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M. Thutewohl, H. Waldmann / Bioorg. Med. Chem. 11 (2003) 2591–2615
2ꢂ1.5 h according to GP P5 variant B followed by
PyAOP (250 mg, 480 mmol), DMAP (15 mg, 120 mmol),
DIEA (205 mL, 1.2 mmol) in 7.5 mL DMF/CH2Cl2 for
24 h According to GP P8. After the liberation of the
C-terminus with Pd(PPh3)4 in 5 mL morpholine/THF
for 16 h according to GP P1 variant C the product was
cleaved from the resin by treatment with 2ꢂ5 mL TFA/
CH2Cl2/H2O 47/47/6 (v/v) for 10 and 20 min and 2ꢂ5
mL TFA/H2O 95/5 (v/v) for 30 min and 180 min
according to GP P5 variant B followed by HPLC pur-
ification affording 35/16 (3 mg, 3%, eight steps) as a
light brown solid as a mixture of E- und Z-isomer
(E/Z=1:1, according to 1H NMR); HPLC (A2,
CH3CN/H2O/TFA, 45/55/0.1–90/10/0.1 in 30 min,
50 ꢁC): Rt=16.19 min; HPLC (A2, CH3CN/H2O/TFA,
HPLC purification affording the appropriate product.
Nꢀ-3-[4-(n-Pentyl)phenyl]propionyl-L-histidyl-L-phenyl-
alanyl-L-tyrosine hydrotrifluoroacetate (35/13). Starting
from tripeptide 34b, reaction according to GP P15
furnished 35/13 (24 mg, 19%, eight steps) as a white
solid; HPLC (A2, CH3CN/H2O/TFA, 20/80/0.1–70/30/
0.1 in 30 min, 50 ꢁC): Rt=20.04 min, purity: 93%;
20
D
mp: >210 ꢁC (decomp.); ½a ꢀ13.2 (c 0.19, MeOH);
1H NMR (400 MHz, CD3OD): d 0.87(t, 3J=6.8 Hz,
3H), 1.25–1.36 (m, 4H), 1.52–1.59 (quint, J=7.6 Hz,
3
3
3
2H), 2.44 (t, J=8.4 Hz, 2H), 2.54 (t, J=7.6 Hz, 2H),
3
2.79 (t, J=7.6 Hz, 2H), 2.81–2.94 (m, 3H), 3.00–3.13
3
(m, 3H), 4.56 (dd, J=5.1 Hz, J=8.2 Hz, 1H), 4.56–
4.62 (m, 2H), 6.67(d, 3J=8.6 Hz, 2H), 6.93 (s, 1H), 7.04
20/80/0.1–90/10/0.1 in 30 min, 50 ꢁC): Rt=13.47min
3
20
D
+6.3 (c 0.24, MeOH); H NMR (400 MHz, CD3OD)
(isomer 1), Rt=15.41 min (isomer 2); mp: 165 ꢁC; ½a
3
1
(d, J=8.6 Hz, 2H), 7.04–7.08 (m, 4H), 7.17–7.25 (m,
+
5H), 8.58 (s, 1H); LC–MS (ESI 2): 668.17[M+H]
,
Z-isomer: d 0.96 (m, 3H), 1.27–1.30 (m, 2H), 1.49–
1.52 (m, 2H), 2.72–2.78 (m, 6H), 4.56–4.72 (m, 3H),
690.15 [M+Na]+, Rt=1.94 min; MS (MALDI-TOF):
668.8 [M+H]+, 690.8 [M+Na]+; HRMS (FAB): calcd
for C38H46N5O6: 668.3448, found: 668.3433.
3
5.93 (d, Jcis=12.7Hz, 1H), 6.30–6.39 (m, 2H), 6.65–
6.68 [6.74–6.77] (m, 2H), 6.89 (m, 1H), 6.99 (d,
3J=8.2 Hz, 1H), 7.15–7.27 (m, 7H), 7.39 (t, 3J=8.8
Nꢀ-3-[4-(n-Pentyl)phenyl]propionyl-L-histidy-N-methyl-
L-phenylalanyl-L-tyrosine hydrotrifluoroacetate (35/14).
Starting from tripeptide 34d, reaction according to GP
P15 furnished 35/14 (38 mg, 30%, eight steps) as a white
solid; HPLC (A2, CH3CN/H2O/TFA, 20/80/0.1–90/10/
0.1 in 40 min, 50 ꢁC) Rt=18.36 min, purity: >95%; mp:
120 ꢁC; ½a2D0, ꢀ12.3 (c, 1.0, MeOH); 1H NMR
(400 MHz, CD3OD): d, 0.87(m, 3H), 1.29 (m, 4H), 1.54
(m, 2H), 2.33–2.45 (m, 4H), 2.52 (s, 3H), 2.65–2.71 (m,
2H), 2.79–2.92 (m, 4H), 3.12–3.23 (m, 2H), 4.55–4.67
(m, 1H), 4.94–5.00 (m, 1H), 5.27–5.39 (m, 1H), 6.67 (m,
2H), 6.94–6.96 (m, 3H), 7.02–7.20 (m, 9H), 8.71 (s, 1H);
LC–MS (ESI-2):, 682.18 [M+H]+, 704.16 [M+Na]+,
Rt=1.99 min; MS (FAB): 682.3 [M+H]+, 704.3
[M+Na]+; HRMS (FAB): calcd for C39H48N5O6
[M+H]+: 682.3605, found: 682.3625.
Hz, 2H), 7.47 (d, J=8.8 Hz, 1H), 8.54 (s, 1H, His);
E-isomer: d 0.96 (m, 3H), 1.27–1.30 (m, 2H), 1.49–
1.52 (m, 2H), 2.72–2.78 (m, 6H), 4.56–4.72 (m, 3H),
3
3
6.30–6.39 (m, 2H), 6.56 (d, Jtrans=15.9 Hz, 1H),
3
6.74–6.77 (m, 2H), 6.93 (m, 1H), 7.04 (d, J=8.2 Hz,
3
1H), 7.15–7.27 (m, 6H), 7.39 (t, J=8.8 Hz, 2H), 7.47
3
3
(d, J=8.8 Hz, 1H), 7.47 (d, J=16.0 Hz, 1H), 8.56
(s, 1H); LC–MS (ESI-1, Grad. A): 664.3 [M+H]+,
686.3 [M+Na]+; Rt=16.06 min (isomer 1), Rt=17.67
min (isomer 2); MS (MALDI-TOF): 664.9 [M+H]+,
686.9 [M+Na]+; HR-MS (FAB, m/z): calcd for
C38H42N5O6 [M+H]+ 664.3135, found: 664.3096.
Nꢀ-3-[3-(1-E-n-Pentenyl)phenyl)]-E-acryloyl-D-histidyl-
L-phenylalanyl-L-tyrosine hydrotrifluoroacetate (35/17).
According to GP P2, the polymer-bound tripeptide 34a
(150 mg, 0.40 mmol/g, 60 mmol) was N-terminally
deblocked by piperidine/DMF treatment for 2ꢂ5 min,
followed by coupling with 30g (39 mg, 180 mmol),
PyAOP (94 mg, 180 mmol), DMAP (7mg, 60 mmol),
HOAt (8 mg, 60 mmol), DIEA (103 mL, 600 mmol) in 3 mL
DMF/CH2Cl2 for 24 h according to GP P8. After the lib-
eration of the C-terminus with Pd(PPh3)4 in 5 mL mor-
pholine/THF for 16 h according to GP P1 variant C the
product was cleaved from the resin by treatment with 2ꢂ5
mL TFA/CH2Cl2/H2O 47/47/6 (v/v) for 10 and 20 min and
2ꢂ5 mL TFA/H2O 95/5 (v/v) for 30 min and 180 min
according to GP P5 variant B followed by HPLC purifica-
tion affording 35/17 (6 mg, 13%, eight steps) as a white
solid. HPLC (A2, CH3CN/H2O/TFA, 45/55/0.1–90/10/0.1
in 30 min, 50 ꢁC); Rt=15.85 min, purity: >95%; mp:
Nꢀ-3-[4-(n-Pentyl)phenyl]propionyl-D-histidyl-N-methyl-
L-phenylalanyl-L-tyrosine hydrotrifluoroacetate (35/15).
Starting from tripeptide 34c, reaction according to GP
P15 furnished 35/15 (27mg, 21%, eight steps) as a
slightly yellow solid. HPLC (A2, CH3CN/H2O/TFA,
20/80/0.1–70/10/0.1 in 30 min, 50 ꢁC) Rt=18.95 min,
20
purity: >95%; mp: 127 ꢁC; ½a ꢀ20.0 (c 0.75, MeOH);
D
1H NMR (400 MHz, CD3OD): d 0.86 (t, 3J=7.0 Hz, 3H),
1.29–1.31 (m, 4H), 1.54 (quint, 3J=7.4 Hz, 2H), 2.32–2.39
(m, 1H), 2.46–2.54 (m, 3H), 2.60 (dd, 2J=15.3 Hz, 3J=6.3
Hz, 2H), 2.70 (s, 3H), 2.78 (t, 3J=7.2 Hz, 2H), 2.86–2.92
(m, 4H), 4.56 (dd, 3J=5.1 Hz, 3J=8.2 Hz, 1H), 4.56–4.62
(m, 2H), 6.67(d, 3J=8.6 Hz, 2H), 6.93 (s, 1H), 7.04 (d,
3J=8.6 Hz, 2H), 7.04–7.08 (m, 4H), 7.17–7.25 (m, 5H),
8.58 (s, 1H); LC–MS (ESI-2): 682.18 [M+H]+, 7 04.16
[M+Na]+, Rt=1.92 min; HRMS (FAB): calcd for
C39H47N5O6 [M+H]+ 682.3605, found: 682.3615.
20
D
144 ꢁC; ½a ꢀ4.2 (MeOH, c 0.33); H NMR (400MHz,
1
CD3OD): d 0.97(t, 3J=7.2 Hz, 3H), 1.49–1.55 (m, 2H),
2
3
2.17–2.23 (m, 2H), 2.81 (dd, J=14.1 Hz, J=10.0, 1H),
2.91–2.97(m, 2H), 3.09–3.19 (m, 3H), 4.48–4.52 (m, 1H),
4.58–4.66 (m, 2H), 6.29–6.34 (m, 1H), 6.41 (d, 3Jtrans=16.0
Hz, 1H), 6.60 (d, J=15.6 Hz, 1H), 6.67(d, 3J=8.6 Hz,
Nꢀ-3-[4-(1-E-n-Pentenyl)phenyl)]-E/Z-acryloyl-D-histidyl-
L-phenylalanyl-L-tyrosine hydrotrifluoroacetate (35/16).
According to GP P2, the polymer-bound tripeptide 34a
(300 mg, 0.40 mmol/g, 120 mmol) was N-terminally
deblocked by piperidine/DMF treatment for 2ꢂ5 min,
followed by coupling with 30h (60 mg, 270 mmol),
3
3
2H), 6.95 (s, 1H), 7.04 (d, J=8.4 Hz, 2H), 7 .16–7 .24 (m,
4H), 7.29–7.33 (m, 1H), 7.37 (m, 2H), 7.50–7.54 (m, 2H),
7.73 (d, 3J=15.8 Hz, 1H), 8.40 (s, 1H); LC–MS (LC–ESI
1): 664.3 [M+H]+, 686.3 [M+Na]+, Rt=16.06 min; MS