close

Вход

Забыли?

вход по аккаунту

?

Stilbene-Based Inhibitors of Estrone Sulfatase with a Dual Mode of Action in Human Breast Cancer Cells.

код для вставкиСкачать
634 von Angerer et al.
Georg Walter,
Renate Liebl,
Erwin von Angerer
Institut für Pharmazie,
Universität Regensburg,
Regensburg, Germany
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Stilbene-Based Inhibitors of Estrone Sulfatase
with a Dual Mode of Action in Human Breast
Cancer Cells
Estrone sulfate (E1S) is an endogenous prodrug that delivers estrone and,
subsequently, estradiol to target cells, after hydrolysis by the enzyme estrone
sulfatase, which is active in various tissues including hormone-dependent breast
cancer. Blockade of this enzyme should reduce the estrogen level in breast
cancer cells and prevent hormonal growth stimulation. In this study, a number
of sulfamoyloxy-substituted stilbenes with side chains that guarantee antiestrogenic activity were synthesized and evaluated as inhibitors of estrone sulfatase.
They inhibited this enzyme in human MDA-MB 231 breast cancer cells, with
IC50 values in the submicromolar range. The effects of both the free hydroxy
derivatives and the sulfamates on gene activation were determined in
transfected MCF-7/2a breast cancer cells stimulated either with estradiol or with
estrone sulfate. The analysis of data revealed a dual mode of action of the
majority of compounds. They blocked gene expression by inhibition of estrone
sulfatase and by antiestrogenic action. This pharmacological profile was also
observed in assays on antiproliferative activity. The most potent derivative 8g
inhibited the growth of wild-type human MCF-7 cells with an IC50 value of 13 nM.
Keywords: Stilbene Sulfamates; Steroid Sulfatase; Antiestrogen; Breast
Cancer Cells
Received: May 5, 2004; Accepted: November 3, 2004 [FP904]
DOI 10.1002/ardp.200400904
Introduction
Since estrogens are known to play a dominant role in
the promotion of breast cancer, several strategies
have been developed to reduce the mitogenic effects
of estrogens on mammary carcinoma cells. Blockade
of the estrogen receptor (ER) by antiestrogens and inhibition of estrogen biosynthesis by aromatase inhibitors are therapeutic options that have proved effective
in the treatment of postmenopausal patients, although
number and duration of remissions are far from being
acceptable. Studies on estrogen metabolism in man
have identified estrone sulfate as the predominant
form of circulating estrogens, which has also been
detected in breast cancer tissue as the main estrogen
[1, 2]. Mammary tumor cells have been shown to be
capable of cleaving this conjugate to free estrone,
which can subsequently be converted by 17β-HSD to
17β-estradiol [3, 4]. It has been demonstrated that the
steroid sulfatase present in breast cancer cells plays
a more important role in the formation of free estrogens than the enzyme aromatase [2, 5]. In mammary
Correspondence: Erwin von Angerer, Institut für Pharmazie,
Universität Regensburg, 93040 Regensburg, Germany;
Phone: +49 941 943-4821, Fax: +49 941 943-4820, e-mail:
erwin.von-angerer@chemie.uni-regensburg.de
tumors, the levels of free and conjugated estrogens,
as well as the sulfatase activity, are significantly higher
than in normal tissues [1, 6]. High expression of
steroid sulfatase mRNA has been associated with
poor prognosis in patients with ER-positive breast cancer [7]. These findings have stimulated the search for
inhibitors of steroid sulfatase as a new therapeutic
option or as co-medication in endocrine therapy of
patients with mammary carcinomas.
The obvious starting point is the natural substrate estrone sulfate, which was first modified chemically in
the 3-position [8⫺10]. The most favorable substituent
in respect to enzyme inhibition proved to be the sulfamoyloxy group, but the inherent estrogenic potency
[11] made this derivative (1, EMATE; Figure 1) unsuitable for further development as a sulfatase inhibitor.
Meanwhile a variety of steroidal [12, 13] and non-steroidal sulfamates [14, 15] have been synthesized and
evaluated as enzyme inhibitors [16]. Examples for
non-steroidal inhibitors are tricyclic coumarin derivatives such as compound 2 (667-COUMATE) [17], 2adamantylthiochromenone sulfamate (3) [18], and the
phenyl sulfamate 4 [19] with a lipophilic residue in the
para-position of the aromatic ring. Recently, we have
shown that 2-phenylindole derivatives such as com-
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Stilbene-Based Inhibitors of Estrone Sulfatase 635
Figure 1. Chemical structures of steroidal and non-steroidal inhibitors of estrone sulfatase.
pound 5, with a sulfamoyl function in the phenyl ring
and a long alkyl chain in the indole moiety, act as sulfatase inhibitors without estrogenic side effects.
Since these sulfamates can act both as inhibitors and
as substrates of steroid sulfatase, the bioactivity of the
free phenols is of great importance and has to be considered in the design of new sulfatase inhibitors for
therapeutic application. Therefore, we developed the
concept of antiestrogen-based inhibitors of the enzyme steroid sulfatase [20]. We were able to demonstrate that non-steroidal antiestrogens of appropriate
structure can be converted into estrone sulfatase inhibitors after modification of the phenolic hydroxy
groups by sulfamoylation. An example for this type of
inhibitors is 6, which derives from the pure antiestrogen ZK 164.015 [21]. An important finding of this earlier study was that the enzyme inhibitory activity of the
2-phenylindole derivatives increases in parallel to the
estrogenic potency of the parent structure. This observation can be rationalized by overlapping structural requirements for binding to the ER and to the active site
of steroid sulfatase. This consideration prompted us to
use diethylstilbestrol (DES) as a non-steroidal substitute of estradiol and modify its structure to implement
both antiestrogenic activity and estrone sulfatase inhibition. In this study, we converted one (7) or both (8)
hydroxy functions into sulfamoyloxy groups, introduced three different side chains and varied the position of the aromatic substituents.
The new sulfamates were tested for enzyme inhibitory
activity in human MDA-MB 231 mammary carcinoma
cells, for estrogenic and antiestrogenic effects as well
as blockade of estrone sulfate action in stably
transfected MCF-7/2a breast cancer cells, and for antiproliferative activity in wild-type MCF-7 cells stimulated with estrone sulfate.
Chemistry
Synthesis of the stilbene derivatives started from the
corresponding desoxyanisoins 9 (Scheme 1). The first
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
636 von Angerer et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Scheme 1. (a) 1. NaH, DMF, 0 °C; 2. R2CH2Br, room temperature, 2 h, 62⫺86 %; (b) R3CH2MgBr, Et2O, reflux,
2 h, 33⫺76 %; (c) BBr3, CH2Cl2, ⫺5 °C to room temperature, 7 h, 52⫺95 %; (d) 5 equiv. H2NSO2Cl, DMF, room
temperature, 12 h, 47⫺97 %.
side chain was introduced by deprotonation and subsequent reaction with ethyl bromide or the bromo alkane with the respective sulfur function to give the ketones 10. The second substituent was introduced by a
Grignard reaction, which led to the formation of a
double bond with orientation towards the side chain
(11). Since the acidity of the sulfone prevented its direct use as the Grignard reagent, the thio ether function had to be oxidized with m-CPBA after the Grignard reaction to give 11h. Cleavage of the methoxy
groups in 11 led to mixtures of the stereoisomeric
phenols 12 and 13, with preference for the stilbene
structure 12.
All of the stilbenes 12 were obtained as E/Z mixtures,
with the E-stereoisomer as the dominant product. The
structural assignment was made on the basis of the
[1H]-NMR spectra recorded in d6-DMSO. Diethylstilbestrol (12b) as the pure E-isomer gave rise to a
broad singlet at δ 9.42 ppm for the two phenolic hydrogens, whereas the regioisomer 12b showed two
singlets at δ 9.31 and 9.32 ppm for the E-stereoisomer
and two for the Z-form at δ 9.00 and 9.08 ppm. The
ratio of both groups was 86:14. Similar chemical shifts
for the phenolic hydrogens were observed for the
other hydroxy derivatives 12. HPLC studies revealed
a ratio of E-12 to Z-12 of approximately 85:15, which
was in accord with the results from the NMR spectra.
When the stereoisomers had been separated by
HPLC, they rapidly isomerized to give the original ratio
of isomers. In the final reaction step, the phenolic compounds were treated with an excess of sulfamoyl chloride to give the corresponding disulfamates 8, generally as the E-isomers. In some cases, the monosulfamates 7 were obtained as byproducts. However, only
two representatives of the monosulfamates (7b, 7h)
were included in this study.
Results and discussion
Since the sulfamates synthesized in this study can act
by a dual mode, either by inhibiting the enzyme estrone sulfatase or by binding to the estrogen receptor
after chemical or enzymatic hydrolysis, a new strategy
for testing was elaborated that allows the quantification of both pharmacological actions. First, all new
sulfamates were tested for inhibitory activity on estrone sulfatase. An appropriate source for this enzyme
are human breast cancer cells, which express this enzyme in sufficient quantities [22, 23] and might be
more relevant than placental microsomes used by
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Stilbene-Based Inhibitors of Estrone Sulfatase 637
Table 1. Inhibition of estrone sulfatase by stilbene-based sulfamates 7a, 7h, and 8a⫺h.
Compound
R1
8a
3-OSO2NH2
8b
4-OSO2NH2
7b
4-OH
8c
3-OSO2NH2
8d
4-OSO2NH2
8e
3-OSO2NH2
8f
3-OSO2NH2
8g
4-OSO2NH2
8h
3-OSO2NH2
7h
3-OH
1 (EMATE)
6 (ZK 164,015-sulfamate)
‡
R2
R3
IC50‡ (nM)
Me
Me
Me
-(CH2)9-S-C5H11
-(CH2)9-S-C5H11
Me
-(CH2)9-SO2-C5H11
-(CH2)9-SO2-C5H11
Me
Me
Me
Me
Me
Me
Me
-(CH2)9-S-C5H11
Me
Me
-(CH2)9-SO2-C5H11
-(CH2)9-SO2-C5H11
1.7
0.67
34
313
23
63
126
12
21
990
0.3
200
Inhibitory effect on the conversion of [3H]estrone sulfate (2 nM) to [3H]estrone by MDA-MB 231 cells; mean
of triplicates.
others. In a previous study, we used estrogen-sensitive MCF-7 cells, which were replaced later by the hormone-independent MDA-MB 231 cell line whose proliferation cannot be influenced by hormonal effects of
the test compounds. Enzyme inhibition was determined in a whole-cell assay [24] because intact cells
were also used in the following assays. Cells were incubated with [3H]estrone sulfate in the presence of an
inhibitor for 20 h. Enzyme activity was estimated by
the amount of [3H]estrone formed during incubation
and extracted with toluene. The extraction yield was
quantified by the addition of 14C-labeled estrone and
measurement of both radioactive nuclei. The inhibitory
activity of the new sulfamates varied over a wide range
(Table 1). As expected, the strongest effect was observed for the bis-sulfamate of diethylstilbestrol, with
an IC50 value close to the reference drug 1 (EMATE).
Obviously, both sulfamoyl groups are required for
strong inhibition. All of the monosulfamates 7 studied
displayed inhibitory effects two orders of magnitude
lower than those of the disulfamates.
Comparison of the six derivatives with sulfur-containing long side chains revealed that para-substitution in
both aromatic rings is most favorable for inhibition.
Shifting the sulfamoyloxy group in one of the rings to
the meta-position had only a major effect when this
modification was performed in the ring vicinal to the
long side chain. The kind of functional group in the
side chain had no pronounced effect on enzyme inhibition, contrary to the binding affinity for the estrogen
receptor (Table 2).
Since sulfamates are susceptible to chemical and
possibly enzymatic hydrolysis [25], the corresponding
hydroxy derivatives might contribute to the biological
activity. Thus, estrogenic and antiestrogenic activities
of the free phenols were detemined in MCF-7/2a cells
stably transfected with a reporter gene under the control of an estrogen-responsive element (ERE) [26]. As
expected, DES and its isomer 12a acted as potent
estrogens, whereas the other stilbene derivatives
behaved as pure antiestrogens without any agonistic
activity. At a concentration of 10⫺6 M, they exhibited
values for luciferase activity below that of control cells
grown in steroid-depleted medium (Table 2). Luciferase activities below baseline are characteristic of pure
antiestrogens and indicate the blockade of ligand-independent activation of the ER, responsible for the basal luciferase activity in control cells [26]. All IC50 values were in the submicromolar range except for 12c
(IC50 = 1.0 µM).
This assay was also applied to study the effect of the
free hydroxy derivatives 12cⴚh on estrone sulfatestimulated gene activation (Table 2). Since the phen-
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
638 von Angerer et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Table 2. Hormonal effects of hydroxylated stilbene derivatives 12a⫺h.
Compound
RBA‡
Estrogenic
activity†
(% of E1)
Antiestrogenic
activity§
IC50 (µM)
Inhibition of E1
sulfate action#
IC50 (nM)
12a
12b (DES)
12c
12d
12e
12f
12g
12h
33
67
0.03
0.11
0.07
0.87
2.44
1.72
98
123
⫺10
⫺11
⫺11
⫺11
⫺11
⫺11
⫺¥
⫺¥
1.00
0.73
0.20
0.37
0.13
0.05
⫺¥
⫺¥
130
110
23
55
18
6
‡
†
§
#
¥
Relative binding affinities for the calf uterine ER, determined by incubation at 4 °C for 20 h. RBA value for 17βestradiol (E2) = 100.
Stimulation of luciferase activity by test compounds (1 µM) in stably transfected MCF-7/2a cells, given in % of
the value for estrone (E1; 1 nM). Basal activity (12 % of E1) was subtracted from all values.
Inhibition of luciferase activiy by test compounds in stably transfected MCF-7/2a cells stimulated with 1 nM E2;
mean values of two independent experiments with six replicates; SD are less than 10 %.
Inhibition of luciferase activity, stimulated with 100 nM estrone sulfate, in stably transfected MCF-7/2a cells;
mean values of two independent experiments with six replicates; SD are less than 10 %.
No inhibition.
ols do not inhibit estrone sulfatase (data not shown)
they can only act as antiestrogens and antagonize estrone and estradiol formed enzymatically from estrone
sulfate. Comparison of IC50 values obtained with estradiol and estrone sulfate as the agonist revealed that
about 0.13 % of the estrone sulfate had been converted to estradiol during incubation. This figure is important for the subsequent evaluation of the stilbenebased sulfamates.
All sulfamates deriving from antiestrogens exhibited
antiestrogenic activity when tested in MCF-7/2a cells
stimulated with estradiol. The effect was lower than
that of the hydroxy analogs and indicates a partial hydrolysis of sulfamates in the range of 5⫺20 % (Table
3). The strong inhibition of estrone sulfate-stimulated
gene expression results from both sulfatase inhibition
and antiestrogenic effects. We analyzed the data to
determine the proportion of each action for the total
effect. Two different hydrolysis reactions have to be
considered: (i) enzymatic cleavage of estrone sulfate
to the active estrogen and (ii) conversion of the sulfamates to the active antiestrogens. Generally, the calculated IC50 values for the antiestrogenic effect of the
sulfamates ⫺ or more precisely of the hydrolyzed fraction ⫺ were higher than the observed values, indicating a contribution of enzyme inhibition. In Table 3, this
contribution is shown as enhancement of the inhibitory
activity and is given as the factor by which the IC50
values decreased in comparison to the calculated antiestrogenic effect. For compounds 8c and 8f, enzyme
inhibition plays hardly a role (1.2-fold enhancement),
whereas all other sulfamates obviously act by a dual
mode of action. The sulfatase-inhibiting activity led to
a decrease of the IC50 values by factors between 2.1
and 3.6. These results are in accord with the inhibitory
activities of the sulfamates on estrone sulfatase (Table
1). The IC50 values of 8c and 8f were by one order of
magnitude higher than those of the other sulfamates.
The primary goal of these investigations is the discovery of new agents with potential for treatment of
estrogen-dependent malignancies. Thus, all sulfamates were tested for antiproliferative activity in estrogen-sensitive MCF-7 breast cancer cells. The experimental setting was similar to that for gene activation,
except that cell mass was used as parameter instead
of gene activation. Under the assumption that there is
no great difference in the rate of hydrolysis between
the two cell lines, the same calculations were performed. As expected, the results from both assays
were rather similar (Table 4). Only a weak contribution
of enzyme inhibition, if any, was noticed for 8c and 8f,
which both possess the same basic structure. The
most potent derivative was 8g with an IC50 value of
13 nM, which is mainly due to the contribution of enzyme inhibition (6.9-fold enhancement of the antiestrogenic effect).
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Stilbene-Based Inhibitors of Estrone Sulfatase 639
Table 3. Antiestrogenic and enzyme inhibitory effects of stilbene-derived sulfamates 8c⫺h in MCF-7/2a cells.
Compound
8c
8d
8e
8f
8g
8h
Antiestrogenic
activity‡
IC50 (µM)
Inhibition of E1
sulfate action†
IC50 (µM)
Calculated antiestrogenic
effect on E1 sulfate action§
IC50 (µM)
Hydrolysis of
sulfamates#
(%)
Calculated effect of
sulfatase inhibition##
(fold enhancement)
5.0¥
4.0¥
3.0¥
2.0¥
1.3
0.88
0.55
0.15
0.12
0.23
0.065
0.055
0.67
0.53
0.40
0.27
0.17
0.12
11
18
6.7
19
10
5.6
1.2
3.6
3.3
1.2
2.7
2.1
‡
Inhibition of luciferase activity by test compounds in stably transfected MCF-7/2a cells stimulated with 1 nM
E2; mean values of two independent experiments with six replicates; SD are less than 10 %.
†
Inhibition of luciferase activity, stimulated with 100 nM estrone sulfate, in stably transfected MCF-7/2a cells;
mean values of two independent experiments with six replicates; SD are less than 10 %.
§
Values of column 2 were multiplied by 0.133 to quantify the inhibitory effect of the sulfamates on the action of
estrogens released from E1 sulfate (see Table 2).
#
Estimated from the values of hydroxy derivatives 12 and sulfamates 8 for antiestrogenic activity.
##
Ratio between corrected IC50 values for antiestrogenic activity and IC50 values for the total effect of the sulfamates (columns 4 and 3) as measure of enhancement of antiestrogenic action.
¥
Values >1.5 µM were estimated by extrapolation.
Table 4. Antiestrogenic and enzyme inhibitory effects of stilbene-derived sulfamates 8c⫺h on the proliferation of
estrogen-sensitive MCF-7 breast cancer cells.
Compound
8c
8d
8e
8f
8g
8h
‡
†
§
#
Antiestrogenic
effect‡
IC50 (µM)
Inhibition of E1
sulfate action†
IC50 (µM)
Calculated antiestrogenic
effect on E1 sulfate action§
IC50 (µM)
Calculated effect of
sulfatase inhibition#
(fold enhancement)
5.9
3.3
3.9
2.0
6.8
0.8
1.8
0.15
0.19
0.18
0.013
0.034
0.79
0.44
0.52
0.27
0.09
0.11
0.44
2.9
2.7
1.5
6.9
3.2
Growth inhibition of wild-type MCF-7 breast cancer cells stimulated with E2 (10⫺9 M).
Growth inhibition of MCF-7 cells stimulated with 100 nM estrone sulfate.
Values of column 2 were multiplied by 0.133 to quantify the inhibitory effect of the sulfamates on the action of
estrogens released from E1 sulfate (see Table 2).
Ratio between corrected IC50 values for antiestrogenic activity and IC50 values for the total effect of the sulfamates (columns 4 and 3).
One of the interesting aspects of sulfamates is their
ability of irreversibly inhibiting estrone sulfatase. This
feature is also found for the stilbene-based sulfamates
8, which completely inactivated the enzyme after a
2⫺4-h preincubation period at a concentration of
1 µM, except for 8c which only gave rise to 80 % inhi-
bition (data not shown). The inhibition of the enzyme
is a long-lasting effect. Even 169 h after the incubation
period, the enzyme had not recovered and no enzyme
activity could be detected, except in the case of 8f
which was one of the weak inhibitors (data not shown).
The results of this study clearly showed that di-
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
640 von Angerer et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Acknowledgments
The authors wish to thank the Deutsche Forschungsgemeinschaft for financial support.
Experimental
General methods
Melting points were determined on a Büchi 510 apparatus
and are uncorrected. NMR spectra were recorded on a
Bruker AC-250 spectrometer with TMS as internal standard
and DMSO-d6 as solvent, and were in accord with the assigned structures. Elemental analyses of crystalline compounds were performed by the Mikroanalytisches Laboratorium (University of Regensburg, Germany) and were within
0.4 % of the calculated values. Purity and ratio of E/Z-isomers
of final products were checked by HPLC (RP-18, 20 ⫻ 4 mm)
with MeCN/buffer mixtures as eluent (buffer: 0.5 % NEt3 adjusted to pH 7.0 with H3PO4) and UV detection at λ = 250 nm.
General procedure for alkylation of desoxyanisoins
Figure 2. Dual mode of action of antiestrogen-derived
inhibitors of estrone sulfatase on the proliferation of
hormone-dependent breast cancer cells.
hydroxystilbenes are appropriate for the synthesis of
new steroid sulfatase inhibitors after conversion of the
two phenolic hydroxy groups to sulfamates. Despite
the presence of a long side chain with a functional
group, they inhibit the human estrone sulfatase with
IC50 values in the nanomolar range. Interestingly, both
sulfamoyloxy groups are required for strong inhibition,
although only one interacts with the active site. This
observation is in accord with our previous findings that
a second polar element is required for strong inhibition
[20]. Careful analysis of the results from the assays
performed showed that four of the sulfamates (8d, e,
g, h) inhibited the growth of estrogen-sensitive MCF7 breast cancer cells by a dual mode of action (Figure
2). Primarily, they inhibited the enzyme estrone sulfatase and thereby blocked the release of active estrogens such as estrone and estradiol. These sulfamates
can also undergo hydrolysis to give the free phenolic
derivatives that bind to the ER and block gene activation. The proportion of these two mechanisms can
vary, depending on the relative concentrations of estrone sulfate and free estrogens. In combination, they
decrease intracellular levels of active estrogens and
block their action via the ER.
Under N2, a solution of 5.0 mmol desoxyanisoin (9) in 25 mL
dry DMF was added slowly, with stirring, to an ice-cold suspension of 7.0 mmol NaH in dry DMF. Stirring was continued
until the gas evolution ceased. Then, 5.0 mmol alkyl halogenide in dry DMF (20 mL) was added dropwise, with cooling in
an ice bath. After addition, the ice bath was removed and
stirring continued for another 2 h at room temperature. The
excess of NaH was destroyed carefully by dropwise addition
of water, followed by the addition of 50 mL water and 50 mL
EtOAc. The organic layer was separated, and the aqueous
layer was extracted three times with EtOAc. The combined
organic layers were washed with water and dried (MgSO4).
The solvent was removed in vacuo and the residue purified
by chromatography (SiO2).
(+/⫺)-1-(4-Methoxyphenyl)-2-(3-methoxyphenyl)butan-1-one
(10a)
Colorless oil, yield 62 %. 1H-NMR δ = 0.82 (t, 3J = 7.3 Hz,
3H, CH2-CH3), 1.71 and 2.02 (AB of quin, 2J = 13.5 Hz, 3J =
7.3 Hz, 2H, CH-CH2-CH3), 3.71 (s, 3H, OCH3), 3.80 (s, 3H,
OCH3), 4.65 (t, 3J = 7.3 Hz, 1H, CH-CH2-CH3), 6.74⫺7.23
(m, 4H, ArH), 6.98 and 8.01 (AA⬘BB⬘, 3J = 8.9 Hz, 4H, ArH).
(+/⫺)-1-(4-Methoxyphenyl)-2-(3-methoxyphenyl)-12-pentylthiododecan-1-one (10c)
Colorless oil, yield 80 %. 1H-NMR δ = 0.85 (t, 3J = 6.9 Hz;
3H, -CH2-CH3), 1.13⫺1.36 (m, 18H, -(CH2)2-CH3, S-(CH2)2(CH2)7-), 1.42⫺1.49 (m, 4H, -CH2-CH2-S-CH2-CH2-),
1.52⫺2.08 (m, 2H, -CO-CH-CH2-), 2.44 (t, 3J = 7.1 Hz, 4H,
-CH2-S-CH2-), 3.70 (s, 3H, OCH3), 3.80 (s, 3H, OCH3), 4.71
(t, 3J = 7.2 Hz, 1H, -CO-CH-CH2-), 6.73⫺7.22 (m, 4H, ArH),
6.98 and 8.01 (AA⬘BB⬘, 3J = 8.9 Hz, 4H, ArH).
(+/⫺)-1,2-Bis-(4-methoxyphenyl)-12-pentylthio-dodecan-1one (10d)
Colorless oil, yield 86 %. 1H-NMR δ = 0.85 (t, 3J = 6.9 Hz,
3H, -CH2-CH3), 1.20⫺1.30 (m, 18H, -(CH2)2-CH3 and
-S-(CH2)2-(CH2)7-), 1.30⫺1.36 (m, 4H, -CH2-CH2-S-CH2CH2-), 1.58⫺2.01 (m, 2H, -CO-CH-CH2-), 2.45 (t, 3J = 7.2
Hz, 4H, -CH2-S-CH2-), 3.68 (s, 3H, OCH3), 3.80 (s, 3H,
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Stilbene-Based Inhibitors of Estrone Sulfatase 641
OCH3), 4.67 (t, 3J = 7.2 Hz, 1H, -CO-CH-CH2-), 6.83 and
7.22 (AA⬘BB⬘, 3J = 8.7 Hz, 4H, ArH), 6.97 and 7.99 (AA⬘BB⬘,
3
J = 8.9 Hz, 4H, ArH).
(+/⫺)-1-(4-Methoxyphenyl)-2-(3-methoxyphenyl)-12-pentylsulfonyldodecan-1-one (10f)
Colorless oil, yield 80 %. 1H-NMR δ = 0.87 (t, 3J = 7.5 Hz,
3H, -CH2-CH3), 1.15⫺1.37 (m, 18H, -(CH2)2-CH3 and -SO2(CH2)2-(CH2)7-), 1.62⫺1.69 (m, 4H, -CH2-CH2-S-CH2-CH2-),
1.62⫺2.07 (m, 2H, -CO-CH-CH2-), 3.03 (t, 3J = 7.7 Hz, 4H,
-CH2-SO2-CH2-), 3.70 (s, 3H, OCH3), 3.80 (s, 3H, OCH3),
4.71 (t, 3J = 7.1 Hz, 1H, -CO-CH-CH2-), 6.74⫺7.22 (m, 4H,
ArH), 6.98 and 8.01 (AA⬘BB⬘, 3J = 8.9 Hz, 4H, ArH).
(+/⫺)-1,2-Bis-(4-methoxyphenyl)-12-pentylsulfonyldodecan1-one (10g)
Colorless oil, yield 86 %. 1H-NMR δ = 0.87 (t, 3J = 6.9 Hz,
3H, -CH2-CH3), 1.15⫺1.38 (m, 18H, -(CH2)2-CH3 and
-S-(CH2)2-(CH2)7-), 1.58⫺1.66 (m, 4H, -CH2-CH2-S-CH2CH2-), 1.58⫺2.06 (m, 2H, -CO-CH-CH2-), 3.03 (t, 3J = 7.8
Hz, 4H, -CH2-S-CH2-), 3.68 (s, 3H, OCH3), 3.80 (s, 3H,
OCH3), 4.67 (t, 3J = 7.2 Hz, 1H, -CO-CH-CH2-), 6.83 and
7.22 (AA⬘BB⬘, 3J = 8.7 Hz, 4H, ArH), 6.97 and 7.99 (AA⬘BB⬘,
3
J = 8.9 Hz, 4H, ArH).
General procedure for introduction of the second side chain
Following start of the reaction by addition of an iodine crystal
or local heating, a solution of 5.0 mmol alkyl halide in dry
Et2O (10 mL) was added dropwise to Mg turnings (5.0 mmol)
in dry Et2O (10 mL). After addition, the mixture was kept
refluxing for 1 h. After cooling, a solution of the ketone 2
(5 mmol) in dry Et2O (10 mL) was added slowly, followed by
heating (2 h) under reflux. After cooling and dropwise addition
of 2N HCl, the mixture was extracted three times with THF.
The combined organic layers were washed with NaHCO3
solution and water, and dried (MgSO4). Following evaporation
of the solvent, the residue was purified by chromatography.
(E/Z)(+/⫺)-3-(4-Methoxyphenyl)-4-(3-methoxyphenyl)hex-2ene (11a)
Colorless oil, yield 58 %. 1H-NMR δ = 0.82 (t, 3J = 7.3 Hz,
3H, -CH-CH2-CH3), 1.47 and 1.87 (d, 3J = 6.7 Hz, 3H, =CHCH3), 1.57⫺1.81 (m, 2H, -CH-CH2-CH3), 3.33⫺3.41 (m, 1H,
-CH-CH2-CH3), 3.68 (s, 3H, OCH3), 3.71 (s, 3H, OCH3),
5.56⫺5.65 (m, 1H, =CH-CH3), 6.61⫺7.21 (m, 8H, ArH).
(E/Z)-3-(4-Methoxyphenyl)-4-(3-methoxyphenyl)-14-pentylthiotetradec-3-ene (11c)
Colorless oil, yield 76 %. 1H-NMR δ = 0.39 (t, 3J = 7.4 Hz)
and 0.69 (t, 3J = 7.1 Hz) (3H, =C-CH2-CH3), 0.85 (t, 3J =
6.8 Hz, 3H, -(CH2)4-CH3), 1.12⫺1.36 (m, 18H, -(CH2)2-CH3,
S-(CH2)2-(CH2)7-), 1.41⫺1.94 (m, 8H, -CH2-CH2-S-CH2-CH2-,
-CH2-C=C-CH2-), 2.44 (t, 3J = 7.1 Hz, 4H, -CH2-S-CH2-), 3.59
and 3.69 (s, 3H, OCH3), 3.73 and 3.75 (s, 3H, -OCH3),
6.38⫺7.36 (m, 8H, ArH).
(E/Z)(+/⫺)-3,4-Bis-(4-methoxyphenyl)-14-pentylthiotetradec2-ene (11d)
Colorless oil, yield 66 %. 1H-NMR δ = 0.85 (t, 3J = 7.0 Hz,
3H, -(CH2)4-CH3), 1.19⫺1.33 (m, 18H, -(CH2)2-CH3, -S(CH2)2-(CH2)7-), 1.43⫺1.74 (m, 4H, -CH2-CH2-S-CH2-
CH2-), 1.44 and 1.84 (d, 3J = 6.6 Hz, 3H, C=CH-CH3),
1.58⫺2.01 (m, 2H, C=C-CH-CH2-), 2.45 (t, 3J = 7.3 Hz, 4H,
-CH2-S-CH2-), 3.67 and 3.72 (3H, OCH3), 3.697 and 3.702
(s, 3H, OCH3), 3.36⫺3.39 (m, 1H, C=C-CH-CH2-), 5.48⫺5.56
(m, 1H, C=CH-CH3), 6.71 and 7.08 (m, 8H, ArH).
(E/Z)(+/⫺)-3-(3-Methoxyphenyl)-4-(4-methoxyphenyl)-14pentylthiotetradec-4-ene (11e)
Colorless oil, yield 33 %. 1H-NMR δ = 0.43 (t, 3J = 7.4 Hz)
and 0.56 (t, 3J = 7.2 Hz) (3H, C=C-CH-CH2-CH3), 0.82 and
0.85 (t, 3J = 7.3 Hz, 3H, -(CH2)4-CH3), 1.14⫺1.36 (m, 18H,
-(CH2)2-CH3, -S-(CH2)2-(CH2)7-), 1.42⫺1.60 (m, 4H, -CH2CH2-S-CH2-CH2-), 1.61⫺2.08 (m, 2H, =C-CH-CH2-CH3),
2.44 and 2.45 (t, 3J = 7.6 Hz, 4H, -CH2-S-CH2-), 3.32⫺3.35
(m, 1H, C=C-CH-CH2-CH3), 3.59 and 3.68 (s, 3H, OCH3),
3.70 and 3.80 (s, 3H, OCH3), 4.48⫺4.67 (m, 1H, C=CH(CH2)9-), 6.37⫺8.04 (m, 8H, ArH).
(E/Z)(+/⫺)-3-(4-Methoxyphenyl)-4-(3-methoxyphenyl)-14pentylsulfonyltetradec-2-ene (11f)
Colorless oil, yield 73 %. 1H-NMR δ = 0.87 (t, 3J = 7.1 Hz,
3H, -(CH2)4-CH3), 1.20⫺1.37 (m, 18H, -(CH2)2-CH3, -S(CH2)2-(CH2)7-), 1.46 (d, 3J = 6.7 Hz) and 1.86 (d, 3J = 6.9
Hz) (3H, C=CH-CH3), 1.63⫺1.69 (m, 4H, -CH2-CH2-SO2CH2-CH2-), 1.63⫺2.10 (m, 2H, -CH2-CH-C=C), 3.03 (t, 3J =
7.7 Hz, 4H, -CH2-SO2-CH2-), 3.56 (t, 3J = 8.4 Hz, 1H, -CH2CH-C=C), 3.68 and 3.71 (s, 3H, OCH3), 3.72 and 3.84 (s,
3H, -OCH3), 5.57⫺5.61 (m, 1H, C=CH-CH3), 6.61⫺7.24 (m,
8H, ArH).
(E/Z)(+/⫺)-3,4-Bis-(4-methoxyphenyl)-14-pentylsulfonyltetradec-2-ene (11g)
Colorless oil, yield 54 %. 1H-NMR δ = 0.87 (t, 3J = 6.9 Hz,
3H, -(CH2)4-CH3), 1.15⫺1.38 (m, 18H, -(CH2)2-CH3, -SO2(CH2)2-(CH2)7-), 1.58⫺1.70 (m, 4H, -CH2-CH2-SO2-CH2CH2-), 1.44 (d, 3J = 6.6 Hz) and 1.86 (d, 3J = 6.8 Hz) (3H,
C=CH-CH3), 1.58⫺1.99 (m, 2H, C=C-CH-CH2-), 3.03 (t, 3J =
7.7 Hz, 4H, -CH2-SO2-CH2-), 3.68 and 3.72 (s, 3H, OCH3),
3.698 and 3.704 (s, 3H, OCH3), 3.33⫺3.43 (m, 1H, C=C-CHCH2-), 5.53⫺5.56 (m, 1H, C=CH-CH3), 6.65 and 7.01 (m,
8H, ArH).
(E/Z)(+/⫺)-3-(3-Methoxyphenyl)-4-(4-methoxyphenyl)-14pentylsulfonyltetradec-4-ene (11h)
Prepared by oxidation of 3e with meta-chloroperbenzoic acid.
Colorless oil, yield 76 %. 1H-NMR δ = 0.43 (t, 3J = 6.2 Hz)
and 0.55 (t, 3J = 7.3 Hz) (3H, C=C-CH-CH2-CH3), 0.87 (t, 3J =
7.1 Hz, 3H, -(CH2)4-CH3), 1.18⫺1.33 (m, 18H, -(CH2)2-CH3,
-SO2-(CH2)2-(CH2)7-), 1.64⫺1.73 (m, 4H, -CH2-CH2-SO2CH2-CH2-), 1.63⫺2.23 (m, 2H, C=C-CH-CH2-CH3),
3.31⫺3.34 (m, 1H, C=C-CH-CH2-CH3), 3.04 (t, 3J = 7.5 Hz,
4H, -CH2-SO2-CH2-), 3.59 and 3.68 (s, 3H, OCH3), 3.73 and
3.75 (s, 3H, OCH3), 4.64 (t, 3J = 7.3 Hz, 1H, C=CH-(CH2)9-),
6.38⫺8.02 (m, 8H, ArH).
The aromatic methoxy groups were cleaved with BBr3, as
described previously [20]. Analytical data are only given for
the main products 12 and not for the isomeric byproducts 13.
(E/Z)-3-(4-Hydroxyphenyl)-4-(3-hydroxyphenyl)hex-3-ene
(12a)
Colorless crystals, mp 128⫺129 °C, yield 78 %. 1H-NMR δ =
0.71 and 0.73 (t, 3J = 7.4 Hz, 3H, C=C-CH2-CH3), 0.88 (t,
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
642 von Angerer et al.
3
J = 7.4 Hz, 3H, C=C-CH2-CH3), 2.07 (q, 3J = 7.4 Hz, 2H,
C=C-CH2-CH3), 2.44 and 2.45 (q, 3J = 7.4 Hz, 2H, =C-CH2CH3), 6.33⫺7.20 (m, 4H, ArH), 6.77 and 6.99 (AA⬘BB⬘, 3J =
8.4 Hz, 4H, ArH), 9.03 and 9.10 (s, 1H, -OH), 9.33 and 9.34
(s, 1H, -OH).
(E/Z)-3-(4-Hydroxyphenyl)-4-(3-hydroxyphenyl)-14-pentylthiotetradec-3-ene (12c)
Colorless oil, yield 43 %. 1H-NMR δ = 0.70 (t, 3J = 7.3 Hz,
3H, C=C-CH2-CH3), 0.85 (t, 3J = 7.0 Hz, 3H, -(CH2)4-CH3),
1.05⫺1.36 (m, 18H, -(CH2)2-CH3, -S-(CH2)2-(CH2)7-),
1.42⫺2.13 (m, 8H, -CH2-CH2-S-CH2-CH2-, -CH2-C=C-CH2-),
2.45 (t, 3J = 7.3 Hz, 4H, -CH2-S-CH2-), 6.57⫺7.17 (m, 4H,
ArH), 6.75 and 6.96 (AA⬘BB⬘, 3J = 8.4 Hz, 4H, ArH), 9.30 (s,
2H, -OH).
(E/Z)-3,4-Bis-(4-hydroxyphenyl)-14-pentylthiotetradec-3-ene
(12d)
Colorless oil, yield 46 %. 1H-NMR δ = 0.69 (t, 3J = 7.3 Hz,
3H, =C-CH2-CH3), 0.85 (t, 3J = 7.1 Hz, 3H, -(CH2)4-CH3),
1.03⫺1.55 (m, 18H, -(CH2)2-CH3, -S-(CH2)2-(CH2)7-),
1.45⫺2.11 (m, 8H, -CH2-CH2-S-CH2-CH2-, -CH2-C=C-CH2-),
2.45 (t, 3J = 7.3 Hz, 4H, -CH2-S-CH2-), 6.44⫺6.97 (m, 8H,
ArH), 9.04 and 9.05 (s, 1H, -OH), 9.27 and 9.28 (s, 1H, -OH).
(E/Z)-3-(3-Hydroxyphenyl)-4-(4-hydroxyphenyl)-14-pentylthiotetradec-3-ene (12e)
Colorless oil, yield 39 %. 1H-NMR δ = 0.72 (t, 3J = 7.5 Hz,
3H, C=C-CH2-CH3), 0.85 (t, 3J = 7.0 Hz, 3H, -(CH2)4-CH3),
1.03⫺1.33 (m, 18H, -(CH2)2-CH3, -S-(CH2)2-(CH2)7-),
1.41⫺2.12 (m, 8H, -CH2-CH2-S-CH2-CH2-, -CH2-C=C-CH2-),
2.45 (t, 3J = 7.3 Hz, 4H, -CH2-S-CH2-), 6.55⫺7.17 (m, 8H,
ArH), 9.01 and 9.07 (s, 1H, -OH), 9.309 and 9.316 (s, 1H,
-OH).
(E/Z)-3-(4-Hydroxyphenyl)-4-(3-hydroxyphenyl)-14-pentylsulfonyl-tetradec-3-ene (12f)
Colorless oil, yield 43 %. 1H-NMR δ = 0.70 (t, 3J = 7.3 Hz,
3H, C=C-CH2-CH3), 0.87 (t, 3J = 7.2 Hz, 3H, -(CH2)4-CH3),
1.05⫺1.38 (m, 18H, -(CH2)2-CH3, -SO2-(CH2)2-(CH2)7-),
1.60⫺2.11 (m, 8H, -CH2-CH2-SO2-CH2-CH2-, -CH2-C=CCH2-), 3.03 (t, 3J = 8.1 Hz, 4H, -CH2-SO2-CH2-), 6.32⫺7.18
(m, 4H, ArH), 6.76 and 6.97 (AA⬘BB⬘, 3J = 8.4 Hz, 4H, ArH),
9.00 and 9.08 (s, 1H, -OH), 9.31 and 9.32 (s, 1H, -OH). HPLC
[MeOH/H2O 80:20 (vol/vol)], RT [min] 10.54 (85 %), 13.77
(15 %).
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
(E/Z)-3-(3-Hydroxyphenyl)-4-(4-hydroxyphenyl)-14-pentylsulfonyltetradec-3-ene (12h)
Colorless oil, yield 39 %. 1H-NMR δ = 0.72 (t, 3J = 7.4 Hz,
3H, =C-CH2-CH3), 0.87 (t, 3J = 7.2 Hz, 3H, -(CH2)4-CH3),
1.03⫺1.38 (m, 18H, -(CH2)2-CH3, -SO2-(CH2)2-(CH2)7-),
1.60⫺2.12 (m, 8H, -CH2-CH2-SO2-CH2-CH2-, -CH2-C=CCH2-), 3.00 (t, 3J = 7.9 Hz, 4H, -CH2-SO2-CH2), 6.46⫺7.18
(m, 8H, ArH), 9.02 and 9.08 (s, 1H, -OH), 9.31 and 9.32 (s,
1H, -OH). HPLC [MeOH/H2O 84:16 (vol/vol)], RT [min] 6.54
(82 %), 10.02 (17 %).
General procedure for the preparation of the sulfamates 8
A solution of the dihydroxystilbene derivative 12 (2.77 mmol)
in 15 mL of dry DMF was cooled to 10⫺15 °C. Sulfamoyl choride [27] (27.8 mmol) was added in portions. After the addition, the mixture was stirred for 12 h under N2. The mixture
was hydrolyzed by treating with 40 mL water, followed by extraction with EtOAc. The combined organic layers were
washed with water and dried (MgSO4). After evaporation of
the solvent, the residue was chromatographed over SiO2 with
CH2Cl2/EtOAc (10:1, vol/vol) as eluent. Sometimes, it was
possible to isolate one of the monosulfamates (7) from the
first fraction as a by-product.
(E/Z)-3-(4-Sulfamoyloxyphenyl)-4-(3-sulfamoyloxyphenyl)hex-3-ene (8a)
Yellow resin, yield 47 %. 1H-NMR δ = 0.74 (t, 3J = 7.5 Hz,
3H, -CH2-CH3), 0.75 (t, 3J = 7.5 Hz, 3H, -CH2-CH3), 2.12 (q,
3
J = 7.5 Hz, 4H, -CH2-CH3), 7.11⫺7.51 (m, 4H, ArH), 7.31 (s,
4H, ArH), 8.04 (s, br, 2H, -OSO2NH2), 8.05 (s, br, 2H,
-OSO2NH2). HPLC [MeCN/buffer 55:45 (vol/vol)], RT [min]
6.39 (95 %), 8.90 (4 %).
(E/Z)-4-(4-Hydroxyphenyl)-3-(4-sulfamoyloxy)hex-3-ene (7b)
Viscous oil, yield 27 %. 1H-NMR δ = 0.72 (t, 3J = 7.5 Hz, 3H,
-CH2-CH3), 0.73 (t, 3J = 7.5 Hz, 3H, -CH2-CH3), 2.05 (q, 3J =
7.5 Hz, 2H, -CH2-CH3), 2.08 (q, 3J = 7.5 Hz, 2H, -CH2-CH3),
6.77 and 7.02 (AA⬘BB⬘, 3J = 8.6 Hz, 4H, ArH), 7.27 and 7.28
(AA⬘BB⬙, 3J = 7.4 Hz, 4H, ArH), 8.01 (s, br, 2H, -OSO2NH2),
9.35 (s, 1H, -OH). HPLC [MeCN/buffer 55:45 (vol/vol)], RT
[min] 6.52 (92 %), 7.12 (6 %).
(E)-3,4-Bis(4-sulfamoyloxyphenyl)hex-3-ene (8b)
Viscous oil, yield 65 %. 1H-NMR δ = 0.74 (t, 3J = 7.5 Hz, 6H,
-CH2-CH3), 2.11 (q, 3J = 7.5 Hz, 4H, -CH2-CH3), 7.30 (s, 8H,
ArH), 8.04 (s, br, 4H, -OSO2NH2). HPLC [MeCN/buffer 55:45
(vol/vol)], RT [min] 6.08 (98 %).
(E/Z)-3,4-Bis-(4-hydroxyphenyl)-14-pentylsulfonyltetradec-3ene (12g)
(E)-3-(4-Sulfamoyloxyphenyl)-4-(3-sulfamoyloxyphenyl)-14pentylthiotetradec-3-ene (8c)
Colorless oil, yield 46 %. 1H-NMR δ = 0.70 (t, 3J = 7.5 Hz,
3H, =C-CH2-CH3), 0.87 (t, 3J = 6.9 Hz, 3H, -(CH2)4-CH3),
1.15⫺1.38 (m, 18H, -(CH2)2-CH3, -SO2-(CH2)2-(CH2)7-),
1.60⫺2.11 (m, 8H, -CH2-CH2-SO2-CH2-CH2-, -CH2-C=CCH2-), 3.03 (t, 3J = 8.0 Hz, 4H, -CH2-SO2-CH2-), 6.75⫺6.97
(m, 8H, ArH), 9.06 (s, 1H, -OH), 9.29 (s, 1H, -OH). HPLC
[MeOH/H2O 80:20 (vol/vol)], RT [min] 10.82 (82 %), 16.12
(18 %).
Colorless oil, yield 48 %. 1H-NMR δ = 0.73 (t, 3J = 7.3 Hz,
3H, C=C-CH2-CH3), 0.85 (t, 3J = 7.3 Hz, 3H, -(CH2)4-CH3),
1.05⫺1.37 (m, 18H, -CH2-(CH2)7-CH2-CH2-S-, -CH2-(CH2)2CH3), 1.41⫺2.16 (m, 8H, -CH2-CH2-S-CH2-CH2-, -CH2-C=CCH2-), 2.44 and 2.45 (t, 3J = 7.3 Hz, 4H, -CH2-S-CH2-),
7.11⫺7.51 (m, 4H, ArH), 7.29 (s, 4H, ArH), 8.03 (s, br, 4H,
-OSO2NH2). HPLC [MeCN/buffer 78 : 22 (vol/vol)], RT [min]
15.43 (99 %).
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
Stilbene-Based Inhibitors of Estrone Sulfatase 643
(E)-3,4-Bis-(4-sulfamoyloxyphenyl)-14-pentylthiotetradec-3ene (8d)
Viscous oil, yield 68 %. 1H-NMR δ = 0.73 (t, 3J = 7.7 Hz, 3H, =
C-CH2-CH3), 0.85 (t, 3J = 7.2 Hz, 3H, -(CH2)4-CH3),
1.03⫺1.34 (m, 18H, -CH2-(CH2)7-CH2-CH2-S-, -CH2-(CH2)2CH3), 1.43⫺2.14 (m, 8H, -CH2-CH2-S-CH2-CH2-, -CH2-C=CCH2-), 2.45 and 2.47 (t, 3J = 7.2 Hz, 4H, -CH2-S-CH2), 7.29
and 7.29 (AA⬘BB⬘, 3J = 8.6 Hz, 8H, ArH), 8.03 (s, br, 4H,
-OSO2NH2). HPLC [MeCN/buffer 78 : 22 (vol/vol)], RT [min]
13.63 (96 %).
(E)-3-(3-Sulfamoyloxyphenyl)-4-(4-sulfamoyloxyphenyl)-14pentylthiotetradec-3-ene (8e)
Colorless oil, yield 66 %. 1H-NMR δ = 0.74 (t, 3J = 7.5 Hz,
3H, =C-CH2-CH3), 0.85 (t, 3J = 7.0 Hz, 3H, -(CH2)4-CH3),
1.05⫺1.34 (m, 18H, -CH2-(CH2)7-CH2-CH2-S-, -CH2-(CH2)2CH3), 1.36⫺2.16 (m, 8H, -CH2-CH2-S-CH2-CH2-, -CH2-C=CCH2-), 2.44 and 2.45 (3J = 7.5 Hz, 4H, -CH2-S-CH2-),
7.10⫺7.51 (m, 4H, ArH), 7.30 (s, 4H, ArH), 8.02 (s, br, 4H,
-OSO2NH2). HPLC [MeCN/buffer 78 : 22 (vol/vol)], RT [min]
15.49 (97 %).
(E)-3-(4-Sulfamoyloxyphenyl)-4-(3-sulfamoyloxyphenyl)-14pentylsulfonyltetradec-3-ene (8f)
Colorless oil, yield 50 %. 1H-NMR δ = 0.73 (t, 3J = 7.5 Hz,
3H, =C-CH2-CH3), 0.87 (t, 3J = 7.1 Hz, 3H, -(CH2)4-CH3),
1.04⫺1.40 (m, 18H, -CH2-(CH2)7-CH2-CH2-SO2-, -CH2(CH2)2-CH3), 1.60⫺2.15 (m, 8H, -CH2-CH2-SO2-CH2-CH2-,
-CH2-C=C-CH2-), 3.00⫺3.05 (m, 4H, -CH2-SO2-CH2-),
6.70⫺7.50 (m, 4H, ArH), 7.29 (s, 4H, ArH), 7.99 (s, br, 4H,
-OSO2NH2). HPLC [MeCN/buffer 55 : 45 (vol/vol)], RT [min]
31.06 (96 %).
(E)-3,4-Bis-(4-sulfamoyloxyphenyl)-14-pentylsulfonyltetradec-3-ene (8g)
Viscous oil, yield 59 %. 1H-NMR δ = 0.73 (t, 3J = 7.5 Hz, 3H, =
C-CH2-CH3), 0.87 (t, 3J = 7.2 Hz, 3H, -(CH2)4-CH3),
1.04⫺1.39 (m, 18H, -CH2-(CH2)7-CH2-CH2-SO2-, -CH2(CH2)2-CH3), 1.60⫺2.14 (m, 8H, -CH2-CH2-SO2-CH2-CH2-,
-CH2-C=C-CH2-), 3.01 and 3.03 (t, 3J = 7.8 Hz, 4H, -CH2SO2-CH2-), 7.30 and 7.30 (AA⬘BB⬘, 3J = 7.9 Hz, 8H, ArH),
8.04 (s, br, 4H, -OSO2NH2). HPLC [MeCN/buffer 55 : 45 (vol/
vol)], RT [min] 28.11 (98 %).
(E/Z)-3-(3-Hydroxyphenyl)-4-(4-sulfamoyloxyphenyl)-14pentylsulfonyltetradec-3-ene (7h)
Colorless oil, yield 32 %. 1H-NMR δ = 0.73 (t, 3J = 7.4 Hz,
3H, =C-CH2-CH3), 0.87 (t, 3J = 7.1 Hz, 3H, -(CH2)4-CH3),
1.05⫺1.38 (m, 18H, -CH2-(CH2)7-CH2-CH2-SO2-, -CH2(CH2)2-CH3), 1.60⫺2.13 (m, 8H, -CH2-CH2-SO2-CH2-CH2-,
-CH2-C=C-CH2-), 3.02 and 3.03 (t, 3J = 7.9 Hz, 4H, -CH2SO2-CH2-), 6.58⫺7.45 (m, 4H, ArH), 7.27 and 7.27 (AA⬘BB⬘,
3
J = 7.7 Hz, 4H, ArH), 7.99 (s, br, 2H, -OSO2NH2), 9.35 (s,
br, 1H, -OH). HPLC (MeCN/buffer 78 : 22 (vol/vol), RT [min]
5.52 (90 %), 9.77 (3 %).
(CH2)2-CH3), 1.56⫺2.15 (m, 8H, -CH2-CH2-SO2-CH2-CH2-,
-CH2-C=C-CH2-), 3.02 (t, 3J = 7.9 Hz, 4H, -CH2-SO2-CH2-),
6.99⫺7.51 (m, 4H, ArH), 7.30 (s, 4H, ArH), 8.01 (s, br, 4H,
-OSO2NH2). HPLC [MeCN/buffer 55 : 45 (vol/vol)], RT [min]
30.31 (99 %).
Materials and reagents for bioassays
[3H]17β-Estradiol, [3H]estrone sulfate (ammonium salt), and
[14C]estrone were purchased from New England Nuclear
(Dreieich, Germany); all other biochemicals were obtained
from Sigma (Munich, Germany). Hormone-sensitive human
MCF-7 breast cancer cells and hormone-independent human
MDA-MB 231 breast cancer cells were obtained from the
American Type Culture Collection (ATCC). MCF-7/2a cells
with the reporter construct integrated in the genome have
been generated in the authors’ laboratory [26]. Reference
compounds EMATE and ZK 164.015-sulfamate were synthesized in the authors’ laboratory [20].
Bioassays
Experimental details for all of the bioassays, including
determination of ER binding affinities [20], estrone sulfatase
inhibition [28], estrogenic and antiestrogenic activities in
transfected MCF-7/2a cells [20], and antiproliferative
activities in wild-type MCF-7 cells [28] have been reported
previously.
References
[1] G. S. Chetrite, J. Cortes-Prieto, J. C. Philippe, F. Wright,
J. R. Pasqualini, J. Steroid Biochem. Mol. Biol. 2000,
72, 23⫺27.
[2] J. R. Pasqualini, G. Chetrite, C. Blacker, M. C. Feinstein,
L. Delalonde, M. Talbi, C. Maloche, J. Clin. Endocrinol.
Metab. 1996, 81, 1460⫺1464.
[3] T. R. Evans, M. G. Rowlands, M. C. Silva, M. Law, R. C.
Coombes, J. Steroid Biochem. Mol. Biol. 1993, 44,
583⫺587.
[4] J. R. Pasqualini, G. Chetrite, B. L. Nguyen, C. Maloche,
L. Delalonde, M. Talbi, M. C. Feinstein, C. Blacker, J.
Botella, J. Paris, J. Steroid Biochem. Mol. Biol. 1995,
53, 407⫺412.
[5] G. S. Chetrite, J. R. Pasqualini, J. Steroid Biochem. Mol.
Biol. 2001, 76, 95⫺104.
[6] T. Utsumi, N. Yoshimura, S. Takeuchi, M. Maruta, K.
Maeda, N. Harada, J. Steroid Biochem. Mol. Biol. 2000,
73, 141⫺145.
[7] Y. Miyoshi, A. Ando, S. Hasegawa, M. Ishitobi, T. Taguchi, Y. Tamaki, S. Noguchi, Clin. Cancer Res. 2003, 9,
2288⫺2293.
[8] K. W. Selcer, S. Jagannathan, M. E. Rhodes, P. K. Li,
J. Steroid Biochem. Mol. Biol. 1996, 59, 83⫺91.
[9] P. K. Li, R. Pillai, L. Dibbelt, Steroids 1995, 60, 299⫺306.
(E)-3-(3-Sulfamoyloxyphenyl)-4-(4-sulfamoyloxyphenyl)-14pentylsulfonyltetradec-3-ene (8h)
[10] N. M. Howarth, A. Purohit, M. J. Reed, B. V. Potter,
J. Med. Chem. 1994, 37, 219⫺221.
Colorless oil, yield 60 %. 1H-NMR δ = 0.74 (t, 3J = 7.5 Hz,
3H, =C-CH2-CH3), 0.87 (t, 3J = 7.2 Hz, 3H, -(CH2)4-CH3),
1.05⫺1.38 (m, 18H, -CH2-(CH2)7-CH2-CH2-SO2-, -CH2-
[11] W. Elger, S. Schwarz, A. Hedden, G. Reddersen,
B. Schneider, J. Steroid Biochem. Mol. Biol. 1995, 55,
395⫺403.
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
644 von Angerer et al.
[12] A. Purohit, K. A. Vernon, A. E. Hummelinck, L. W. Woo,
H. A. Hejaz, B. V. Potter, M. J. Reed, J. Steroid Biochem.
Mol. Biol. 1998, 64, 269⫺275.
[13] L. C. Ciobanu, R. P. Boivin, T. V. Luu, F. Labrie,
D. Poirier, J. Med. Chem. 1999, 42, 2280⫺2286.
[14] L. W. Woo, N. M. Howarth, A. Purohit, H. A. Hejaz,
M. J. Reed, B. V. Potter, J. Med. Chem. 1998, 41,
1068⫺1083.
[15] B. Malini, A. Purohit, D. Ganeshapillai, L. W. Woo, B. V.
Potter, M. J. Reed, J. Steroid Biochem. Mol. Biol. 2000,
75, 253⫺258.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 634−644
[20] T. Golob, R. Liebl, E. von Angerer, Bioorg. Med. Chem.
2002, 10, 3941⫺3953.
[21] C. Biberger, E. von Angerer, J. Steroid Biochem. Mol.
Biol. 1996, 58, 31⫺43.
[22] T. R. Evans, M. G. Rowlands, Y. A. Luqmani, S. K.
Chander, R. C. Coombes, J. Steroid Biochem. Mol. Biol.
1993, 46, 195⫺201.
[23] A. Purohit, C. V. de Giovani, M. J. Reed, J. Steroid Biochem. Mol. Biol. 1999, 68, 129⫺135.
[24] L. Duncan, A. Purohit, N. M. Howarth, B. V. Potter, M. J.
Reed, Cancer Res. 1993, 53, 298⫺303.
[16] D. Poirier, L. C. Ciobanu, R. Maltais, Exp. Opin. Ther.
Patents 1999, 9, 1083⫺1099.
[25] P. Nussbaumer, A. P. Winiski, A. Billich, J. Med. Chem.
2003, 46, 5091⫺5094.
[17] L. L. Woo, A. Purohit, B. Malini, M. J. Reed, B. V. Potter,
Chem. Biol. 2000, 7, 773⫺791.
[26] F. Hafner, E. Holler, E. von Angerer, J. Steroid Biochem.
Mol. Biol. 1996, 58, 385⫺393.
[18] P. Nussbaumer, P. Lehr, A. Billich, J. Med. Chem. 2002,
45, 4310⫺4320.
[27] S. Schwarz, I. Thieme, M. Richter, B. Undeutsch,
H. Henkel, W. Elger, Steroids 1996, 61, 710⫺717.
[19] L. C. Ciobanu, V. Luu-The, D. Poirier, J. Steroid Biochem. Mol. Biol. 2002, 80, 339⫺353.
[28] G. Walter, R. Liebl, E. von Angerer, J. Steroid Biochem.
Mol. Biol. 2004, 88, 409⫺420.
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Документ
Категория
Без категории
Просмотров
0
Размер файла
156 Кб
Теги
base, sulfatase, mode, cancer, dual, inhibitors, action, human, breast, stilbene, cells, estrone
1/--страниц
Пожаловаться на содержимое документа