Synthesis and HPLC-analysis of N-(2-phenyl-cyclopropyl)-substituted chain elongated nucleoside analogues

Article abstract of DOI:10.1515/znb-1999-1120

A novel class of nucleoside analogues containing adenine, uracil, thymine and 5-fluorouracil as the heterocyclic moieties have been prepared from 2-phenylcyclopropane carboxylic acid. These compounds showed weak antitumor activity. The resolution of the r

Full text of DOI:10.1515/znb-1999-1120

Synthesis and HPLC-Analysis of N-(2-Phenyl-cyclopropyl)-Substituted
Chain Elongated Nucleoside Analogues
Rene Csuk, Anja Kern, and Kristina Mohr
Institut für Organische Chemie. Martin-Luther-Universität Halle-Wittenberg,
Kurt-Mothes-Str. 2, D-06120 Halle (Saale), Germany
Reprint requests to Prof. Dr. R. Csuk. E-mail: csuk@chemie.uni-halle.de
Z. Naturforsch. 54b, 1463-1468 (1999); received July 9, 1999
Nucleoside Analogues, HPLC
A novel class of nucleoside analogues containing adenine, uracil, thymine and 5-fluoro-
uracil as the heterocyclic moieties have been prepared from 2-phenylcyclopropane carboxylic
acid. These compounds showed weak antitumor activity. The resolution of the racemates on
an analytical and semipreparative scale was performed by HPLC using chiral stationary
phases.
Introduction
(±)-6 [8]; this sequence provides a shortened ac-
cess to the central intermediate (±)-6.
In the course of ongoing QSAR studies of anti-
tumor active cyclopropanoid nucleoside analogues
we became interested in the synthesis and biologi-
cal evaluation of N-(2-phenyl-cyclopropyl)-substi-
tuted nucleoside analogues [1]. Since these com-
pounds showed only weak antitumor activity and
preliminary tests revealed no anti-H IV -activity
associated with these compounds, it was assumed
that the reduced bioactivity might be due to a de-
creased conformational flexibility as a conse-
quence of the presence of a rather rigid cyclopro-
pyl ring [2]. Rigidity seems to be unfavorable
either for the interaction of the corresponding tri-
phosphate with viral DNA-polymerases or in a
primary step for the interaction with phosphory-
lating enzymes [3], To avoid lengthy routes a strat-
egy had to be developed that should allow in a
forthcoming project the chosen strategy to be eas-
ily transferred into a combinatorial approach using
parallel synthesis on beads.
Mitsunobu reaction [9, 10] of (±)-6 in the pres-
ence of adenine, triphenylphosphane (TPP) and
diethyl azodicarboxylate (DEAD) gave the ade-
nine analogue (±)-7; under similar conditions
from (±)-6 in the presence of N3-benzoylthymine
[10] the protected thymine analogue (±)-8 was ob-
tained. Deprotection of (±)-8 with aqueous am-
monium hydroxide gave the target compound (±)-
9. In an analogous manner from (±)-6 and N3-
benzoyluracil [10] (±)-10 was obtained which gave
upon deprotection with ammonium hydroxide the
cyclopropanoid uracil derivative (± )-ll.
From (±)-6 and N3-benzoyl-5-fluorouracil [10]
compound (±)-12 was obtained in 61% yield. (±)-
12 is characterized in its I9F NMR spectrum by the
presence of a singulet at d = -166.79 ppm. Depro-
tection of (±)-12 finally gave the 5-fluorouracil an-
alogue (±)-13.
Preliminary screening of racemic 7, 9, 11 and 13
revealed weak antitumor and cytotoxic activity for
several of these compounds. Since it is well estab-
lished that the biological activity of many nucleo-
side analogues resides only in one of the enanti-
Results and Discussion
Lithium aluminumhydride reduction of trans-2-
phenyl-cyclopropane carboxylic acid (1) gave the omers [11], a chromatographic resolution of the
corresponding alcohol (±)-2 [4, 5] which was chlo- racemates was undertaken. According to our pre-
rinated with thionyl chloride in the presence of liminary experiences in this field, HPLC seemed
tributylamine to afford 86% of the chloride (±)-3 to be the method of choice. The results of these
[6], Nucleophilic substitution [6] using dry tetrabu-
tylammonium cyanide [7] gave the nitrile (±)-4
which was hydrolyzed to the acid (±)-5 whose lith-
ium alanate reduction finally afforded the alcohol
separations on analytical columns using chiral
phases are enlisted in Table I. Best results for the
thymine, uracil and 5-fluorouracil analogue were
achieved using a Daicel Chiralpak AD® column
0932-0776/99/1100-1463 $06.00 © 1999 Verlag der Zeitschrift für Naturforschung, Tübingen ■www.znaturforsch.com
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1464
R. Csuk et al. • HPLC of Nucleoside Analogues
3 R = Cl
4 R = CN
5 R = COOH
Experimental
General
Table I. HPLC conditions for the separation of the en-
antiomers (UV and chiral detection [ChiralyserTM],
column: 0.46 x 25 cm).
-
Melting points are uncorrected
(Leica hot stage microscope), optical rotations
were obtained using a Perkin-Elmer 341 polarime-
ter (1 cm micro cell), NMR spectra (internal
Me4Si) were recorded using the Varian spectrome-
ters XL300, Gemini 200, Gemini 2000 or Unity
500 (d given in ppm, J in Hz, internal Me4Si for
JH and 13C NMR spectra, internal CC13F was used
for 19F NMR spectra, the designation C’ corres-
ponds to the atoms of the heterocycle), CD
spectra were recorded on a JASCO J710 spec-
trometer, IR spectra (film or KBr pellet) on a Per-
kin-Elmer FT-IR spectrometer Spectrum 1000, MS
spectra were taken on a Intectra GmbH AMD 402
(electron impact, 70 eV) or on a Finnigan MAT
LCQ 7000 (electrospray, voltage 4.5 kV, under ni-
trogen) instrument; for elemental analysis a Foss-
Heraeus Vario EL instrument was used; TLC was
performed on silica gel (Merck 5554, detection by
UV absorption or by treatment with a solution of
10% sulfuric acid, ammonium molybdate and ceri-
um(IV) sulfate followed by gentle heating). For the
HPLC either a Merck LaChrom L7100/L7250/
L7450/D7000 system or a Merck-Hitachi L6200A/
L4000/D-2500 system was used.
Flow
[ml/min]
Column
Temper- /R [min]
ature
[°C]
Comp.
Eluent
7
7
AD
OD
methanol
0.5
1.0
10
20
(+): 26.05
(-): 26.83
(-): 188.61
(+): 345.07
hexane/2-
propanol
(90/10)
acetonitrile/
water
7
OD-R
0.5
20
(-): 10.47
(+): 11.39
(90/10)
methanol
9
11
13
AD
AD
AD
0.5
0.5
0.5
20
20
20
(+): 18.67
(-): 23.89
(+): 26.24
(-): 40.19
(+): 23.36
(-): 36.00
methanol
methanol
with methanol as the eluent. For the adenine ana-
logue (±)-7, however, no baseline separation
could be achieved using these conditions; better
results were obtained using a Daicel Chiralcel
OD® column that gave a clean separation of the
corresponding enantiomers. These conditions
could be used for a semipreparative resolution of
(± )-ll leading to the pure enantiomers (+)-ll and
(-)-ll, respectively. The resolution of the other ra-
cemates on a (semi)-preparative scale and their bi-
ological testing is now investigated in our
laboratories.
(±)-[(l RS, 2 SR)-2-Phenylcyclopropyl]methanol
f(±)-2J
Reduction of /ra«s-2-phenylcyclopropanecar-
boxylic acid (5.12 g, 31.57 mmol) with lithium alu-
minum hydride (2.55 g. 63.93 mmol. 95%) in anhy-
drous tetrahydrofuran (150 ml) [4, 5] gave after
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1465
R. Csuk et al. ■HPLC of Nucleoside Analogues
bulb-to-bulb distillation (120 °C, 0.1 mbar) the de- (20 ml), extracted with diethyl ether (5 x 50 ml),
sired product 2 (4.40 g, 94%) as a yellowish oil; the combined organic phases were dried (M gS04)
Rf (ethyl acetate/hexane 2:7) 0.3; UV (methanol): and the solvent was removed. Work-up was per-
Amax= 226 nm (log
e
= 3.90); IR (film):
v
3335brm, formed by fractional bulb-to-bulb distillation
1605m, 1498m, 1091m, 10335, 10215; lH NMR (500 (160 °C, 0.4 mbar) to afford 4 (1.27 g, 90%) [6].
MHz, CDCI3): <3 7.30-7.08 (m, 5 H, H-phenyl), R f (ethyl acetate/hexane 2:7) 0.4; UV (methanol):
3.62 (dd, J = 14.5, 11.2, 2 H, CH.-O), 1.83 (ddd,
J = 8.9, 4.7, 4.7, 1 H, 2-H), 1.47-1.44 (m , 1 H, 1-
Amax = 272 nm (log e = 3.95); IR (film): v 3062m,
3028m, 2249m, 16055, 15005, 1463m, 1446m,
H), 0.95 (ddd, J = 18.7, 8.8, 5.0, 2 H, 3-HAB); 13C 1420m, 1087m, 1031m, 1006m; JH NMR (400
NMR (125 MHz, CDC13): (3 142.41 (5, Cq phenyl), MHz, CDCI ): (5 7.29-7.08 (m, 5 H, H-C phenyl),
3
128.27, 125.79, 125.57 (d, phenyl), 66.37 (t, CH?- 2.53 (dd, J = 9.09, 6.16, 2 H, CH2-CN), 1.91 (ddd,
O), 25.16 (d, C-2), 21.24 (d, C-l), 13.71 (t, C-3); J = 9.13, 4.74, 4.74, 1 H, 2-H), 1.36-1.30 (m, 1 H,
MS (EI, 70 eV): m/z 148 (38), 130 (21), 129 (12), 1-H), 1.09 (ddd, J = 8.45, 5.52, 5.52, 1 H, 3-HA),
118 (22), 117 (100); HRMS calcd. for C10H 12O:
148.08881; found: 148.08880.
1.01 (ddd, J = 8.98, 5.57, 5.46, 1 H, 3-HB); 13C
NMR (100 MHz, CDCI ): (3 141.04 (5, C^ phenyl),
3
128.39, 162.05, 125.91 (d, phenyl), 118.17 (5, CN),
22.55 (d, C-2), 21.08 (d, C-l), 17.26 (t, CH2-CN),
14.55 (t, C-3); MS (EI, 70 eV): m/z 157 (38), 156
(7), 130 (23), 129 (13), 118 (10), 117 (100); HRMS
calcd. for Cn H n N: 157.08914; found: 157.08915.
Analysis for C]0H 12O (148.20)
Calcd
C 81.05 H 8.16%,
Found C 80.81 H8.13%.
(±)-[(l RS, 2 SR)-2-Phenylcyclopropyl]methyl
chloride [(± )-3]
Analysis for Cn H n N (157.22)
Calcd
C 84.04 H 7.05 N8.91%,
The reaction of 2 (3.81 g, 25.72 mmol) with tri-
butylamine (7.72 g, 41.65 mmol) and thionylchlor-
ide (6.38 g, 53.63 mmol) according to [6] at -10 °C
gave after bulb-to-bulb distillation 3 (3.67 g, 86%)
as a yellowish oil; RF (ethyl acetate/hexane 2:7)
0.69; UV (methanol): Amax= 224 nm (log e = 4.02);
IR (film): v 1606m, 1497m, 1265m, 1228w; JH
NMR (400 MHz, CDC13): <57.37-7.11 (m, 5 H, H-
phenyl), 3.60 (dd, J = 13.28, 7.23, 2 H, CH2-C1),
1.96 (ddd,J = 9.23, 4.74, 4.74, 1 H, 2-H), 1.61-1.54
(m, 1 H, 1-H), 1.15 (ddd, J = 8.40, 5.47, 5.47, 1 H,
3-HA), 1.05 (ddd, J = 9.03, 5.32, 5.32, 1 H, 3-HB);
13C NMR (100 MHz, CDC13): (3 141.58 (5, Cq phe-
nyl), 128.43, 126.16, 126.00 (d, phenyl), 49.15 (t,
CH2-C1), 24.75 (d, C-2), 23.94 (d, C-l), 15.70 (t, C-
3); MS (EI, 70 eV): m/z 168 (9), 166 (30), 131 (45),
130 (22), 129 (28), 128 (15), 125 (32), 117 (100);
HRMS calcd. for C10H„C1: 166.05492; found:
166.05492.
Found C 83.96 H 7.01 N8.71%.
(±)-[(l RS, 2 RS)-2-Phenylcyclopropyl] acetic acid
f(±) 57
A solution of 4 (1.17 g, 7.47 mmol), NaOH
(0.60 g, 14.94 mmol) and water (2 ml) was heated
under reflux for 4 h. After cooling to room tem-
perature the pH was adjusted by the addition of
HC1 (20%) to 3 and the mixture was extracted
with diethyl ether (5 x 50 ml). The combined or-
ganic layers were dried (M gS04), the solvent was
removed under reduced pressure and 5 (1.2 g,
91%) was obtained as a yellowish oil [13]; RF
(ethyl acetate/hexane 2:7) 0.07; UV (methanol):
imax= 264 nm (log £ = 3.93); IR (film): v 3062m,
30275, 2929m, 17095, 1605m, 1498m, 1415m,
1300m, 1227m, 1207m, 1031m; *H NMR (400
MHz, CDCI3): (3 11.00 (brs, 1 H, OH), 7.40-7.09
(m, 5 H, H-phenyl), 2.53-2.37 (m, 2 H, CH2), 1.79
(ddd, J = 9.03, 4.64, 4.64, 1 H, 2-H), 1.43-1.38 (m,
1 H, 1-H), 1.03 (ddd, J = 8.55, 5.23, 5.23, 1 H, 3-
H a ), 0.89 (ddd, J = 8.79, 5.37, 5.37, 1 H, 3-HB);
13C NMR (100 MHz, CDC13): <3 179.20 (5, CO),
142.43 (5, C^ phenyl), 128.38, 126.11, 125.77 (d,
phenyl), 38.65 (t, CH2), 22.76 (d, C-2), 17.87 (d, C-
1), 15.10 (t, C-3); MS (EI, 70 eV): m/z 176 (82),
157 (3), 131 (11), 130 (23), 117 (100); HRMS calcd.
for C nH 120 2: 176.08372; found: 176.08371.
Analysis for CioHu Cl (166.65)
Calcd
C 72.07 H 6.65%,
Found C 72.00 H 6.70%.
(± )-(] RS, 2 RS)-2-(Phenylcyclopropyl)acetonitrile
f(±HJ
To a solution of 3 (1.5 g, 9.00 mmol) in dry tetra-
hydrofuran (20 ml) a solution of tetrabutylam-
monium cyanide (2.42 g, 9 mmol) in dry tetrahy-
drofuran (20 ml) was added dropwise with stirring
under argon. After stirring the mixture for 4 h at Analysis for Cn H 120 2 (176.22)
60-70 °C, the solvent was removed under reduced
pressure. The residual oil was suspended in water
Calcd
C 74.98" H6.86%,
Found C 75.03 H7.03%.
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R. Csuk et al. ■HPLC of Nucleoside Analogues
1466
(5, C-4'), 143.56 (d, C-8'), 141.67 (5, Cq phenyl),
133.20, 129.51, 126.61 (d, phenyl), 120.82 (5, C-5'),
44.80 (t, CH2N), 35.37 (t, CH2-ethyl), 23.70 (d, C-
2), 21.50 (d, C-l), 16.30 (t, C-3); MS (EI, 70 eV):
m/z 279 (32), 278 (7), 264 (1), 250 (1), 224 (5), 188
(18), 176 (19), 175 (100); HRMS calcd. for
C16H 17N5: 279.1484; found: 279.1483.
(±)-2-[(l SR, 2 SR)-2-Phenylcyclopropyl]-1-
ethanol [(±) 6]
Following the procedure given for the prepara-
tion of compound 2, reaction of
5.67 mmol) in anhydrous tetrahydrofuran (10 ml)
with lithium aluminum hydride (0.44 g,
5
(1.0 g,
11.01 mmol, 95%) in dry THF (20 ml) afforded 6
(0.76 g, 83%) after bulb-to-bulb distillation
(130 °C, 0.1 mbar) [8]; RF (ethyl acetate/hexane
2:7) 0.28; UV (methanol): Amax= 227 nm (log e =
4.00); IR (film): v 33465, 3063m, 30265, 29995,
29275, 28745, 16045, 14975, 1462m, 1445m, 1382m,
1223m, 1183m, 10545; >H NMR (400 MHz,
CDC13): (3 7.28-7.05 (m, 5 H, H-phenyl), 3.78-
3.74 (m, 2 H, CH20 ), 1.73-1.62 (m, 3 H, 2-H) and
CH2-ethyl), 1.11-1.08 (m, 1 H, 1-H), 0.95 (ddd,
J = 8.45, 4.93, 4.93, 1 H, 3-HA), 0.83 {ddd, J =
8.20, 5.19, 5.18, 1 H, 3-HB); 13C NMR (100 MHz,
CDCI3): (3 144.55 (5, Cq phenyl), 129.42, 126.73,
126.50 (d, phenyl), 63.75 (t, CH20 ), 38.26 (t, CH2-
ethyl), 23.71 (d, C-2), 21.13 {d, C-l), 16.50 (t, C-
3); MS (EI, 70 eV): m/z 162 (28), 144 (29), 143
(26), 133 (12), 129 (100); HRMS calcd. for
Cn H 140: 162.10446; found: 162.10446.
Analysis for C16H 17N5 (279.34)
Calcd
C 68.80 H 6.13 N 25.07%,
Found C 68.55 H 6.02 N 25.19%.
(± )-3-Benzoyl-5-methyl-1-{2-[(1 RS, 2 RS)-2-
phenylcyclopropyl]ethyl}-1,2,3,4-tetrahydro-2,4-
pyrimidinedione [( +) 8/
Following the procedure given for the prepara-
tion of compound 7, reaction of
6
(1.0 g,
(3.31 g,
(2.85 g,
6.16 mmol),
12.61 mmol),
triphenylphosphane
N3-benzoylthymine
12.37 mmol) in dry 1,4-dioxane (40 ml) and
DEAD (2.15 g, 12.34 mmol) in 1,4-dioxane (20 ml)
gave 8 (1.53 g, 66%) after column chromatography
(1. silica gel, ethyl acetate/hexane 1:1; 2. silica gel
RP-18, methanol/water 8:3) as a white solid; mp:
174.8-176.4 °C; RF(ethyl acetate/hexane 1:1) 0.51;
UV (methanol): Amax = 258 nm (log e = 4.24); IR
(KBr): v 34405, 2921m, 1743m, 1695m, 16485; !H
NMR (400 MHz, CDC13): (3 7.92-7.90 (m, 2 H,
or/7zo-H2-phenyl (Bz)), 7.66-7.62 (m, 1 H,para-H-
phenyl (Bz)), 7.50-7.46 (m, 2 H, meta-H2-phenyl
(Bz)), 7.28-7.24 (m, 2 H, orr/zo-H2-phenyl), 7.17-
7.13 (m, 1 H, para-H-phenyl), 7.02-6.97 (m, 2 H,
meta-H2-phenyl), 6.97 (5,1 H, 6'-H), 3.87-3.51 (m,
2 H, CH2N), 1.95-1.90 (m, 1 H, ethyl-HA), 1.74
(5, 3 H, CH3), 1.70-1.65 (m, 2 H, ethyl-HB, 2-H),
1.03-0.90 (m, 2 H, 1-H, 3-HA), 0.86 (ddd, J = 8.45,
4.25, 4.25, 1 H, 3-HB); 13C NMR (100 MHz,
Analysis for Cn H 140 (162.23)
Calcd
C 81.44 H8.70%,
Found C 81.16 H 8.52%.
(±)-9-{2-[(l RS, 2 RS)-2-Phenylcyclopropyl]-
ethyl}-9H-6-purinamine [(±)-l]
To a mixture of 6 (1.0 g, 6.16 mmol), triphenyl-
phosphane (3.29 g, 12.54 mmol) and adenine
(1.68 g, 12.43 mmol) in dry 1,4 -dioxane (40 ml) a
solution of DEAD (2.15 g, 12.34 mmol) in dry 1,4-
dioxane (20 ml) was added dropwise at room tem-
perature over a period of 2.5 h. The solution was
stirred overnight, the solvent was removed in
vacuo and the remaining yellowish oil was purified
by column chromatography (silica gel, ethyl ace-
tate/methanol 10:1) to afford 6 (1.21 g, 71%) as
a yellowish solid; m.p. 157.5-158.3 °C; R F (ethyl
acetate/methanol 10:1) 0.38; UV (methanol):
Amax = 267 nm (log £ = 4.15); IR (KBr): v 32945,
3111m, 16735, 16025, 1572m, 1481m, 1415m,
CDCI ): d 169.26 (5, CO benzoyl), 163.34 (5, C-
3
2'), 149.84 (5, C-4'), 142.60 (d, C-6'), 140.86 (5, Cq
phenyl), 134.98 (5, Cq phenyl (Bz)), 131.87 (d,
C p a r a phenyl (Bz)), 130.46 (d, Cortho phenyl (Bz)),
129.18 (d, Cmeta phenyl (Bz)), 128.55 (d, Cortho
phenyl), 125.76 (d, Cmeta phenyl), 125.48 (d, Cpara
phenyl), 110.39 (5, C-5'), 49.19 (t, CH2N), 32.91 (t,
CH2-ethyl), 22.88 (d, C-2), 20.46 (d, C-l), 15.89 (q,
CH3), 11.93 (t, C-3); MS (EI, 70 eV): m/z 374 (29),
283 (3), 270 (33), 253 (1), 241 (5.), 214 (1), 165 (2),
143 (11), 129 (11), 115 (3), 105 (100); HRMS calcd.
for C23H22N20 3: 374.16303; found: 374.16303.
1310m, 1241m; ]H NMR (400 MHz, CDCI ): 6
3
8.29 (5, 1 H, 8'-H), 7.71 (5, 1 H, 2'-H), 7.21-6.89
(m, 5 H, H-phenyl), 6.04 (5, 2 H, NH2), 4.30 (ddd,
J = 13.62, 10.20, 6.88, 2 H, CH,N), 2.02-1.90 (m,
2 H, CH2-ethyl), 1.54 (ddd, J = 8.79, 4.49, 4.49, 1
H, 2-H), 1.28-1.21 (m, 1 H, 1-H), 0.87 (ddd, J =
8.79, 4.59, 4.59, 1 H, 3-HA), 0.70 (ddd, J = 9.09,
4.79, 4.79, 1 H, 3-HB); 13C NMR (100 MHz,
CDCI3): d 156.75 (5, C-6'), 154.06 (d, C-2'), 151.20
Analysis for C23H22N20 3 (374.44)
Calcd
C 73.78 H 5.92 N 7.48%,
Found C 73.51 H 6.01 N7.56%.
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R. Csuk et al. ■HPLC of Nucleoside Analogues
1467
C-4'), 144.49 (d, C-6 '), 142.33 ( 5, Cq phenyl),
135.01 ( 5, Cq phenyl (Bz)), 131.50 (d, Cpara phenyl
(Bz)), 130.33 (d, Cortho phenyl (Bz)), 129.12 (d,
Cmeta phenyl (Bz)), 128.36 (d, Cortho phenyl),
125.66 (d, Cmeta phenyl), 125.46 (d, Cpara phenyl),
101.72 ( 5, C-5'), 49.42 (t, CH2N), 33.06 (t, CH2-
ethyl), 22.90 (d, C-2), 20.28 (d, C-l), 15.75 (t, C-
3); MS (EI, 70 eV): m!z 360 (45), 346 (1), 269 (3),
256 (41), 248 (39), 227 (3), 199 (1), 143 (13), 129
(14), 115 (3), 105 (100); HRMS calcd. for
C22H20N2O3: 360.14738; found: 360.14739.
(±)-5-Methyl-l-{2-[(lRS, 2 RS)-2-phenylcyclo-
propyl]ethyl}-1,2,3,4-tetrahydro-2,4-
pyrimidinedione [(+)-9J
A suspension of compound 8 (1.16, 3.10 mmol)
in methanol (100 ml) was treated with ammonium
hydroxide (20 ml) at room temperature for 24 h.
The volatiles were evaporated and the remaining
oil was subjected to column chromatography (sil-
ica gel, ethyl acetate/hexane 2:1) to afford 9
(0.49 g, 59%) as a white solid; m.p. 161.8-
162.3 °C; Rf (ethyl acetate/hexane 2:1) 0.38; UV
Analysis for C22H20N2O 3 (360.41)
Calcd C 73.32 H 5.59 N 7.77%,
Found C 73.11 H 5.47 N 8.02%.
(methanol): Amax = 279 nm (log
e
= 3.73); IR
(KBr): v 1686s, 16635; :H NMR (200 MHz,
CDCI3): (3 8.87 (s, 1 H, NH), 7.24-6.94 (m , 5 H,
H-phenyl), 6.82 (s, 1 H, 6 '-H), 3.78 (ddd, J = 10.40,
6.88, 3.86, 2 H, CH2N), 1.96-1.78 (m, 1 H, ethyl-
(±)-l-{2-[(l RS, 2 RS)-2-Phenylcyclopropyl]-
Ha ), 1.69-1.53 (m, 2 H, ethyl-HB, 2-H), 1.68 (s, 3 ethyl}-l,2,3,4-tetrahydro-2,4-pyrimidinedione
H, CH3)), 0.99-0.88 (m, 2 H, 1-H, 3-HA), 0.80
(ddd, J = 8.30, 4.00, 4.00, 1 H, 3-HB); 13C NMR
(50 MHz, CDCL): <5 164.19 (s, C-2'), 150.71 (s, C-
4'), 142.52 (d, C-6 '), 141.04 (s, Cq phenyl), 128.40,
125.61, 125.31 (d, phenyl), 110.26 (s, C-5'), 48.88
(t, CH2N), 33.08 (t, CH2-ethyl), 22.98 (d, C-2),
20.62(d, C-l), 15.98 (q, CH3), 11.99 (t, C-3); MS
(EI, 70 eV): m/z 270 (100), 179 (8), 166 (10), 144
(36), 140 (18), 129 (34), 127 (20), 115 (6), 96 (9),
91 (9); HRMS calcd. for C16H 18N20 2: 270.13682;
found: 270.13683.
[(±)-ll]
According to the preparation of 9 compound 10
(1.60 g, 4.44 mmol) was dissolved in methanol
(100 ml) and treated with ammonium hydroxide
(20 ml) for 3 h. Purification by column chromatog-
raphy (silica gel, ethyl acetate/hexane 1:1 —* 2 :1 )
afforded 11 (1.09 g, 96%) as a colorless oil; semi-
preparative HPLC (Chiralpak AD 1 x 25 cm, 2 ml/
min, methanol, 15 kg.cm-2) gave (+)-ll ( +73.8 (c
1, methanol) and ( - ) - l l ( -72.6 (c 1, methanol);
Rf (ethyl acetate/hexane 2:1) 0.24; UV (metha-
nol): /lmax- 272 nm (log e = 3.97); IR (film): v
1678m; !H NMR (400 MHz, CDC13): (3 9.30 (s, 1
H, NH), 7.26-6.98 (m, 6 H, H5-phenyl, H-C(6 ')),
5.59 (d, J = 7.81, 1 H, 5'-H), 3.85 (ddd, J = 26.61,
13.53, 6.59, 2 H, CH2N), 1.86-1.73 (m, 2 H, CH2-
ethyl), 1.65 (ddd, J = 8.79, 4.49, 4.49, 1 H, 2-H),
I.32-1.24 (m, 1 H, 1-H), 0.97 (ddd, J = 17.48, 9.08,
4.20, 1 H, 3-HA), 0.81 (ddd, J = 9.03, 4.64, 4.64, 1
H, 3-Hb); 13C NMR (100 MHz, CDC13): <3 164.90
( 5, C-2'), 151.92 ( 5, C-4'), 145.85 (d, C-6 '), 143.55
( 5, Cq phenyl), 129.48, 126.77, 126.57 (d, phenyl),
Analysis for C16H 18N20 2 (270.33)
Calcd
C 71.09" H 6.71 N 10.36%,
Found C 70.88 H 6.52 N 10.09%.
(±)-3-Benzoyl-l-{2-[(l RS, 2 RS)-2-phenylcyclo-
propyljethyl}-1,2,3,4-tetrahydro-2,4-
pyrimidinedione [(±J-10/
Similarly as described for compound 7, the reac-
tion of 6 (1.0 g, 6.16 mmol) with triphenylphos-
phane (3.31 g, 12.62 mmol) and N3-benzoyluracil
(2.62 g, 12.10 mmol) in dry 1,4-dioxane (40 ml) 102.97 ( 5, C-5'), 50.04 (t, CH2N), 34.17 (t, CH2-
ethyl), 23.92 (d, C-2), 21.28 (d, C-l), 16.63 (t, C-
(20 ml) afforded 10 (1.93 g, 87%) as a colorless oil 3); MS (EI, 70 eV): m/z 256 (80), 165 (8), 149 (8),
144 (7 5 ), 143 (24), 129 (100); HRMS calcd. for
and DEAD (2.1 g, 12.06 mmol) in dry 1,4-dioxane
after column chromatography (silica gel, ethyl ace-
tate/hexane 1:1); Rf (ethyl acetate/hexane 1:1) C15H 16N20 2: 256.12117; found: 256.12116.
0.30; UV (methanol): /lmax = 258 nm (log e = 4.27);
IR (film): v 2999m, 17475, 17045, I66O5, 15995,
1497m, 14355, 13855, 13455, 12535, 1178m, 1159m,
1093m, 1046m, 1001m; *H NMR (200 MHz,
Analysis for C15H 16N20 2 (256.30)
Calcd
C 70.29” H 6.29 N 10.93%,
Found C 70.01 H 5.99 N 10.66%.
CDCI ): d 7.93-6.98 (m, 11 H, H 10-phenyl, 6 '-H),
3
(±)-3-Benzoyl-5-fluoro-l-f2-[(l RS, 2 RS)-2-
phenylcyclopropyljethyl}-1,2,3,4-tetrahydro-2,4-
pyrimidinedione [(± >-12 /
5.66 (d, J = 7.82, 1 H, 5'-H), 3.88-3.82 (m, 2 H,
CH2N), 1.84-1.63 (m, 3 H, CH2-ethyl, 2-H), 1.01-
0.95 (m, 2 H, 1-H), 3-HA), 0.82 (ddd, J = 5.52, 2.88,
2.88, 1 H, 3-Hb-); 13C NMR (50 MHz, CDC13): d
The reaction was performed under the condi-
168.80 ( 5, CO benzoyl), 162.42 ( 5, C-2'), 149.66 ( 5, tions as described for 7, using 6 (1.0 g, 6.16 mmol)
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1468
R. Csuk et al. ■HPLC of Nucleoside Analogues
in dry 1,4-dioxane (20 ml), triphenylphosphane
(3.32 g, 12.65 mmol), N3-benzoyl-5-fluorouracil
(±)-5-Fluoro-l-{2-[(lRS, 2 RS)-2-phenylcyclo-
propyljethyl}-! ,2,3,4-tetrahydro-2,4-
(2.83 g,
12.09 mmol)
and
DEAD
(2.10 g, pyrimidinedione [(±j-13/
12.07 mmol) in dry 1,4-dioxane (20 ml). After
evaporation of the solvents and purification by
column chromatography (1 . silica gel, ethyl ace-
tate/hexane 1:1; 2. silica gel RP-18, methanol/
water 8:3) 12 (1.42 g, 61%) was obtained as a yel-
lowish oil. Rf (ethyl acetate/hexane 1:1) 0.68; UV
(methanol): i max = 259 nm (log e = 4.23); IR (film):
v 3070m, 3027m, 3000m, 2916m, 17485, 17145,
I 6685, 1616m, 16035, 1585m, 1557m, 1496m, 14505,
13555, 1316m, 12415, 1180m, 1161m, 1121m,
1094m, 1066m, 1029m, 1018m, 1001m; JH NMR
(400 MHz, CDC13): 6 7.95-7.89 (m, 2 H, ortho-
H2-phenyl (Bz)), 7.69-7.65 (m, 1 H, para-H-phe-
nyl (Bz)), 7.52-7.46 (m, 2 H, meta-H2-phenyl
(Bz)), 7.31-7.01 (m, 6 H, Hs-phenyl, H-C(6 ')),
3.85 (ddd, J = 6.93, 6.93, 2.44, 2 H, CH2N), 1.84-
1.79 (m, 2 H, CH2-ethyl), 1.69 (ddd, J = 8.94, 4.54,
4.54, 1 H, 2-H), 1.02-0.96 (m, 2 H, 1-H, 3-HA),
0.83 (ddd, J = 5.76, 5.76, 2.84, 1 H, 3-HB); 13C
NMR (100 MHz, CDCI3): (5 167.50 ( 5, CO ben-
zoyl), 156.41 (d, JCF = 27, C-4', 148.30 ( 5, C-2'),
142.29 ( 5, Cq phenyi), 139.73 (d, / c , f = 240, C-5'),
135.42 ( 5, Cq phenyl (Bz)), 130.46 (d, Cpara phenyl
(Bz)), 129.28 (d, Corlho phenyl (Bz)), 128.94 (d,
Cmeta phenyl (Bz)), 128.43 (d, Cortho phenyl),
128.27 (dd, / C,F = 32, C-6 '), 125.73 (d, Cmeta phe-
nyl), 125.48 (d, Cpara phenyl), 49.42 (t, CH2N),
32.72 (t, CH2-ethyl), 22.76 (d, C-2), 19.97 (d, C-l),
15.45 (t, C-3); 19F NMR (188 MHz, CDC13):<3 -
166.79 ( 5); MS (EI, 70 eV): m/z 378 (39), 287 (2),
274 (26), 257 (6), 248 (3), 190 (4), 184 (4), 170 (2),
162 (6), 144 (14), 129 (8), 115 (4), 105 (100);
HRMS calcd. for C22H 19FN20 3: 378.13795;
found: 378.13794.
According to the preparation of compound 9
compound 12(0.95 g, 2.50 mmol) was dissolved in
methanol (100 ml) and treated with ammonium
hydroxide (20 ml) at room temperature for 12 h
and 13(0.5 g, 72%) was obtained after purification
by column chromatography (silica gel, ethyl ace-
tate/hexane 1:1) as a colorless oil. RF (ethyl ace-
tate/hexane 1:1) 0.46; UV (methanol): i max =
280 nm (log e = 3.94); IR (film): v 17055, 16745,
1370m, 1235m; 'H NMR (400 MHz, CDC13): <3
9.90 ( 5, 1 H, NH), 7.26-6.98 (m, 6 H, H5-phenyl,
6 '-H), 3.82 (ddd, J = 13.43, 6.79, 6.79, 2 H, CH2N),
1.86-1.72 (m, 2 H, CH2-ethyl), 1.64 (ddd, J = 8.89,
4.59, 4.59 1 H, 2-H), 1.03-0.94 (m, 2 H, 1-H, 3-
HA, 0.81 (ddd, J = 8.94, 4.54, 4.54, 1 H, 3-HB); 13C
NMR (100 MHz, CDCI3): Ö 158.49 (d, JCF = 25,
C-4'), 150.72 ( 5, C-2'), 143.39 ( 5, Cq phenyl)', 141.44
(d, JCF = 238, C-5'), 130.04 (dd, JcF = 32, C-6 '),
129.53,126.82, 126.51 (d, phenyl), 50.26 (t, CH2N),
34.03 (t, CH2-ethyl), 23.99 (d, C-2), 21.19 (d, C-l),
16.55 (t, C-3); 19F NMR (188 MHz, CDC13):<3 -
167.66 ( 5); MS (EI, 70 eV): m/z 274 (76), 254 (2),
183 (5), 170 (10), 144 (75), 143 (27), 131 (196), 129
(100); HRMS calcd. for C15H 15FN20 2: 274.11175;
found: 274.11175.
Analysis for C15H 15FN20 2 (274.29)
Calcd
C 65.68 H5.51 N 10.21%,
Found C 65.55 H 5.31 N 10.00%.
Acknowledgements
Financial support by the European Community
(SCl*-CT92-0780) and the Fonds der Chem-
ischen Industrie is gratefully acknowledged; we
are indebted to Dr. P. Rosyk for his help in the
preparation of the manuscript and to Mrs. R.
Ziehn for her help with the HPLC separations.
Analysis for C22H 19FN20 3 (378.40)
Calcd
C 69.83 H 5.06 N7.40%,
Found C 69.67 H 5.21 N7.21%.
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