SYNTHESIS AND ANTITUMOR PROPERTIES OF CARBORANE CONJUGATES
95
Table. (Contd.)
UV/Vis, λmax
1H NMR (
CDCl3)
δ
, ppm,
11B NMR (
CDCl3)
δ
, ppm,
MS,
m/z
,
Comꢀ
pound
IR (КBr, ν )
, cm–1
nm (ε × )
10–3
XVII 646.0 (4.75),
591.0 (5.61),
551.0 (8.20),
3318 (NH), 3060 10.20 (s, 1H, NH), 8.84 (s, 8H,
β
ꢀpyrꢀ –4.00 (d, 3B,
J
J
J
J
151 Hz),
927
(carborane CH),
2594 (BH),
1678–1599
role), 8.22 (m, 8H, Рh), 7.95 (m, 2H, –8.79 (d, 1B,
154 Hz), [M+1]+
Рh), 7.75 (m, 9H, Рh), 6.34 (d, 1H, –11.47 (d, 1B, 183 Hz),
13.6 Hz, СH–СH–N), 6.21 (d, 1H, –12.94 (d, 4B, 179 Hz),
13.6 Hz, СH–СH–N), 4.51 (s, 2H, –14.44 (d, 1B, 180 Hz)
CH2), 3.48 (s, 1H, carborane CH),
2.90–0.88 (m, 10H, BH)
515.0 (14.6),
J
J
419.0 (354.16) (CO– in COOH,
amide I), 1512
(amide II)
J
XVIII 619.0 (2.31),
3318 (NH), 3060
(carborane CH),
2594 (BH), 1728
8.89 (s, 2H, ꢀpyrrole), 8.88 (s, 2H,
ꢀpyrrole), 8.85 (s, 4H, ꢀpyrrole),
8.32 (m, 2H, Рh), 8.21 (m, 6H, Рh), –4,23 (d, 4B,
β
–0.37 (s, 1B),
–0.77 (s, 1B),
1043
[M+1]+
592.0 (2.91),
551.0 (2.96),
514.0 (5.02),
419.0 (111.0) 1626 (amide I),
1592 (amide II)
β
β
J
160 Hz),
(CO– in COOH), 7.76 (m, 11H, Рh), 4.68 (s, 1H, NH), –9.54 (d, 2B,
3.08 (s, 1H, NH), 3.59 (dd, 1H, 18.6, –13.92 (d, 2B,
9.42 Hz CH2–СH–S), 3,44 (s, 4H,
carborane CH), 3.10 (d.d, 2H,
CH2–СH–S, 18.6, 9.42 Hz),
–2.80 (2H, porphyrin NH)
J 163 Hz),
J
J
J
J
169 Hz),
170 Hz),
180 Hz),
161 Hz)
–14.63 (d, 4B,
–15.32 (d, 4B,
–18.40 (d, 2B,
J
J
Notes: I, 5ꢀ(pꢀAminophenyl)ꢀ10,15,20ꢀtriphenylporphyrin; IV, 5ꢀ[4'ꢀ(3''ꢀ(carboxypropanoyl)ꢀ aminophenyl]ꢀ10,15,20ꢀtriphenylporphyrin;
V, 5ꢀ[4'ꢀ(3''ꢀcarboxypropenoyl)aminophenyl]ꢀ10,15,20ꢀtriphenylporphyrin; VIII, 5ꢀ[4'ꢀ(Nꢀcarboranꢀ3ꢀyl)succinamidophenylꢀ10,15,20ꢀ
triphenylporphyrin; IX, 5ꢀ[4'ꢀNꢀ(propargyl)succinamidophenylꢀ10,15,20ꢀtriphenylporphyrin; X, zinc 5ꢀ[4'ꢀNꢀ(propargyl) succinamidopheꢀ
nylꢀ10,15,20ꢀtriphenylporphyrin; XII, zinc 5ꢀ{4'ꢀNꢀ(oꢀcarboranꢀ1''ꢀyl)methyl]triazolyl}succinamidophenylꢀ10,15,20ꢀtriphenylporphyrin;
XIII, 5ꢀ{[4'ꢀNꢀ(oꢀcarboranꢀ1''ꢀyl)methyl]triazolyl}succinamidophenylꢀ10,15,20ꢀtriphenylporphyrin; XIV, 5ꢀ{4'ꢀ[Nꢀ(oꢀcarboranꢀ3''ꢀyl)maleinꢀ
amido]phenyl}ꢀ10,15,20ꢀtriphenylporphyrin, XVI, 5ꢀ{4'ꢀ[3''ꢀ(mꢀthiocarboranꢀ9''ꢀyl)carboxypropanoyl]ꢀaminophenyl}ꢀ10,15,20ꢀtripheꢀ
nylporphyrin; XVII, 5ꢀ{4'ꢀ[5''ꢀcarboxyꢀ1ꢀ(oꢀcarboranꢀ1''ꢀyl)methyl](triazolinꢀ4''ꢀyl)carbonylamino]phenyl}ꢀ10,15,20ꢀtriphenylporphyrin;
4
1
XVIII, 5ꢀ{4'ꢀ[2''ꢀ(mꢀcarboranꢀ9''ꢀyl)thio)ꢀN ꢀ(oꢀcarboranꢀ3''ꢀyl)succinamidoꢀN ] phenyl}ꢀ10,15,20ꢀtriphenylporphyrin. For compounds I,
VII, and VIII, see the text.
The structures of all of the synthesized compounds iodide and counting of the percentage of colored cells
1
were confirmed by UV/Vis, IR, H and 11B NMR
using a flow type cytofluorimeter [9]. After illuminaꢀ
tion of the cells incubated with compound XIV, the
cell death was slower and the morphology of the damꢀ
aged cells was indicative of apoptosis; the cells
rounded and gradually detached from the substrate.
spectroscopy and mass spectrometry (table). The IR
spectra of carboraneꢀcontaining porphyrins (VIII
,
XII, XIII, XIV–XVIII) show an absorption band at
2580–2600 cm–1 corresponding to –BH stretching
vibrations. In addition, amide I and amide II bands
occur at 1627 and 1598 cm–1, respectively.
Within 24 h after the end of light exposure, ~90% of
the cells were detached from the solid substrate; howꢀ
ever, the morphological pattern of the death differed
from that of photoinduced necrosis. Finally, illuminaꢀ
tion of the cells incubated with compound XVIII
induced a much slower damage: after 24–48 h many
cells changed insignificantly and remained linked to
the substrate.
The antitumor characteristics of porphyrins
V,
XIV, and XVIII were assessed by measuring the dark
toxicity and the ability to cause lightꢀinduced death of
cultured human tumor cells (HCT116 colon cancer
cells). The dark cytotoxicity was determined in the
MTT test [15] after cell incubation with compounds
V
,
XIV, and XVIII for 72 h. The 50% inhibition of cell
growth was observed at rather high concentrations, the
IC50 values being >12.5 M for , >10 M for XIV
and >25 M for XVIII. The low dark cytotoxicity
Thus, compounds
dark toxicity. The action of light on the cells loaded
with nonꢀboronated porphyrin caused fast (during
V, XIV, and XVIII showed no
µ
V
µ
,
V
µ
the first minutes) necrosis. In the presence of monoꢀ
carboranylporphyrin XIV, the death developed more
slowly and by a mechanism other than PDTꢀinduced
necrosis, while in the presence of dicarboranylporphyꢀ
rin XVIII, the cell damage was even less efficient. The
results show that the boron polyhedra change the phoꢀ
tosensitizer properties of porphyrins. The addition of
one boron polyhedron to the macrocycle can switch
the death mechanism from necrosis to apoptosis,
which is significant for the selection of the therapy
enables the use of these compounds in the PDT of
cancer.
The experimental PDT was carried out after 30 to
60 min incubation of the cells with 10 µM of V, XIV,
or XVIII and removal of the compounds from the culꢀ
ture medium [9]. The illumination of cells loaded with
compound
within 5 to 7 min after the end of light exposure. The
necrosis was detected based on typical morphology
V caused necrosis of 100% of the cells
(cell shadows) and on the inclusion of propidium protocol. The addition of the second boron polyheꢀ
DOKLADY CHEMISTRY Vol. 443
Part 2
2012