The synthesis and anticancer activities of peptide 5-fluorouracil derivatives

Article abstract of DOI:10.3184/030823409X431364

A new series of peptide 5-fluorouracil derivatives was designed and synthesised in order to test in vitro anticancer activities. The results indicated that peptide 5-fluorouracil derivatives possessed anticancer activities against human HL-60 and Bel-7402

Full text of DOI:10.3184/030823409X431364

JOURNAL OF CHEMICAL RESEARCH 2009
APRILꢃꢀꢄꢇꢅ±264
RESEARCH PAPER 261
The synthesis and anticancer activities of peptide 5-fluorouracil derivatives
Xiaowei Yan, Maolin Hu*, Qian Miao, Shun Wang and Kejian Zhao
College of Chemistry and Materials Engineering, Wenzhou University, Wenzhou, 325027, P. R. China
A new series of peptide 5-fluorouracil derivatives was designed and synthesised in order to test in vitro anticancer
activities. The results indicated that peptide 5-fluorouracil derivatives possessed anticancer activities against human
HL-60 and Bel-7402 cell lines. The structures of the compounds were determined by means of 1H NMR, 13C NMR, IR,
mass spectra and elemental analyses.
Keywords: SHSWLGHꢀꢁꢂÀXRURXUDFLOꢃꢀDQWLFDQFHUꢀDFWLYLWLHVꢃꢀFU\VWDOꢀVWUXFWXUH
Despite major breakthroughs in many areas of modern
medicine over the past 100 years, the successful treatment
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selective, and less toxic anticancer agents remains one of the
most pressing health problems.1,2
5-Fluorouracil (5-FU) has been used in treating cancer and
squamous cell carcinoma of the head and neck.3,4 However,
the clinical dosage of (5-FU) is very close to its toxic one
when given intravenously, resulting in strong toxicities to the
gastric and intestinal mucosa and the bone marrow.5 Novel
prodrugs of 5-FU possessing a broader spectrum of antitumour
activity and less toxicity have been sought diligently by
many researchers.6-8 The common feature of these prodrugs
is that they are all N1ꢂPRGL¿HGꢀGHULYDWLYHVꢀWKURXJKꢀGLIIHUHQWꢀ
biodegradable linkages.
antibiotics, driving considerable pharmacological interests
in the design and application of new drugs involving
peptides.9,10
Based on the mutual prodrug concept, in continuation of
our interest in the synthesis of 5-FU derivatives,11-13 we have
synthesised some peptide derivatives of 5-FU with the aim of
¿QGLQJꢀDSSURSULDWHꢀELRGHJUDGDEOHꢀOLQNDJHVꢀDQGꢀRIꢀLPSURYLQJꢀ
WXPRXUꢀVHOHFWLYLW\ꢃꢀHI¿FLHQF\ꢀDQGꢀVDIHW\ꢆ
As shown in Scheme 1, compound 1 was prepared from
5-FU and 2-bromoacetic acid according to the literature.14
Compound
2 was obtained by the reaction of amino
acid hydrochlorides with compound 1 in the presence of
dicyclohexyl carbodiimide (DCC) and 1-hydroxybenzotriazole
(HOBt) at room temperature. Hydrolysis of 2 in aqueous
sodium hydroxide and subsequent treatment with concentrated
HCl afforded compound 3. Compound 4 was prepared from
3ꢀ E\ꢀ Dꢀ VLPLODUꢀ PHWKRGꢀ WRꢀ WKHꢀ WUDQVHVWHUL¿FDWLRQꢀ RIꢀ DPLQRꢀ
acid hydrochlorides used for the preparation of compound 2
(Scheme 1).
Peptides play important roles in the metabolism of human
beings and other organisms. They can function as hormones,
enzyme inhibitors/substrates, growth promoters, inhibitors,
neurotransmitters and immunomodulating agents as well as
R2
COOR1
-
*
F
F
NH3+Cl
KOH/H2O
+
BrCH2COOH
O
NH
O
O
N
O
CH2COOH
NEt3, DCC, HOBT
HN
HN
1
5-FU
F
F
COOR1
COOH
O
O
(1) 2 mol/L NaOH
(2) HCl, pH = 5-6
*
*
O
N
O
CH2CNH
O
N
O
CH2CNH
HN
R2
HN
R2
2
3
S-2a: R1 = Me; R2 = Ph
R-2b: R1 = Et; R2 = Ph
S-2c: R1 = Me; R2 = Me
S-2e: R1 = Me; R2 = CH2CO2Me
R-2f: R1 = Et; R2 = CH2CO2Et
S-3a: R2 = 4-CH2C6H4
R-3b: R2 = 4-CH2 C6H4OH
3c: R2 = H
OH
S-2g: R1 = Me; R2 = 4-CH2
C
6H4OH
R-2d: R1 = Me; R2 = Me R-2h: R1 = Et; R2 = 4-CH2 C6H4OH
2i: R2 = H; R1 = Me
R3
COOR4
-
*
O
CNH
F
R3
O
*
NH3+Cl
O
N
O
CH2CNH
COOR4
NEt3, DCC, HOBT
HN
R2
4
S-4a: R2 = H; R3 = CH2CO2Me; R4 = Me
4 = Me
S-4b: R2 = H; R3 = 4-CH2 C6H4OH; R
S-4c: R2 = H; R3 = CHMe2; R4 = Me
Scheme 1
* Correspondent. E-mail: hu403cn@yahoo.com.cn

262 JOURNAL OF CHEMICAL RESEARCH 2009
Compounds 2aíh and 4aíc were evaluated for percentage
of inhibition of HL-60 and BEL-7402 cell growth. The
antitumour activities in vitro for these compounds were
evaluated by the SRB method for BEL-7402 cells and the
MTT method for HL-60 cells. The activity data are presented
in Tables 1 and 2. The results indicated that among the tested
compounds, a majority of the newly prepared compounds
showed a moderate inhibiting effect on the growth of the
HL-60 and BEL-7402 cells at different concentrations of 10-4
mol L-1, 10-5 mol L-1, 10-6 mol L-1, 10-7 mol L-1, and 10-8
mol L-1. The inhibiting effect of all the compounds against
the HL-60 cells was different from that against the BEL-7402
cell lines, which suggested that these compounds possibly had
different inhibition mechanisms against various tumor cells.15
The anticancer activities of the SꢂFRQ¿JXUDWLRQꢀ FRPSRXQGVꢀ
were different from the Ríanalogues, which indicated that the
DQWLFDQFHUꢀ DFWLYLW\ꢀ ZDVꢀ UHODWHGꢀ WRꢀ WKHꢀ FRQ¿JXUDWLRQꢆ16 Also,
inspection of the chemical structure and anticancer activity of
2e, 2g, 4a and 4b showed that the activities of longer peptide
chain compounds (4a and 4bꢈꢀZHUHꢀVLJQL¿FDQWO\ꢀORZHUꢀWKDQꢀ
those of the shorter ones (2e and 2g).
All the products have been characterised by elemental
analysis, IR, 1H NMR, 13C NMR and MS. Meanwhile, in order
WRꢀFRQ¿UPꢀIXUWKHUꢀWKHꢀVWUXFWXUHꢀRIꢀFRPSRXQGVꢀ3, the crystal
structure of 3b as a representative example was determined
by X-ray crystallography and an ORTEP view of the molecule
together with the atomic numbering is shown in Fig. 1.
In conclusion, 15 5-FU derivatives (2a±i, 3a±c, and
4a±c) were synthesised and the crystal structure of 3b was
determined by X-ray diffraction analysis. The results of assay
for the anticarcinogenic activity against the HL-60 and BEL-
7402 cell lines indicated that some of the target compounds
SRVVHVVHGꢀ VLJQL¿FDQWꢀ DQWLFDQFHUꢀ DFWLYLWLHVꢆꢀ )XUWKHUꢀ VWXG\ꢀ LQꢀ
this area is underway in our laboratory.
Fig. 1
WRS-1B and are uncorrected. TLC was performed using precoated
silica gel 60 GF254 and column chromatography was performed using
VLOLFDꢀJHOꢀꢉꢊꢋꢋ±ꢌꢋꢋꢀPHVKꢈꢆꢀ,5ꢀVSHFWUDꢀZHUHꢀWDNHQꢀRQꢀDQꢀ(48,12;ꢂ
55 instrument. 1H NMR and 13C NMR spectra were recorded on a
AVANCE-300 instrument using tetramethylsilane (TMS) as an
internal standard and DMSO-d6 as the solvent at room temperature.
Chemical shifts are given in G relative to TMS, coupling constants
(J) are expressed in Hz. Mass spectra were obtained on a DECAX-
3000LCQ DecaXPPlus. The crystallographic data were collected on
a Bruker Smart-Apex CCD diffractometer. Antimicrobial activities of
the title compound were evaluated at The National Centre for Drug
Screening in Shanghai, China.
General method for the synthesis of compound 2
7KHꢀ VWDUWLQJꢀ PDWHULDOꢃꢀ ꢁꢂÀXRURXUDFLOꢂꢅꢂDFHWLFꢀ DFLGꢀ ꢉꢁꢂÀXRURꢂꢄꢃꢌꢂ
dioxo-1,2,3,4-tetrahydropyrimidine-1-acetic acid) was prepared from
5-FU and bromoacetic acid, in line with the literature method.14 2a±i
ZHUHꢀV\QWKHVLVHGꢀIURPꢀꢁꢂÀXRURXUDFLOꢂꢅꢂDFHWLFꢀDFLGꢃꢀ'&&ꢃꢀ+2%WꢀDVꢀ
follows: A solution of DCC (6.18 g, 30 mmol) in DMF (30 mL)
ZDVꢀDGGHGꢀGURSZLVHꢀWRꢀDꢀ'0)ꢀVROXWLRQꢀꢉꢁꢋꢀP/ꢈꢀRIꢀꢁꢂÀXRURXUDFLOꢂ
1-acetic acid (3.76 g, 20 mmol) and HOBt (2.97 g, 22 mmol) at
0°C over a period of 40 min. The resulting solution was stirred at
room temperature for 5 h. The appropriate ester of the amino acid
hydrochloride (20 mmol) and triethylamine (2.8 mL, 20 mmol) was
added to the above mixture. After stirring 4 hours, a white solid was
REWDLQHGꢆꢀ$IWHUꢀ¿OWUDWLRQꢃꢀWKHꢀ¿OWUDWHꢀZDVꢀFRQFHQWUDWHGꢀXQGHUꢀUHGXFHGꢀ
Experimental
All chemicals and solvents were purchased from Aldrich, and Fluka.
Melting points were recorded on a Digital Melting Point Apparatus
Table 1 Percentage of inhibition of HL-60 cell growth
Entry
Compound
Concentration/mol L-1
10-4
10-5
10-6
10-7
10-8
1
2
2a
2b
2c
2e
2f
2g
2h
4a
4b
4c
63.1
65.1
59.1
67.2
55.8
58.3
68.3
3.7
0
5.4
4.6
0
23.2
12.8
6.3
9.8
6.3
9.5
6.3
9.7
13.0
0
11.0
5.4
6.2
12.7
8.2
4.0
12.5
2.3
0.3
1.2
3.9
5.9
8.1
8.9
8.7
7.9
8.2
0
3
0
0
4
5
2.7
2.7
25.5
5.9
6.5
10.8
6
7
8
9
11.0
3.5
10
Table 2 Percentage of inhibition of BEL-7402 cell growth
Entry
Compound
Concentration/mol L-1
10-6
10-4
10-5
10-7
10-8
1
2
2a
2b
2c
2e
2f
2g
2h
4a
4b
4c
37.2
46.4
41.7
47.5
34.0
34.2
40.6
19.9
1.2
10.4
7.7
15.8
16.1
9.2
9.7
2.4
0
3.4
6.9
16.8
0
2.4
9.8
13.0
11.8
3.5
7.3
7.3
7.4
0
2.2
9.6
16.3
5.6
5.9
7.3
4.8
0
3
4
5
4.1
2.3
2.1
5.5
0
6
7
8
9
0
0
10
11.4
6.5
0
0
0

JOURNAL OF CHEMICAL RESEARCH 2009 263
pressure and the residue was separated by column chromatography to
afford compound 2.
3270 (NíH), 1738 (C=O), 1661 (C=C), 1552 (C=N), 1515, 1450
(íCH2í), 1386 (CíH), 1230 (CíF), 1164 (CíOíC), 778. 1H NMR
(300 MHz, DMSO-d6): G = 11.84 (s, 1H), 9.28 (s, 1H), 8.67 (d,
J = 7.8 Hz, 1H), 7.94 (d, J = 7.2 Hz, 1H), 6.99 (d, J = 8.4 Hz, 2H),
6.66 (d, J = 8.4 Hz, 2H), 4.30í4.40 (m, 3H), 3.58 (s, 3H), 2.76í2.92
(m, 2H). 13C NMR (75 MHz, DMSO-d6): G = 171.7, 166.7, 157.7,
156.1, 149.6, 139.2 (d, JC-F = 226.9 Hz), 131.0 (d, JC-F = 32.2 Hz),
130.1, 126.8, 115.1, 54.2, 51.9, 49.3, 36.1. ESI-MS (m/z): 364.6 (M+,
100). Elemental anal. Calcd for C16H16N3FO6: C, 52.60; H, 4.38; N,
11.51. Found: C, 52.73; H, 4.26; N, 11.38%.
ꢀ6ꢁꢂ0HWK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
yl)acetamido)-2-phenyl acetate (2a): Yield: 56%, m.p. 184í186°C.
[D]D20ꢀ ꢀ ±ꢅꢋꢁꢆꢌꢀ ꢉFꢀ ꢅꢆꢋꢃꢀ '0)ꢈꢆꢀ Qmax (KBr)/cmíꢅ 3262 (NíH), 3054
(=CíH), 1671 (C=O), 1549 (C=N), 1378, 1243 (CíOíC), 975, 700.
1H NMR (300 MHz, DMSO-d6): G = 11.85 (d, J = 3.9 Hz, 1H), 9.16 (d,
J = 7.2 Hz, 1H), 8.03 (d, J = 6.6 Hz, 1H), 7.36í7.39 (m, 5H), 5.46 (d,
J = 7.2 Hz, 1H), 4.45 (d, J = 16.8 Hz, 1H), 4.38 (d, J = 16.8 Hz, 1H),
3.64 (s, 3H). 13C NMR (75 MHz, DMSO-d6): G = 170.6, 166.6, 157.4
(d, JC-F = 25.7 Hz), 149.6,139.1 (d, JC-F = 226.7 Hz), 136.0, 131.0 (d,
ꢀ5ꢁꢂ(WK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
yl)acetamido)-3-(4-hydroxyphenyl) propanoate (2h): Yield: 66%,
J
C-F = 33.9 Hz,), 128.7, 128.3, 127.5, 56.2, 52.3, 49.2. ESI-MS (m/z):
334.1 ([M-1]+, 100). Elemental anal. Calcd for C15H14N3FO5: C, 53.73;
H, 4.18; N, 12.54. Found: C, 53.42; H, 4.14; N, 12.58%.
m.p. 159í160°C. [D]D ꢀ±ꢄꢍꢆꢎꢀꢉc 1.0, DMF).Qmax (KBr)/cmíꢅ 3336
20
(NíH), 3064 (=CíH), 2850 (íCH2í), 1690 (C=O), 1532 (C=N), 1430
(íCH2í), 1380 (CíH), 1220 (CíOíC), 972 (CíH). 1H NMR (300
MHz, DMSO-d6): G = 11.83 (s, 1H), 9.24 (s, 1H), 8.64 (d, J = 7.5
Hz, 1H), 7.95 (d, J = 6.9 Hz, 1H), 6.95 (d, J = 8.1 Hz, 2H), 6.66 (d,
J = 8.4 Hz, 2H), 4.31í4.41 (m, 3H), 4.03 (q, J = 6.9 Hz, 2H),
2.82í2.87 (m, 2H), 1.10 (t, J = 6.9 Hz, 3H). 13C NMR (75 MHz,
DMSO-d6): G = 171.2, 166.7, 157.5 (d, JC-F = 25.7 Hz), 156.1, 149.6,
139.2 (d, JC-F = 227.0 Hz), 131.0 (d, JC-F = 33.3 Hz), 130.1, 126.8,
115.1, 60.5, 54.2, 49.3, 36.2, 13.9. ESI-MS (m/z): 378.3 (M+, 100).
Elemental anal. Calcd for C17H18N3FO6: C, 53.83; H, 4.75; N, 11.08.
Found: C, 53.91; H, 4.51; N, 10.86%.
ꢀ5ꢁꢂ(WK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
yl)acetamido)-2-phenylacetate (2b): Yield: 67%, m.p. 186í187°C.
[D]D = + 72.4 (c 1.0, DMF). Qmax (KBr)/cmíꢅ 3327 (NíH), 3075
20
(=CíH), 1708(C=O), 1545 (C=N), 1445 (íCH2í), 1377 (CíH), 1233
1
(CíOíC), 977, 700. H NMR (300 MHz, DMSO-d6): G = 11.83 (s,
1H), 9.12 (d, J = 7.2 Hz, 1H), 8.03 (d, J = 6.9 Hz, 1H), 7.36í7.41
(m, 5H), 5.42 (d, J = 7.2 Hz, 1H), 4.44 (d, J = 16.5 Hz, 1H), 4.38 (d,
J = 16.5 Hz, 1H), 4.02í4.19 (m, 2H,), 1.13 (t, J = 7.2 Hz, 3H). 13C
NMR (75 MHz, DMSO-d6): G = 170.1, 166.6, 157.4 (d, JC-F = 25.4
Hz), 149.5, 139.1 (d, JC-F = 227.0 Hz), 136.0, 131.0 (d, JC-F = 31.8
Hz), 128.7, 128.3, 127.5, 61.0, 56.3, 49.2, 13.8. ESI-MS (m/z): 348.0
([M-1]+, 100). Elemental anal. Calcd for C16H16N3FO5: C, 55.01; H,
4.58; N, 12.03. Found: C, 54.96; H, 4.64; N, 12.12%.
0HWK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ\Oꢁ
acetamido)acetate (2i): Yield: 83%, m.p. 192í195°C. Qmax (KBr)/
cmíꢅ 3362 (NíH), 3042 (=CíH), 2833 (CíH), 1735 (C=O), 1678
(C=C), 1383 (CíH), 1236 (CíOíC). 1H NMR (300 MHz, DMSO-
d6): G = 11.87 (s, 1H), 8.67 (t, J = 5.9 Hz, 1H), 8.04 (d, J = 6.9 Hz,
1H), 4.34 (s, 2H), 3.90 (d, J = 6.0 Hz, 2H), 3.63 (s, 3H). 13C NMR
(75 MHz, DMSO-d6): G = 170.1, 167.4, 157.7 (d, JC-F = 25.8 Hz),
149.8, 139.5 (d, JC-F = 228.4 Hz), 131.0 (d, JC-F = 33.9 Hz), 51.9,
49.5, 40.7. ESI-MS (m/z): 258.0 (M+, 100). Elemental anal. Calcd for
C9H10N3FO5: C, 41.71; H, 3.86; N, 16.22. Found: C, 41.43; H, 3.84;
N, 16.07%.
ꢀ6ꢁꢂ0HWK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
20
yl)acetamido) propanoate (2c): Yield: 58%, m.p. 300í302°C. [D]D
= -12.7 (c 1.0, DMF). Qmax (KBr)/cmíꢅ 3437 (NíH), 3081 (=CíH),
2930 (CíH), 1709 (C=O), 1662 (C=C), 1379 (CíH), 1232 (CíOíC).
1H NMR (300 MHz, DMSO-d6): G = 11.85 (d, J = 5.1 Hz, 1H), 8.67
(d, J = 7.2 Hz, 1H), 8.02 (d, J = 6.6 Hz, 1H), 4.28í4.32 (m, 3H), 3.63
(s, 3H), 1.28 (d, J = 7.2 Hz, 3H). 13C NMR (75 MHz, DMSO-d6):
G = 172.7, 166.5, 157.5 (d, JC-F = 25.1 Hz), 149.6, 139.2 (d, JC-F
=
226.7 Hz), 131.0 (d, JC-F = 34.1 Hz), 51.9, 49.2, 47.6, 17.1. ESI-MS
(m/z): 272.1 (M+, 100). ESI-MS (m/z): 272.1 (M+, 100). Elemental
anal. Calcd for C10H12N3FO5: C, 43.96; H, 4.40; N, 15.38. Found: C,
43.93; H, 4.42; N, 15.31%.
General method for the synthesis of compound 3
2 (5 mmol) was dissolved in 2 mol L-1 NaOH (10 mL) (the solution
maintains the pH at 10 to 11) and the solution was stirred at room
temperature until 2 was fully hydrolysed (as monitored by TLC).
7KHꢀVROXWLRQꢀZDVꢀWKHQꢀDFLGL¿HGꢀZLWKꢀK\GURFKORULFꢀDFLGꢀWRꢀS+ꢀ ꢀꢄꢆꢀ
7KHꢀ UHVXOWLQJꢀ SUHFLSLWDWHꢀ ZDVꢀ ¿OWHUHGꢃꢀ ZDVKHGꢀ ZLWKꢀ HWK\Oꢀ DFHWDWHꢃꢀ
dried, and recrystallised from ethanol.
ꢀ5ꢁꢂ0HWK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
20
yl)acetamido) propanoate (2d). Yield: 49%, m.p. 301í303°C. [D]D
= + 12.7 (c 1.0, DMF). Qmax (KBr)/cmíꢅ 3437 (NíH), 3081 (=CíH),
2930 (CíH), 1709 (C=O), 1662 (C=C), 1379 (CíH), 1232 (CíOíC)
1H NMR (300 MHz, DMSO-d6): G = 11.84 (d, J = 5.1 Hz, 1H), 8.67
(d, J = 7.2 Hz, 1H), 8.02 (d, J = 6.6 Hz, 1H), 4.29í4.32 (m, 3H), 3.63
(s, 3H), 1.28 (d, J = 7.2 Hz, 3H). 13C NMR (75 MHz, DMSO-d6):
G = 172.7, 166.5, 157.5 (d, JC-F = 25.1 Hz), 149.6, 139.2 (d, JC-F = 226.7
Hz), 131.0 (d, JC-F = 34.7 Hz), 51.9, 49.2, 47.6, 17.1. ESI-MS (m/z):
272.1 (M+, 100). Elemental anal. Calcd for C10H12N3FO5: C, 43.96;
H, 4.40; N, 15.38. Found: C, 43.87; H, 4.45; N, 15.20%.
(S)-2-(2-(5-Fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)
acetamido)-3-(4-hydroxyphenyl) propanoic acid (3a): Yield: 86%,
m.p. 251í252°C. [D]D = + 97.7 (c 1.0, DMF).Qmax (KBr)/cmíꢅ
20
3372 (NíH), 3308 (NíH), 3213 (OíH), 1716 (C=O), 1663 (C=C),
1548 (C=N), 1515, 1482 (íCH2í), 1381 (COOH), 1258 (CíF), 671.
1H NMR (300 MHz, DMSO-d6): G = 12.77 (s, 1H), 11.82 (d, J = 5.1 Hz,
1H), 9.23 (s, 1H), 8.51 (d, J = 8.1 Hz, 1H), 7.94 (d, J = 6.9 Hz,
1H), 7.01 (d, J = 8.4 Hz, 2H), 6.66 (d, J = 8.7 Hz, 2H), 4.25í4.39
(m, 3H), 2.92 (dd, J = 13.8 Hz, J = 4.8 Hz 1H), 2.78 (dd, J = 13.8 Hz,
J = 8.7 Hz 1H). 13C NMR (75 MHz, DMSO-d6): G = 172.5, 166.5,
157.5 (d, JC-F = 25.7 Hz), 156.0, 149.6, 139.1 (d, JC-F = 225.8 Hz),
131.0 (d, JC-F = 34.2 Hz), 130.1, 127.2, 115.0, 54.0, 49.3, 36.0. ESI-
MS (m/z): 350.1 (M+, 100). Elemental anal. Calcd for C15H14N3FO6:
C, 51.28; H, 3.99; N, 11.97. Found: C, 51.53; H, 4.07; N, 11.808%.
(R)-2-(2-(5-Fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)
acetamido)-3-(4-hydroxy phenyl) propanoic acid (3b): Yield: 88%,
m.p. 251í252°C. [D]D20ꢀ ꢀ ±ꢍꢎꢆꢎꢀ ꢉc 1.0, DMF). Qmax (KBr)/cmíꢅ
ꢊꢊꢎꢄꢀꢉ1±+ꢈꢃꢀꢊꢄꢅꢋꢀꢉ2íH), 3092 (=CíH), 1724 (C=O), 1662 (C=C),
1550 (C=N), 1429 (íCH2í), 1375 (COOH), 1256 (CíF), 825, 669.
1H NMR (300 MHz, DMSO-d6): G = 12.76 (s, 1H), 11.82 (d, J = 5.1 Hz,
1H), 9.22 (s, 1H), 8.51 (d, J = 7.5 Hz, 1H), 7.94 (d, J = 6.9 Hz, 1H),
7.00 (d, J = 8.1 Hz, 2H), 6.66 (d, J = 8.1 Hz, 2H), 4.30í4.39 (m, 3H),
2.92 (dd, J = 13.8 Hz, J = 8.4 Hz 1H), 2.78 (dd, J = 13.8 Hz, J = 8.1
Hz 1H). 13C NMR (75 MHz, DMSO-d6): G = 172.6, 166.5, 157.5 (d,
JC-F = 33.8 Hz), 156.0, 149.6, 139.2 (d, JC-F = 227.3 Hz), 131.0 (d,
ꢀ6ꢁꢂ'LPHWK\OꢃꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁ
20
-yl)acetamido) succinate (2e): Yield: 69%, m.p. 109í110 °C. [D]D
ꢀ±ꢅꢌꢆꢌꢀꢉc 1.0, DMF). Qmax (KBr)/cmíꢅ 3412 (NíH), 3046 (=CíH),
2849 (íCH2í), 1753 (C=O), 1690 (C=O), 1522 (C=N), 1438 (íCH2í),
1385 (CíH), 1249 (CíF), 1222 (CíOíC). 1H NMR (300 MHz,
DMSO-d6): G = 11.85 (d, J = 4.8 Hz, 1H), 8.75 (d, J = 8.1 Hz, 1H),
8.01 (d, J = 6.9 Hz, 1H), 4.67 (dd, J = 6.3 Hz, J = 13.8 Hz, 1H), 4.33
(s, 2H), 3.61 (s, 3H), 3.64 (s, 3H), 2.69í2.85 (m, 2H). 13C NMR (75
MHz, DMSO-d6): G = 170.7, 170.2, 166.7, 157.5 (d, JC-F = 25.0 Hz),
149.6, 139.2 (d, JC-F = 226.2 Hz), 130.9 (d, JC-F = 35.6 Hz), 52.2,
51.7, 49.3, 48.6, 35.7. ESI-MS (m/z): 330.2 (M+, 100). Elemental
anal. Calcd for C12H14N3FO7: C, 43.50; H, 4.32; N, 12.69. Found: C,
43.44; H, 4.64; N, 12.85%.
ꢀ5ꢁꢂ'LHWK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
20
yl)acetamido) succinate (2f): Yield: 70%, m.p.109í110 °C. [D]D
=
+ 16.0 (c 1.0, DMF). Qmax (KBr)/cmíꢅ 3313.58 (NíH), 3075 (=CíH),
2989 (íCH2í), 1720 (C=O), 1549 (C=N), 1467 (íCH2í), 1377
(CíH), 1239 (CíF), 1167 (CíOíC). 1H NMR (300 MHz, DMSO-
d6): G = 11.83 (d, J = 4.5 Hz, 1H), 8.72 (d, J = 7.8 Hz, 1H), 7.99 (d,
J = 6.6 Hz, 1H), 4.64 (dd, J = 6.6 Hz, J = 14.1 Hz, 1H), 4.33 (s, 2H),
4.04í4.13 (m, 4H), 2.66í2.82 (m, 2H), 1.18 (t, J = 7.2 Hz, 3H), 1.17
(t, J = 6.9 Hz, 3H). 13C NMR (75 MHz, DMSO-d6): G = 170.2, 169.8,
166.7, 157.5 (d, JC-F = 24.5 Hz), 149.6, 139.2 (d, JC-F = 227.0 Hz),
130.9 (d, JC-F = 35.1 Hz), 61.0, 60.4, 49.3, 48.7, 35.9, 14.0, 13.9. ESI-
MS (m/z): 358.2 (M+, 100). Elemental anal. Calcd for C14H18N3FO7:
C, 46.80; H, 5.01; N, 11.70. Found: C, 46.60; H, 4.94; N, 11.68%.
ꢀ6ꢁꢂ0HWK\Oꢃ ꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
yl)acetamido)-3-(4-hydroxyphenyl) propanoate (2g): Yield: 73%,
J
C-F = 35.1 Hz), 130.1, 127.2, 115.0, 54.1, 49.3, 36.0. ESI-MS (m/z):
350.3 (M+, 100). Elemental anal. Calcd for C15H14N3FO6: C, 51.28;
H, 3.99; N, 11.97. Found: C, 51.30; H, 3.96; N, 12.07%.
2-(2-(5-Fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)
acetamido)acetic acid (3c): Yield: 86%, m.p. 257í259°C.Qmax
(KBr)/cmíꢅ 3490 (NíH), 3412 (OíH), 3070 (=CíH), 1701 (C=O),
1662 (C=O), 1541 (NíC=O), 1383 (COOH). 1H NMR (300 MHz,
DMSO-d6): G = 12.64 (s, 1H), 11.83 (d, J = 4.5 Hz, 1H), 8.52 (t,
J = 6.0 Hz, 1H), 8.02(d, J = 6.9 Hz, 1H), 4.34 (s, 2H), 3.80 (d, J =
6.0 Hz, 2H). 13C NMR (75 MHz, DMSO-d6): G = 170.8, 166.9, 157.5
m.p. 156í157°C. [D]D = + 38.4 (c 1.0, DMF). Qmax (KBr)/cmíꢅ
(d, JC-F = 24.8 Hz), 149.6, 139.2 (d, JC-F = 226.8 Hz), 130.9 (d, JC-F =
20

264 JOURNAL OF CHEMICAL RESEARCH 2009
34.0 Hz), 49.3, 40.6. ESI-MS (m/z): 244.0 (M+, 100). Elemental anal.
Calcd for C8H8N3FO5: C, 39.18; H, 3.27; N, 17.14. Found: C, 39.13;
H, 3.14; N, 17.11%.
V 1922.4(2) Å3, Dc (Z 4) 1.466 g cm-3, F (000) 888, ȝ 0.120 mm-1,
R1 0.0991, wR2 0.2955.
&U\VWDOꢃVWUXFWXUHꢃGHWHUPLQDWLRQꢃDQGꢃUH¿QHPHQW
General method for the synthesis of compound 4
Diffraction data were collected at 25(2)°C using
a graphite
A solution of DCC (3.09 g, 15 mmol) in DMF (15 mL) was added
dropwise to a DMF solution (25 mL) of 3c (2.45 g, 10 mmol) and
HOBt (1.49 g, 11 mmol) at 0°C over a period of 40 min. The resulting
solution was stirred at room temperature for 5 h. The appropriate
ester of the amino acid hydrochloride (10 mmol) and triethylamine
(1.4 mL, 10 mmol) were added to the above mixture. After 4 h
VWLUULQJꢃꢀDꢀZKLWHꢀVROLGꢀZDVꢀREWDLQHGꢆꢀ$IWHUꢀ¿OWUDWLRQꢃꢀWKHꢀ¿OWUDWHꢀZDVꢀ
concentrated under reduced pressure and the residue was separated
by column chromatography to afford compound 4.
monochromated Mo KD (O = 0.071073 nm) radiation with an Z-scan
technique. Determination of the crystal class, orientation matrix,
and cell dimensions was performed according to the established
procedures. Lorentz polarisation and absorption corrections
were applied. Empirical absorption corrections were performed
with SADABS program. Most of the non-hydrogen atoms were
located using direct methods, and those remaining were derived by
VXEVHTXHQWꢀ)RXULHUꢀV\QWKHVHVꢆꢀ$OOꢀQRQꢂK\GURJHQꢀDWRPVꢀZHUHꢀUH¿QHGꢀ
anisotropically, and all hydrogen atoms were held stationary and
LQFOXGHGꢀ LQꢀ WKHꢀ ¿QDOꢀ VWDJHꢀ RIꢀ IXOOꢂPDWUL[ꢀ OHDVWꢂVTXDUHVꢀ UH¿QHPHQWꢀ
based on F2 using SHELXS-97 and SHELXL-97 program packages.
CCDC contains the supplementary crystallographic data for
this paper. The data can be obtained free of charge from the
Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.uk/
data_request.cif.
ꢀ6ꢁꢂ'LPHWK\OꢂꢄꢂꢀꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂ
1(2H)-yl)acetamido)acetamido) succinate (4a): Yield: 48%, m.p.
195í196°C. [D]D ꢀ ±ꢅꢌꢆꢋꢀ ꢉc 1.0, DMF).Qmax (KBr)/cmíꢅ 3365
20
(NíH), 1662 (C=O), 1543 (C=N), 1429 (íCH2í), 1382 (CíH); 1237
1
(CíOíC). H NMR (300 MHz, DMSO-d6): G = 11.80 (s, 1H), 8.45
(t, J = 5.6 Hz, 1H), 8.39 (d, J = 7.8 Hz, 1H), 7.99 (d, J = 6.6 Hz, 1H),
4.67 (dd, J = 14.7 Hz, J = 6.9 Hz, 1H), 4.36 (s, 1H), 3.78 (d, J = 6.0
Hz, 2H), 3.63 (s, 3H), 3.61 (s, 3H), 2.67í2.85 (m, 2H). 13C NMR (75
Supplementary data (product NMR spectra) are available directly
from the correspondent.
MHz, DMSO-d6): G = 170.9, 170.3, 168.4, 166.8, 157.5 (d, JC-F
=
16.4 Hz), 149.6, 139.2 (d, JC-F = 227.6 Hz), 130.9 (d, JC-F = 33.1 Hz),
52.1, 51.6, 49.5, 48.4, 41.6, 35.6. ESI-MS (m/z): 387.1 (M+, 100).
Elemental anal. Calcd for C14H17N4FO8: C, 43.30; H, 4.38; N, 14.43.
Found: C, 43.52; H, 4.14; N, 14.18%.
:Hꢀ DFNQRZOHGJHꢀ WKHꢀ ¿QDQFLDOꢀ VXSSRUWꢀ RIꢀ WKHꢀ 1DWLRQDOꢀ
Natural Science Foundation of China (No. 20571057) and
the Wenzhou Technology Project Foundation of China
(No. S20060026) and the National Centre for Drug Screening,
Shanghai, China, for evaluation of antitumour activities.
ꢀ6ꢁꢂ0HWK\OꢂꢄꢂꢀꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ\Oꢁ
acetamido)acetamido)-3-(4-hydroxyphenyl)propanoate (4b): Yield:
20
55%, m.p. 168í169°C. [D]D = + 16.7 (c 1.0, DMF). Qmax (KBr)/
cmíꢅ 3366 (NíH), 3025( =CíH), 1662 (C=O), 1543 (C=N), 1429
(íCH2í), 1382 (CíH), 1238 (CíOíC), 672. 1H NMR (300 MHz,
DMSO-d6): G = 11.84 (s, 1H), 9.23 (s, 1H), 8.32(t, J = 5.6 Hz,
1H), 8.31 (d, J = 6.9 Hz, 1H), 8.00 (d, J = 6.9 Hz, 1H), 6.98 (d,
J = 8.4 Hz, 2H), 6.65 (d, J = 8.4 Hz, 2H), 4.33í4.43 (m, 3H),
3.66í3.83 (m, 2H), 3.58 (s, 3H), 2.89 (dd, J = 13.5 Hz, J = 6.0 Hz,
1H), 2.78 (dd, J = 13.5 Hz, J = 8.7 Hz, 1H). 13C NMR (75 MHz,
DMSO-d6): G = 171.9, 168.4, 166.8, 157.5 (d, JC-F = 25.1 Hz), 156.0,
149.7, 139.3 (d, JC-F = 227.0 Hz), 130.9 (d, JC-F = 33.5 Hz), 129.9,
126.9, 115.1, 53.9, 51.8, 49.5, 41.5, 36.1. ESI-MS (m/z): 421.2 (M+,
100). Elemental anal. Calcd for C18H19N4FO7: C, 51.18; H, 4.50; N,
13.27. Found: C, 51.26; H, 4.64; N, 12.92%.
Received 27 November 2008; accepted 22 February 2009
Paper 08/0309 doi: 10.3184/030823409X431364
Published online: 29 April 2009
References
1
2
3
C. Maria, R.S. Fernando, A. Juan, C. Octavio, A. Antonia, A. Miguel,
E. Antonio and M. Joaquin, Tetrahedron Lett., 2007, 63, 183.
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1116.
ꢀ6ꢁꢂ0HWK\OꢂꢄꢂꢀꢄꢂꢀꢄꢂꢀꢅꢂÀXRURꢂꢄꢆꢇꢂGLR[RꢂꢈꢆꢇꢂGLK\GURS\ULPLGLQꢂꢉꢀꢄ+ꢁꢂ
yl)acetamido) acetamido)-3-methyl butanoate (4c): Yield: 45%,
4
5
H.J. Lehmler and S. Parkin, Acta. Cryst., 2000, C56, e518.
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5063.
20
m.p. 200í201°C. [D]D = + 4.4 (c 1.0, DMF).Qmax (KBr)/cmíꢅ 3359
(NíH), 2967 ( =CíH), 1743 (C=O), 1665 (C=C), 1430 (íCH2í), 1383
(CíH), 1241 (CíOíC). 1H NMR (300 MHz, DMSO-d6): G = 11.86
(s, 1H), 8.46 (t, J = 5.7 Hz, 1H), 8.19 (d, J = 8.4 Hz, 1H), 8.03
(d, J = 6.9 Hz, 1H), 4.33 (s, 2H), 4.17 (dd, J = 7.5 Hz, J = 7.5 Hz, 1H),
3.83 (d, J = 5.7 Hz, 2H), 3.63 (s, 3H), 1.97í2.06 (m, 1H), 0.86 (m, 6H).
13C NMR (75 MHz, DMSO-d6): G = 171.9, 168.6, 166.8, 157.5 (d,
JC-F = 24.8 Hz), 149.7, 139.2 (d, JC-F = 227.0 Hz), 130.9 (d,
6
7
8
9
P.M. Moyle, C. Olive, M.F. Ho, M.F. Good and I. Toth, J. Med. Chem.,
2006, 49, 6364.
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11 M.L. Hu, J.X. Yuan and A. Morsali, Solid State Sci., 2006, 8, 981.
12 P. Yin, M.L. Hu and L.C. Hu, J. Mol. Struct., 2008, 882, 75.
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25, 417.
14 X. Liu, R. Chen and Y. Liu, Chem. J. Chin. U., 2002, 23, 1299.
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16 C. Chevrier, A. Defoin and C. Tarnus, Bioorg. Med. Chem., 2007, 15,
4125.
J
C-F = 35.2 Hz), 57.4, 51.6, 49.5, 41.6, 29.9, 18.9, 18.1. ESI-MS (m/z):
357.1 (M+, 100). Elemental anal. Calcd for C14H19N4FO6: C, 46.92; H,
5.31; N, 15.64. Found: C, 47.12; H, 5.24; N, 15.58%.
Crystal data
Compound (3b). C18H21FN4O7, M 424.39, T 298(2) K, orthorhombic,
Space group P 212121, a 4.8774(3), b 15.4880(10), c 25.4480(17) Å.