Inorganic Chemistry
Article
3
1
1
53.5 (s, CH3). 31P{ H} NMR (162 MHz): δ = +32.1 (s). HRMS:
1
CH ), 14.1 (s, br, CH ). P{ H} NMR (162 MHz): δ = +7.7 (s).
3
3
HRMS: calcd. for C H NPCl Ru ([M] + NEt ), 670.1704; found,
calcd. for calcd. for C H Cl NO PRu ([M] + NH ), 712.0507;
3
4
45
2
3
36 31
2
2
4
6
70.1711. C, H Anal.: cald. for C H Cl PRu, C 59.16%, H 5.14%;
found, 712.0499. Anal.: cald. for C H Cl O PRu, C 62.25%, H
2
8
29
2
36 27 2 2
found, C 57.89%, H 5.32%.
3.92%; found, C 59.25%, H 3.95%.
6
6
[
Ru(η -methyl benzoate)Cl (L1)] (2). Procedure A: L1 (200 mg,
[Ru(η -methyl benzoate)Cl (S-L3)] (6). The procedure A applied
2
2
0
[
.76 mmol) was dissolved in 10 mL of dichloromethane and
RuCl(μ-Cl)(η -methyl benzoate)] (DB; 196 mg, 0.32 mmol) was
to prepare 2 was followed (see above) using 388 mg (1.20 mmol) of
S-L3 and 308 mg (0.50 mmol) of DB. Pure 6 was obtained as a dark-
red solid (571 mg, 90%).
6
2
subsequently added. The resulting red solution was stirred for 1 h
protected from light and subsequently filtered; the solvent was then
removed under reduced pressure. The solid residue was recrystallized
from dichloromethane/hexane to give pure 2 as a dark-red solid (280
mg, 77%).
Procedure B: the method applied for 1 was followed (see above)
using 1.69 g (6.00 mmol) of 1-bromopyrene, 3.6 mL (5.7 mmol) of
−
1
IR (KBr, cm ): 3052, 2949, 1727 ν(C = O), 1581, 1520, 1486,
1434, 1293, 1275, 1109, 895, 851, 768, 749, 720, 691, 629, 602, 499.
1
H NMR (CDCl , 400 MHz): δ = 8.59 (t, J = 7.6, 1H), 8.43 (d, J =
3
9
1
7
.2, 1H), 8.37 (d, J = 7.2, 1H), 8.30−8.17 (m, 4H), 8.09 (d, J = 7.6,
H), 8.04 (d, J = 9.6, 1H), 7.66 (t, J = 8.4, 1H), 7.35 (t, J = 6.4, 1H),
.26 (m, 2H), 6.47 (s, br, 1H), 6.09 (s, br, 1H), 5.43 (s, br, 1H), 5.36
1
.6 M n-BuLi in hexane, 0.4 mL (500 mg, 5.20 mmol) of
2
(
s, br, 1H), 4.55 (s, br, 1H), 3.86 (s, 3H), 2.36 (d, J = 10.8, 3H).
C{ H} NMR (101 MHz): δ = 164.8 (s, C = O), 133.6−124.2 (C,
HP
chlorodimethylphosphane and 388 mg (0.63 mmol) of DB. After
workup and recrystallization, pure 2 was obtained a dark-red solid
13
1
CH, Ar), 97.2 (s, CH), 91.5 (s, CH), 90.7 (s, CH), 87.7 (s, CH), 84.0
(
(
(
680 mg, 94%).
1
31
1
s, CH), 53.2 (s, CH ), 16.0 (d, J = 36.1, CH3). P{ H} NMR
−
1
3 CP
IR (KBr, cm ): 3081, 2953, 1701 ν(C = O), 1592, 1518, 1422,
162 MHz): δ = +16.6 (s). HRMS: calcd. for calcd. for
1
1
292, 1272, 1110, 949, 917, 850, 773, 720. H NMR (CDCl , 400
3
C H NO PCl Ru ([M] + NH ), 650.0349; found, 650.0354.
3
1
29
2
2
4
MHz): δ = 9.11 (d, J = 9.2, 1H), 8.49 (dd, J = 10.0, 8.0, 1H), 8.35−
Anal.: cald. for C H Cl O PRu, C 58.87%, H 3.98%; found, C
3
1
25
2
2
8
8
.30 (m, 4H), 8.24 (d, J = 8.8, 1H), 8.17 (d, J = 8.8, 1H), 8.12 (d, J =
.1, 1H), 6.25 (s, br, 2H), 5.47 (t, J = 5.2, 1H), 3.88 (s, 3H), 2.06 (s,
5
7.86%, H 4.12%.
X-ray Structure Determination. Single crystals, suitable for X-
13
1
br, 6H). C{ H} NMR (101 MHz): δ = 165.5 (s, C = O), 133.5−
ray diffraction studies, of 1, 2, and 4 were obtained by slow diffusion
of hexane into a dichloromethane solution of each complex, at 4 °C.
Crystallographic data for 1, 2, and 4 were collected on a Bruker APEX
II QUAZAR diffractometer equipped with a microfocus multilayer
monochromator with Mo Kα radiation (λ = 0.71073 Å), at the Group
of Magnetism and Functional Molecules (GMMF) of the Universitat
de Barcelona. Data reduction and absorption corrections were
performed by using SAINT and SADABS (Bruker AXS Inc.,
Madisson, Wisconsin, USA), respectively. The structures were solved
1
(
24.1 (C, CH, Ar), 92.4 (s, 2CH), 79.8 (s, CH), 53.2 (s, CH ), 13.5
3
m, br, 2CH3). 31P{ H} NMR (162 MHz): δ = +9.6 (s). HRMS:
1
calcd. for C H NO PCl Ru ([M] + NEt ), 672.1133; found,
32
39
2
2
3
6
72.1143. Anal.: cald. for C H Cl O PRu, C 54.75%, H 4.06%;
26 23 2 2
found, C 52.99%, H 4.35%.
6
[
Ru(η -p-cymene)Cl (L2)] (3). Procedure A: the synthetic proce-
2
dure used to prepare 1 was followed with 173 mg (0.45 mmol) of L2
and 114 mg (0.19 mmol) of DC. Pure 3 was obtained as a dark-red
solid (237 mg, 90%).
51,52
53
using SHELXT
and refined with OLEX2 suite. Crystallographic
Procedure B: the synthetic procedure applied for 1 was used with
00 mg (1.79 mmol) of 1-bromopyrene, 1.0 mL (1.6 mmol) of 1.6 M
5
n-BuLi in hexane, 0.2 mL (290 mg, 1.36 mmol) of chlorodimethyl-
phosphane and 420 mg (0.69 mmol) of DC. Pure 3 was obtained as a
dark-red solid (890 mg, 94%).
(
2), and 1863808 (4) that contain the supplementary crystallographic
−
1
IR (KBr, cm ): 3041, 2958, 2923, 2869, 1581, 1481, 1468, 1434,
374, 1207, 1189, 1159, 1091, 1028, 857, 751, 741, 720, 692, 634,
06, 582, 540, 516, 465, 425. H NMR (CDCl , 400 MHz): δ = 8.90
1
6
1
Agarose Gel Electrophoresis. Twenty microliters of cacodylate-
buffered solutions of pBR322 plasmid DNA (15 μM, in base pair)
containing 0.42, 0.83, 1.67, and 3.33 equiv of the complexes (6.25−50
μM, obtained from DMSO stock solutions) were incubated for 24 h
at 37 °C. Control samples of free DNA and DNA bound to cisplatin
(0.5 and 1 equiv) were also prepared. Following the incubation, 4 μL
of a xylene cyanol 0.25% aqueous solution (containing 30% glycerol)
was added to all samples, which were subsequently electrophoretized
in agarose gel (1% in TBE buffer) for 1 h at 6.25 V cm , using a Bio-
Rad horizontal tank connected to a Consort EV231 variable potential
power supply. The gel was then treated with SYBR Safe DNA gel stain
and consequently photographed with a Bio-Rad Gel Doc EZ imager.
Ethidium Bromide Displacement Assays. Samples containing
ct-DNA (15 μM, in base pair) and ethidium bromide (ethidium
bromide, EB; 75 μM) in cacodylate buffer (1 mM sodium cacodylate,
3
(
(
d, J = 9.6, 1H), 8.29 (t, J = 8.0, 2H), 8.23 (d, J = 9.2, 1H), 8.14−8.08
m, 4H), 7.74 (t, J = 10.4, 1H), 7.66 (t, J = 8.8, 4H), 7.37−7.26 (m,
3
6
3
1
H), 5.29 (s, 2H), 4.47 (s, br, 2H), 3.12 (h, J = 6.8, 1H), 1.62 (s,
HH
3
13
1
H), 1.26 (d, J = 6.8, 6H). C{ H} NMR (101 MHz): δ =
HH
2
35.7−123.7 (C, CH, Ar), 113.6 (d, J = 6.5, C), 98.3 (s, C), 88.6
CP
(
s, 2CH), 86.3 (s, 2CH), 30.5 (s, CH), 22.1 (s, 2CH ), 18.2 (s, CH ).
3 3
31
1
P{ H} NMR (162 MHz): δ = +32.1 (s). HRMS: calcd. for calcd. for
−
1
C H NPCl Ru ([M] + NH ), 710.1078; found, 710.1073. Anal.:
38
37
2
4
cald. for C H Cl PRu, C 65.90%, H 4.80%; found, C 65.00%, H
38
33
2
5
.11%.
6
[Ru(η -methyl benzoate)Cl (L2)] (4). Procedure A: 200 mg (0.52
2
mmol) of ligand L2 was dissolved in 10 mL of dichloromethane, and
85 mg (0.30 mmol) of DB was added. The resulting red solution was
1
stirred for 1 h protected from light and subsequently filtered, and the
solvent was removed under reduced pressure. The solid residue was
recrystallized from dichloromethane/hexane, giving pure 4 as a dark-
red solid (300 mg, 83%).
Procedure B: the method applied for 1 was followed (see above)
using 500 mg (1.79 mmol) of 1-bromopyrene, 1.0 mL (1.60 mmol) of
20 mM NaCl, pH = 7.2) were incubated at 37 °C for 1 h. Then, the
samples were treated with increasing amounts of 250 μM stock
solutions of the complexes freshly prepared in DMSO (final complex
concentrations of 1, 2, 3, 6, 9, 12, 15, 20, and 25 μM were used for the
experiments). The optimum DNA/EB ratio of 1:5 was determined
experimentally, by fluorescence spectroscopy. This ratio corresponds
to the saturation of the emission signal, illustrated by a plateau and is
indicative of the occupation of all possible intercalation sites by EB.
The samples, containing 1−25 μM of the complexes investigated and
up to a maximum of 5% DMSO in a final volume of 3 mL, were
subsequently incubated at 37 °C for 24 h. The fluorescence emission
spectra of all samples were then recorded at room temperature in the
1
.6 M n-BuLi in hexane, 0.2 mL (290 mg, 1.36 mmol) of
chlorodimethylphosphane and 420 mg (0.68 mmol) of DB. Pure 4
was obtained as a dark-red solid (900 mg, 95%).
−
1
IR (KBr, cm ): 3059, 2949, 1717 ν(C = O), 1657, 1556, 1451,
271, 1106, 843, 856, 704. H NMR (CDCl , 400 MHz): δ = 8.81 (d,
1
1
3
J = 9.2, 1H), 8.31 (d, J = 8.0, 1H), 8.29 (d, J = 8.0, 1H), 8.24 (d, J =
8
.8, 1H), 8.14 (d, J = 2.8, 2H), 8.12 (d, J = 2.8, 2H), 7.61−7.56 (m,
5H), 7.41 (s, br, 3H), 7.31 (s, br, 3H), 6.44 (d, J = 6.0, 2H), 4.89 (s,
range 520−700 nm, applying an excitation wavelength λ = 514 nm.
Blank experiments were also carried out with cacodylate-buffered
solutions of ct-DNA/EB (namely, without complex).
exc
1
3
1
br, 3H), 4.75−4.71 (m, 2H), 3.91 (s, 3H). C{ H} NMR (101
MHz): δ = 164.2 (s, C = O), 135.6−124.0 (C, CH, Ar), 89.6 (s, CH),
I
Inorg. Chem. XXXX, XXX, XXX−XXX