M. Balaz et al. / Bioorg. Med. Chem. 13 (2005) 2413–2421
2419
8
ꢁ
.19 (m, 6H, CH), 7.74 (m, 9H, CH), 4.10(s, 3h, Me),
2.76 (s, 2H, NH).
4.7. N-[2-[Bis-(4-methoxy-phenyl)-phenyl-methoxymethyl]-
5-(5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-
tetrahydro-furan-3-yl]-4-(10,15,20-triphenyl-porphyrin-5-
yl)-benzamide (4b)
4.4. 4-(10,15,20-Tri-pyridin-2-yl-porphyrin-5-yl)-benzoic
acid, TPyrPhP-COOH (2a)
1
H NMR (400 MHz, CDCl ) ppm (d) 8.84 (m, 6H, CH),
3
8
.75 (d, 2H, CH), 8.21 (m, 10H, CH), 7.74 (m, 10H,
CH), 7.43 (m, 3H, CH), 7.33 (m, 3H, CH), 7.25 (m,
H, CH), 7.13 (m, 1H, CH), 6.78 (m, 3H, CH), 6.61
m, 1H, CH), 5.05 (m, 1H, CH), 4,31 (br s, 1H, NH),
.69 (m, 1H, CH), 3.62 (s, 3H, CH ), 3.58 (m, 1H,
The methyl ester from the previous reaction (140mg,
0
1
2
.21 mmol) was dissolved in 5.6 mL of ethanol and
1 mL of 2 M NaOH. The solution was refluxed for
h. The reaction was then allowed to cool to room tem-
4
(
3
3
perature. Once cooled, an equivalent amount of 1 M
HCl (22 mL) was added. The resulting solution
CH), 3.56 (s, 3H, CH ), 2.68 (m, 2H, CH), 1.50(s,
3
3
H, CH ), ꢁ2.78 (s, 2H, NH). FAB-HRMS calcd for
3
(
pH = 5) was then extracted several times with ethyl ace-
+
C H N O (MH ) 1183.4632, found 1183.4644.
7
6
61
7
7
tate. The product was pure according to TLC and was
used in the next reaction without further purification.
A total of 120mg of TPyrPhP-COOH were obtained
with an 87% yield.
4
.8. N-[2-Hydroxymethyl-5-(5-methyl-2,4-dioxo-3,4-
dihydro-2H-pyrimidin-1-yl)-tetrahydro-furan-3-yl]-4-
10,15,20-trispyridin-2-yl-porphyrin-5-yl)-benzamide,
(
1
3
TPyrPhP-CONHThyOH (5a)
H NMR (400 MHz, CDCl ) ppm (d) 8.95 (d, J = 4 Hz,
3
3
8
8
H, pyrrole CH), 8.75 (broad band, 8H, pyrrole CH),
0
3
.36 (d, J = 8 Hz, 2H, CH), 8.20(m, 3H, CH),
5 -DMT–porphyrin–thymidine conjugate (60mg) was
3
.16 (d, J = 7.9 Hz, 2H, CH), 8.13 (m, 3H, CH), 7.82
dissolved in 1.3 mL of 60/40 (by volume) mixture of
DCM/MeOH. The solution was placed in an ice bath
and a 4% solution of p-TsOH in an 80/20 solution of
DCM/MeOH was added drop wise. The solution turned
from dark red to dark green in color. Once the reaction
was complete (followed by TLC), 2.5 mL of a saturated
solution of Na CO were added. The product was ex-
tracted using DCM. The crude product was purified
via column chromatography over silica gel using a 90/
/1 mixture of DCM/MeOH/TEA (R = 0.35). The ob-
f
(m, 3H, CH), ꢁ2.91 (s, 2H, NH). FAB-HRMS
calcd for C H N O (MH ) 661.2226, found
+
42 27 7 2
6
61.2262.
4
(
.5. 4-(10,15,20-Triphenyl-porphyrin-5-yl)-benzoic acid
2b)
2
3
1
H NMR (400 MHz, CDCl ) ppm (d) 8.83 (m, 6H, CH),
3
9
8
.76 (d, 2H, CH), 8.41 (d, 2H, CH), 8.28 (d, 2H, CH),
tained product was dissolved in DCM and was washed
three times with water (to remove inorganic impurities).
After evaporation, 42 mg of TPyrPhP-COOThyOH was
obtained with a 95% yield.
8.19 (m, 6H, CH), 7.74 (m, 9H, CH), ꢁ2.76 (s, 2H,
NH). FAB-HRMS calcd for C H N O (MH )
+
4
5
30
4
2
6
58.2369, found 658.2367.
1
4
5
.6. N-[2-[Bis-(4-methoxy-phenyl)-phenyl-methoxymethyl]-
-(5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-
H NMR (400 MHz, CDCl ) ppm (d) 9.10(m, 3H, CH),
3
8.84 (s, 4H, CH), 8.79 (d, 2H, CH), 8.68 (d, 2H, CH),
8.23 (m, 3H, CH), 8.06 (m, 6H, CH), 8.18 (d, 1H,
CH), 7.88 (s, 1H, NH), 7.64 (s, 1H, OH), 7.59 (s, 1H,
CH), 6.96 (s, 1H, CH), 5.80(s, 1H, CH), 4.30(s, 1H,
CH), 3.65 (m, 2H, CH), 3.54 (s, 1H, NH), 1.89 (s, 3H,
tetrahydro-furan-3-yl]-4-(10,15,20-trispyridin-2-yl-por-
phyrin-5-yl)-benzamide, TPyrPhP-CONHThyDMT
(
4a)
Porphyrin acid 2 (46 mg) was dissolved, under argon, in
mL of dry DCM. To the red solution, DMAP
CH ), ꢁ2.88 (s, 2H, NH). FAB-HRMS calcd for
3
+
C H N O (MH ) 884.3183, found 884.3220.
2
5
2
49 10
5
(
9.34 mg, 0.0765 mmol), EDC (39 mg, 0.073 mmol),
0
and 3 -aminothymidine 3 (39 mg, 0.073 mmol) were
added successively. The solution was stirred at room
temperature overnight. The crude reaction mixture was
loaded directly onto a silica gel column and eluted using
a mixture of DCM/MeOH = 95/5 with 0.5% of triethyl
amine (TEA). A total of 60mg of TPyrPhP-CON-
4.9. N-[2-Hydroxymethyl-5-(5-methyl-2,4-dioxo-3,4-
dihydro-2H-pyrimidin-1-yl)-tetrahydro-furan-3-yl]-4-
(10,15,20-triphenyl-porphyrin-5-yl)-benzamide, TPhP-
CONHThy-OH (5b)
1
H NMR (400 MHz, CDCl ) ppm (d) 8.82 (br s, 6H,
3
HThyDMT (R = 0.22, DCM/MeOH = 9/1) were ob-
f
tained with a 73% yield.
CH), 8.74 (m, 2H, CH), 8.26 (s, 4H, CH), 8.17 (m,
6H, CH), 7.65–7.7.80(m, 11H, CH), 6.39 (t, J = 6 Hz,
1
(m, 1H, CH), 4,02 (m, 1H, CH), 2.70 (m, 1H, CH),
3
1
H, H ), 5.40(br s, 1H, NH), 4.83 (m, 1H, CH), 4.13
1
H NMR (400 MHz, CDCl ) ppm (d) 9.07 (m, 3H, CH),
3
8
8
7
7
4
3
.88–8.71 (m, 8H, pyrrole CH), 8.12–8.19 (m, 6H, CH),
.00 (m, 3H, CH), 7.92 (s, 1H, NH), 7.64 (m, 4H, CH),
.40(d, 2H, CH), 7.28 (m, 4H, CH), 7.20(m, 2H, CH),
.09 (s, 1H, CH), 6.73 (m, 4H, CH), 5.00 (s, 1H, NH),
.25 (s, 1H, CH), 3.59 (s, 3H, CH ), 3.55 (s, 3H, CH ),
2.46 (m, 1H, CH), 1.98 (s, 3H, CH ), 1.24 (t, 1H,
3
OH), ꢁ2.79 (s, 2H, NH). FAB-HRMS calcd for
+
C H N O (MH ) 881.3326, found 881.3422.
5
5
43
7
5
4.10. TPyrPhP-CONHThy-Phosphoramidite (6a)
3
3
.59 (s, 3H, CH ), 3.20(q, 1H, CH), 2.54 (s, 2H,
3
CH ), ꢁ2.81 (s, 2H, NH). FAB-HRMS calcd for
TPyrPhP-CONHThyOH (8.9 mg, 0.01017 mmol) was
dissolved in anhydrous DCM under argon and at room
2
+
C H N O (MH ) 1186.4490, found 1186.4586.
7
3
58 10
7