Substituted phenanthrenes with basic amino side chains: A new series of anti-breast cancer agents

Article abstract of DOI:10.1016/j.bmc.2005.10.002

In the course of our search for new anti-breast cancer agents, substituted phenanthrenes with basic amino side chains were synthesized and some of them showed remarkable antiproliferative activity against ER +ve MCF-7 cell line with IC₅₀ in the

Full text of DOI:10.1016/j.bmc.2005.10.002

Bioorganic & Medicinal Chemistry 14 (2006) 1497–1505
Substituted phenanthrenes with basic amino side chains:
A new series of anti-breast cancer agentsq
Shagufta,a Ajay Kumar Srivastava,a Ramesh Sharma,b Rajeev Mishra,c
Anil Kumar Balapure,b Puvvada S. R. Murthyc and Gautam Pandaa,*
aMedicinal and Process Chemistry Division, Central Drug Research Institute, Lucknow-226001, UP, India
bTissue Culture Laboratory, National Laboratory Animal Center (NLAC), Central Drug Research Institute,
Lucknow-226001, UP, India
cCarcinogenicity Unit, Toxicology Division, Central Drug Research Institute, Lucknow-226001, UP, India
Received 20 April 2005; revised 1 October 2005; accepted 3 October 2005
Available online 24 October 2005
Abstract—In the course of our search for new anti-breast cancer agents, substituted phenanthrenes with basic amino side chains
were synthesized and some of them showed remarkable antiproliferative activity against ER +ve MCF-7 cell line with IC50 in
the range of 3.53–22.25 lM. One of the compounds 15ca showed anti-breast cancer activity in 7,12-dimethylbenz[a]anthracene
(DMBA) induced hormone-dependent mammary tumor in rat and the activity was comparable to that shown by tamoxifen.
Ó 2005 Elsevier Ltd. All rights reserved.
1. Introduction
antiestrogens (2, 3, and 4 in Fig. 1) which compete with
endogenous estradiol for ER binding and directly inter-
act with growth factors, thereby inhibiting estrogen
action.
Stimulation of estrogen receptors (ER) by endogenous
estrogens plays an important role in both male and fe-
male physiology.1 Estrogen stimulation is implicated in
the development of breast cancer.2 The blockade of
estrogen action is a major approach for the treatment
of estrogen-dependent breast cancer.3 Consequently,
many estrogen receptor ligands with antiestrogenic effect
are being developed with the aim of preventing estrogen
mediated growth. Triarylethylenes such as tamoxifen 1
(TAM) are the first successful series of potent selective
estrogen receptor modulators (SERMs) with antiestro-
genic activity that was developed for the treatment and
prevention of ER positive breast cancer.4 However, be-
cause these antiestrogens generally possess partial estro-
genic activity in some estrogen target tissues such as
bone, uterus, and liver,5 considerable efforts have been
made to the search of pure antiestrogens without
estrogenic activity.6 This approach has resulted in the
development of steroidal and non-steroidal pure
Phenanthrenes and their substituted analogs are well
known to exhibit various biological activities such as
OH
OCH2CH2N(CH3)2
X
HO
(CH2)nR
2
ICI 164, 384; X=H, n=10, R= CON(CH )C H
4 9
3
EM 139; X=Cl, n=10, R= CON(CH )C H
4 9
3
Tamoxifen
ICI 182, 780; ; X=H, n=9, R= SO(CH ) C F
2 5
2 3
1
OCH3
H3CO
H3CO
OR
OCH3
RO
O
N
Keywords: Substituted phenanthrenes; Anti-breast cancer agents; Basic
amino side chain.
q
Check CDRI communication No. 6754.
O
O
NRR
5, R=CH , C H
2 5
R-R= (CH ) , ( CH )
3, R=H, EM-652
4, R=Pivaloyl, EM-800
3
*
Corresponding author. Tel.: +91 522 2364635; fax: +91 522
2223938; e-mail addresses: gautam_panda@lycos.com; bapi.rm@
yahoo.com
2 4
2 5
Figure 1. Structures of the estrogen receptor ligands.
0968-0896/$ - see front matter Ó 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2005.10.002

1498
Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
antimicrobial,7a cytotoxic,7b antifungal,7c and antima-
larial.7d Recently, diaryloxy methano phenanthrene
derivatives having antitubercular activity are reported
from our laboratory.8 Particularly, we have been inter-
ested in the relationship between substituted phenanth-
renes and anti-breast cancer activities as some of the
phenanthrene derivatives are known to exhibit antipro-
liferative effect against breast cancer cells.9 It is well
known that most of the compounds having high relative
binding affinity to estrogen receptor possess at least one
or two strategically placed hydroxy or methoxy groups
in their structure.10 On the other hand, there are consid-
erable literature reports that 7a substituted estradiol
derivatives especially ICI 164, 182, 384, 780, and EM-
139 are having antiestrogenic activity on breast cancer
cells.11 Thus, we presume that methoxy substituted
phenanthrene will mimic the skeleton of estradiol for
receptor binding and substitution at position nine of
phenanthrene nucleus might act as 7a substituent in 2
and thus will exert antiestrogenic effect on breast cancer
cells. Thus, we have designed the compound 5 as our
target molecule with various amines as side chains and
evaluated their antiproliferative activity against estrogen
responsive MCF-7 cell lines.
was followed to synthesize the trimethoxy-phenan-
threne-9-carboxylic acid 6.12 Treatment of 6 with anisole
in the presence of PPA gave the methanone derivative 7
(58%) which on reduction with LAH furnished the car-
binol 8 in 83% yield. Friedel–Crafts arylation with phe-
nol in the presence of concd. H2SO4 and benzene
furnished 9 as major product and 10 as minor one by
nucleophilic attack of phenol through para and ortho
position on the carbinol carbon atom of 8, respectively.
The reaction between 9 and 10 with alkylaminohydro-
chloride chains in the presence of K2CO3 and acetone
led to the formation of 11a–d and 12a–b, respectively,
in good yields, Scheme 1. Since all the six compounds
were crystallizable solids, they were used as such
for evaluation of their anti-breast cancer activity, Tables
1 and 2.
Out of six compounds 11a–d, 12a–b synthesized, only
one compound 11a was showing antiproliferative activ-
ity with IC50 21.80 against ER +ve MCF-7 cell line. To
rationalize our pharmacological results, we performed
comparative docking of compound 11a with 4-hydroxy-
tamoxifen (OHT) in estrogen receptor (ERa) using
MOE software. From Figure 2, it is evident that the
compound 11a does not fit well into the ERa ligand
binding pocket. The improper orientation of 11a in
ERa may be due to the steric hindrance of methoxy
functionality of phenanthrene. Thus, we presumed that
the compounds lacking methoxy on phenanthrene may
be suitable for binding with ERa ligand binding pocket.
The comparative docking study of compound 15bc and
OHT with ERa shows encouraging results and as
2. Results and discussion
2.1. Chemistry
To synthesize the target molecule as described in
Figure 1, 11 and 12 were selected. Literature procedure
H CO
3
H CO
3
H CO
3
OH
O
CO H
2
a
b
OCH
OCH
3
3
H CO
3
H CO
3
H CO
3
OCH
8
OCH
7
OCH
3
3
3
OH
6
c
11
N(R)2
%
H CO
3
N(CH3)2
H CO
3
87
82
OH
a
b
N(CH2CH3)2
78
81
+
c
d
N
OCH
3
OCH
3
N
H CO
3
H CO
3
OCH
3
OCH
3
N(R)2
%
12
a
9
10
d
d
N(CH2CH3)2
N
99
90
OCH CH NRR
2
2
b
H CO
3
H CO
3
OCH CH NRR
2
2
OCH
3
OCH
3
H CO
3
H CO
3
OCH
3
OCH
3
12
11
Scheme 1. Synthesis of (2-{4-[(4-methoxy-phenyl)-(2,3,6-trimethoxy-phenanthren-9-yl)-methyl]-phenoxy}-ethyl)-alkylamine derivatives 11 and 12.
Reagents and conditions: (a) anisole, PPA, 58%; (b) LAH, THF, 83%; (c) Phenol, concd. H2SO4, benzene, 65%; (d) ClCH2CH2NRRÆHCl, K2CO3,
acetone, 78–99%.

Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
1499
Table 2. In vitro anti-breast cancer activities of 11a–d, 12a–b, 15ÆHCl,
16ÆHCl, and 18ÆHCl, against ER Àve MDA MB-453 cell line
Table 1. In vitro anti-breast cancer activities of 11a–d, 12a–b, 15ÆHCl,
16ÆHCl, and 18ÆHCl, against human breast cancer cell line ER +ve
MCF-7
Serial No.
Compound
IC50 (lM) (MDA MB-453)a
Serial No.
Compound
IC50 (lM) (MCF-7)a
1
2
11a
11b
2.75
3.50
3.50
5.75
5.80
NA
NA
NA
6.2
1
2
11a
11b
21.80
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
3.53
5.97
3.88
4.29
6.50
7.91
NA
NA
NA
22.25
NA
NA
9.97
4.94
NA
NA
20.19
12.48
3
11c
11d
4
3
11c
11d
5
12a
12b
4
6
5
12a
12b
7
15ad
15ca
15cb
15cd
16cd
18c
6
8
7
15aa
15ab
15ac
15ad
15ba
15bb
15bc
15bd
15ca
15cb
15cc
15cd
16ca
16cb
16cc
16cd
18a
9
8
10
11
12
13
14
15
16
10.0
7.33
6.30
2.75
NA
NA
13.50
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
18d
18e
18f
Tamoxifen
a IC50 values for anti-breast cancer activity of ligands using ER +ve
and Àve cell lines viz. MCF-7 and MDA MB-453, respectively, using
tamoxifen as a positive control. The results are representative of one
of three similar experiments each performed in triplicate. Each datum
depicts the IC50 for each ligand. The deviation in each case was less
than 5% and ꢀNAꢁ represents ligands that were found to be inactive at
25 lM in either of the two cell types in which the SRB assays were
conducted.
18b
18c
18d
18e
effect of ether side chain containing various aminoalkyl
groups on the pharmacophore.
18f
18g
Tamoxifen
Toward this objective, 13c was treated with epichlorohy-
drin in the presence of K2CO3 to furnish epoxides 17 in
good yield (86%). The epoxide 17, in turn, reacted with
commercially available different amines to afford a vari-
ety of 1-aminopropan-2-ol derivatives 18 (64–96%,
Scheme 3) through nucleophilic reaction of amines onto
less hindered side of the epoxide. By treatment of 18
with ethanolic hydrochloride, the corresponding salts
18ÆHCl were prepared and tested against MCF-7 and
MDA MB-453 cell lines in vitro (Tables 1 and 2).
a IC50 values for anti-breast cancer activity of ligands using ER +ve
and Àve cell lines viz. MCF-7 and MDA MB-453, respectively, using
tamoxifen as a positive control. The results are representative of one
of three similar experiments each performed in triplicate. Each datum
depicts the IC50 for each ligand. The deviation in each case was less
than 5% and ꢀNAꢁ represents ligands that were found to be inactive at
25 lM in either of the two cell types in which the SRB assays were
conducted.
expected 15bc orients well in pocket through hydrogen
bonding with ARG 394 and GLU 353 residues of
ERa (Fig. 3). Thus, the feasible synthetic strategies to-
ward compounds 15 and 16 were undertaken.
2.2. Biology
2.2.1. Determination of anti-breast cancer activity in vitro.
The activities of the compounds against MCF-7 and
MDA MB-453 cell lines were determined through SRB
(Sulforhodamine B) antiproliferative activity assay and
the results are shown in Tables 1 and 2.
Toward the objective, the compounds 13a–c8 and 14a–c8
were reacted with different alkylamine hydrochlorides in
the presence of K2CO3 and acetone to furnish 15 and16
in good yield (Scheme 2). By treatment of amines 15 and
16 with ethanolic hydrogen chloride the corresponding
salts 15ÆHCl and 16ÆHCl were prepared. The salts
15ÆHCl and 16ÆHCl were tested and found to be active
against MCF-7 cell lines in vitro with IC50 in the range
of 3.53–22.25 lM (Table 1). Therefore, 4-[(2 or 3 or
4-methoxy-phenyl)-phenanthren-9-yl-methyl]-phenol deriv-
atives were selected as active pharmacophores and fur-
ther synthetic transformations were performed. From
close analysis of the structures of 15 and 16, it is clear
that the aminoalkyl containing basic ether side chain is
responsible for its modulating influence on antagonistic
efficacy among diaryloxymethanophenanthrene phar-
macophore. Thus, we became interested to study the
2.2.2. Structure–activity relationship. From the compar-
ison of the data of compounds (Table 1), it is evident
that compounds lacking methoxy groups on the phenan-
threne nucleus gave lower IC50 agsinst ER +ve MCF-7
cell line. Compounds containing an ortho-methoxy
group were found to be inactive, whereas molecules con-
taining meta-methoxy and para-methoxy groups except
for 15ba and 15bb, were active. The contribution of
the basic ether chain to estrogen antagonistic activity
of the phenanthrene is structure as well as position spe-
cific. The protype is active with the chain at para posi-
tion and inactive with chain at ortho position of the
phenyl ring. The activity increased with the decrease of

1500
Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
Figure 2. Comparative docking orientation and predicted receptor interactions for compound 11a (gray), 4-hydroxytamoxifen (OHT, green), and
ERa (red) (PDB ID3ERT).
Figure 3. Comparative docking orientation and predicted receptor interactions for compound 15bc (gray), 4-hydroxytamoxifen (OHT, green), and
ERa (red) (PDB ID3ERT).
the length of alkyl group on nitrogen. This was evident
from MIC values of piperidino ether 15bd and 15cd
(5.91 and 7.91 lM, respectively), pyrolidone ether 15bc
and 15cc (3.53 and 6.50 lM, respectively), diethylamino
ether 15cb (4.29 lM), and dimethylamino ether 15ca
(3.88 lM). In 1-aminopropan-2-ol derivatives (18a–g),
compounds having N-methylpiperazine, cyclohexyl-
amine or cyclopropyl amine in the side chain impart
better antagonistic activity than the compounds having
pyrrolidine, piperidine, morpholine or N-benzylpiper-
azine amino group.
It is also interesting to note from IC50 values of 11a–d,
12a–b, 15ad, 15cb, 15cd, 16cd, and 18c–f against ER
Àve MDA MB-453 (Table 2) that 11a–d and 12a–b con-
taining methoxy groups on phenanthrene gave lower
IC50 against ER Àve MDA MB-453 in comparison of
IC50 (Table 1) against ER +ve MCF-7. The specificity

Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
1501
R1
N(R2)2
%
15
aa
1
1
R
R
67
62
2-OMe
2-OMe
N(CH3)2
+
N(CH2CH3)2
ab
72
70
ac
N
N
2-OMe
2-OMe
OH
R1
N(R2)2
ad
%
16
ca
cb
80
92
4-OMe
4-OMe
N(CH3)2
ba
bb
77
77
3-OMe
3-OMe
3-OMe
N(CH3)2
14 a-c
OH
N(CH2CH3)2
N(CH2CH3)2
13a-c
a
78
65
cc
cd
N
N
4-OMe
4-OMe
a
bc
bd
72
70
N
N
1
1
R
R
3-OMe
ca
cb
93
83
80
4-OMe
4-OMe
15,16
N(CH3)2
HCl / ethanol
2
2
N(CH2CH3)2
OCH CH NR R
2
2
15 HCl
16 HCl
cc
cd
4-OMe
4-OMe
N
N
77
2
2
OCH CH NR R
16
2
2
15
Scheme 2. Synthesis of 4-[(2 or 3 or 4-methoxy-phenyl)-phenanthren-9-yl-methyl]-phenols 13a–c, 14a–c, 1-(2-{4-[2 or 3 or 4-methoxy-phenyl)-
phenanthren-9-yl-methyl]-phenoxy}-ethyl)-N,N-dialkylamines 15, 16, and their hydrochlorides 15ÆHCl and 16ÆHCl. Reagents and conditions:
(a) ClCH2CH2N(R2)2ÆHCl, K2CO3, acetone, 62–93%.
3
4
18
a
NR R
%
75
OCH3
OCH3
b
OCH3
N
96
70
b
c
d
e
N
a
N
N-Me
O
84
72
NH
N
OH
13 c
O
O
O
HO
18
f
66
64
HN N CH Ph
2
HCl / ethanol
NR3R4
18.HCl
18
17
g
NH
Scheme 3. Synthesis of 1-amino-3-{4-[(4-methoxy-phenyl)-phenanthren-9-yl-methyl]-phenoxy}-propan-2-ols 18a–g. Reagents and conditions:
(a) epichlorohydrin, K2CO3, 86%; (b) HNR3R4, ethanol, 64–96%.
may be due to the presence of methoxy groups of 11a–d
and 12a–b, Figures 2 and 3. Compounds 15cb, 15cd, and
16cd containing para-methoxy groups gave better anti-
proliferative activity than 15ad with ortho-methoxy
functionality. 18c–d containing N-methylpiperazine
and cyclohexyl amine showed enhanced activity than
18e–f containing morpholine and N-benzylpiperazine
rings, Table 2.
400
300
200
Untreated
100
15ca (10mg/kg)
Tamoxifen (10mg/kg)
50
0
-50
-100
Since the objective was to develop estrogen responsive
anti-breast cancer agents, 15ca was selected to find its
activity in vivo because of a lower IC50 value against
the ER +ve MCF-7 cell line in vitro. From Figure 4, it
is evident that 15ca at 10 mg/kg dose level effectively
inhibited the growth and induced significant regression
of mammary tumors. By the end of third week of treat-
ment, the tumors regressed to almost 50% of their 0-day
size. The compound was however ineffective at 20 mg/kg
dose (data not shown). Also, all animals of both the
dose groups died in the fourth week of treatment. The
mortality largely appeared to be due to anorexia, which
was evident in the third week of treatment.
0
1
2
3
4
5
Treatment Duration (Weeks)
Figure 4. The data of compound 15ca in vivo.
this class of compounds appears to have anti-breast can-
cer activity. The compound 15ca seems to protect the
mice from the challenge of tumor and this protection
is dose dependent. We are reporting for the first time
that diaryloxymethano phenanthrenes might be a suit-
able pharmacophore for developing novel anti-breast
cancer agents. A rational and logical design of a com-
pound retaining the anti-breast cancer activity without
toxicity may be a favorable molecule. Synthesis of the
compounds and their biological evaluation toward this
direction are currently underway.
3. Conclusion
The analysis of in vitro data of the compounds 11a–d,
12a–b, 15ÆHCl, 16ÆHCl, and 18ÆHCl clearly suggests that

1502
Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
4. Experimental details
J = 8 Hz), 6.88 (d, 1H, J = 7 Hz), 6.85 (s, 1H), 6.82 (d,
2H, J = 8 Hz), 6.77 (d, 1H, J = 7 Hz), 6.08 (s, 1H),
4.95 (br s, 1H), 4.07 (s, 3H), 3.93 (s, 6H), 3.78 (s, 3H);
MS: 480 (M+). Anal. Calcd for C31H28O5): C, 77.48;
H, 5.87. Found: C, 77.41; H, 5.94.
4.1. (4-Methoxy-phenyl)-(2,3,6-trimethoxy-phenanthren-
9-yl)-methanone (7)
A mixture of trimethoxy-phenanthrene-9-carboxylic
acid 6 (5 g, 16.02 mmol), PPA (50 g),and anisole (2.5 g,
23.14 mmol) was taken and heated for 2 h at 100 °C.
The reaction mixture was poured into ice-cold water
and extracted with ethyl acetate, washed with NaHCO3,
and dried over anhydrous Na2SO4. Column chromatog-
raphy over silica gel and elution with 40% ethyl acetate
in hexane furnished the methanone 7: white solid
(3.73 g, 58%), mp 147 °C, IR (KBr): 2934, 1600, 1511,
4.5. 2-[(4-Methoxy-phenyl)-(2,3,6-trimethoxy-phenan-
thren-9-yl)-methyl]-phenol (10)
Pale yellow solid (340 mg), mp 150 °C, IR (KBr): 2935,
1
1614, 1510, 1460, 1245, 1033 cmÀ1, H NMR (CDCl3,
200 MHz): d 7.92 (d, 1H, J = 7 Hz), 7.89 (s, 1H), 7.86 (s,
1H), 7.18 (d, 1H, J = 7 Hz), 7.14 (s, 1H), 7.11 (d, 1H,
J = 8 Hz), 7.10 (d, 2H, J = 7 Hz), 7.09 (d, 2H, J = 8 Hz),
6.87 (s, 1H), 6.83 (d, 1H, J = 8 Hz), 6.80 (d, 2 H,
J = 8Hz), 6.31 (s, 1H), 4.97 (br s, 1H), 4.09 (s, 3H), 3.95
(s, 6H), 3.79 (s, 3H); MS: 480 (M+). Anal. Calcd
C31H28O5: C, 77.48; H, 5.87. Found: C, 77.53; H, 5.83.
1
1425, 1248 cmÀ1, H NMR (CDCl3, 200 MHz): d 8.01
(d, 1H, J = 7 Hz), 7.89 (s, 1H), 7.86 (s, 1H), 7.85 (d,
2H, J = 8 Hz), 7.58 (s, 1H), 7.14 (s, 1H), 7.13 (d, 1H,
J = 7.8 Hz), 6.89 (d, 2H, J = 7 Hz), 4.08 (s, 3H), 3.96
(s, 6H), 3.81 (s, 3H); MS: 402 (M+). Anal. Calcd for
C25H22O5: C, 74.61; H, 5.51. Found: C, 74.57; H, 5.45.
4.6. Representative chain reaction
4.2. (4-Methoxy-phenyl)-(2,3,6-trimethoxy-phenanthren-
9-yl)-methanol (8)
A mixture of compounds 9 or 10 (300 mg, 0.625 mmol),
anhydrous K2CO3 (431 mg, 3.12 mmol), aminohydro-
chloride chain (0.93 mmol), and dry acetone (10 mL)
was refluxed for 7 h. K2CO3 was filtered off and acetone
was removed. The residue was diluted with water and
extracted with ethyl acetate. The organic layer was
washed with water, brine, and dried over anhydrous
Na2SO4. Column chromatography over silica gel and
elution with 30% ethyl acetate in hexane furnished the
compound. The product was dissolved in absolute etha-
nol (10 mL) and ethanolic HCl was added dropwise un-
till the pH of the mixture became acidic. Ethanol was
removed in vacuo. The residue was recrystallized from
a mixture of absolute ethanol and dry ether to give the
compound as hydrochloride salt.
The methanone 7 (2.0 g, 4.97 mmol) was taken in dry
THF (60 mL) at 0 °C and LAH (380 mg, 10.27 mmol)
was added in portions into it and the reaction mixture
was stirred for 2 h. Excess LAH was destroyed by slow
addition of ethyl acetate and solvent was distilled off.
Extraction with ethyl acetate and elution of 40% ethyl
acetate in hexane over silica gel furnished the methanol
8: white solid (1.67 g, 83%), mp 176 °C, IR (KBr): 2929,
1611, 1511, 1464, 1248, 1160, 1034, 760 cmÀ1, 1H NMR
(CDCl3, 200 MHz): d 7.90 (d, 1H, J = 8.4 Hz), 7.85 (s,
1H), 7.79 (s, 1H), 7.64 (s, 1H), 7.31 (d, 2H,
J = 8.6 Hz), 7.15 (s, 1H), 7.07 (dd, 1H, J1 = 8.4 Hz,
J2 = 2.6 Hz), 6.82 (d, 2H, J = 8.6 Hz), 6.37 (s, 1H),
4.07 (s, 3H), 3.99 (s, 3H), 3.95 (s, 3H), 3.75 (s, 3H),
1.80 (br s, 1H); MS: 404 (M+). Anal. Calcd for
C25H24O5: C, 74.24; H, 5.98. Found: C, 74.28; H, 6.03.
4.7. (2-{4-[(4-Methoxy-phenyl)-(2,3,6-trimethoxy-phe-
nanthren-9-yl)-methyl]- phenoxy}-ethyl)-dimethyl-amine
(11a)
4.3. Representative Friedel–Crafts alkylation
Pale yellow solid (300 mg, 87%), mp 100 °C, IR (KBr):
2934, 1614, 1509, 1462, 1244, 1032 cmÀ1 1H NMR
,
To a solution of carbinol 8 (1.5 g, 3.71 mmol) and phe-
nol (523 mg, 5.57 mmol) in dry benzene (15 mL), cata-
lytic amount of concd. H2SO4 was added and the
reaction mixture was refluxed at 80 °C for 1 h. It was
cooled to room temperature, treated with saturated
NaHCO3, and extracted with ethyl acetate. The organic
layer was washed with water and dried over anhydrous
Na2SO4. Column chromatography over silica gel and
elution with 15% ethyl acetate in hexane furnished the
desired compound (1.15 g, ortho and para together,
65%).
(CDCl3, 200 MHz): d 7.94 (d, 1H, J = 8 Hz), 7.89 (s,
1H), 7.87 (s, 1H), 7.10 (d, 1H, J = 7.8 Hz), 7.07 (s,
1H), 7.02 (d, 4H, J = 8.2 Hz), 6.89 (s, 1H), 6.82 (d,
4H, J = 8.2 Hz), 4.09 (s, 3H), 4.05 (t, 2H, J = 7 Hz),
3.97 (s, 6H), 3.78 (s, 3H), 2.71 (t, 2H, J = 7 Hz), 2.32
(s, 6H); MS: 552 (M+), Anal. Calcd for C35H37NO5:
C, 76.20; H, 6.76; N, 2.54. Found: C, 77.29; H, 6.69;
N, 2.57.
4.8. Diethyl-(2-{4-[(4-methoxy-phenyl)-(2,3,6-trimethoxy-
phenanthren-9-yl)-methyl]-phenoxy}-ethyl)-amine (11b)
4.4. 4-[(4-Methoxy-phenyl)-(2,3,6-trimethoxy-phenan-
thren-9-yl)-methyl]-phenol (9)
Pale yellow solid (300 mg, 82%), mp 105 °C, IR (KBr):
2962, 1612, 1509, 1463, 1246, 1034 cmÀ1 1H NMR
,
(CDCl3, 200 MHz): d 7.95 (d, 1H, J = 8 Hz), 7.91 (s,
1H), 7.88 (s, 1H), 7.10 (d, 1H, J = 7.8 Hz), 7.05 (d,
4H, J = 8.4 Hz), 7.02 (s, 1H), 6.92 (s, 1H), 6.85 (d, 4H,
J = 8.4 Hz), 6.11 (s, 1H), 4.11 (s, 3H), 4.06 (t, 2H, J =
7 Hz), 3.99 (s, 6H), 3.80 (s, 3H), 2.90 (t, 2H, J = 7 Hz),
2.67 (q, 4H, J = 7 Hz), 1.09 (t, 2H, J = 7 Hz); MS: 580
Pale yellow solid (810 mg), mp 175 °C, IR (KBr): 2929,
1611, 1465, 1243, 1033 cmÀ1
,
1H NMR (CDCl3,
200 MHz): d 7.93 (d, 1H, J = 7 Hz), 7.90 (s, 1H), 7.88
(s, 1H), 7.10 (d, 1H, J = 7 Hz), 7.07 (s, 1H), 7.04 (d,
2H, J = 8 Hz), 7.02 (d, 1H, J = 7 Hz), 6.99 (d, 1H,

Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
1503
(M1+), Anal. Calcd for C37H41NO5: C, 76.66; H, 7.13;
N, 2.42. Found: C, 76.71; H, 7.19; N, 2.39.
Syntheses of 15 and 16 are reported earlier (Ref. 8).
4.13. 2-{4-[(4-Methoxy-phenyl)-phenanthren-9-yl-meth-
yl]-phenoxymethyl}-oxirane (17)
4.9. 1-(2-{4-[(4-Methoxy-phenyl)-(2,3,6-trimethoxy-phe-
nanthren-9-yl)-methyl]-phenoxy}-ethyl)-pyrrolidine (11c)
The compound 13c (2.0 g, 5.12 mmol) and anhydrous
K2CO3 (2.52 g, 18.23 mmol) were taken in epichloro-
hydrin (75 mL) and refluxed for 12 h. K2CO3 was fil-
tered off and epichlorohydrin was removed in vacuo.
The residue was diluted with water and extracted with
ethyl acetate. The organic layer was washed with
water, brine, and dried over anhydrous Na2SO4. Col-
umn chromatography over silica gel and elution with
20% ethyl acetate in hexane furnished the epoxide
17. Pale yellow solid (1.97 g, 86%), mp 90 °C, IR
(KBr): 3017, 1608, 1508, 1456, 1246, 1178,
Light brown solid (270 mg, 78%), mp 173 °C, IR (KBr):
1
2933, 1612, 1509, 1247, 1036 cmÀ1, H NMR (CDCl3,
200 MHz): d 7.94 (d, 1H, J = 8 Hz), 7.90 (s, 1H), 7.85
(s, 1H), 7.10 (d, 1H, J = 7.8 Hz), 7.02 (s, 1H), 6.90 (s,
1H), 6.82 (d, 4H, J = 8.4 Hz), 6.08 (s, 1H), 4.08 (s,
3H), 4.04 (t, 2H, J = 6 Hz), 3.96 (s, 6H), 3.77 (s, 3H),
2.88 (t, 2H, J = 6 Hz), 2.61–2.59 (m, 4H), 1.82–1.76
(m, 4H); MS: 578 (M1+), Anal. Calcd for C37H39NO5:
C, 76.92; H, 6.80; N, 2.42. Found: C, 76.99; H, 6.89;
N, 2.49.
1
1037 cmÀ1 , H NMR (CDCl3, 200 MHz): d 8.71 (d,
4.10. 1-(2-{4-[(4-Methoxy-phenyl)-(2,3,6-trimethoxy-phe-
nanthren-9-yl)-methyl]-phenoxy}-ethyl)-piperidine (11d)
1H, J = 8.2 Hz), 8.64 (d, 1H, J = 8 Hz), 8.02 (d, 1H,
J = 8 Hz), 7.69–7.47 (m, 5H), 7.13 (s, 1H), 7.05 (d,
4H, J = 8.2 Hz), 6.83 (d, 4H, J = 8.2 Hz), 6.14 (s,
1H), 4.16 (dd, 1H, J1 = 10 Hz, J2 = 3.2 Hz), 3.93 (dd,
1H, J1 = 10 Hz, J2 = 5.6 Hz), 3.77 (s, 3H), 3.33–3.20
(m, 1H), 2.90–2.85 (m, 1H), 2.74–2.71 (m, 1H); MS:
446 (M+). Anal. Calcd for C31H26O3: C, 83.38; H,
5.87. Found: C, 83.42; H, 5.83.
Pale yellow solid (300 mg, 81%), mp 107 °C, IR (KBr):
2930, 1612, 1509, 1463, 1245, 1036 cmÀ1 1H NMR
,
(CDCl3, 200 MHz): d 7.93 (d, 1H, J = 8 Hz), 7.90 (s,
1H), 7.86 (s, 1H), 7.10 (d, 2H, J = 7.8 Hz), 7.06 (d,
4H, J = 8.2 Hz), 6.90 (s, 1H), 6.82 (d, 4H, J = 8.2 Hz),
4.08 (t, 2H, J = 7 Hz), 4.07 (s, 3H), 3.96 (s, 6H), 3.78
(s, 3H), 2.75 (t, 2H, J = 7 Hz), 2.49 (t, 4H, J = 7 Hz),
1.62–1.40 (m, 6H); MS: 592 (M1+), Anal. Calcd for
C38H41NO5: C, 77.13; H, 6.98; N, 2.37. Found: C,
77.09; H, 6.91; N, 2.32.
4.14. Representative epoxide opening reactions with
amines
The compound 17 (300 mg, 0.67 mmol) and amine
(1.05 mmol) were dissolved in ethanol (20 mL) and re-
fluxed for 7 h. The ethanol was removed and the res-
idue was diluted with water and extracted with ethyl
acetate. The organic layer was washed with water,
brine, and dried over anhydrous Na2SO4. Column
chromatography over silica gel and elution with 5%
methanol in chloroform furnished the title compound
given below. The aminopropan-2-ol derivatives were
dissolved in absolute ethanol (15 mL) and ethanolic
HCl was added dropwise untill the pH of the mixture
became acidic. Ethanol was removed in vacuo. The
residue was recrystallized from a mixture of absolute
ethanol and dry ether.
4.11. Diethyl-(2-{2-[(4-methoxy-phenyl)-(2,3,6-trimeth-
oxy-phenanthren-9-yl)-methyl]-phenoxy}-ethyl)-amine
(12a)
Pale yellow solid (370 mg, 99%), mp 175 °C, IR (KBr):
2964, 1613, 1510, 1244, 1043, 757 cmÀ1 1H NMR
,
(CDCl3, 200 MHz): d 7.93 (d, 1H, J = 8 Hz), 7.90 (s,
1H), 7.87 (s, 1H), 7.21 (m, 1H), 7.10 (d, 1H,
J = 7.8 Hz), 7.09 (d, 1H, J = 7.8 Hz), 7.03 (d, 2H,
J = 8.4 Hz), 6.93 (s, 1H), 6.89 (d, 1H, J = 8 Hz), 6.83
(d, 1H, J = 7.8 Hz), 6.82 (s, 1H), 6.81 (d, 2H,
J = 8.4 Hz), 6.48 (s, 1H), 4.09 (s, 3H), 3.97 (t, 2H, J =
6 Hz), 3.96 (s, 6H), 3.79 (s, 3H), 2.58 (t, 2H, J = 6 Hz),
2.39 (q, 4H, J = 7 Hz), 0.85 (t, 6H, J = 7.8 Hz); MS:
580 (M1+). Anal. Calcd for C37H41NO5: C, 76.66; H,
7.13; N, 2.42. Found: C, 76.60; H, 7.07; N, 2.38.
4.15. 1-{4-[(4-Methoxy-phenyl)-phenanthren-9-yl-meth-
yl]-phenoxy}-3-pyrrolidin-1-yl-propan-2-ol (18a)
Pale yellow solid, mp 110 °C (260 mg, 75%), IR (KBr):
4.12. 1-(2-{2-[(4-Methoxy-phenyl)-(2,3,6-trimethoxy-phe-
nanthren-9-yl)-methyl]-phenoxy}-ethyl)-piperidine (12b)
3011, 1606, 1508, 1457, 1246, 1038, 754 cmÀ1 1H
,
NMR (CDCl3, 200 MHz): d 8.70 (d, 1H, J = 8 Hz),
8.64 (d, 1H, J = 8.2 Hz), 8.01 (d, 1H, J = 8.2 Hz),
7.64–7.47 (m, 5H), 7.13 (s, 1H), 7.04 (d, 4H,
J = 8.4 Hz), 6.81 (d, 4H, J = 8.4 Hz), 6.13 (s, 1H), 5.76
(br s, 1H), 4.25 (m, 1H), 4.02–3.87 (m, 2H), 3.77 (s,
3H), 3.11–2.92 (m, 6H), 2.03–1.93 (m, 4H); MS: 518
(M1+). Anal. Calcd for C35H35NO3: C, 81.21; H, 6.81;
N, 2.71. Found: C, 81.25; H, 6.87; N, 2.78.
Pale yellow solid, mp 171 °C (270 mg, 90%), IR (KBr):
2933, 1614, 1511, 1244, 1036, 754 cmÀ1 1H NMR
,
(CDCl3, 200 MHz): d 7.93 (d, 1H, J = 8 Hz), 7.90 (s,
1H), 7.86 (s, 1H), 7.20 (m, 1H), 7.10 (d, 1H,
J = 7.8 Hz), 7.09 (d, 1H, J = 7.8 Hz), 7.03 (d, 2H,
J = 8.4 Hz), 6.92 (s, 1H), 6.86 (d, 1H, J = 8 Hz), 6.83
(d, 1H, J = 7.8 Hz), 6.82 (s, 1H), 6.80 (d, 2H,
J = 8.2 Hz), 6.48 (s, 1H), 4.09 (s, 3H), 4.03 (t, 2H,
J = 7 Hz), 3.97 (s, 6H), 3.78 (s, 3H), 2.52 (t, 2H,
J = 6 Hz), 2.19–2.16 (m, 4H), 1.35–1.22 (m, 6H); MS:
592 (M1+). Anal. Calcd for C38H41NO5: C, 77.13; H,
6.98; N, 2.37. Found: C, 77.17; H, 7.02; N, 2.43.
4.16. 1-{4-[(4-Methoxy-phenyl)-phenanthren-9-yl-meth-
yl]-phenoxy}-3-piperidin-1-yl-propan-2-ol (18b)
Pale yellow solid, mp 105 °C (340 mg, 96%), IR (KBr):
2934, 1607, 1507, 1246, 1177, 1037, 755 cmÀ1 1H
,

1504
Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
NMR (CDCl3, 200 MHz): d 8.71 (d, 1H, J = 8.2 Hz),
8.64 (d, 1H, J = 8 Hz), 8.02 (d, 1H, J = 8.2 Hz), 7.69–
7.47 (m, 5H), 7.14 (s, 1H), 7.05 (d, 4H, J = 8.4 Hz),
6.82 (d, 4H, J = 8.4 Hz), 6.14 (s, 1H), 4.11 (m, 1H),
3.96–3.92 (m, 2H), 3.78 (s, 3H), 3.51 (br s, 1H), 2.69–
2.42 (m, 6H), 1.51–1.45 (m, 6H); MS: 532 (M1+). Anal.
Calcd for C36H37NO3: C, 81.32; H, 7.01; N, 2.63.
Found: C, 81.39; H, 7.08; N, 2.68.
3H), 3.51 (s, 2H), 3.01–2.50 (m, 10H); MS: 623 (M1+).
Anal. Calcd for C42H42N2O3: C, 81.00; H, 6.80; N,
4.50. Found: C, 81.09; H, 6.86; N, 4.55.
4.21. 1-Cyclopropylamino-3-{4-[(4-methoxy-phenyl)-phe-
nanthren-9-yl-methyl]-phenoxy}-propan-2-ol (18g)
Pale yellow solid, mp 115 °C (216 mg, 64%), IR (KBr):
3015, 1607, 1508, 1456, 1243, 1178, 1037, 743 cmÀ1
,
8.71 (d, 1H,
4.17. 1-{4-[(4-Methoxy-phenyl)-phenanthren-9-yl-meth-
yl]-phenoxy }-3-(4-methyl-piperazin-1-yl)-propan-2-ol
(18c)
1H NMR (CDCl3, 200 MHz):
d
J = 8.2 Hz), 8.64 (d, 1H, J = 8.2 Hz), 8.02 (d, 1H,
J = 8.2 Hz), 7.69–7.44 (m, 5H), 7.14 (s, 1H), 7.05 (d,
4H, J = 8.4 Hz), 6.82 (d, 4H, J = 8.4 Hz), 6.14 (s, 1H),
4.07–4.03 (m, 1H), 3.95–3.88 (m, 2H), 3.78 (s, 3H),
2.94–2.82 (m, 2H), 2.38 (br s, 1H), 0.47–0.36 (m, 4H);
MS: 504 (M1+). Anal. Calcd for C34H33NO3: C, 81.08;
H, 6.60; N, 2.78. Found: C, 81.01; H, 6.54; N, 2.68.
Pale yellow solid, mp 195 °C (256 mg, 70%), IR (KBr):
3016, 1668, 1508, 1455, 1245, 1219, 768 cmÀ1 1H
,
NMR (CDCl3, 200 MHz): d 8.71 (d, 1H, J = 8.4 Hz),
8.65 (d, 1H, J = 8.4 Hz), 8.03 (d, 1H, J = 8 Hz), 7.69–
7.47 (m, 5H), 7.14 (s, 1H), 7.05 (d, 4H, J = 8.4 Hz),
6.83 (d, 4H, J = 8.4 Hz), 6.14 (s, 1H), 4.08–4.04 (m,
1H), 3.96–3.94 (m, 2H), 3.78 (s, 3H), 2.75–2.34 (m,
10H), 2.29 (s, 3H); MS: 547 (M1+). Anal. Calcd for
C36H38N2O3: C, 79.01; H, 7.01; N, 5.12. Found: C,
79.05; H, 7.11; N, 5.17.
4.22. Test procedure for the evaluation of antiproliferative
cytotoxic activity in vitro
The procedure is based on the following methods: new
colorimetric assay for anticancer drug screening, Skehan
et al.13 and feasibility of high-flux anticancer drug screen
using a diverse panel of cultured human tumor cell lines
Saxena et al.14
4.18. 1-Cyclohexylamino-3-{4-[(4-methoxy-phenyl)-phe-
nanthren-9-yl-methyl]-phenoxy}-propan-2-ol (18d)
Pale yellow solid, mp 155 °C (307 mg, 84%), IR (KBr):
Briefly, a fully confluent flask of MCF-7 cells was tryp-
sinized and 104 cells/well were plated in a 96-well flat-
bottomed plate in 200 ll minimum essential medium
(MEM), pH 7.4, and allowed to attach for 24 h at
37 °C in a humidified CO2 incubator. Subsequently,
the compound of invention dissolved in DMSO or eth-
anol was added at specified concentration and further
incubated for 48 h as before. The cells were then fixed
in 50 ll cold 50% TCA and incubated for 1 h at 4 °C.
The supernatant was discarded and the plate was
washed five times with deionized water and air-dried.
Hundred microliters of 0.4% (w/v) Sulforhodamine B
(SRB) in 1% acetic acid was added to each well and
incubated at room temperature for 30 min. Unbound
SRB was removed by five washes with chilled 1% acetic
acid and the plate was air-dried. Two hundred microli-
ters of unbuffered 10 mM Tris base was added to solubi-
lize the bound stain for 5 min at room temperature and
OD was read at 560 nm in a plate reader. The graph is
plotted between OD and concentration, and IC50 is cal-
culated with respect to tamoxifen that is used as positive
control.
2939, 1608, 1509, 1408, 1246, 1219, 1040, 766 cmÀ1
,
8.71 (d, 1H,
1H NMR (CDCl3, 200 MHz):
d
J = 8.2 Hz), 8.65 (d, 1H, J = 7.8 Hz), 7.99 (d, 1H,
J = 8.2 Hz), 7.65–7.47 (m, 5H), 7.13 (s, 1H), 7.04 (d,
4H, J = 8.4 Hz), 6.84 (d, 4H, J = 8.4 Hz), 6.13 (s, 1H),
4.20 (br s, 1H), 4.07–4.04 (m, 1H), 3.95–3.88 (m, 2H),
3.78 (s, 3H), 2.95–2.86 (m, 3H), 1.86–1.64 (m, 4H),
1.32–1.13 (m, 6H); MS: 546 (M1+). Anal. Calcd for
C37H39NO3): C, 81.43; H, 7.20; N, 2.57. Found: C,
81.37; H, 7.26; N, 2.49.
4.19. 1-{4-[(4-Methoxy-phenyl)-phenanthren-9-yl-meth-
yl]-phenoxy}-3-morpholin-4-yl-propan-2-ol (18e)
Pale yellow solid, mp 120 °C (257 mg, 72%), IR (KBr):
3417, 1609, 1504, 1449, 1243, 1113, 1032, 747 cmÀ1
,
8.71 (d, 1H,
1H NMR (CDCl3, 200 MHz):
d
J = 8.2 Hz), 8.65 (d, 1H, J = 8.2 Hz), 8.03 (d, 1H, J =
8 Hz), 7.69–7.44 (m, 5H), 7.14 (s, 1H), 7.05 (d, 4H,
J = 8.4 Hz), 6.84 (d, 4H, J = 8.4 Hz), 6.14 (s, 1H),
4.13–4.06 (m, 1H), 3.97–3.95 (m, 2H), 3.78 (s, 3H),
3.74–3.70 (m, 4H), 2.68–2.42 (m, 6H); MS: 534 (M1+).
Anal. Calcd for C35H35NO4: C, 78.77; H, 6.61; N,
2.62. Found: C, 78.72; H, 6.66; N, 2.68.
4.23. In vivo evaluation of anti-breast cancer activity
The Huggins15 rat model of 7,12-dimethylbenz[a]anthra-
cene induced hormone responsive breast cancer was
used for evaluating the anti-cancer potential of the
compound. Groups of rats (n = 3/per group) with palpa-
ble (0.5 cm diameter) tumor were given by oral gavage
the compound 15ca (10 and 20 mg/kg dose levels),
tamoxifen (10 mg/kg) or the vehicle (1% gum acacia)
only (untreated control), once daily for 4 weeks. Tamox-
ifen was used as a standard SERM for comparison.
Tumor dimensions were measured at least once weekly
during the period of treatment and the tumor volume
4.20. 1-(4-Benzyl-piperazin-1-yl)-3-{4-[(4-methoxy-phen-
yl) -phenanthren-9-yl-methyl]-phenoxy}-propan-2-ol (18f)
Pale yellow solid, mp 175 °C (250 mg, 60%), IR (KBr):
3020, 1216, 758, 669 cmÀ1
,
1H NMR (CDCl3,
200 MHz): d 8.71 (d, 1H, J = 8.2 Hz), 8.64 (d, 1H,
J = 8.2 Hz), 8.02 (d, 1H, J = 8.2 Hz), 7.64–7.47 (m,
5H), 7.30–7.25 (m, 5H), 7.13 (s, 1H), 7.05 (d, 4H,
J = 8.4 Hz), 6.82 (d, 4H, J = 8.4 Hz), 6.14 (s, 1H),
4.09–4.05 (m, 1H), 3.94 (d, 2H, J = 4.8 Hz), 3.78 (s,

Shagufta et al. / Bioorg. Med. Chem. 14 (2006) 1497–1505
1505
37, 1115–1125; (e) Akama, T.; Shida, Y.; Sugaya, T.;
Ishida, H.; Gomi, K.; Kasai, M. J. Med. Chem. 1996, 39,
3461–3469; (f) Lubczyk, V.; Bachmann, H.; Gust, R. J.
Med. Chem. 2003, 46, 1484–1491; (g) Gauthier, S.; Caron,
B.; Cloutier, J.; Dory, Y. L.; Favre, A.; Larouche, D.;
Mailhot, J.; Ouellet, C.; Schwerdtfeger, A.; Leblanc, G.;
Martel, C.; Simard, J.; Merand, Y.; Belanger, A.; Labrie,
C.; Labrie, F. J. Med. Chem. 1997, 40, 2117–2122; (h)
Poirier, D.; Auger, S.; Merand, Y.; Simard, J.; Labrie, F.
J. Med. Chem. 1994, 37, 1115–1125; (i) Sharma, A. P.;
Saeed, A.; Durani, S.; Kapil, R. S. J. Med. Chem. 1990, 33,
3216–3222.
was calculated using the formula for ellipsoids. The
effect of the treatment was expressed as % change in
tumor volume.
Acknowledgments
We thank Mr. Pramod Kumar for his technical assistance
in this project. Shagufta, Ajay and Rajeev thank CSIR for
providing fellowships. This research project was support-
ed by Department of Science and Technology (SR/FTP/
CSA-05/2002), New Delhi, India. We thank the referees
for their valuable comments and suggestions which
helped us a great deal to improve the quality of the paper.
7. (a) Boger, D. L.; Mitscher, L. A.; Mullican, M. D.; Drake,
S. D.; Kitos, P. J. Med. Chem. 1985, 28, 1543–1547; (b)
Miles, D. H.; Bhattacharya, J.; Mody, N. V.; Atwood, J.
L.; Black, S.; Hedin, P. A. J. Am. Chem. Soc. 1977, 99,
618; (c) Coxon, D. T.; Ogundana, S. K.; Dennis, C.
Phytochemistry 1982, 21, 1389–1392; (d) Ridley, R. G.
Nature 2002, 415, 686–693.
References and notes
8. Panda, G.; Shagufta; Mishra, J. K.; Chaturvedi, V.;
Srivastava, A. K.; Srivastava, R.; Srivastava, B. S. Bioorg.
Med. Chem. 2004, 12, 5269–5276.
9. Schmidt, J. M.; Mercure, J.; Tremblay, G. B.; Page,
M.; Kalbakji, A.; Feher, M.; Dunn-Dufault, R.; Peter,
M. G.; Redden, P. R. J. Med. Chem. 2003, 46, 1408–
1418.
10. Jordan, V. C. J. Med. Chem. 2003, 46, 883–908.
11. (a) Wakeling, A. E. Breast Cancer Res. Treat. 1993, 25,
1–9; (b) Wakeling, A. E.; Bowler, J. J. Steroid Biochem.
1988, 30, 141–147; (c) Wakeling, A. E.; Dukes, M.;
Bowler, J. J. Cancer Res. 1991, 51, 3867–3873; (d)
Wakeling, A. E.; Bowler, J. J. Steroid Biochem. 1992, 43,
170–177; (e) de Launoit, Y.; Dauvois, S.; Dufour, M.;
Simard, J.; Labrie, F. Cancer Res. 1991, 51, 2797–2802; (f)
Levesque, C.; Merand, Y.; Dufour, J. M.; Labrie, C.;
Labrie, F. J. Med. Chem. 1991, 34, 1624–1630; (g) Labrie,
C.; Martel, C.; Dufour, J. M.; Levesque, C.; Merand, Y.;
Labrie, F. Cancer Res. 1992, 52, 610–615.
1. (a) Jordan, V. C. Breast Cancer Res. Treat. 1995, 36,
267–285; (b) Gradishar, W. J.; Jordan, V. C. J. Clin.
Oncol. 1997, 15, 840–852; (c) Ray, S.; Dwivedy, I. Adv.
Drug Res. 1997, 29, 171–270, and references cited
therein.
2. Jordan, V. C. Breast Cancer Res. Treat. 1994, 31, 41–52.
3. (a) McGuire, W. L. Semin. Oncol. 1978, 5, 428–433; (b)
Miller, W. R. J. Steroid Biochem. Mol. Biol. 1990, 37, 467–
480; (c) Jordan, V. C. Pharmacol. Rev. 1984, 36(4), 245–
276; (d) Magarian, R. A.; Overacre, L. B.; Singh, S.;
Meyer, K. L. Curr. Med. Chem. 1994, 1, 61–104.
4. (a) Jordan, V. C. Cancer 1992, 70, 977–982; (b) Meegan,
M. J.; Hughes, R. B.; Lloyd, D. G.; Williams, D. C.;
Zisterer, D. M. J. Med. Chem. 2001, 44, 1072–1084; (c)
Jordan, V. C.; Collins, M. M.; Rowsby, L.; Prestwich, G.
A. J. Endocrinol. 1977, 75, 305–316; (d) Jordan, V. C.
J. Med. Chem. 2003, 46, 1081–1111; (e) Lerner, L. J.;
Jordan, V. C. Cancer Res. 1990, 50, 4177–4189.
5. (a) MacGregor, J.; Jordan, V. C. Pharmacol. Rev. 1998,
50, 151–196; (b) Mitlak, B. H.; Cohen, F. J. Horm. Res.
12. Bradsher, C. K.; Berger, H. J. Am. Chem. Soc. 1958, 80,
930–932.
13. Skehan, P.; Storeng, R.; Scudiero, D.; Monks, A.;
Mcmahon, J.; Vistica, D.; Warren, J. T.; Bokesch, H.;
Kenney, S.; Boyd, M. R. J. Natl. Cancer Inst. 1990, 82,
1107–1112.
14. Saxena, A. K.; Ramchandani, S.; Dwivedi, A.;
Sharma, R.; Bajpai, V. K.; Bhardwaj, K. R.; Bala-
pure, A. K. In vitro Cell. Dev. Biol. Anim. 1995, 31,
326–329.
15. (a) Huggins, C. G.; Brillantes, F. P. Nature 1961, 189,
204–207; (b) Huggins, C.; Yang, N. C. Science 1962,
137, 257–262.
´
1997, 48, 155–163; (c) Smith, C. L.; O Malley, B. W.
Trends Endocrinol. Metab. 1999, 10, 299–300; (d) McDon-
nell, D. P. Trends Endocrinol. Metab. 1999, 10, 301–311.
6. (a) Thompson, E. W.; Katz, D.; Shima, T. B.; Wakeling,
A. E.; Lippman, M. E.; Dickson, R. B. Cancer Res. 1989,
49, 6929–6934; (b) Jones, C. D.; Blaszczak, L. C.; Goettel,
M. E.; Suarez, T.; Crowell, T. A.; Mabry, T. E.; Ruenitz,
P. C.; Srivatsan, V. J. Med. Chem. 1992, 35, 931–938; (c)
Angerer, E. V.; Knebel, N.; Kager, M.; Ganss, B. J. Med.
Chem. 1990, 33, 2635–2640; (d) Poirier, D.; Auger, S.;
Merand, Y.; Simard, J.; Labrie, F. J. Med. Chem. 1994,