Triarylcyclopropanes as antiestrogens and antitumor agents

Triarylcyclopropane derivatives in which one or more of the aryl groups includes a hydroxy as a substituted or unsubstituted or alkoxy or side chain substituent. The compounds are useful as antiestrogens and anti-tumor agents.

FIELD OF THE INVENTION 
This invention relates to triarylcyclopropane compounds and their use in 
mammals for producing anti-estrogenic activity in the mammal, and for 
inhibiting the development of an estrogen-dependent tumor in the mammal. 
SUMMARY OF THE INVENTION 
The present invention comprises a compound having the formula: 
##STR1## 
or any pharmaceutically acceptable salt thereof, in which R.sub.1 is 
hydrogen, a hydroxyl group, an alkoxy group or a substituted or 
unsubstituted arylalkoxy group (with the proviso that R.sub.1, R.sub.2 and 
R.sub.3 are not simultaneously arylalkoxy). Any substituent of the aryl 
group comprises an alkyl group. R.sub.2 is hydrogen, a hydroxyl group 
(with the proviso that R.sub.1 and R.sub.3 are not simultaneously alkoxy 
when R.sub.2 is hydroxyl), an alkoxy group or a substituted alkoxy group. 
The substituent of the alkoxy group comprises either a dialkylamino group, 
a heterocycle containing between about 5 and 6 members, at least one of 
which is nitrogen, or a substituted or unsubstituted aryl group, in which 
any substituent of the aryl group comprises an alkyl group. R.sub.3 is 
hydrogen, a hydroxyl group, an alkoxy group or a substituted alkoxy group 
and in which the substituent of the alkoxy group comprises either a 
dialkylamino group, or a heterocycle containing between about 5 and 6 
members, at least one of which is nitrogen, or a substituted or 
unsubstituted aryl group, in which any substituent of the aryl group 
comprises an alkyl group. R.sub.4 is hydrogen, and X is a halogen or 
hydrogen. The compounds of the present invention may be combined with a 
pharmaceutically acceptable carrier to form pharmaceutical compositions. 
The present invention further comprises a method of inducing antiestrogenic 
activity in a mammal in need of such therapy comprising administering to 
the mammal an antiestrogenically effective amount of one or more compounds 
having the above-described formula. 
The present invention also comprises a method of inhibiting the development 
of an estrogen-dependent tumor in a mammal in need of such therapy 
comprising administering to the mammal an effective amount of one or more 
compounds having the above-described formula. 
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
The present invention comprises a compound having the formula: 
##STR2## 
or any pharmaceutically acceptable salt thereof. R.sub.1 represents a 
hydrogen, a hydroxyl group, an alkoxy group, preferably containing from 1 
to about 3 carbon atoms, or a substituted or unsubstituted arylalkoxy 
group (with the proviso that R.sub.1, R.sub.2 and R.sub.3 are not 
simultaneously arylalkoxy). The alkyl group attached to the oxygen atom in 
such an arylalkoxy group preferably contains from 1 to about 3 carbon 
atoms, and any substituent of the aryl group preferably comprises an alkyl 
group, preferably containing from 1 to about 3 carbon atoms. 
In the compounds of the present invention R.sub.2 is hydrogen, a hydroxyl 
group (with the proviso that R.sub.1 and R.sub.3 preferably are not 
simultaneously alkoxy when R.sub.2 is a hydroxyl group), an alkoxy group, 
preferably containing from 1 to about 3 carbon atoms, or a substituted 
alkoxy group in which the alkyl group attached to the oxygen atom 
preferably contains from 1 to about 3 carbon atoms. The substituent of the 
alkoxy group preferably comprises either a dialkylamino group in which 
each alkyl substituent thereof preferably contains from 1 to about 3 
carbon atoms, a heterocycle preferably containing between about 5 and 6 
members, at least one of which is nitrogen, or a substituted or 
unsubstituted aryl group, in which any substituent of the aryl group 
preferably comprises an alkyl group preferably containing from 1 to about 
3 carbon atoms. 
R.sub.3 in the compounds of the present invention is either hydrogen, a 
hydroxyl group, an alkoxy group, preferably containing from 1 to about 3 
carbon atoms, or a substituted alkoxy group in which the alkyl group 
attached to the oxygen atom preferably contains from 1 to about 3 carbon 
atoms and in which the substituent of the alkoxy group preferably 
comprises either a dialkylamino group in which each alkyl substituent 
thereof preferably contains from 1 to about 3 carbon atoms, or a 
heterocycle preferably containing between about 5 and 6 members, at least 
one of which is nitrogen, or a substituted or unsubstituted aryl group, in 
which any substituent of the aryl group preferably comprises an alkyl 
group preferably containing from 1 to about 3 carbon atoms. 
R.sub.4 is hydrogen, and X is a halogen or hydrogen. In a preferred 
embodiment R.sub.4 is a halogen, and more preferably, is chlorine. 
Compounds within the scope of the present invention include both the (R) 
and (S) optical isomers having the formula described above, and racemic 
mixtures of these optical isomers. 
One particularly preferred compound of the present invention comprises 
(R/S)-(Z)-1,1-dichloro-2,3-diphenyl-2-(4-methoxyphenyl)cyclopropane 
including each of its (R) and (S) optical isomers: 
##STR3## 
Another preferred compound is 
(Z)-1,1-dichloro-2,3-diphenyl-2-[4[2-(dimethylamino)-ethoxy]phenyl]cyclopr 
opane, citrate salt: 
##STR4## 
including each of its (R) and (S) optical isomers and racemic mixtures 
thereof. 
Another preferred compound is 
(Z)-1,1-dichloro-2-[4-[2-(dimethylamino)-ethoxy]phenyl]-2-(4-methoxyphenyl 
)-3-phenylcyclopropane: 
##STR5## 
including each of its (R) and (S) optical isomers and racemic mixtures 
thereof. 
Another preferred compound is 
(Z)-1,1-dichloro-2-(4-benzyloxyphenyl)-2-(4-methoxyphenyl)-3-phenylcyclopr 
opane: 
##STR6## 
including each of its (R) and (S) optical isomers and racemic mixtures 
thereof. 
Yet another preferred compound is (E) and 
(Z)-1,1-dichloro-2-(4-benzyloxyphenyl)-2,3-bis-(4-methoxyphenyl)cyclopropa 
ne: 
##STR7## 
including each of its (R) and (S) optical isomers and racemic mixtures 
thereof. 
Preferably, the compounds of the present invention are combined with a 
pharmaceutically acceptable carrier to form a pharmaceutical composition 
appropriate for therapeutic delivery to a mammal. The pharmaceutically 
acceptable carrier should not substantially interfere with the 
anti-estrogenic and anti-tumor activities of the compound, and may be a 
solid or liquid in which the compound is solubilized, suspended or 
dispersed in any manner. The compounds of the present invention may be 
administered orally in solid dosage forms, such as tablets, and powders, 
or in liquid dosage forms, such as elixirs, syrups and suspensions; they 
may also be administered parenterally, in sterile liquid dosage forms. 
Such parenteral administration may include intravenous, intramuscular, 
subcutaneous, intra-arterial, and direct tumor perfusion techniques. 
If the compound is to be injected, the pharmaceutical carrier should 
preferably be isotonic, and have about a physiological pH. Suitable 
pharmaceutical carriers for parenteral administration may be any suitable 
oil, saline, aqueous dextrose or related sugar solutions, or glycols such 
as propylene glycol or polyethylene glycols. Solutions for parenteral 
administration contain preferably a water soluble salt of the active 
ingredient, suitable stabilizing agents, and if necessary, buffer 
substances. Additionally, parenteral solutions can contain preservatives. 
Other suitable pharmaceutical carriers are described in Remington's 
Pharmaceutical Sciences, Mack Publishing Co., and similar reference texts. 
The present invention further comprises a method of inducing antiestrogenic 
activity in a mammal, such as a human, in need of such therapy comprising 
administering to the mammal an antiestrogenically effective amount of one 
or more compounds having the formula, preferably in the form described 
above. The dosage of the compounds of the present invention may vary due 
to the therapeutically desired result which is affected by the type of 
disease or condition in the mammal; the age, weight and health of the 
recipient; the severity of the condition or disease in the mammal; the 
kind of concurrent treatment, if any, being administered to the mammal; 
and the frequency of treatment. Generally, a therapeutically effective 
dosage is less than about 0.5 mg to about 2 mg per kilogram of body weight 
of the mammal over a 24 hour period. The method of administration of the 
compound of the present invention can be by any suitable method as 
previously described. 
The present invention also comprises a method of inhibiting the development 
of an estrogen-dependent tumor in a mammal, such as a human, in need of 
such therapy comprising administering to the mammal a therapeutically 
effective amount of one or more compounds having the formula described 
above, preferably in the form of a pharmaceutical composition comprising 
at least one of the compounds combined with a pharmaceutically acceptable 
carrier. "Inhibiting the development of an estrogen-dependent tumor" means 
either slowing the growth of a tumor, diminishing the size of a tumor, or 
preventing the formation of a tumor from cells having the potential of 
developing into a tumor wherein the tumor requires the presence of an 
estrogenic substance for the growth, development and/or metastatic 
involvement of the tumor. 
The compounds previously described may be administered to the mammal to 
inhibit the development of the estrogen-dependent tumor by an 
administration method of the type previously described. The dosage may 
vary according to the type of the disease; the size of the tumor or 
tumors, if present; and the quantity of tumors as well as other factors 
previously described. Generally, a daily dosage of less than about 0.5 mg 
to about 2 mg/kg of body weight of the mammal will suffice. 
The following examples illustrate the practice of the present invention.

EXAMPLE 1 
Synthetic Preparation of the Compounds of the Present Invention 
Exemplary compounds of the present invention include compounds 4a-8 
described in Table I. 
TABLE I 
______________________________________ 
Physical Characteristics of 1,1-Dichloro-2,2,3- 
triarylcyclopropanes. 
##STR8## 
Compound 
No. R.sub.1 R.sub.2 R.sub.3 
Formula.sup.b 
______________________________________ 
4a H H H C.sub.21 H.sub.16 Cl.sub.2.sup.d 
4d* OCH.sub.3 
OCH.sub.2 Ph 
OCH.sub.3 
C.sub.30 H.sub.26 Cl.sub.2 O.sub.3.sup. 
f 
4e OCH.sub.3 
OCH.sub.3 OCH.sub.3 
-- 
5a H OCH.sub.3 H C.sub.22 H.sub.18 Cl.sub.2 O.sup.f 
5b OCH.sub.3 
H H C.sub.22 H.sub.18 Cl.sub.2 O.sup.f 
5c OCH.sub.3 
OCH.sub.2 Ph 
H C.sub.29 H.sub.24 Cl.sub.2 O.sub.2.sup. 
f 
5d OCH.sub.2 Ph 
OCH.sub.3 H C.sub.29 H.sub.24 Cl.sub.2 O.sub.2.sup. 
f 
6a H OH H C.sub.21 H.sub.16 Cl.sub.2 O 
6b OCH.sub.3 
OH H C.sub.22 H.sub. 18 Cl.sub.2 O.sub.2 
6c OH OCH.sub.3 H C.sub.22 H.sub.18 Cl.sub.2 O.sub.2 
7a H O(CH.sub.2).sub.2 NMe.sub.2 
H C.sub.31 H.sub.35 Cl.sub.2 NO.sub.8 
7b OCH.sub.3 
O(CH.sub.2).sub.2 NMe.sub.2 
H C.sub.26 H.sub.27 Cl.sub.2 NO.sub.2 
8* OCH.sub.3 
OH OCH.sub.3 
C.sub.23 H.sub.20 Cl.sub.2 O.sub.3.sup. 
f 
______________________________________ 
.sup.b All compounds gave combustion elemental analysis for C, H, and Cl 
that were within 0.4% of theoretical values, or formulas were confirmed b 
FABMS. 
.sup.c Petroleum ether. 
.sup.d See Dehmlow, E.V.; Schonefeld, J. Liebigs Ann. Chem. 1971, 744, 42 
.sup.e Et.sub.2 O/petroleum ether. 
.sup.f Determined as a 1:1 mixture of (E) and (Z)isomers. 
.sup.g Me.sub.2 CO/petroleum ether. 
.sup.h SiO.sub.2 chromatography. 
*Mixture of (E) and (Z)isomers. 
The general synthetic route to the compounds of the present invention is 
the addition of para-methoxy or para-benzyloxy substituted Grignard 
reagents to para-methoxy or para-benzyloxy protected or unsubstituted 
deoxybenzoins, followed by acid-catalyzed dehydration of the resulting 
carbinols to give a mixture of (E)- and (Z)-olefins as described in the 
Examples. These olefins reacted with dichlorocarbene to give fully 
protected compounds as described in greater detail in Examples 2-18. 
The structures of all compounds were supported by their proton NMR spectra, 
which were performed on either a Varian EM-360A or an XL-300 spectrometer. 
The spectra are reported in parts per million in CDCl.sub.3 with TMS, 
tetramethylsilane, as the internal standard. .sup.13 C-NMR were determined 
on a Varian XL-300 spectrometer at 75.5 MHz, referenced by the CDCl.sub.3 
signal. Gas chromatography/mass spectral analyses were performed in the EI 
mode on a Hewlett-Packard 5995 GC/MS system using a 30 m.times.0.25 mm 
DB-1 fused silicon capillary column (J and W scientific). Positive ion 
FAB-MS were determined on a VG analytical ZAB-E spectrometer at 0.degree. 
C. in a 3-nitrobenzyl alcohol matrix ionized at 0.95 to 2 volts with the 
source operated at 8 kV using xenon as the discharge gas. High resolution 
spectra were calibrated with CsNaRbI. Infrared spectra were obtained from 
KBr pellets on a Beckman Acculab 1 spectrometer. Silica gel (J. T. Baker) 
of approximately 40 .mu.m diameter was used for flash chromatography, 
which was performed at 5-10 psi. Petroleum ether was of bp 
30.degree.-60.degree. C. 
All organic starting materials and reagents were obtained from Aldrich 
Chemical Co. and were used without additional purification, unless 
otherwise indicated. Inorganic reagents were obtained from Fisher 
Scientific. Solvents were of the highest available grade and were obtained 
from Aldrich, Fisher, or J. T. Baker Chemical Co. Absolute ethanol was 
purchased from U.S. Industrial Chemicals Co. When necessary, solvents or 
reagents were dried by appropriate methods. Evaporations were carried out 
in vacuo on a rotary evaporator or under a stream of dry N.sub.2. Melting 
points were taken on a Thomas-Hoover capillary melting point apparatus and 
are uncorrected. Reaction progress and product purity were monitored by 
analytical TLC on strips of Eastman Kodak plastic-backed SiO.sub.2 60 
F.sub.254 or Al.sub.2 O.sub.3 F.sub.254. Developed strips were viewed 
under light of 254- and 366-nm wavelengths. Elemental analyses were done 
by Midwest Microlab Ltd., Indianapolis, Ind. Analytical results were 
within .+-.0.4% of theoretical values. Acceptable elemental results are 
denoted by the formula, followed by the elements analyzed. Yields are 
based on pure samples, unless otherwise noted. 
EXAMPLE 2 
Preparation of Benzyl 4-bromophenyl ether 
4-Bromophenol (17.3 g, 0.1 mol), 19.03 g (0.15 mol) benzyl chloride, and 
27.64 g (0.2 mol) pulverized K.sub.2 CO.sub.3 were suspended in 150 mL 
Me.sub.2 CO and heated to reflux 2 h. The resulting suspension was 
concentrated on a rotary evaporator. The mixture was then diluted with 200 
mL 5% NaOH and extracted with 100 mL Et.sub.2 O. The organic layer was 
washed with an additional 100 mL 5% NaOH, dried (MgSO.sub.4), filtered, 
and concentrated to give a yellow oil, which was crystallized and 
recrystallized from boiling EtOH to yield 17 g (65%) of Benzyl 4-bromo 
phenyl ether as white needles, mp: 60.degree.-61.degree. C. NMR .delta. 
7.35 and 6.82 (AA'BB', c, 4H, C.sub.6 H.sub.4), 7.36 (s, 5H, C.sub.6 
H.sub.5), 5.0 (s, 2H, OCH.sub.2). 
EXAMPLE 3 
Preparation of Benzyl 4-Benzyloxyphenyl Ketone 
Benzyl 4-hydroxyphenyl ketone (5.1 g; 24 mmol), 3.8 g (3.5 mL, 30 mmol) of 
benzyl chloride, and 16.6 g (120 mmol) of pulverized and flame dried 
K.sub.2 CO.sub.3 were slurried in 75 mL dry Me.sub.2 CO under Ar. The 
mixture was heated to reflux 16 h, cooled to room temperature, diluted 
with CH.sub.2 Cl.sub.2 (100 mL), and filtered. The resulting filter cake 
was washed with C.sub.6 H.sub.6 (50 mL) and EtOAc (50 mL). The combined 
filtrates were concentrated and the resulting orange oil was dissolved in 
C.sub.6 H.sub.6, washed with 10% K.sub.2 CO.sub.3 (100 mL), 5% NaOH 
(2.times.50 mL), and H.sub.2 O (100 mL). The organic layer was dried 
(MgSO.sub.4), filtered, and concentrated to give 10 g of a light orange 
solid, which was purified on 60 g flash SiO.sub.2 (1:1 petroleum 
ether-CH.sub.2 Cl.sub. 2) to give Benzyl 4-benzyloxphenyl ketone as a 
clear oil which was crystallized from C.sub.6 H.sub.6 -EtOH to yield 3.1 g 
of white needles (43%). mp: 134.5.degree.-136.degree. C. NMR .delta. 8.03 
and 7.03 (AA'BB', c, 4H, ArH), 7.43 (s, 5H, ArH), 7.31 (s, 5H, ArH), 5.15 
(s, 2H, PhCH.sub.2 O), 4.25 (s, 2H, COCH.sub.2). 
EXAMPLE 4 
Preparation of 1,2-Diphenyl-1-(4-Methoxyphenyl)Ethene 
To the Grignard reagent prepared from 4-bromoanisole (10 g, 53.5 mmol) and 
Mg turnings (1.32 g, 54.3 mg-atom) in 25 mL Et.sub.2 O was added 
deoxybenzoin (9.81 g, 50 mmol) in 75 mL Et.sub.2 O dropwise under reflux. 
After stirring at reflux 3 h, the mixture was carefully poured onto 1N 
H.sub.2 SO.sub.4 and ice. The layers were allowed to separate and the 
aqueous layer was extracted with 25 mL Et.sub.2 O. The ether layers were 
combined, washed with H.sub.2 O (2.times.80 mL), dried (Na.sub.2 
SO.sub.4), and concentrated to give a yellow oil which solidified at room 
temperature (14.2 g, 93.3% crude yield). Recrystallization from 95% EtOH 
gave 1,2-diphenyl-1-(4-methoxyphenyl)ethanol as a white powder mp: 
111.degree.-112.degree. C. (lit. 112.degree.-113.degree. C.); NMR .delta. 
7.2 (m, 14H, ArH), 3.75 (s, 3H, OCH.sub.3), 3.63 (s, 2H, CH.sub.2), 2.35 
(brs, 1H, OH). The tertiary carbinol (14 g, 46 mmol) and p-TsOH.H.sub.2 O 
(1 g, 5.31 mmol) were dissolved in 40 mL dry C.sub.6 H.sub.6, heated to 
reflux, and stirred 2 h. The red mixture was cooled to room temperature 
and washed with 5% NaHCO.sub.3 (2.times.75 mL). The combined aqueous 
layers were extracted with 25 mL C.sub.6 H.sub.6. The organic layers were 
combined, washed with 100 mL H.sub.2 O, dried (Na.sub.2 SO.sub.4), and 
concentrated to give a yellow oil. The (E)- and (Z)-olefin mixture was 
purified on 60 g flash SiO.sub.2 (petroleum ether) and 
1,2-Diphenyl-1-(4-methoxyphenyl)ethene was obtained as a clear oil (8 g, 
61%); NMR .delta. 7.15 (m, 10H, ArH), 6.9 (s, 1H, C.dbd.CH), 6.8 (AA'BB', 
c, 4H, MeOC.sub.6 H.sub. 4), 3.71 (d, 3H, (E)- & (Z)-OCH.sub.3). 
EXAMPLE 5 
Preparation of 1-(4-Benzyloxyphenyl)-1-(4-methoxyphenyl)-2-phenylethene 
To the Grignard reagent prepared from 1.52 g (63 mg-atom) Mg turnings and 
13.45 g (9 mL, 72 mmol) 4-bromoanisole (initiated with one I.sub.2 crystal 
and one drop EtBr.sub.2) in 25 mL THF was added 3 g (9.9 mmol) of Benzyl 
4-benzyloxyphenyl ketone, prepared in Example 3. The reaction mixture was 
stirred at reflux for 48 h and then treated with 10 mL aqueous NH.sub.4 
Cl. After cooling to room temperature, the slurry was filtered, the filter 
cake washed with 50 mL THF, and the filtrates concentrated to give an 
orange oil. The oil was chromatographed over 60 g flash SiO.sub.2 
(petroleum ether followed by Me.sub.2 CO). The Me.sub.2 CO fractions were 
combined, concentrated, dissolved in 100 mL 1:1 95% EtOH-2N H.sub.2 
SO.sub.4, and refluxed with stirring for 8 h. The solvents were removed to 
give a brown gum, which was dissolved in 100 mL Et.sub.2 O, washed with 
saturated aqueous NaHCO.sub.3 (100 mL), brine (100 mL), and H.sub.2 O (100 
mL). The organic layer was dried (MgSO.sub.4), filtered, and concentrated 
to give a brown oil. The oil was purified on 60 g flash SiO.sub.2 (9:1 
petroleum ether-CH.sub.2 Cl.sub.2) to give 3.1 g of 
1-(4-Benzyloxyphenyl)-1-(4-methoxyphenyl)-2-phenylethene (80%) as a clear 
oil. NMR .delta.7.41 (brs, 5H, O-benzyl C.sub.6 H.sub.5), 7.3 to 6.75 (m, 
9H, substituted rings' ArH and C.dbd.CH), 7.11 (s, 5H,.dbd.C--C.sub.6 
H.sub.5), 5.05 (s, 2H, OCH.sub.2 Ph), 3.79 (s, 3H, OCH.sub.3). 
EXAMPLE 6 
Preparation of 1-(4-Benzyloxyphenyl)-1,2-bis(4-methoxyphenyl)ethene 
Benzyl 4-bromophenyl ether (13.155 g; 50 mmol), prepared in Example 2, in 
20 mL THF was added to Mg turnings (1.17 g; 48 mg-atom) in 20 mL THF. 
Grignard reagent formation was facilitated by addition of a few drops of 
EtBr.sub.2, one crystal of I.sub.2, and heat. This mixture was stirred and 
heated to reflux for 2 h. 11.54 g (45 mmol) desoxyanisoin was added under 
reflux as a slurry in 60 mL THF. The resulting mixture was stirred at 
reflux for 18 h, cooled to room temperature, poured onto 200 g 2N H.sub.2 
SO.sub.4 and ice (1:1), and stirred at room temperature until the ice 
melted. The resulting mixture was placed in a separatory funnel and 
extracted with 200 mL Et.sub.2 O. The organic layer was washed with 
aqueous Na.sub.2 SO.sub.5 (100 mL), and brine (2.times.100 mL). The 
combined aqueous layers were extracted with Et.sub.2 O (50 mL). The 
combined organic layers were dried (MgSO.sub.4), filtered, and 
concentrated to give 19 g of a wine colored oil, which was dissolved in 
Et.sub.2 O and refrigerated to precipitate any unreacted deoxyanisoin. 
After filtration and concentration of the solution, the resulting oil was 
purified on 60 g flash SiO.sub.2 (7:3 petroleum ether-CH.sub.2 Cl.sub.2) 
to yield 12.4 g (65%) of 
1-(4-Benzyloxyphenyl)-2,2-bis(4-methoxyphenyl)ethene as a yellow oil. NMR 
.delta.7.41 (s, 5H, O-benzyl C.sub.6 H.sub.5), 7.35 to 6.57 (m, 13H, ArH 
and C.dbd.CH), 5.07 (s, 2H, PhCH.sub.2 O), 3.78 (s, 1.5H, geminal ring 
OCH.sub.3), 3.77 (s, 1.5H, central ring OCH.sub.3), 3.72 (s, 3H, vicinal 
ring OCH.sub.3). 
EXAMPLE 7 
Preparation of 1,1,2-Tris(4-methoxyphenyl)ethene 
To the Grignard reagent prepared from 20 g 4-bromoanisole (107 mmol) and 
2.43 g Mg turnings (107 mg-atom) in 20 mL Et.sub.2 O was added 19.25 g dry 
(P.sub.2 O.sub.5) desoxyanisoin (76 mmol) as a solid in four portions, 
each followed by a 50 mL wash of Et.sub.2 O. The mixture was stirred at 
reflux 17 h, cooled to room temperature, poured onto 200 g ice and 40 mL 
1N H.sub.2 SO.sub.4, and filtered into a separatory funnel. 100 mL 
Et.sub.2 O was added and the organic layer was washed with 100 mL H.sub.2 
O and saturated NaHCO.sub.3 (2.times.100 mL). The organic layer was dried 
(MgSO.sub.4), filtered, and the Et.sub.2 O was removed under reduced 
pressure to give crude 1,1,2-Tris(4-methoxyphenyl)ethanol as an orange oil 
which crystallized (25 mL EtOH with a trace of NH.sub.4 OH) as an orange 
solid, 18.6 g (67% crude yield), mp: 109.degree.-127.degree. C.; NMR 
.delta.7.47 to 6.63 (m, 12H, ArH), 3.73 (3 overlapping s, 9H, OCH.sub.3), 
3.48 (s, 2H, CH.sub.2), 2.27 (s, 1H, OH). p-TsOH.H.sub.2 O (2 g; 10.5 
mmol) and 18.5 g of the crude carbinol (50.8 mmol) were dissolved in 50 mL 
C.sub.6 H.sub.6, heated to reflux 2 h, cooled to room temperature, and 
poured onto 75 mL 5% NaHCO.sub.3. The organic layer was washed with 75 mL 
5% NaHCO.sub.3. The combined aqueous layers were extracted with 25 mL 
C.sub.6 H.sub.6. The combined organic layers were washed with 100 mL 
H.sub.2 O, dried (Na.sub.2 SO.sub.4), filtered, and the solvent removed to 
give a dark oil. The oil was purified on 80 g flash SiO.sub.2 (gradient 
elution with petroleum ether-CH.sub.2 Cl.sub. 2) yielding an amber oil 
which solidified at room temperature to yield 
1,1,2-Tris(4-methoxyphenyl)ethene as white cubes (11.3 g, 64%), mp: 
100.degree.-101.degree. C. (lit. 100.degree.-101.degree. C.). NMR 
.delta.7.29 to 6.55 (m, 13H, ArH and C.dbd.CH), 3.78 (s, 3H, geminal ring 
OCH.sub.3), 3.75 (s, 3H, vicinal ring OCH.sub.3), 3.69 (s, 3H, central 
ring OCH.sub.3). 
EXAMPLE 8 
Preparation of 1,1-Dichloro-2,2,3-triphenylcyclopropane (Compound 4a) 
Prepared in 64% yield from 3.2 g (12.5 mmol) of triphenylethylene (3.2 g; 
12.5 mmol) and 0.25 g (1.1 mmol) benzyltriethylammonium chloride (TEBA) by 
dissolving in 50 g (419 mmol) CHCl.sub.3, stirring rapidly at room 
temperature, and treating with 27 mL of chilled 50% aqueous NaOH dropwise 
according to the method of Dehmlow, E. V., et al., S.S. Phase Transfer 
Catalysis, Verlag Chemie: Deerfield Beach 1980. After stirring 75 h, the 
resulting mixture was poured onto 150 mL H.sub.2 O and extracted with 
CH.sub.2 Cl.sub.2 (2.times.100 mL). The combined organic layers were 
washed with 100 mL H.sub.2 O, dried (Na.sub.2 SO.sub.4), filtered and 
concentrated. The resulting brown oil was dissolved in boiling petroleum 
ether, quickly filtered, and allowed to stand at room temperature, 
yielding 2.7 g (64%) of Compound 4a as a white powder, mp: 
106.degree.-107.degree. C. (petroleum ether) (lit. 105.degree.- 
107.degree. C.); NMR .delta.7.23 (m, 15H, ArH), 3.57 (s, 1H, cyclopropyl 
H); IR (CM.sup.-1) 3020, 3000, 1600, 1490, 1435, 860, 775 765, 750, 695; 
MS (m/z, % base) 342 (M+4), 3.2; 340 (M+2), 6.3; 338 (M+), 10.23; 303 
(--Cl), 100; 267 (--Cl, --HCl), 80.7. 
EXAMPLE 9 
Preparation of (R/S) (E)-and 
(Z)-1,1-Dichloro-2-(4-benzyloxyphenyl)-2,3-bis-(4-methoxyphenyl)cyclopropa 
ne (Compound 4d) 
1-(4-Benzyloxyphenl)-1,2-bis(4-methoxyphenyl)ethene (20 g; 47.4 mmol); 
prepared in Example 6, and 1 g TEBA (4.39 mmol) in 100 mL CHCl.sub.3 were 
treated with 80 mL chilled 50% NaOH with rapid stirring for 30 h. This 
reaction was monitored by NMR (appearance of cyclopropyl H signal) since 
no suitable TLC solvent system was found. The dark emulsion was poured 
into a separatory funnel containing 100 mL H.sub.2 O and 100 mL CH.sub.2 
Cl.sub.2. The aqueous layer was extracted with CH.sub.2 Cl.sub.2 
(2.times.50 mL). The combined organic layers were washed with H.sub.2 O 
(3.times.100 mL), dried (K.sub.2 CO.sub.3), filtered, and concentrated to 
yield a dark oil, which was dissolved in C.sub.6 H.sub.6 and 
chromatographed on 200 g Al.sub.2 O.sub.3 (Act. I, 1:1 C.sub.6 H.sub.6 
-petroleum ether) under 5 psi. Fractions 2-6 (100 mL ea.) were 
concentrated to give an orange oil, which was dissolved in 200 mL 
petroleum ether containing ca. 10 mL Et.sub.2 O. After standing at room 
temperature 4 days, Compound 4d precipitated as a white powder. Four crops 
yielded 6.2 g (26%), mp: 127.degree.-128.degree. C. NMR (300 MHz) 
.delta.7.39 and 6.82 (AA'BB', c, 4H, J=8.5 Hz, ArH), 7.36 to 7.25 (m, 5H, 
C.sub.6 H.sub.5), 7.19 and 6.80 (AA'BB', c, 4H, J=8.5 Hz, ArH), 6.91 and 
6.74 (AA'BB', c, 4H, J=8.5 Hz, ArH), 4.92 (s, 2H, OCH.sub.2 Ph), 3.70 (s, 
3H, OCH.sub.3), 3.69 (s, 3H, OCH.sub.3), 3.47 (s, 1H, cyclopropyl H). 
Anal. (C.sub.30 H.sub.26 Cl.sub.2 O.sub.3) C, H, Cl. This compound readily 
decomposes in solution with evolution of HCl gas on contact with 
scratched, sintered, or ground glass. 
EXAMPLE 10 
Preparation of 1,1-Dichloro-2,2,3-tris(4-methoxyphenyl)cyclopropane 
(Compound 4(e)) 
1,1,2-Tris(4-methoxyphenyl)ethene (10 g; 29 mmol), prepared in Example 7, 
and 0.5 g of TEBA (2.2 mmol) were dissolved in 68 mL CHCl.sub.3 and 
treated dropwise with 53 mL of chilled 50% NaOH. The mixture was stirred 
at room temperature 48 h. 100 mL CH.sub.2 Cl.sub.2 was added and the 
aqueous layer was removed. The aqueous layer was taken to pH 7 with 1N HCl 
and extracted with 50 mL CH.sub.2 Cl.sub.2. The combined organic layers 
were washed with 100 mL H.sub.2 O, dried (MgSO.sub.4), filtered, and 
concentrated to give crude Compound 4e as a black oil (11.5 g, 92% crude 
yield). NMR .delta.7.47 to 6.62 (m, 12H, ArH), 3.8 (s, 3H, geminal ring 
OCH.sub.3), 3.73 (s, 3H, vicinal ring OCH.sub.3), 3.69 (s, 3H, central 
ring OCH.sub.3 ), 3.5 (s, 1H, cyclopropyl H). 
EXAMPLE 11 
Preparation of (R/S) 
(Z)-1,1-Dichloro-2,3-diphenyl-2-(4-methoxy-phenyl)cyclopropane (5a), and 
(E)-isomer (Compound 5b) 
The mixture of (E)- and (Z)-olefins 1,2-Diphenyl-1-(4-methoxyphenyl)ethene 
(5 g, 17.5 mmol), prepared in Example 4, and TEBA (0.25 g, 1.1 mmol) were 
dissolved in CHCl.sub.3 (50 g, 419 mmol). Chilled 50% NaOH (26.7 mL) was 
added dropwise. The two phases were rapidly stirred at room temperature 
for 96 h. The resulting orange emulsion was poured onto 150 mL H.sub.2 O 
and extracted with CH.sub.2 Cl.sub.2 (2.times.100 mL). The organic layers 
were combined, washed with 100 mL H.sub.2 O, dried (Na.sub.2 SO.sub.4), 
and concentrated to give a dark oil. The mixture of cyclopropanes was 
isolated by flash chromatography (4:1 petroleum ether-CH.sub.2 Cl.sub.2) 
as a white powder. Recrystallization from petroleum ether or pentane gave 
a mixture of 5a and 5b as a white amorphous solid (2.53 g, 38%): mp: 
118.degree.-123.degree. C.; NMR .delta.7.15 (m, 14H, ArH), 3.76 (s, 3H, 
OCH.sub.3), 3.52 (s, 1H, cyclopropyl H). IR (CM.sup.-1) 3030, 3015, 3000, 
2980, 2965, 2920, 1605, 1510, 1450, 1440, 1250, 1175, 1025, 870, 830, 770, 
700; MS (m/z, % base) 334 (--Cl), 7.09; 297 (--Cl, --HCl), 100. Anal. 
(C.sub.22 H.sub.18 Cl.sub.2 O) C, H, Cl. The (Z)-isomer, Compound 5a, was 
isolated by fractional crystallization (petroleum ether/Me.sub.2 CO), 1.8 
g (27%), mp: 132.degree.-133.5.degree. C.; NMR .delta.7.33 (s, 10H, ArH), 
6.72 and 7.52 (AA'BB', c, 4H, central ring), 3.71 (s, 3H, OCH.sub.3), 
3.52 (s, 1H, cyclopropyl H). The (E)-isomer, Compound 5b, was obtained as 
clear crystals by thin layer chromatography (SiO.sub.2, petroleum ether) 
as the more mobile compound, 0.7 g (11%), mp: 114.5.degree.-116.5.degree. 
C.; NMR .delta.7.33 (s, 10H, ArH), 6.92 and 7.33 (AA'BB', c, 4H, geminal 
ring), 3.79 (s, 3H, OCH.sub.3), 3.52 (s, 1H, cyclopropyl H). 
EXAMPLE 12 
Preparation of (R/S) 
(Z)-1,1-Dichloro-2-(4-benyloxyphenyl)-2-(4-methoxyphenyl)-3-phenylcyclopro 
pane (5c) and (E)-isomer (Compound 5d) 
1-(4-Benzyloxyphenyl)-1-(4-methoxyphenyl)-2-phenylethene (6 g; 15.3 mmol), 
prepared in Example 5, was dissolved in 100 mL CHCl.sub.3 along with 0.5 g 
TEBA. The resulting solution was treated with 30 mL chilled 50% NaOH and 
rapidly stirred at room temperature 96 h. The resulting dark mixture was 
poured onto 200 mL CHCl.sub.3 in a separatory funnel and the aqueous layer 
was carefully adjusted to pH 7 with 2N HCl. An additional 100 mL H.sub.2 O 
was used to wash the organic layer. The combined aqueous layers were 
extracted with 50 mL CHCl.sub.3. The combined organic layers were dried 
(MgSO.sub.4), filtered, and concentrated to give an orange oil. The oil 
was treated with Et.sub.2 O/petroleum ether to give a 1:1 mixture of (E) 
and (Z) isomers 4c: mp 129.degree.-134.degree. C.; Anal. C.sub.29 H.sub.24 
Cl.sub.2 O.sub.2 (C, H, Cl). The (Z)-isomer, Compound 5c, precipitated 
rapidly at room temperature from 75 mL boiling butanone as a white powder 
(two crops, 2.3 g, 32%); mp: 169.degree.-171.degree. C. NMR .delta.7.42 to 
7.17 (m, 12H, ArH), 7.01 (m, 2H, ArH), 6.84 (d, 4H, J=8.7 Hz, ortho 
signals), 4.98 (s, 2H, OCH.sub.2), 3.75 (s, 3H, OCH.sub.3), 3.5 (s, 1H, 
cyclopropyl H). Recrystallization from boiling EtOAc raised the melting 
point to 173.5.degree.-175.degree. C. The (E)-isomer, Compound 5d, could 
be obtained by concentration of the mother liquor and purification over 60 
g flash SiO.sub.2 (4:1 petroleum ether-CH.sub.2 Cl.sub.2), followed by 
recrystallization from boiling EtOH containing a trace of Et.sub.2 O (0.8 
g, 11%); mp: 133.degree.-134.degree. C. NMR .delta. 7.43 to 6.95 (m, 9H, 
ArH), 7.36 (s, 5H, O-benzyl C.sub.6 H.sub.5), 7.1 (d, 2H, J=8.7 Hz, meta 
signals), 6.78 (d, 2H, J=8.7 Hz, meta signals), 5.03 (s, 2H, OCH.sub.2), 
3.73 (s, 3H, OCH.sub.3), 3.5 (s, 1H, cyclopropyl H). 
EXAMPLE 13 
Preparation of (R/S) 
(Z)-1,1-Dichloro-2,3-diphenyl-2-(4-hydroxyphenyl)cyclopropane (Compound 
6a) 
To cooled n-BuSH (0.63 g, 7 mmol) in 6 mL CH.sub.2 Cl.sub.2 was added 
AlCl.sub.3 (0.75 g, 6 mmol) under Ar. The methoxy Compound 5a (0.5 g, 1.35 
mmol), prepared in Example 11, in 15 mL CH.sub.2 Cl.sub.2 was added 
dropwise at 0.degree. C. After stirring 2 h at room temperature the 
reaction was quenched with 20 mL H.sub.2 O. The yellow mixture was 
extracted with Et.sub.2 O (2.times.40 mL). The combined organic layers 
were washed with 30 mL 0.1N H.sub.2 SO.sub.4, dried (Na.sub.2 SO.sub.4), 
and concentrated to give 475 mg of an amber oil. Flash chromatography on 
20 g SiO.sub.2 (1:1 CH.sub.2 Cl.sub.2 -petroleum ether) yielded 400 mg of 
an off-white glass. Crystallization from EtOH yielded 400 mg of Compound 
6a as white needles (84%): mp: 135.5.degree.-136.degree. C.; NMR 
.delta.7.38 and 6.58 (AA'BB', c, 4H, central ring), 7.02 (m, 10H, ArH), 
4.75 (brs, 1H, OH), 3.53 (s, 1H, cyclopropyl H). IR (CM.sup.-1) 3305, 
3030, 3010, 1600, 1505, 1490, 1450, 1440, 1210, 1155, 865, 820, 765, 720. 
MS (m/z, % base) 320 (--Cl), 6.96; 319 (--HCl, 6.03); 318 (--H, --HCl, 
27.05); 283 (-2HCl, 100). Anal. (C.sub.21 H.sub.16 Cl.sub.2 O) C, H, Cl. 
EXAMPLE 14 
Preparation of (R/S) 
(E)-1,1-Dichloro-2-(4-hydroxyphenyl)-2-(4-methoxyphenyl)-3-phenylcycloprop 
ane (Compound 6b) 
Benzyl ether 5c (1 g; 2.1 mmol), prepared in Example 12, was dissolved in 
50 mL dry THF and hydrogenolyzed in the presence of 50 mg 5% Pd/C at one 
atmosphere and room temperature for 8 h. The resulting clear solution was 
filtered, concentrated to give a red oil, and purified on 20 g flash 
SiO.sub.2 (1:1 petroleum ether-CH.sub.2 Cl.sub.2) to yield 0.8 g (99%) of 
Compound 6b as an orange semi-solid. NMR .delta.7.31 to 6.94 (m, 7H, ArH), 
7.3 (d, 2H, J=8.7 Hz, meta signals), 6.83 (d, 2H, J=8.7 Hz, ortho 
signals), 6.65 (d, 2H, J=8.7 Hz, ortho signals), 5.75 (brs, 1H, OH), 3.7 
(s, 3H, OCH.sub.3), 3.5 (s, 1H, cyclopropyl H). An analytical sample was 
obtained by drying over P.sub.2 O.sub.5 at 60.degree. C. 6 h. Anal. 
C.sub.22 H.sub.18 Cl.sub.2 O.sub.2 (C, H, Cl). 
EXAMPLE 15 
Preparation of (R/S) 
(Z)-1,1-Dichloro-2-(4-hydroxyphenyl)-2-(4-methoxyphenyl)-3-phenylcycloprop 
ane (Compound 6c) 
Benzyloxy ether 5d (2 g; 4.2 mmol), prepared in Example 12, was 
hydrogenolyzed in 60 mL dry THF with H.sub.2 in the presence of 110 mg 5% 
Pd/C for 4 h. The resulting clear solution was filtered, concentrated, and 
purified on 45 g flash SiO.sub.2 (1:1 petroleum ether-CH.sub.2 Cl.sub.2) 
to yield 1.6 g (99%) of Compound 5c as a red oil. A small amount 
crystallized after dissolving in toluene and treating the solution with 
hexane, mp: 133.degree.-135.degree. C. NMR .delta.7.55 to 6.83 (m, 8H, 
ArH), 7.3 (s, 5H, C.sub.6 H.sub.5), 5.53 (brs, 1H, OH), 3.69 (s, 3H, 
OCH.sub.3), 3.5 (s, 1H, cyclopropyl H). Anal. C.sub.22 H.sub.18 Cl.sub.2 
O.sub.2 (C, H, Cl). 
EXAMPLE 16 
Preparation of (R/S) (Z)-1,1-Dichloro-2,3-diphenyl-2-[4 
-[2-(dimethylamino)ethoxy]-phenyl]cyclopropane, dihydrogen citrate salt 
(Compound 7a) 
Phenol 6a (1.25 g; 3.52 mmol), prepared in Example 13, 0.58 g of 
N,N-dimethylaminoethyl chloride hydrochloride (4 mmol), and 1.38 g 
pulverized and flame-dried K.sub.2 CO.sub.3 (10 mmol) were slurried in 20 
mL dry Me.sub.2 CO under Ar and heated to reflux overnight. The resulting 
brown slurry was cooled to room temperature, filtered, and concentrated to 
give a brown oil. This was dissolved in 30 mL Et.sub.2 O and washed with 
1N NaOH (2.times.40 mL). The combined aqueous layers were extracted with 
25 mL Et.sub.2 O. The combined organic layers were washed with 40 mL 
brine, H.sub.2 O (2.times.20 mL), dried (MgSO.sub.4), filtered, and 
concentrated to give a brown gum. The crude material was dissolved in 
MeOH-Et.sub.2 O, 2 mL 30% HCl was added, and the mixture was stirred at 
room temperature 0.5 h. The solvents were removed to give a brown glass. 
The glass was treated with 30% NaOH up to pH 11 and 50 mL Et.sub.2 O was 
added. The organic layer was washed with 10% NaOH (2.times.50 mL), 50 mL 
brine, and 50 mL H.sub.2 O. The organic layer was dried (K.sub.2 
CO.sub.3), filtered, and concentrated to give a brown oil. This was loaded 
on a 20 mL basis Al.sub.2 O.sub.3 (activity I) column and eluted with 
CH.sub.2 Cl.sub.2. A yellow oil was collected which had spectral features 
of the desired product: NMR .delta.7.73 to 7.02 (m, 12H, ArH), 6.85 (d, 
J=9 Hz, central ring 2H ortho to O), 4.03 (t, J=5.5 Hz, 2H, OCH.sub.2), 
3.52 (s, 1H, cyclopropyl H), 2.69 (t, J=5.5 Hz, 2H, CH.sub.2 N), 2.3 (s, 
6H, NMe.sub.2). The oil was dissolved in hot EtOH and treated with excess 
anhydrous citric acid in hot EtOH. Treatment of the cooled solution with 
Et.sub.2 O yielded 0.11 g (5%) of Compound 7a in two crops as a white 
powder mp: 140.degree.-142.degree. C. Anal. (C.sub.25 H.sub.25 Cl.sub.2 
NO.C.sub.6 H.sub.8 O.sub.7) C, H, Cl, N. 
EXAMPLE 17 
Preparation of (R/S) 
(Z)-1,1-Dichloro-2-[4-[2-(dimethylamino)ethoxy]phenyl]-2-(4-methoxyphenyl) 
-3-phenylcyclopropane (Compound 7b) 
Phenol 6b (1.45 g; 3.78 mmol), prepared in Example 14, 1.63 g (11.3 mmol) 
of N,N-dimethylaminoethyl chloride hydrochloride, and 5.2 g (38 mmol) of 
dry, pulverized K.sub.2 CO.sub.3 in 60 mL of dry Me.sub.2 CO were stirred 
and heated to reflux under Ar 24 h. The resulting slurry was cooled to 
room temperature, filtered, and concentrated. The resulting oil was 
dissolved in 60 mL EtOAc and washed with 10% NaOH (2.times.50 mL). The 
combined aqueous layers were extracted with 25 mL EtOAc. The combined 
organic layers were washed with H.sub.2 O (2.times.50 mL), dried (K.sub.2 
CO.sub.3), filtered, and concentrated to give a dark oil, which was 
initially purified on 20 g flash SiO.sub.2 (9:1 C.sub.6 H.sub.6 -Et.sub.3 
N), followed by two further purifications on flash SiO.sub.2 (20 g each, 
Me.sub.2 CO) to give 225 mg (13%) of Compound 7b as a yellow gum. NMR 
.delta.7.57 to 6.78 (m, 13H, ArH), 4.08 (t, 2H, J=5 Hz, OCH.sub.2), 3.8 
(s, 3H, OCH.sub.3), 3.53 (s, 1H, cyclopropyl H), 2.75 (t, 2H, J=5 Hz, 
CH.sub.2 N), 2.37 (s, 6H, N(CH.sub.3).sub.2). FAB-MS (m/z, % base): 458 
(M+H+2), 25.8; 456 (M+H), 50.3; 421 (M+H-Cl), 100. High resolution FAB-MS: 
C.sub.26 H.sub.27 Cl.sub.2 NO.sub.2 calculated for 455.1419, found 
455.1496. 
EXAMPLE 18 
Preparation of (R/S) (E)- and 
(Z)-1,1-Dichloro-2-(4-hydroxyphenyl)-2,3-bis-(4-methoxyphenyl)cyclopropane 
(Compound 8) 
Benzyl ether 4d (0.5 g; 0.99 mmol), prepared in Example 9, in 20 mL THF was 
treated with H.sub.2 in the presence of 150 mg 5% Pd/C at one atmosphere 
and ambient temperature for 4.5 h. The mixture was filtered, concentrated, 
and crystallized from Et.sub.2 O/petroleum ether to give white solid that 
tenaciously held Et.sub.2 O, even after drying (P.sub.2 O.sub.5, 0.1 mm 
Hg, 40.degree. C., 18 h) in an Abderhalden apparatus, mp: 
105.degree.-106.degree. C. (dec., with prior softening). Elemental 
analysis and NMR showed 1.8 molecules of Et.sub.2 O per molecule of 8. 
Several recrystallizations from C.sub.6 H.sub.6 /hexane yielded 3.9 g 
(95%) of the product as a white powder, mp: 96.degree.-97.degree. C. 
(dec.). NMR .delta.7.45 (d, 2H, meta signals), 7.3 to 6.64 (m, 11H, 
ArH+OH), 3.81 (s, 3H, OCH.sub.3), 3.79 (s, 3H, OCH.sub.3), 3.49 (s, 1H, 
cyclopropyl H). This powder would not give a satisfactory combustion 
elemental analysis, and was subjected to FAB-MS (m/z, % base): 419 
(M+H+4), 1.1; 417 (M+H+2), 2.1; 415 (M+H), 3.1; 381 (M+2-Cl), 33.6; 379 
(M-Cl), 100; 343 (--HCl, --Cl), 34. 
EXAMPLE 19 
Biological Testing 
The biological evaluation of the test compounds consisted of the in vitro 
rat cytosolic estradiol receptor binding assay, the in vivo immature mouse 
uterotrophic (estrogenic) assay, and the in vivo immature mouse and rat 
antiuterotrophic (antiestrogenic) assay and in the in vitro suppression of 
the proliferation of the MCF-7 human breast cancer cell line (Table II). 
All assays contained estradiol, TAM, MER 25, and 
1-1-dichloro-2-3-cis-diphenylcyclopropane (Analog II) as standards. 
Biological Assays. Tamoxifen was obtained from Stuart Pharmaceutical, 
Division of ICI Americas, Inc., Wilmington, Del. MER 25 was obtained from 
Merrell Dow Research Institute, Division of Merrell Dow Pharmaceuticals, 
Inc., Cincinnati, Ohio. Absolute ethanol was obtained from U.S. Industrial 
Chemicals Co. Hormones and biochemicals were purchased from Sigma Chemical 
Co. Animals and Housing. Viral-free immature female Swiss-Webster mice 
were obtained at 17-19 days of age from Sasco (Omaha, Nebr.) weighing 8-10 
g, and were used in the uterotrophic and antiuterotrophic assays. Immature 
female Sprague-Dawley rats, obtained also at 17-19 days of age from Sasco, 
weighing 28-33 g, were used in the estradiol receptor binding assay. 
Animals were housed in wire topped polycarbonate cages with six animals 
per cage. Environment was controlled at 25.degree. C. with a 12-hour 
light/dark cycle. The animals received a diet of Wayne Lab Blox rodent 
chow and tab water ad libitum. 
EXAMPLE 20 
Receptor Binding Assay 
The receptor binding activies of the test compounds for the estrogenic 
receptors were determined by displacement of [.sup.3 H]-estradiol from rat 
uterine cytosol in vitro. Female Sprague-Dawley rats (17-19 days old) were 
treated with 0.53 .mu.g of estradiol in 0.1 mL sesame oil for three 
consecutive days (total dose 1.6 .mu.g). On the fourth day the rats were 
anesthetized with Et.sub.2 O and sacrificed by cervical dislocation. A 
modification of Korenman's receptor binding assay method (Korenman, S. G., 
Steroids 13: 163 (1969)) was used. Uteri were removed, cleaned of adhering 
connective tissue and fat, weighed (avg. wt.=83 mg/animal), and 
homogenized (Polytron PT-10 stainless steel homogenizer, rheostat setting 
7, five ten-second bursts with a ten-second pause between bursts) at 
0.degree.-4.degree. C. in five volumes (w/v) of TEDM buffer (10 mM 
Tris-HCl, 1.5 mM disodium ethylenediamine tetraacetic acid, 1.0 mM 
dithiothreitol, 10.0 mM sodium molybdate, pH adjusted to 7.4 with 5M 
NaOH). The resulting homogenate was centrifuged at 2000 g for 15 min 
(4.degree. C.). The supernatant was then centrifuged at 104,000 g for 1 h 
(4.degree. C.). The supernatant from the high speed centrifugation 
(cytosol) was carefully decanted and used immediately. The protein content 
of the cytosol was determined and adjusted to 4-5 mg protein/mL. The test 
compounds were dissolved in EtOH or DMSO and diluted with TEDM so that the 
final EtOH concentration was less than 2% or the final DMSO concentration 
was less than 10%. Neither of these concentrations of the organic solvents 
affected the binding of the tritiated estradiol to the cytosolic receptor 
or the amount of non-specific binding seen as determined by parallel 
incubations. Duplicate incubations were conducted at 4.degree. C. for 24 h 
in a total volume of 0.5 mL containing: 200 .mu.L cytosol; 100 .mu.L 
(0.218 .mu.Ci) of 2,4,6, 7(n)-[.sup.3 H]-17.beta.-estradiol (93.35 
mCi/mmole); 100 .mu.L of the test compounds at concentrations ranging from 
10.sup.-4 to 10.sup.-6 M, or unlabelled estradiol at concentrations 
ranging from 10.sup.-6 to 10.sup.-8 M; and sufficient TEDM to obtain a 
final volume of 0.5 mL. Single parallel incubations at each concentration 
of test compound and estradiol contained 100 .mu.L of 2.times.10.sup.-5 M 
DES in the final TEDM addition to distinguish between specific receptor 
binding and non-specific protein binding of the compounds. 
After incubation, 0.5 mL of a dextran-coated charcoal (DCC) solution (0.5% 
activated charcoal and 0.05% Dextran T-70, w/v, in TEDM buffer) was added 
and the tubes were gently vortexed at 4.degree. C. 15 min. The tubes were 
centrifuged at 2000 g 15 min (4.degree. C.) to remove the unbound [.sup.3 
H]-estradiol. A 0.5 mL aliquot of the supernatant was added to 10 mL 
Beckmann Ready-Solv VI scintillation cocktail in subdued light and the 
tritium content of each vial was determined by liquid scintillation 
spectrometry. The radioactivity was plotted as a function of the log 
concentration of competing ligand and subjected to linear regression 
analysis. Relative binding affinity of each compound was determined by the 
method of Bliss, C.I., The Statistics of Bioassay, Academic Press (New 
York 1952). The observed receptor binding activities are set out in Table 
II. 
TABLE II 
______________________________________ 
Estrogen Receptor Binding Affinities.sup.a and Antiestrogenic 
Activities.sup.b of 1,1-Dichloro-2,2,3-triarylcyclopropanes 
Antiestrogenic 
No. RBA.sup.c, % 
dose (.mu.g) 
% Reduction.sup.d 
______________________________________ 
4a 0.13 30 11 
150 3 
750 13 
4d.sup.e 0.08 10 2 
30 15 
150 49 
750 2 
5a 0.25 30 22 
150 48 
750 33 
5c 0.09 30 37 
150 47 
750 22 
6a 0.69 30 23 
150 12 
750 25 
6b 1.70 30 15 
150 -7 
750 40 
6c 2.40 30 28 
150 18 
750 22 
7a.sup.f 0.87 4 -6 
10 10 
30 37 
150 30 
750 18 
7b 0.73 4 -10 
10 6 
30 33 
150 42 
750 53 
8.sup.e 3.62 30 19 
150 10 
750 26 
Tamoxifen 0.87 30 4 
150 0 
750 - 13 
MER 25 0.0016 30 1 
150 30 
750 82 
Analog II 0.0086 200 24 
400 36 
800 49 
Estradiol 100 -- -- 
______________________________________ 
.sup.a Determined by competitive radiometric binding assay with rat 
uterine cytosol as a source receptor, [.sup.3 H]estradiol as tracer, and 
dextrancoated charcoal as absorbant for free ligand. 
.sup.b Determined as the decrease in the estradiolstimulated (0.03 .mu.g 
total dose) uterine weight of immature (17-19 day old) female mice. 
.sup.c Binding affinities are expressed relative to that of estradiol = 
100% (RBA = relative binding affinity) and are the average of duplicate 
determinations minus nonspecific binding. 
.sup.d Calculated by: 100 - {(mean uterine weight of test compound treate 
animals - mean uterine weight of control animals)/(mean uterine weight of 
estradiolstimulated animals - mean uterine weight of controls)} .times. 
100. 
.sup.e Determined as a 1:1 mixture of isomers. 
.sup.f Dihydrogen citrate salt. 
EXAMPLE 21 
Uterotrophic Assay 
Estrogenic activity of the compounds was determined using a modification of 
the method of Rubin, B. L., et al., Endocrinology 49: 429 (1951) (see 
Pento, J. T., et al.; l J. Endocrinol. 61: 1216 (1978)) using immature 
(17-19 days old) female Swiss-Webster mice. The test compounds were 
dissolved separately in a minimum amount of isopropyl myristate (IPM), and 
diluted serially with sesame oil to the proper concentrations (final 
concentration of IPM&lt;5%). Solutions were shaken at 25.degree. C. for 
several hours to ensure complete dissolution. The mice were randomly 
separated into groups of six animals, weighed, and the compounds were 
administered by s.c. injection of 0.1 mL of the oil solutions into the 
nape of the neck for 3 consecutive days. The solutions were periodically 
checked by TLC to insure homogeneity. A control group received 0.1 mL 
sesame oil alone. 
The animals were anesthetized with Et.sub.2 O and sacrificed by cervical 
dislocation 24 h after the last injection. Body weights were determined 
and the uteri were removed, cleaned of adhering connective tissue and fat, 
blotted to remove tissue fluid, and weighed to the nearest 0.1 mg. The new 
compounds showed no significant uterotrophic activity at doses of 10, 50, 
and 250 .mu.g (6a-b, 7a-b, 8, 10), or at doses of 30, 150, and 750 .mu.g 
(4a, 4d, 5a, 5c, 6b, 9a-b). Tamoxifen elicited a significant estrogenic 
response at a total dose of 1 .mu.g, while MER 25 was slightly estrogenic 
only at the high total dose of 750 .mu.g. 
EXAMPLE 22 
Antiuterotrophic Assay 
Antiestrogenic activity of the compounds was determined by inhibition of 
the estradiol-induced uterotrophic activity in immature female 
Swiss-Webster mice. Animals were distributed into groups of six animals. A 
modification of the uterotrophic assay described in Example 21 was used 
(Dorfman, R. I., et al., Endocrinology 68: 17 (1960)). Estradiol was 
dissolved in sesame oil (0.1 .mu.g/mL). The test compounds were dissolved 
in sesame oil and diluted with sesame oil to achieve desired 
concentrations. The solutions were periodically checked by TLC to insure 
homogeneity. Injections were made in the nape of the neck for 3 
consecutive days. The unstimulated control group received vehicles alone 
(0.05 mL IPM and 0.1 mL sesame oil each day), while the stimulated control 
group received 0.1 mL of the estradiol solution (total dose 0.03 .mu.g). 
All test groups received 0.1 mL of the stimulating dose of estradiol (0.01 
.mu.g) plus 0.05 mL of the test compounds solutions each day. The IPM and 
oil injections were made at separate sites to minimize possible physical 
or chemical interactions or reduced absorption of either compound. 
Antiestrogenic activity was measured as a decrease from the 
estradiol-induced increase in uterine weight seen in the test compound 
groups versus the estradiol-stimulated group alone. The observed 
antiestrogenic activities are set out in Table II. 
Tamoxifen elicited no antagonistic action at any of the doses tested, while 
MER 25 and Analog II yielded a dose-dependent decrease in uterine weight. 
None of the new compounds active in the mouse antiuterotrophic assay 
showed activity in the immature rat antiuterotrophic assay at doses up to 
500 .mu.g (data not shown). 
All U.S. patent applications and publications cited herein are hereby 
incorporated by reference. 
Changes may be made in the embodiments of the invention described herein or 
in parts or elements of the embodiments described herein or in the steps 
or in the sequence of steps of the methods described herein without 
departing from the spirit and scope of the invention as defined in the 
following claims.