Synthesis of lipid derivatives of pyrrole polyamide and their biological activity

Article abstract of DOI:10.1271/bbb.60650

Novel fatty acyl and phospholipid derivatives of pyrrole polyamide were synthesized. Their cytotoxicity against a cancer cell line of MT-4 cells and those infected by human immunodeficiency virus (HIV) was examined. Although no anti-HIV activity was found, their cytotoxicitty against the cancer cells was significantly enhanced by introducing a lipophilic group into the pyrrole polyamide.

Full text of DOI:10.1271/bbb.60650

Biosci. Biotechnol. Biochem., 71 (4), 1078–1082, 2007
Note
Synthesis of Lipid Derivatives of Pyrrole Polyamide
and Their Biological Activity
1
2
2
2
1
1
Masahiko YAMAMOTO, Changjin ZHU, Lui YI, Zheng RONG, Yoshie MIURA, Minoru IZUMI,
1
3
4
1;y
Shuhei NAKAJIMA, Ken-ichi TANAMOTO, Sakayu SHIMIZU, and Naomichi BABA
1
Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology,
Okayama University, 1-1-1 Tsushimanaka, Okayama, Okayama 700-8530, Japan
2
School of Chemical Engineering & Environment, Beijing Institute of Technology,
No. 5 South Zhongguan Street, Haidian District, P. O. Box 327, Beijing 100081, P.R.C., China
3
National Institute of Health Sciences, Division of Food Additives,
1
-18-1 Kamiyoga, Setagaya, Tokyo 158-8501, Japan
4
Department of Applied Bioscience, Graduate School of Agriculture, Kyoto University,
Sakyo-ku, Kyoto 606-8502, Japan
Received November 17, 2006; Accepted January 15, 2007; Online Publication, April 7, 2007
[doi:10.1271/bbb.60650]
Novel fatty acyl and phospholipid derivatives of
compounds, Zerouga et al. have reported that metho-
trexate, a cytotoxic drug, conjugated to phosphatidyl-
choline (PC) having a docosahexaenoyl group showed
higher anti-proliferative activity against murine leuke-
pyrrole polyamide were synthesized. Their cytotoxicity
against a cancer cell line of MT-4 cells and those
infected by human immunodeficiency virus (HIV) was
examined. Although no anti-HIV activity was found,
their cytotoxicitty against the cancer cells was signifi-
cantly enhanced by introducing a lipophilic group into
the pyrrole polyamide.
1
4)
mia cells than one having a stearoyl group. In our
previous study, conjugates of qunine with fatty acid
were found to show higher cytotoxicity against tumor
1
5)
cell line FM3A than qunine itself. Parang et al. have
extensively reviewed the relationship between the lipid
0 0 0
Key words: pyrrole polyamide; lipid; phospholipid; can-
cer cell; human immunodeficiency virus
modifications of 3 -azido-2 ,3 -dideoxythymidine (AZT)
1
6)
and anti-HIV activity. In the present study, novel fatty
acyl derivatives of pyrrole polyamide were synthesized
by using stearic acid (3, a typical saturated fatty acid
rich in mammal fats), linoleic acid (4, a typical n-6 type
of dienoic acid rich in plant lipids) and icosapentaenoic
acid (5, a typical n-3 type of pentaenoic acid rich in fish
oil), and the cytotoxicity of the conjugates was exam-
ined by using MT-4 cells.
(HIV)-II
Since the naturally occurring pyrrole polyamide,
netropsin, as an antibiotic was reported by Kopka et
1
,2)
al.,
been synthesized so far. Subsequently, their highly
a number of pyrrole polyamide analogues has
3
)
sequence-specific binding to DNA has intensively
triggered the design of novel functional pyrrole poly-
amides,4 and the search for new biological functions
including anti-cancer and anti-HIV activities.
As the major components of polyunsaturated fatty
acids (PUFA) in marine fish oil, docosahexaenoic acid
Pyrrole polyamide has so far been synthesized by
a reaction sequence involving the nitration of pyrrole,
reduction of the nitro group to an amino group and
condensation of the amine with nitro-pyrrole carboxylic
)
5
)
6)
1
7)
acid. This route, however, involves some intermedi-
ates having a nitro group that is known to cause allergic
symptoms. To minimize the number of these intermedi-
ates, a different approach was investigated to obtain
a key intermediate (2). Briefly, the route involves
trichloroacetylation at the 2-position of N-methylpyr-
role, nitration at the 4-position of the pyrrole nucleus,
conversion of the trichloroacetyl group to a methoxy-
carbonyl group, reduction of the nitro group to an amino
group, condensation of the amine with N-methylpyrrole
(
DHA) and icosapentaenoic acid (EPA) have a non-
conjugated all-cis-polyunsaturated olefinic structure.
They are known to exhibit a variety of biochemical
and physiological functions including enhanced cell
7
)
membrane permeability, growth regulation and apop-
8
–10)
tosis-inducing capability to cancer cells, cytotoxicity
enhancement for some anti-cancer drugs against cancer
cells1
ing the effect of the lipid modification of bioactive
1,12)
and potential anti-malarial activity. Regard-
13)
y
To whom correspondence should be addressed. Tel: +81-86-251-8292; Fax: +81-86-251-8388; E-mail: babanaom@cc.okayama-u.ac.jp

Synthesis of Lipid Derivatives of Pyrrole Polyamide]
1079
H
H
N
N
N
N
NaOH
H
H
O
O
N
N
CH OH
N
N
3
O
O
N
N
2
COOH
1
COOCH3
O
O
DIC, HOBt, H N
NH2
2
2, DIC, HOBt
CHCl3, DMF
R
OH
R
N
NH2
DMF
H
3
4
5
: Stearic acid
: Linoleic acid
: Icosapentaenoic acid
6* (58%)
7 (38%)
8 (32%)
H
N
N
H
N
O
O
O
N
N
N
H
N
H
O
O
O
N
N
N
N
H
N
O
O
O
N
O
9
: Stearoyl amide (45%)
H
N
H
N
H
N
H
N
N
O
H
N
1
0: Linoleoyl amide (75%)
H
N
H
N
O
1
1: Icosapentaenoyl amide (60%)
Scheme 1. Synthesis of Fatty Acyl Derivatives (9)–(11).
ꢁ
Ref. 21. DIC, Diisopropylcarbodimide; HOBt, Hydroxybenzotriazole
2
-carboxylic acid, hydrolysis of the methyl ester to a
enzyme as a molecular switch to liberate the pyrrole
polyamide at the right time when it should play some
roles in biological systems.
carboxylic acid, condensation of this acid with N-
methyl-4-aminopyrrole 2-carboxylic acid methyl ester
that had been prepared as already described to afford an
intermediate 1,1 and finally hydrolysis of the methyl
ester to afford 2.
Acylated pyrrole polyamide (9–11) were synthesized
by condensation of fatty acid half amides (6–8) with the
carboxylic acid (2) respectively (Scheme 1) as described
in the experimental section.
A phospholipid derivative (14) was synthesized by
using the same intermediate 2 (Scheme 2). In this case,
methyl ꢀ-aminobutylate was introduced into 2 as a
spacer giving ester 12, which was hydrolyzed to 13. Due
to its instability, a product 13 was submitted as such to
condensation with a lyso PC having a stearoyl group at
sn-1 position. Silica gel chromatography afforded the
desired compound 14 whose structural integrity was
4
)
As a preliminary test of biological activity in the
present study, lipid derivatives 9–11 and 14 were
examined for their in vitro cytotoxicity against cultured
MT-4 cells. The result showed that the order of their
concentration for complete growth inhibition of cultured
MT-4 cells was stearoyl derivative 9 (45 mM) < ico-
sapentaenoyl derivative 11 (176 mM) < linoleoyl deriv-
ative 10 (ꢀ181 mM) < pyrrole amide methyl ester 1
(ꢀ1304 mM) as a control. This experiment demonstrated
for the first time that lipid modification of a pyrrole
polyamide remarkedly enhanced its cytotoxicity, and
derivative 9 with a saturated acyl group appeared to be
more active than the unsaturated type. The same
tendency was also observed for the concentration range
of partially inhibitive and non-inhibitive cases. Although
phospholipid derivative (14) enhanced the cytotoxicity
(ꢀ260 mM) to some extent, the activity was lower than
those by acyl derivatives 9–11. Combining all the
results, lipid modification of the pyrrole polyamide was
found to significantly enhance the cytotoxicity against
cancer cell line MT-4 cells, and the effect appeared to be
higher with the saturated acyl derivative than with the
8)
1
confirmed by H-NMR and ESI MS data. For further
structural confirmation, hog pancreatic phospholipase
A2 was applied in an acetate buffer (pH 8.4) to substrate
1
4 at room temperature for 12 h. A TLC analysis showed
that the lyso PC and pyrrole polyamide 13 having the
spacer had been liberated by the enzymatic reaction.
This finding might give an opportunity to use this

1080
M. YAMAMOTO et al.
H
N
N
HCl ·
H2N
O
H
O
NaOH
MeOH, H O
N
OMe, DIC, HOBt, DIEA
N
2
O
H
N
2
DMF
O
N
1
2 (71%)
OMe
O
H
N
N
H
N
O
N
DIC, HOBt, Lyso PC, DIEA
O
H
N
O
O
CHCl , DMF
3
N
OH
1
3 (40%)
O
O
H
N
N
H
N
O
N
O
O
H
N
O
O
N
O
P
O
N
O
1
4 (10%)
O
Scheme 2. Synthesis of Phospholipid Derivatives (14).
DIEA, Diisopropylethylamine
unsaturated types. No anti-HIV effect was, however,
protons on the pyrrole rings). ESI MS m=z: found,
þ
1
9)
appearent by the microplate method and Magic 5
method2 for any of the pyrrole polyamide derivatives
synthesized in the present study. The preliminary
biological results described here constitute an additional
example of the effect of lipid modification for bio-
logically active compounds.
(M þ H ) 337.2; C21H40N2O requires 336.6.
0)
Synthesis of N-icosapentaenoylpropane-1,3-diamine
(8). This intermediate was prepared under the same
conditions as those used for the preparation of 7. R ¼
f
1
0:67 (CHCl :CH OH: NH aq, 80:20:5). H-NMR
3
3
3
(CDCl3): ꢁ(ppm): 0.95 (3H, t, J ¼ 7:5, –CH2–CH3),
1
.60 (2H, d, J ¼ 6:6 Hz, –NH–CH2–CH2–CH2–NH–),
Experimental
1
1.75 (2H, m, –C(O)–CH2–CH2–), 2.10 (4H, m, –CH2–
CH=CH–, –CH2CH3), 2.30 (2H, t, J ¼ 8:0, –C(O)–
CH2–), 2.65 (2H, d, J ¼ 6:9 Hz, –CH2–NH2), 2.80 (8H,
m, –(CH=CH–CH ) –), 3.16 (2H, d, J ¼ 6:3 Hz, –NH–
H- and 13C-NMR spectra were recorded by a Varian
Mercury 300 or VXR 500 using CDCl3, and ESI MS
data were recorded by API III (Perkin Elmer) by direct
infusion, using a mixture of THF/CH OH/H O (15:4:1)
2
4
CH –), 3.90–4.00 (9H, s, 3 ꢃ N–CH ), 5.35 (10H, m,
3
2
2
3
ꢂ
þ
with 0.1% HCOOH or 0.1% HCOO NH4 as a solvent
in the positive mode.
–(CH=CH–CH ) –), 6.60–6.90 (6H, m, protons on the
2 5
þ
pyrrole rings). ESI MS m=z: found, (M þ H ) 359.2;
Synthesis of N-linoleoylpropane-1,3-diamine (7). To a
solution of linoleic acid (1.89 g, 6.75 mmol) in ethanol-
free chloroform (25 ml) were added HOBt (0.95 g, 7.0
mmol) and DIC (0.84 g, 7.0 mmol), and the solution was
stirred at r.t. overnight. A solution of 1,3-propanedi-
amine (1.0 g, 13.5 mmol) in ethanol-free chloroform (7
ml) was added dropwise to the reaction mixture which
was stirred at r.t. overnight. After evaporating the
solvent under reduced pressure, the residue was chro-
matographed on silica gel, eluting with a mixture of
cloroform/methanol/aq.NH3 (80:20:5) to afford half
amide 7. Rf ¼ 0:65 (CHCl3:CH3OH:NH3aq, 80:20:5).
C23H38N2O requires 358.6.
Synthesis of the stearoyl derivative (9). To a solution
of 2 (55.2 mg, 0.15 mmol) and HOBt (24.4 mg,
0.18 mmol) in DMF (3 ml) was added DIC (22.2 mg,
0.18 mmol). The reaction mixture was stirred at r.t.
for 24 h under N . Amide 6 (45.6 mg, 0.15 mmol) and
2
distilled ethanol-free chloroform (3 ml) were added to
ꢄ
this reaction mixture which was stirred at 50 C for 24 h
under N2. The solvent was evaporated under reduced
pressure. The resulting residue was chromatographed
in a silica gel column (CHCl3/MeOH, 95:5) to give
final compound 9. Rf ¼ 0:57 (CHCl3:CH3OH, 95:5).
1
1
H-NMR (CDCl )ꢁ(ppm): 0.78 (3H, t, J ¼ 7:8, –CH –
H-NMR (CDCl ) ꢁ(ppm): 0.80 (3H, t, J ¼ 7:0, –CH –
3
2
3
2
CH ), 1.16 (14H, m, –(CH ) –CH –(CH=CH–CH ) –
2 2
CH ), 1.30 (28H, m, –(CH ) –CH ), 1.65 (4H, m,
3 2 14 3
3
2 4
2
(
(
(
(
CH ) –CH ), 1.50 (2H, m, –C(O)–CH –CH –), 1.60
2
–C(O)–CH –CH –, –NH–CH –CH –CH –NH–), 2.20
2 2 2 2 2
2
3
3
2
2H, d, J ¼ 6:6 Hz, –NH–CH2–CH2–CH2–NH–), 2.00
4H, m, –CH2–CH=CH–CH2–CH=CH–CH2–), 2.08
2H, t, J ¼ 7:8, –C(O)–CH2–), 2.65 (2H, d, J ¼ 6:9
(2H, t, J ¼ 8:0, –C(O)–CH2–), 3.35 (4H, m, –NH–
CH2–CH2–CH2–NH–), 3.90–4.00 (9H, s, 3 ꢃ N–CH3),
6.15 (1H, m, –CH=CH–CH=), 6.60–6.90 (6H, m,
protons on the pyrrole rings) ESI MS m=z: found,
Hz, –CH2–NH2), 2.75 (2H, t, J ¼ 6:5, =CH–CH2–
þ
CH=), 3.16 (2H, d, J ¼ 6:3 Hz, –NH–CH2–), 3.90–4.00
(M þ H ) 692.4; C39H61N7O4 requires 691.5.
(
6
9H, s, 3 ꢃ N–CH ), 5.35 (4H, m, –(CH=CH–CH ) –),
Synthesis of the linoleoyl derivative (10). This product
was prepared under the same conditions as those used
3
2 2
.13 (1H, m, –CH=CH–CH=), 6.60–6.90 (6H, m,

Synthesis of Lipid Derivatives of Pyrrole Polyamide]
1081
for the synthesis of 9. TLC (Silica gel) R ¼ 0:38
Lyso-PC (136.2 mg, 0.26 mmol) and DIEA (100 ml)
were added to this reaction mixture which was stirred
f
1
(
CHCl :CH OH, 95:5); H-NMR (CDCl ) ꢁ(ppm): 0.85
3
3
3
(
3H, t, J ¼ 7:5, –CH –CH ), 1.30 (14H, m, –(CH ) –
at room temperature for 24 h under N . The solvent
2
2
3
2 4
CH2–(CH=CH–CH2)2–(CH2)3–CH3), 1.60 (4H, m,
C(O)–CH2–CH2–, –NH–CH2–CH2–CH2–NH–), 2.00
4H, q, J ¼ 7:5, –CH2–CH=CH–CH2–CH=CH–CH2–),
.20 (2H, t, J ¼ 8:0, –C(O)–CH2–), 2.75 (2H, t, J ¼ 6:5,
CH–CH2–CH=), 3.35 (4H, m, –NH–CH2–CH2–CH2–
NH–), 3.90–4.00 (9H, s, 3 ꢃ N–CH ), 5.35 (4H, m,
was evaporated under reduced pressure. The resulting
residue was separated by silica gel column chroma-
tography (CHCl3/MeOH, 6:4) monitored by prepar-
ative TLC (CHCl3/MeOH/NH3aq. 65:35:5) to yield
final compound 13 as an yellow oil. TLC (silica gel)
–
(
2
=
1
R ¼ 0:6 (CHCl :CH OH:NH aq, 60:30:5); H-NMR
3
f
3
3
3
–
(CH=CH–CH ) –), 6.60–6.90 (6H, m, protons on the
(CDCl :CD OD, 8:2) ꢁ(ppm): 0.80 (3H, t, J ¼ 7:5,
2
2
3
3
þ
pyrrole rings); ESI MS m=z: found, ðM þ HÞ 688.5;
–CH –CH ), 1.05 (2H, m, –CH –CH ), 1.20 (26H, m,
2 3 2 3
C39H57N7O4 requires 687.5.
–(CH2)13–CH2–CH3), 1.50 (2H, –C(O)–CH2–CH2–),
1.90 (2H, m, –NH–CH2–CH2–), 2.20 (2H, m, –NH–
CH2–CH2–CH2–), 2.40 (2H, m, –CH2–O–C(O)–CH2–),
2.90 (2H, m, –NH–CH2–), 3.30 (2H, m, –CH2–N–
(CH3)3), 3.50 (9H, s, –N–(CH3)3), 3.70–3.90 (9H, s,
Synthesis of the icosapentaenoyl derivative (11). This
product was prepared under the same conditions as those
used for the synthesis of 9. Rf ¼ 0:32 (CHCl3:CH3OH,
1
9
CH –CH ), 1.75 (4H, m, –C(O)–CH –CH –, –NH–
5:5). H-NMR (CDCl3) ꢁ(ppm): 0.95 (3H, t, J ¼ 7:5, –
3 ꢃ N–CH on the pyrrole rings), 4.00 (2H, m, –CH–
2
3
2
2
3
CH –CH –CH –NH–), 2.10 (4H, m, –CH –CH –
2
CH –O–P–), 4.10 (2H, m, –CH –CH –O–P–), 4.20 (2H,
2
2
2
2
2
2
2
CH=CH–, –CH –CH ), 2.30 (2H, t, J ¼ 8:0, –C(O)–
m, –CH–CH –O–C(O)–), 5.10 (1H, m, –CH –CH–
2 2
2
3
CH2–), 2.80 (8H, m, –(CH=CH–CH2–CH=CH)4), 3.40
4H, m, –NH–CH2–CH2–CH2–NH–), 3.90–4.00 (9H, , s,
CH2–), 6.00 (1H, m, –CH=CH–CH=), 6.60–6.90 (6H,
m, protons on the pyrrole rings). ESI MS m=z: found,
ðM þ HÞþ 960.6; C48H78N7O11P requires 959.6.
(
3
ꢃ N–CH3), 5.35 (10H, m, –(CH=CH–CH2)5–), 6.13
(
1H, m, –CH=CH–CH=), 6.60–6.90 (6H, m, protons on
þ
In vitro cytotoxicity and anti-HIV assays were
respectively conducted by the microplate method and
the pyrrole rings). ESI MS m=z: found, ðM þ HÞ 710.4;
1
9)
C H N O requires 709.4.
4
magic-5 method reported by Otake et al. and Kimpton
2
4
1
55
7
0)
Synthesis of methyl ꢀ-aminobutylate derivative of
pyrrole polyamide (12). A mixture of intermediate acid
et al.
Microplate method. Sample solutions (100 ml) were
sequentially diluted at 1:2 or 1:5 with an RPMI1640
medium containing 10% FCS in a 96-well plate. For
the cytotoxicity experiment, 100 ml of cell a suspension
2
, (312 mg, 0.8 mmol), HOBt (137 mg, 1.03 mmol) and
DIC (130 mg, 1.03 mmol) in DMF (0.68 ml) was stirred
at r.t. for 24 h. To this solution were added methyl 4-
aminobutylate hydrochloride (130 mg, 0.85 mmol) and
DIEA (260 ml, 1.49 mmol), and stirred at r.t. for further
5
of MT-4 cells (2 ꢃ 10 /ml) in a stage of exponential
growth was added to each well. For the anti-HIV
6
2
4 h. After an addition of deionized water (10 ml), the
experiment, MT-4 cells (2 ꢃ 10 ) were infected by the
product was extracted with chloroform. The product was
purified by silica gel chromatography (hexane/ethyl
acetate, 3:7) affording an unstable oil. TLC(Silica gel)
addition of a stock solution of HTLV-BIII to a con-
centration suitable as an infectious dose (100TCID50) to
ꢄ
the tissue culture, which was incubated at 37 C for 1 h.
1
Rf ¼ 0:3 (Hexane:EtOAc, 2:8); H-NMR (CDCl3)
The cells were resuspended in 10 ml of the medium, and
the suspension (100 ml) was added to all the wells in the
96-well plate. After incubating for 5 days, the cytotox-
icity and cytopathic effect (CPE) were evaluated by
counting the cells by optical microscopic observation.
ꢁ(ppm): 1.91 (2H, m, –NH–CH2–CH2–), 2.40 (2H, m,
–
3
6
NH–CH2–CH2–CH2–CO), 3.39 (2H, m, –NH–CH2–),
.80 (3H, s, O–CH3), 3.78–4.00 (9H, s, 3 ꢃ N–CH3),
.60–6.90 (6H, m, protons on the pyrrole rings). ESI MS
þ
4
m=z: Found, ðM þ NH Þ 486.3; C H N O requires
Magic-5 method. Magic-5 cells (10 cells) per one
ꢄ
4
23 28
6
5
4
86.0.
Synthesis of ꢀ-aminobutylate derivative of the pyrrole
polyamide (13). A solution of the ester 11 (200 mg,
well of a 96-well plate were cultured at 37 C to the
stage at which the cells were allowed to adhere to the
plastic surface of the plate. After removing the culture
medium, a sample solution of the pyrrole polyamide
diluted 2 times with the medium was added, this being
followed by the addition of HIV-1 Ba-L strain prepared
to a concentration of 100–200 BFU/50 ml by using the
0
.43 mmol) and 2N–NaOH (10 ml) in methanol (10 ml)
ꢄ
was stirred at 60 C for 12 h. After removing the solvent
under reduced pressure, the residue was acidified with
1N-HCl, and the acid was extracted with a mixture of
chloroform/methanol (2:1). Silica gel chromatography
medium containing DEAE-dextran. The cells were
ꢄ
(
chloroform/methanol, 9:1 ! 5:5) afforded acid 13 as
incubated at 37 C for 48 h in a CO incubator. After
2
an unstable oil which was used as such for the next
reaction.
removing the medium, 1%-formaldehyde and 0.2%-
glutaraldehyde in PBS were added, and the mixture
incubated at r.t. for 5 min. After washing the cells, 4 mM-
potassium ferrocyanide, 4 mM-potassium ferricyanide,
2 mM MgCl2 and 400 mg/ml of X-gal were added, and
Synthesis of the phospholipid derivative (14). To a
solution of 13 (100 mg, 0.22 mmol) and HOBt (34.4 mg,
0
.26 mmol) in a mixed solvent of CHCl3 (1 ml) and
ꢄ
DMF (1 ml) was added DIC (32.8 mg, 0.26 mmol). The
reaction mixture was stirred at r.t. for 24 h under N2.
the mixture incubated at 37 C for 1 h. The staining
solution was removed and the cells were washed. The

1082
M. YAMAMOTO et al.
cells stained blue were counted by using optical micro-
scopic observation. In this experiment, TAK-779 and
AZT were used as controls for the anti-HIV activity.
292 (1998).
1
1
1
1) Das, U. N., Madhavi, N., Kumar, G. S., Padma, M., and
Sangeetha, P., Can tumor cell drug resistance be
reversed by essential fatty acids and their metabolites?
Prostaglandins, Leucotrienes and Essential Fatty Acids,
Acknowledgments
5
8, 39–54 (1998).
2) Germain, E., Chajes, V., Cognault, S., Lhuillery, C., and
Bougnoux, P., Enhancement of doxorubicin cytotoxicity
by polyunsaturated fatty acids in the human breast tumor
cell line MDA-MB-231. Int. J. Cancer, 75, 578–583
(1998).
3) Binh, T. Q., Ilett, K. F., Davis, T. M., Hung, N. C.,
Powell, S. M., Thu, L. T., Thien, H. V., Phuong, H. L.,
and Phuong, V. D. B. P., Oral bioavailability of
dihydroartemisinin in Vietnamese volunteers and in
patients with Falciparum malaria. Brit. J. Clin. Phar-
macol., 51, 541–546 (2001).
14) Zerouga, M., Sillwell, W., and Jenski, L. J., Synthesis of
a novel phosphatidylcholine conjugated to docosahexa-
enoic acid and methotrexate that inhibits cell prolifer-
ation. Anti-Cancer Drugs, 13, 301–311 (2002).
15) Kumura, N., Izumi, M., Nakajima, S., Shimizu, S., Kim,
H.-S., Wataya, Y., and Baba, N., Synthesis and bio-
logical activity of fatty acid derivatives of quninine.
Biosci. Biotechnol. Biochem., 69, 2250–2253 (2005).
16) Parang, K., Wiebe, L. I., and Knaus, E. E., Novel
We are obliged to the laboratory of SC-NMR and
Mass Spectrometry (API III) of Okayama University
and to Ikeda Industries Co., Hiroshima, Japan for
presnting of DHA and EPA.
References
1)
2)
3)
Kopka, M. L., Yoon, C., Goodsell, D., Pjura, P., and
Dickerson, R. E., The molecular origin of DNA-drug
specificity in netropsin and Distamycin. Proc. Natl.
Acad. Sci., 82, 1376–1380 (1985).
Kopka, M. L., Yoon, C., Goodsell, D., Pjura, P., and
Dickerson, R. E., Binding of an antitumor drug to DNA,
netropsin and C-G-C-G-A-A-T-T-BrC-G-C-G. J. Mol.
Biol., 183, 553–563 (1985).
White, S., Baird, E. E., and Dervan, P. B., Orientation
preferences of pyrrole-imidazole polyamides in the
minor groove of DNA. J. Am. Chem. Soc., 119, 8756–
0
0
8765 (1997).
approaches for designing 5 -O-ester prodrugs of 3 -
0
0
4
)
)
Gottesfeld, J. M., Neely, L., Trauger, J. W., Baird, E. E.,
and Dervan, P. B., Regulation of gene expression by
small molecules. Nature, 387, 202–205 (1997).
Sugiyama, H., Lian, C., Isomura, M., Saito, L., and
Wang, H.-J., Distamycin A modulates the sequence
specificity of DNA alkylation by duocarmycin A. Proc.
Natl. Acad. Sci., 93, 14405–14410 (1996).
azido-2 ,3 -dideoxythymidine (AZT). Curr. Med. Chem.,
7, 995–1039 (2000).
17) Kumar, R., and Lown, J. W., Synthesis and cytotoxicity
evaluation of novel C7–C7, C7–N3 and N3–N3 dimers
of 1-chloromethyl-5-hydroxy-1, 2-dihydro-3H-ben-
zo[e]indole (seco-CBI) with pyrrole and imidazole
polyamide conjugates. Org. Biomol. Chem., 3, 2630–
2647 (2003).
18) Thomas, M., Varshney, U., and Bhattacharya, S.,
Distamycin analogues without leading amide at their
N-termini—Comparative binding properties to AT- and
GC-rich DNA sequences. Eur. J. Org. Chem., 2002,
3604–3615 (2002).
19) Otake, T., Mori, H., Morimoto, M., Ueba, N., Sutardjo,
S., Kusumoto, T., Hattori, M., and Mamba, T., Screening
of indonesian plant extracts for anti-human immunode-
ficiencyvirus-type 1 (HIV-1) activity. Phytother. Res., 9,
6–10 (1995).
20) Kimpton, J., and Emerman, M., Detection of replication-
competent and pseudotyped human immunodeficiency
virus with a sensitive cell line on the basis of activation
of an integrated ꢂ-galactosidase gene. J. Virol., 66,
2232–2239 (1992).
5
6)
Dickinson, L. A., Gulizia, R. J., Trauger, J. W., Baird, E.
E., Mosier, D. E., Gottesfeld, J. M., and Dervan, P. B.,
Inhibition of RNA polymerase II transcription in human
cells by synthetic DNA-binding ligands. Proc. Natl.
Acad. Sci. USA, 95, 12890–12895 (1998).
7
8
)
)
Stillwell, W., Ehringer, W., and Jenski, L. J., Docosa-
hexaenoic acid increases permeability of lipid vesicles
and tumor cells. Lipids, 28, 103–108 (1993).
Begin, M. E., Ellis, G., and Horrobin, D. F., Polyunsa-
turated fatty acid-induced cytotoxicity against tumor
cells and its relationship to lipid peroxidation. J. Natl.
Cancer Inst., 80, 188–194 (1988).
9)
Smets, L. A., Rooij, H. V., and Salomons, G. S.,
Signaling steps in apoptosis by ether lipids. Apoptosis, 4,
421–427 (1999).
1
0) N o¨ ding, R., Schonberg, S. A., Krokan, H. E., and Bjerve,
K. S., Effects of polyunsaturated fatty acids and their n-6
hydroperoxides on growth of five malignant cell lines
and the significance of culture media. Lipids, 33, 285–
21) Hepburn, C., and Mahdi, M. S., Amine bridged
amides(aba s). Plast. Rub. Proces. Appl., 4, 343–348
(1984).