in methanol (2 mL). Drops of N(eth)3 were added to the
solution till the apparent pH value was ~7. The reaction mixture was
stirred for 4 h at room temperature and then it was left to the
refrigerator overnight. The green powder was filtered and washed
with cold MeOH and dried in vacuo to afford (3), yield 69%. D.p. 190–
192 °C. IR (cm−1): 3312 m, ν(NH); 3070 m 2923 m, ν(CH3); 1618 s,
2.3. X-ray crystallography
A green needle crystal of [Cu4(meclo)6(OH)2(DMSO)2]•2DMSO (3a)
and a colourless plate crystal of [Cd(meclo)2(DMSO)3] (5a) were
mounted on a glass fiber and used for data collection. Crystal data were
collected at 100(1) K, using a Bruker X8 KappaAPEXII diffractometer and
a Bruker SMART CCD 1000 diffractometer for 3a and 5a respectively.
Graphite monochromated MoK(alpha) radiation was used throughout.
The data were processed with APEX2 and with SAINT for 3a and for 5a
respectively. The data were corrected for absorption using SADABS
(transmissions factors: 1.000–0.0.904) and (transmissions factors:
1.000–0.887) for 3a and for 5a respectively [24]. The structures were
solved by direct methods using the program SHELXS-97 and refined by
full-matrix least-squares techniques against F2 using SHELXL-97 [25].
Positional and anisotropic atomic displacement parameters were
refined for nonhydrogen atoms. Hydrogen atoms were located in
difference maps and included as fixed contributions riding on attached
atoms with isotropic thermal parameters 1.2 times those of their carrier
atoms. The H atoms of methyl groups and isotropically refined atoms
were included in geometrically idealized positions. Criteria of a
satisfactory complete analysis were the ratios of “rms” shift to standard
deviation less than 0.001 and no significant features in final difference
maps. The C atoms of the methyl groups in each ligand for 3a showed
disorder and was refined using a split with 50% occupancy for each
“meta” position (C24/C25, C44/C45 and C64/C65). The lowest
(−1.68 e Å−3) and highest (2.44 e Å−3) peaks for 5a in the final
difference Fourier map are located close to the Cl23 atom at distances of
0.76 and 0.79 Å, respectively. Molecular graphics were performed from
PLATON [26]. A summary of the crystal data, experimental details and
refinement results is listed in Table 1.
νasym(COO); 1391 s, νsym(COO); 480 ms, ν(Cu-OH2O); 375 m, 327 m,
245 ms, ν(Cu-Ooco). UV–vis: λ, nm (ε/logε) in DMF: 733 (2.25), 321
(4.13), 281 (4.16); in CHCl3: 682 (1.83), 339 (4.07), 381 (4.06), 283
(3.85). μeff =1.94 MB.Mass spectrum, MS (Electrospray Ionization,
ESI, m/z): 652 [3-2H3O]−, 295 [1-H]−. Anal. calc. for C28H24Cl4CuN2O6
(689.8 g mol–1): C, 48.9; H, 3.5; N, 4.1; Cu, 9.2 found: C, 49.1, H, 3.2; N,
4.2; Cu, 8.9%. The complex is soluble in DMSO, DMF, MeOH, EtOH,
MeCN, benzene, and toluene and slightly soluble in CH2Cl2 and CHCl3.
Recrystallization of 3 from DMSO solution gives green single-crystals
of the tetramer complex [Cu4(meclo)6(OH)2(DMSO)2]•2DMSO (3a)
suitable for X-ray structure determination.
2.2.4. Synthesis of [Zn(meclo)2(H2O)2]; (4)
Zinc(II) acetate dihydrate ([Zn(CH3COO)2(H2O)2]) (0.0274 g,
0.125 mM) was dissolved in methanol (2 mL) and this solution was
added to a solution of meclofenamic acid (0.0740 g, 0.25 mM) in
methanol (2 mL). The same procedure as in Section 2.2.2 was
followed. Yellow powder (4), yield 29%. M.p. 140–141 °C. IR
(cm−1): 3295 m, ν(NH); 3068 m 2986 m, ν(CH3); 1618 s, νasym
(COO); 1397 s, νsym(COO); 467 ms, ν(Zn-OH2O); 380 m, 303 m,
270 ms, ν(Zn-Ooco). 1H NMR (DMSO): 10.23 (s, NH); 8.00 (d, H–C(3),
JH(3)–H(4)=9.0); 6.67 (t, H–C(4), JH(4)–H(5)=7.4,); 7.16 (m, H–C(5)),
6.12 (d, H–C(6), JH(6)–H(5)=8.2); 7.19 (d, H4′), JH(4′)–H(5′) =8.3);
7.38 (d, H5′), JH(5′)–H(4′)=8.3); 2.30 (s, 3′ Me); 13C NMR: 173.6
(COOH); 146.2 (C(1)); 132.0 (C(3)); 116.7 (C(4)); 135.5 (C(5)); 112.3
(C(6)); 133.3 (C(1′)); 132.1 (C(2′)); 130.2 (C(3′)); 128.5 (C(4′)); 127.9
(C(5′)); 136.2 (C(6′)); 20.1 (3′ Me); 1H NMR (CDCl3): 9.42 (s, NH);
8.00 (d, H–C(3)); 6.63 (t, H–C(4), JH(4)–H(5) =7.2); 7.17 (d, H–C(5),
J(5–4)=8.1), 6.25 (d, H–C(6), JH(6)–H(5) = 8.3); 6.97 (d, H4′),
2.4. Antiproliferative assay in vitro
JH(4′)–H(5′) = 8.3); 7.21 (d, H5′), JH(5′)–H(4′)=8.2); 2.28 (s, 3′ Me); 13
C
Compounds: Test solutions of the compounds tested (1 mg/mL)
were prepared by dissolving the substance in 100 μL of DMSO
completed with 900 μL of tissue culture medium. Afterwards, the
tested compounds were diluted in culture medium to reach the final
concentrations of 100, 50, 10, 1 and 0.1 ng/μL. The solvent (DMSO) in
the highest concentration used in test did not reveal any cytotoxic
activity.
NMR: 175.2 (COOH); 147.25 (C(1)); 113.6 (C(2)); 131.1 (C(3)); 117.1
(C(4)); 135.9 (C(5)); 115.2 (C(6)); 134.3 (C(1′)); 133.3 (C(2′)); 129.1
(C(3′)); 127.0 (C(4′)); 127.9 (C(5′)); 136.2 (C(6′)); 20.5 (3′ Me); Mass
spectrum, MS (APCI, m/z): 728 [4+2HO]−, 295 [1-H]−. Mass spectrum,
MS (Electrospray Ionization, ESI, m/z): 753 [4+2CH3O− −
]
, 1345 [42-
2H3O]− 569 [4-2Cl–CH3]+
, . Anal. calc. for C28H24Cl4N2O6Zn
(691.6 g mol–1): C, 48.6; H, 3.5; N, 4.1; Zn, 9.5 found: C, 48.4, H, 3.4; N,
4.2; Zn, 9.6%. The complex is soluble in DMSO and DMF and slightly
soluble in CH2Cl2, CHCl3.
2.4.1. Cells
The cell lines are maintained in the Cell Culture Collection of the
University of Ioannina. Twenty-four hours before addition of the
tested agents, the cells were plated in 96-well plates at a density of
104 cells per well. The MCF-7 cells were cultured in the D-MEM
(Modified Eagle's Medium) medium supplemented with 1% antibiotic
and 10% fetal calf serum. L-929 cells were grown in Hepes-buffered
RPMI 1640 medium supplemented with 10% fetal calf serum,
2.2.5. Synthesis of [Cd(meclo)2(H2O)2]; (5)
Cadmium(II) chloride dihydrate (CdCl2•2H2O) (0.1097 g, 0.25 mM))
was dissolved in methanol (2 mL) and this solution was added to a
solution of meclofenamic acid (0.0740 g, 0.25 mM) in methanol (2 mL).
The same procedure as in Section 2.2.3 was followed. Yellow powder (5),