AU712711B2 - Method of using triaryl-ethylene derivatives in the treatment and prevention of osteoporosis - Google Patents

Description

AUSTRALIA

Patents Act COMPLETE SPECIFICATION

(ORIGINAL)

Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: Hoechst Marion Roussel, Inc.

Actual Inventor(s): Alan J. Bitonti Address for Service: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: METHOD OF USING TRIARYL-ETHYLENE DERIVATIVES TREATMENT AND PREVENTION OF OSTEOPOROSIS IN THE Our Ref 571540 POF Code: 356471/1432 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1- -2- METHOD OF USING TRIARYL-ETHYLENE DERIVATIVES IN THE TREATMENT AND PREVENTION OF OSTEOPOROSIS This application is a divisional application of Australian Patent application 41967/96 the entire contents of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION The present invention relates to a method of using triaryl-ethylene derivatives in the treatment or prevention osteoporosis or bone tissue loss.

Osteoporosis is a significant problem for the developed nations. The term :osteoporosis is often used to describe different clinical situations. Osteoporosis was first used to describe the syndrome in which post-menopausal women tended to suffer vertebral fractures. F. Albright et al., J. Am. Med. Assoc. 116, 2465-2474 (1941). To avoid ambiguity, the terms bone tissue loss or osteopenia are used to describe the clinical situation in which loss of bone mass or density has occurred in the absence of a fracture.

o• Osteoporosis is most commonly associated with post-menopause and 20 age related bone tissue loss. Osteoporosis or bone tissue loss can also occur secondarily to various drugs and diseases, including: corticosteroids, anticonvulsants, alcohol, malabsorption syndromes, primary biliary cirrhosis, myeloma, thalassemia, thyrtoxicosis, Cushing's syndrome, Turner's syndrome, and primary hyperparathyroidism.

SUMMARY OF THE INVENTION The present invention relates to a method of using triaryl-ethylene derivatives in the treatment or prevention bone tissue loss or osteoporosis.

C-IWINVVRDEN14YM SPECNKIA1967D0V-DO -3- In one aspect, the present invention provides a method of treating or preventing bone tissue loss or osteoporosis in a patient, comprising administering an effective antiosteoporosis amount of a compound of the Formula III:

X

Formula I wherein A is a radical of the formula -N- G

S

wherein R and R, are each independently hydrogen or C,-C 4 alkyl; and G is HN, H 3 CN, CH 2 or O; w is an integer from 2 to 3;

R

2 is hydrogen, C,-C 4 alkyl, C 1

-C

4 alkoxy, halogen, or hydroxy;

R

3 is hydrogen, CI-C 4 alkyl, C 1

-C

4 alkoxy, halogen, hydroxy, or -Y(CH 2 )zA 1 in which A 1 is a radical of the formula C:\WINWORDUENNYMISP6CNKIM17DIV.DOC R4

N-

R

5 wherein Y is -HN-;

R

4 and R 5 are each independently hydrogen or C 1

-C

4 alkyl; G is HN, H 3 CN, CH 2 or 0; and z is an integer from 2 to 3; a X is chloro or bromo; or a pharmaceutically acceptable salt thereof.

In a further aspect, the present invention provides a method of treating or preventing bone tissue loss or osteoporosis in a patient, comprising administering an effective antiosteoporosis amount of a compound of the Formula IV: A-(CH2)w-NH Formula IV wherein A is a radical of the formula C:WNORDUENNYMSPECNVJ1g701VOO

R

RN- wherein R and R, are each independently hydrogen or C1-C4 alkyl; and G is HN, H 3 CN, CH 2 or O; w is an integer from 2 to 3;

R

2 is hydrogen, C,-C4 alkyl, Ci-C4 alkoxy, halogen, or hydroxy; S• R 3 is hydrogen, C1-C4 alkyl, C1-C4 alkoxy, halogen, hydroxy, or -Y(CH 2 )zA 1 in which A, is a radical of the formula R4

N--

Yis-HN-;

R

4 and R 5 are each independently hydrogen or C 1

-C

4 alkyl; G, is HN, H 3 CN, CH 2 or 0; and z is an integer from 2 to 3; X is chloro or bromo; or a pharmaceutically acceptable salt thereof.

In addition the present invention provides a pharmaceutical composition for oral administration comprising an effective antiosteoporosis amount of a compound of Formula III or Formula IV in admixture or otherwise in association with one or more pharmaceutically acceptable carriers or excipients.

C:%WNV-'OMUENNYMPECCrKl957tXV DO DETAILED DESCRIPTION OF THE INVENTION As used herein, the term "C 1

-C

4 alkyl" refers to a saturated, straight or branched chain, hydrocarbon radical of one to four carbon atoms and includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, t-butyl, and the like.

As used herein, the designation refers to a bond o for which the stereochemistry is not designated.

As used herein, the term "halogen" refers to a fluorine, 15 chlorine, bromine, or iodine atom.

As used herein, the term "C 1

-C

4 alkoxy" refers to a C 1

-C

4 alkyl bearing an oxy group and includes methoxy, ethoxy, npropoxy, n-butoxy, iso-propoxy, iso-butoxy, t-butoxy, and the like.

As used herein, the term "hydroxy" or "hydroxy group" refers to a -OH radical.

As used herein, the term "(CH 2 refers to a straight chain alkylene radical of from 2 carbon atoms to 12 carbon atoms for example; ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, and dodecyl.

-7- As used herein, the term "(CH 2 refers to a straight chain alkylene radical of from 2 carbon atoms to 3 carbon atoms for example; ethyl and propyl.

As used-herein, the term "(CH 2 refers to a straight chain alkylene radical of from 2 carbon atoms to 3 carbon atoms for example; ethyl and propyl.

As used herein, the term "pharmaceutically acceptable addition salt refers to either an acid addition salt or a basic addition salt.

*wo The expression "pharmaceutically acceptable acid 15 addition salts" is intended to apply to any non-toxic organic or inorganic acid addition salt of the base compounds represented by Formula III or IV or any of their intermediates. -Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric 20 and phosphoric acid and acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate.

Illustrative organic acids which form suitable salts include the mono-, di- and tri-carboxylic acids. Illustrative of such acids are, for example, acetic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, salicylic,.

2 -phenoxybenzoic, and sulfonic acids such as ptoluenesulfonic acid, methanesulfonic acid and 2hydroxyethane sulfonic acid. Either the mono- or di-acid salts may be formed, and such salts may exist in either a hydrated or substantially anhydrous form.

Compounds of Formula III, and Formula IV exist as geometric isomers.

Any reference in this application to one of the compounds represented by Formula III or Formula IV is meant to encompass each of the specific geometric isomers. The specific geometric isomers can be separated and recovered by techniques known in the art such as chromatography on silica gel, chromatography on a reverse-phase adsorbent, or fractional recrystallization.

As is well known by one of ordinary skill in the art the Cahn-lngold-Prelog designation of and for isomers of compounds of Formula III, and Formula IV depends on the nature of Y, X, m, w, p, z, A, R 2 and R 3 Compounds of Formula III or IV in which the substituent R 3 is -NH(CH 2 and w=z and A=Aj do not exist as geometrical isomers.

Illustrative Examples of compounds encompassed by the present -invention include: 4(-itylmnehlmn9*ey]12dpey-2clr-tyee 4 2 -Diethylaminoethylamino)phenyl]-1,2-diphenyl-2-chloro-ethylene; 4 3 -Diethylaminopropylamino)phenyl] 1 ,2-d iphenyl-2-chloro-ethylene; 4 3 -Diethylaminopropylamino)phenyl]-1,2-diphenyl-2-chloro-ethylene; 4 2 -Diethylaminoethylamino)phenyl]l ,2-diphenyl-2-bromo-ethylene; 4 2 -Diethylaminoethylamino)phenyl]-1, 2 -diphenyl-2-bromo-ethylene; 4 3 -Diethylaminopropylamino)phenyl] 1 2 -diphenyl-2-bromo-ethylene; 1 4 3 -Diethyla min opropyla mino) phenyl]- 1 ,2-d iphenyl-2-bromo-ethyle ne.

C %MNWVORDUENNYMSPECNKI41W7DIV -9- The compounds of Formula III and IV can be prepared as described in Scheme C. In Scheme C, compounds which include the alkylene group (CH 2 n wherein n is an integer from 2 to 3 encompass the compounds of Formula III, and IV. All substituents, unless otherwise indicated, are previously defined.

The reagents and starting materials are readily available to one of ordinary skill in the art.

S*

0* w

S

C:IWIW40RDUENWW,-,PECWPAI9157~DIV.DO SCHEME C 59.5 5@ 5* 5 @5 0 S S

S.

*5

S

ACYLATIO

step a AMI NATI step b 0 (CH2)q

H

S. S S S S5 55@b 0 S S *554

S

S

0 (C H2) q\4 H -11- SCHEME C (cont.) 0 (CH2)q

NH

S

S. S

S

555* 5555e*

HALOGENATION

step c 0 (CH2)q H foe* o

REDUCTION

step d

(CH

2 )n A' NH -12- (C H2) n A* NH SCHEME C (cont.) 6 DEP ROTE CTION Optional step e

(CH

2 )n A* 'NH a a a a.

a a a ISOMER SEPARATION step f (C H2) n A' NH

(CH

2 )n A- NH -13- In Scheme C, step a, an appropriate o-haloalkylacid halide, Z-(CH2)q-C(O)Zi, is added to an appropriate aminotriaryl-ethylene of structure 9 in an acylation reaction to give a o-haloalkylamido-triaryl-ethylene of structure An appropriate o-haloalkylacid halide, Z-(CH2)q-C(O)Zi, is one in which q is 1 less than n, an integer of 2 or 3 as desired in the final product of Formula III, or IV, and Z and ZI may each independently be a chlorine atom or a bromine atom. An appropriate amino-triaryl-ethylene of the structure 9 is one in which R 6 is R 2 as defined above, or is a suitably protected hydroxy which after deprotection provide compounds of Formula III, or IV in which R 2 is a hydroxy group; Rg is R 3 as defined above, or is a suitably protected hydroxy which after deprotection providescompounds of Formula III, or IV in which R 3 is a hydroxy group, or Rg is an amino group, a protected amino group, ora group which gives rise to an amino group, such as a nitro group. Appropriate amino-triaryl-ethylenes of the structure 9 are readily prepared by methods analogous to those used to prepare triaryl-ethylene of structure 1 described in U. S. Patent No. 2,914,563, R. E.

Allen etal; U. S. Patent No. 2,429,556, C. F. Longfellow et al; and Syn. Comm. 17, 1787-1796 (1987), M. I. Al-Hassan.

For example, a slight molar excess of a c-haloalkylacid halide is contacted with a amino-triaryl-ethylene of the S..structure 9 in a suitable solvent, such as pyridine, dimethylformamide, acetonitrile, or tetrahydrofuran. The reaction is carried out in the presence of a suitable base, such as pyridine, triethylamine, sodium carbonate, or sodium bicarbonate. The reaction may be carried out in the presence of a catalyst, such as 4-dimethylaminopyridine.

The reaction is stirred for from 1-72 hours. The production of a compound of Formula III or IV in which

R

3 is -NH(CH2)zA 1 wherein z=w and A=A 1 -14requires the use of a compound of structure 1 in which R9 is amino and slightly more than two molar equivalents of a ohaloalkylacid halide and gives rise to a compound of structure 10 which is a bis-o-haloalkylamido-triarylethylene. The product can be isolated and purified by techniques well known in the art. For example, the reaction mixture can be concentrated in vacuo to give a residue. The residue can be chromatographed on silica gel using a suitable organic eluent. The material obtained from chromatography can be recrystallized to give a ohaloalkylamido-triaryl-ethylene of structure In Scheme C, step b, a c-haloalkylamido-triarylethylene of structure 10 is contacted in an amination reaction with an appropriate amine,

HNRR

1 in which R and RI are as defined above, morpholine, piperidine, piperazine, 4 -methylpiparizine, or pyrrolidine to give coaminoalkylamido-triaryl-ethylene of structure 11.

20 For example, a o-haloalkylamido-triaryl-ethylene of structure 10 is contacted with a large molar excess of an appropriate amine. A large molar excess of amine is used so that the amine also acts as a base to take up the acid liberated in the reaction. The reaction is carried out in a suitable solvent, such as ethanol, methanol, water, ethanol/water mixtures, or methanol/water mixtures. The reaction may be carried out in the presence of a suitable catalyst, such as potassium iodide. The reaction vessel may be sealed to prevent the escape of volatile amines.

The reaction mixture is heated to temperatures of from to 100 0 C. For compounds of structure 10 in which R9 is a o-haloalkylamido group the use of an additional portion of an appropriate amine gives a bis-c-aminoalkylamido-triarylethylene which gives rise to a compound of Formula

III

or IV in which R 3 is -NH(CH 2 )zA 1 wherein z=w and A=A 1 The product is isolated from the reaction zone by evaporation or extraction and is purified by chromatography or salt formation and recrystallization to give a oaminoalkylamido-triaryl-ethylene of structure 11.

In Scheme C, step c, a o-aminoalkylamido-triarylethylene of structure 11 are chlorinated or brominated to give c-aminoalkylamido-triaryl-ethylene of structure 12.

For example, a o-aminoalkylamido-triaryl-ethylene of structure 11 is contacted with a molar excess of chlorine, bromine, N-chlorosuccinimide, or N-bromosuccinimide in a solvent, such as chloroform or dichloromethane. The reaction is carried out at temperatures from ambient temperature to the reflux temperature of the solvent.

After stirring for from 12-72 hours the product can be isolated and purified by techniques well known in the art.

SFor example, the reaction mixture can be concentrated in vacuo and the product purified by chromatography or by recrystallization to give a o-aminoalkylamido-triaryl- S* 20 ethylene of structure 12.

In Scheme C, step d, a o-aminoalkylamido-triarylethylene of structure 12 is contacted with an appropriate reducing agent in a reduction reaction to give a oaminoalkylamino-triaryl-ethylene of structure 13.

An appropriate reducing agent is one that will reduce the amido group of a o-aminoalkylamido-triaryl-ethylene of structure 12 without effecting the other groups present in the compound. The selection and use of such reducing agents is well known and appreciated in the art.

For example, a o-aminoalkylamido-triaryl-ethylene of structure 12 is contacted with a molar excess of an appropriate reducing agent, such as lithium aluminum hydride, borane, or borane complexes. The reaction is carried out in a solvent, such as diethyl ether or -16tetrahydrofuran when the appropriate reducing agent is lithium aluminum hydride, or dichloromethane or chloroform when the appropriate reducing agent is borane. The reaction is carried out at temperatures from ambient temperature to the refluxing temperature of the solvent.

For compounds of structure 12 in which R9 is a oaminoalkylamido group the use of an additional portion of the appropriate reducing agent gives a bis-oaminoalkylamino-triaryl-ethylene which gives rise to a compound of Formula III or IV in which R 3 is -NH(CH 2 )zAi wherein z=w and A=AI. The product can be isolated from the reaction zone by techniques well known in the art, such as quenching, extraction, and evaporation; and may be purified by methods well known in the art, such as chromatography and recrystallization to give a o-aminoalkylamino-triaryl-ethylene of structure 13.

In Scheme C, optional step e, for o-aminoalkylaminotriaryl-ethylene of structure 13 in which R 6 or R9 are a protected hydroxy group the protecting group is removed in a deprotection step to provide o-aminoalkylamino-triarylethylene of structure 14 in which either, R 2 or R 3 or R 2 and R 3 are hydroxy as desired in the final product of Formula I or Formula II. Additionally, for oaminoalkylamino-triaryl-ethylene of structure 13 in which R9 is a protected amino group is deprotected to provide oaminoalkylamino-triaryl-ethylene of structure 14 in which R9 is an amino group can be, by sequentially performing the steps of Scheme C, used as an intermediate for the preparation of a compound of Formula I or II in which R3 is

-NH(CH

2 )pAi wherein p*n and either A=A 1 or A*AI or in which R3 is -NH(CH 2 )pAI wherein p=m and A*A 1 or a compound of Formula III or IV in which R 3 is -NH(CH 2 )zAl wherein z*w and either A=A 1 or AAI 1 or in which R 3 is -NH(CH 2 2 Ai wherein z=w and AA

I

The removal of amine protecting groups utilizing suitable protecting groups such as those described in -17- Protecting Groups in Organic Synthesis by T. Greene is well known and appreciated by those skilled in the art.

The selection, use, removal, and sequential removal of suitable hydroxy protecting groups, such as benzyl, pmethoxybenzyl, methyl, t-butyldimethylsilyl, and acetyl, is well known and appreciated in the art and described in Protecting Groups in Organic Synthesis by T. Greene.

In Scheme C, step f, the isomers of a oaminoalkylamino-triaryl-ethylene of structure 13 or 14 are separated to give a (E)-c-aminoalkylamino-triaryl-ethylene and the (Z)-c-aminoalkylamino-triaryl-ethylene.

For example, the isomers of compounds of structure 13 or 14 can be separated and purified by high-performance liquid chromatography or fractional recrystallization of salt to give a (E)-c-aminoalkylamino-triaryl-ethylene and the (Z)-ca-aminoalkylamino-triaryl-ethylene.

Pharmaceutically acceptable salts of a aminoalkylamino-triaryl-ethylene and of a aminoalkylamino-triaryl-ethylene can be formed in an additional step as is well known and practiced in the art.

-18- Scheme D H2N ACETYLATI ON step a

HALOGENATION

step b/ 0 HN, CH 3 p.

p p p p p. p p. p a.

p.

p a p p (16) -19- Scheme D (cont.)

ALKYLATION

stp c 0

A(CH

2 )n)K

H

nfl.

fi. S fi. fi

S

S. fi a S fi *5 5*5*

S

HYDROLYSI

step d

(CH

2 )n A' NH In Scheme D, step a, an appropriate amino-triarylethylene of structure 9 is acetylated to give an acetamidotriaryl-ethylene of structure An appropriate amino-triaryl-ethylene of structure 9 is as defined above in Scheme C.

For example, an amino-triaryl-ethylene of the structure 9 is contacted with a suitable acetyling reagent, such as acetyl chloride or acetic anhydride. The reaction is carried out in a suitable solvent, such as pyridine, dichloromethane, dimethylformamide, acetonitrile, or tetrahydrofuran. The reaction is carried out in the presence of a suitable base, such as pyridine, triethylamine, sodium carbonate, or sodium bicarbonate.

The reaction is stirred for from 1-72 hours. The product can be isolated and purified by techniques well known in :the art, such as extraction, evaporation, chromatography, oo recrystallization, and trituration.

As is appreciated by one of ordinary skill in the art an alcohol precursor to an appropriate amino-triarylethylene of structure 9 can be dehydrated and acetylated in one step to give an acetamido-triaryl-ethylene of structure 25 In Scheme D, step b, an acetamido-triaryl-ethylene of structure 15 is chlorinated or brominated as generally taught in Scheme C, step c, to give acetamido-triarylethylene of structure 16.

For example, an acetamido-triaryl-ethylene of structure is contacted with a molar excess of chlorine, bromine, N-chlorosuccinimide, or N-bromosuccinimide in a solvent, such as chloroform or dichloromethane. The reaction is carried out at temperatures from ambient temperature to the reflux temperature of the solvent. After stirring for from -21- 12-72 hours the product can be isolated and purified by techniques well known in the art. For example, the reaction mixture can be concentrated invacuo and the product purified by chromatography or by recrystallization to give a o-aminoalkylamido-triaryl-ethylene of structure 12.

In Scheme D, step c, an acetamido-triaryl-ethylene of structure 16 is alkylated with an appropriate o-aminoalkyl halide or a salt thereof to give a aminoalkyl)acetamido-triaryl-ethylene of structure 17.

An appropriate o-aminoalkyl halide, A(CH 2 )nZ, is one in which Z is a chlorine atom, a bromine atom, or a iodine atom; A and n, an integer as desired in the final product of Formula III, or IV.

S. For example, an acetamido-triaryl-ethylene of structure **16 is contacted with from 1.0 to 10 molar equivalents of an appropriate o-aminoalkyl halide. The reaction is carried 20 out in the presence of a suitable base, such as sodium ethoxide, sodium methoxide, potassium hydroxide, sodium hydroxide, potassium hydroxide, and sodium carbonate. The reaction is carried out in a solvent, such as ethanol, methanol, tetrahydrofuran, acetone, or butanone. The 25 product may be isolated from the reaction zone by evaporation and extraction and may be purified by methods well known in the art, such as chromatography and recrystallization.

In Scheme D, step d, the acetyl group of a N-(oaminoalkyl)acetamido-triaryl-ethylene of structure 17 is hydrolyzed to give an o-aminoalkylamino-triaryl-ethylene of structure 13. The hydrolysis of acetamido compounds using either basic or acidic conditions is well known and appreciated in the art.

-22- As generally taught in Scheme C, optional step e, an oaminoalkylamino-triaryl-ethylene of structure 13 can be deprotected and further modified as required to give a oaminoalkylamino-triaryl-ethylene of structure 14.

As generally taught in Scheme C, step f, the isomers of a o-aminoalkylamino-triaryl-ethylene of structure 13 or 14 are separated to give a (E)-o-aminoalkylamino-triarylethylene and the (Z)-)-aminoalkylamino-triaryl-ethylene.

The following examples present typical syntheses as described in Scheme D. These examples are understood to be illustrative only and are not intended to limit the scope :of the invention in any way. As used in the following 15 examples, the following terms have the meanings indicated: refers to grams, "mg" refers to milligrams, "mmol" *o refers to millimoles, "mol" refers to moles, "mL" refers to milliliters, "mm" refers to millimeters, "OC" refers to degrees Celsius, "Rf" refers to retention factor, "mp" 20 refers to melting point, "HPLC" refers to high performance liquid chromatography.

EXAMPLE 1 (E and 4 -Acetamidophenyl)-1,2-diphenyl-ethylene 25 Combine 4 -aminobenzophenone (50 g, 0.25 mol) and diethyl ether (500 mL). Slowly, add benzylmagnesium chloride (1 L of a IM solution in diethyl ether over 1.5 hours. After 18 hours, pour the reaction onto ice and an aqueous solution of ammonium chloride. Separate the layers, extract the organic layer with water, dry over MgSO 4 filter, and evaporate in vacuo to give a residue. Recrystallize from isopropanol to give l-( 4 -aminophenyl)-l,2-diphenyl-ethanol: mp; 105-107 0

C.

Combine l-( 4 -aminophenyl)-1,2-diphenyl-ethanol (40 g, 0.138 mol) and pyridine (75 mL). Slowly add acetic anhydride (50 mL). Heat on a steam bath. After 20 hours, cool and evaporate invacuo to give a residue. Partition the residue between diethyl ether and water. Separate the -23layers, extract the organic layer with water, dry over MgSO 4 filter, and evaporate invacuo to give the title compound.

EXAMPLE 2 (E and Z)--(4-Acetamidophenyl)-1,2-diphenyl-2-chloroethylene Combine (E and Z)-l-(4-acetamidophenyl)-l,2-diphenylethylene and acetic acid (250 mL). Slowly add chlorine (350 mL of a 0.46M solution in carbon tetrachloride).

After the addition, stir the reaction at ambient temperature for 1 hour and then heat on a steam bath.

After 2 hours, cool and evaporate invacuo to give a residue.

Recrystallize the residue from 95% ethanol to give the 15 title compound: mp; 179-185 0

C.

EXAMPLE 3 (E and 4 -(N-(2-Diethylaminoethyl)acetamidophenyll-l,2diphenyl-2-chloro-ethylene 20 Combine (E and Z)-l-(4-acetamidophenyl)-1,2-diphenyl-2chloro-ethylene (17.4 g, 0.05 mol), 2-diethylaminoethyl chloride hydrochloride (10 g, 0.058 mol) and powdered potassium hydroxide (6.7 g, 0.12 mol) in acetone (150 mL).

Heat to reflux. After 2 hours, filter, and evaporate to 25 give a residue. Partition the residue between diethyl ether and water. Separate the layers, extract the organic layer with water, dry over MgSO 4 filter, and evaporate in vacuo to give the title compound.

-24- EXAMPLE 4 (E and 4 2 -Diethylaminoethylamino)Dhenvll-l-2diphenyl-2-chloro-ethylene citrate salt N- (CH 2 2

CH

2

CO

2

H

C(OH)CO

2

H

C CH 2

CO

2

H

Combine (E and diethylaminoethyl)acetamidophenyl ]-1,2-diphenyl-2-ch-loroethylene, aqueous 10% hydrochloric acid (200 mL), and aqueous concentrated hydrochloric acid (10 mL). Heat on a steam bath. After 6 hours, cool to ambient temperature.

After 18 hours, add aqueous 10% sodium hydroxide until the solution becomes basic. Extract the basic solution with diethyl ether. Extract the organic layer with water, dry :over MgSO 4 filter and evaporate invacuo to give a residue.

Combine the residue and butanone. Add citric acid (4.3 g) to give a solid. Filter and recrystallize twice from butanone to give the title compound: mp; 121-125 0

C.

The compounds of Formula III and IV can be prepared as described in Scheme E. In Scheme E, compounds which include the alkylene group (CH 2 )n wherein n is an integer from 2 to 3 encompass the compounds of Formula III, and IV.

All substituents, unless otherwise indicated, are previously defined. The reagents and statrting materials are readily available to one of ordinary skill in the art.

SCHEME E H 2

N

ACETYLATI ON step a (18) 0 HN,

CH

3

ALKYLATION

step b/ 0

(CH

2 )n A' N) CH 3 (18a) G6

M

OLEFINATION

step c (19) -26- SCHEME E (cont.)

OLEFINATION

step c

HYDROLYSIS

step d a a a.

a a a.

(17)

(CH

2 )n A' NH In Scheme E, step a, an appropriate 4 -aminobenzophenone of structure 18 is acetylated to give an N-acetyl-4aminobenzophenone of structure 18a. As is appeciated by those of ordniary skill in the art groups other than acetyl may be used, such as trifluoroacetyl, benzoyl, methanesulfonyl, or trifluoromethanesulfonyl. An appropriate 4 -aminobenzophenone of structure 18 is one in which Rg is as defined above in Scheme C, step a.

For example, an appropriate 4 -aminobenzophenone of structure 18 is contacted with a suitable acetyling reagent, such as acetyl chloride or acetic anhydride. The reaction is carried out in a suitable solvent, such as -27pyridine, dichloromethane, dimethylformamide, acetonitrile, toluene, or tetrahydrofuran. The reaction is carried out in the presence of a suitable base, such as pyridine, triethylamine, sodium carbonate, or sodium bicarbonate.

The reaction is stirred for about 1-72 hours. Generally, the reaction is carried out at temperatures of from about 0 C to the refluxing temperature of the solvent. The product can be isolated and purified by techniques well known in the art, such as extraction, evaporation, chromatography, recrystallization, and trituration.

In Scheme E, step b, the N-acetyl-4-aminobenzophenone of structure 18a is alkylated with an appropriate oaminoalkyl halide to give an N-c-aminoalkyl-N-acetyl-4- 15 aminobenzophenone of structure 19. An appropriate oaminoalkyl halide, A(CH 2 )nZ, is one as described in Scheme D, step c.

For example, a N-acetyl-4-aminobenzophenone of 20 structure 18a is contacted with from 1.0 to 10 molar equivalents of an appropriate o-aminoalkyl halide. The reaction is carried out in the presence of a suitable base, such as sodium ethoxide, sodium methoxide, potassium hydroxide, sodium hydroxide, potassium carbonate, and 25 sodium carbonate. The reaction is carried out in a solvent, such as ethanol, methanol, tetrahydrofuran, acetone, or butanone. The product may be isolated from the reaction zone by evaporation and extraction and may be purified by methods well known in the art, such as chromatography and recrystallization.

In Scheme E, step c, a N-&-aminoalkyl-N-acetyl-4aminobenzophenone of structure 19 is olefinated using an appropriate a-chloro or a-bromo phosphorous reagent to give a N-(o-aminoalkyl)acetamido-triaryl-ethylene of structure 17.

-28- An appropriate a-chloro or a-bromo phosphorous reagent is one which R 6 is as defined in Scheme C, step a, X is chloro or bromo, M is lithium, sodium, or potassium, and G is P(Ph) 3 P(O)(OPh) 2

P(O)(OCH

2

CH

3 2

P(O)(OCH

3 2 or P(0)(OCH(CH 3 2 2 The preparation and use of appropriate achloro or a-bromo phosphorous reagents well known and appreciated in the art, such as described in K. Lee et al., Syn. Comm. 22 649-655 (1992); J. Petrova et al., Synthesis 658-660 (1975); M. D. Crenshaw and H. Zimmer J. Org. Chem.

48 2782-2784 (1983); T. Gajda, Synthesis 717-718 (1990); T.

Gajda, Phosphorous and Sulfur 53 327-331 (1990); and S. K.

Chakoborty and R. Engle, Syn. Comm. 21 1039-1046 (1991).

An appropriate a-chloro or a-bromo phosphorous reagent can be prepared and isolated or isolated and purified before 15 use or can be prepared and used without isolation.

For example, a N-o-aminoalkyl-N-acetyl-4aminobenzophenone of structure 19 is contacted with a slight excess of lithium diethyl a-chloro 20 benzylphosphonate. The reaction is carried out in a suitable solvent, such as THF, benzene or toluene. The reaction is performed at temperatures from -78 0 C to the refluxing temperature of the solvent. The product can be isolated as generally taught in Scheme D, step c.

"o Alternately, a N-c-aminoalkyl-N-acetyl-4aminobenzophenone of structure 19 is contacted with a slight excess of an anion derived from a a-chloro benzyltriphenylphosphonium salt. The reaction is carried out in a suitable solvent, such as THF, or toluene. The reaction is performed at temperatures from -78 0 C to the refluxing temperature of the solvent. The product can be isolated as generally taught in Scheme D, step c.

As generally taught in Scheme D, step d, the acetyl group of a N-(o-aminoalkyl)acetamido-triaryl-ethylene of -29structure 17 is hydrolyzed to give an o-aminoalkylaminotriaryl-ethylene of structure 13.

As generally taught in Scheme C, optional step e, an oaminoalkylamino-triaryl-ethylene of structure 13 can be deprotected and further modified as required to give a caminoalkylamino-triaryl-ethylene of structure 14.

As generally taught in Scheme C, step f, the isomers of a o-aminoalkylamino-triaryl-ethylene of structure 13 or 14 are separated to give a (E)-o-aminoalkylamino-triarylethylene and the (Z)-a-aminoalkylamino-triaryl-ethylene.

EXAMPLE 15 N-Acetyl-4-aminobenzophenone Combine 4 -aminobenzophenone (10.0 g, 50.8 mmol), acetic anhydride (5.74 mL, 60.9 mmol), and triethylamine (9.5 mL, 68.5 mmol) in toluene (30 mL). Heat to reflux. After 2 hours, cool to ambient temperature and pour the reaction 20 mixture into water. Stir to give a solid. Collect the solid by filtration, rinse with water, and dry.

Recrystallize from acetonitrile to give the title compound.

S. Rf=0.35 (silica gel, ethyl acetate).

EXAMPLE 6 N-Acetyl-4-aminobenzophenone Combine 4 -aminobenzophenone (500 g, 2.54 mol) and triethylamine (307 g,3.03 mol) in dichloromethane (2.54 L).

Add acetic anhydride (313.9 g, 3.07 mol). After 18 hours, add methanol (100 mL) and evaporate invacuo to give a residue. Combine the residue and water (8 L) and stir to give a solid. Collect the solid by filtration, rinse repeatedly with water, and dry to give the title compound.

EXAMPLE 7 N-(2-Diethylaminoethyl)-N-acetyl-4-aminobenzophenone Combine N-acetyl-4-aminobenzophenone (4.0 g, 16.7 mmol), 2-diethylaminoethyl chloride hydrochloride (3.97 g, 23.1 mmol), and powdered potassium hydroxide (2.81 g, 50.1 mmol) in acetone (30 mL). Heat to reflux. After 2 hours, cool and decant the solvent.' Evaporate the solvent invacuo to give a residue. Partition the residue between water and ethyl acetate. Separate the organic layer and extract with water. Dry the organic layer over MgSO 4 filter, and evaporate invacuo to give the title compound: Rf=0.0-0.14 (streaks on silica gel, ethyl acetate).

EXAMPLE 8 N-(2-Diethylaminoethyl)-N-acetyl-4-aminobenzophenone Combine N-acetyl-4-aminobenzophenone (250 g, 1.05 mol), 15 2-diethylaminoethyl chloride hydrochloride (207.5 g, 1.20 mol), and 85% potassium hydroxide (140 g, 2.12 mol) in acetone (3 Heat to reflux. After 2 hours, cool and filter. Evaporate the filtrate invacuo to give a residue.

Combine the residue and t-butyl methyl ether (1 L) and water 20 (2 Add aqueous IM hydrochloric acid solution until the pH is about 2.5. Separate the aqueous layer and extract several times with t-butyl methyl ether. Adjust the pH of the aqueous layer to 10 using aqueous 50% sodium hydroxide solution. Extract the aqueous layer twice with 25 dichloromethane. Dry the combined dichlormethane layers over MgSO 4 filter, and evaporate invacuo to give a residue.

Chromatograph the residue on silica gel eluting sequentially with ethyl acetate, 5/95 methanol/ethyl acetate, 10/90 methanol/ethyl acetate to give the title compound: Rf=0.36 (silica gel, 2/8/0.1 methanol/ethyl acetate/triethylamine).

EXAMPLE 9 (E and 4 -N-Acetyl-N-(2-diethylaminoethylamino)phenyl]- 1,2-diphenyl-2-chloro-ethylene Combine diethyl benzylphosphonate (1.23 mL, 5.91 mmol) and anhydrous tetrahydrofuran (9.0 mL). Cool to -78 0 C using a dry-ice-acetone bath. Add dropwise over 5 minutes a -31solution of n-butyllithium (4.73 mL, 2.5 M in hexanes, 11.8 mmol). After 30 minutes, add benzenesulfonyl chloride -0.76 mL, 5.9 mmol). Add N-(2-diethylaminoethyl)-N-acetyl-4aminobenzophenone (1.0 g, 2.96 mmol) in anhydrous tetrahydrofuran (6.0 mL). After 5 minutes at -78 0 C, warm to ambient temperature. After 30 minutes, add a saturated aqueous solution of ammonium chloride (20 mL) and extract with ethyl acetate. Separate the organic layer and extract with a saturated aqueous solution of sodium bicarbonate, dry the organic layer over MgSO 4 filter, and evaporate invacuo to give a residue. Chromatograph the residue on silica gel eluting with 5/94.9/0.1 methanol/ethyl acetate/ triethylamine to give the title compound

E/Z):

Rf=0.27 (silica gel, 20/80/0.05 methanol/ethyl 15 acetate/triethylamine).

4.

4 *0e* EXAMPLE (E and Z)-l-[4-N-Acetyl-N-(2-diethylaminoethylamino)phenyl1- 1, 2 -diphenyl-2-chloro-ethylene 20 Combine diethyl benzylphosphonate (42.9 g, 187 mmol) and anhydrous tetrahydrofuran (350 mL). Cool to -600C. Add dropwise a solution of n-butyllithium (150 mL, 2.5 M in hexanes, 375 mmol) while maintaining the temperature at less than about -55 0 C. After 30 minutes, add a solution of 25 benzenesulfonyl chloride (32.8 g, 187 mmol) in tetrahydrofuran (90 mL) while maintaining the temperature at less than about -55 0 C. After 15 minutes, warm the reaction mixture to -30 0 C. After 15 minutes, cool to -55C. Add by cannula to a cooled (-55 0 C) solution of N-(2diethylaminoethyl)-N-acetyl-4-aminobenzophenone (55.0 g, 163 mmol) in anhydrous tetrahydrofuran (280 mL). The addition by cannula is carried out while maintaining the temperature of the reaction mixture at less than about -450C. After minutes, warm slowly to ambient temperature. After minutes, add a saturated aqueous solution of ammonium chloride (500 mL) and extract with ethyl acetate (2.5 L).

Separate the organic layer and extract with a saturated

I

-32aqueous solution of sodium bicarbonate, dry over MgSO 4 filter, and evaporate invacuo to give a residue.

Chromatograph the residue on silica gel eluting sequentially with 99.9/0.1 ethyl acetate/triethylamine and 5/94.9/0.1 methanol/ethyl acetate/triethylamine to give the title compound., EXAMPLE 11 (E and Z-1- 2-Diethylaminoethylamino)phenvl]-1,2diphenyl-2-chloro-ethylene -,,(CH2)2-NH Combine (E and Z)-l-[4-N-acetyl-N-(2diethylaminoethylamino)phenyl-1,2-diphenyl-2-chloroethylene (0.1 g, 0.22 mmol), potassium t-butoxide (0.16 g, 1.45 mmol), water (8.1 pL, 0.45 mmol), and diethyl ether (1 mL). Heat to reflux. After 16 hours, cool to ambient temperature, pour into ice-water and extract with ethyl acetate. Separate the organic layer and extract with water .j and saturated aqueous sodium chloride. Dry the organic layer over MgSO 4 filter, and evaporate invacuo to give a residue. Chromatograph the residue on silica gel eluting with 5/94.9/0.1 methanol/ethyl acetate/triethylamine to give the title compound Rf=0.18 (silica gel, 20/80/0.05 methanol/ethyl acetate/triethylamine).

EXAMPLE 12 E and 2-Diethylaminethylamino)phen -1,2diphenyl-2-chloro-ethylene

I

-33- Combine (E and Z)-l-[4-N-acetyl-N-(2diethylaminoethylamino)phenyl]-1,2 -diphenyl-2-chloroethylene (59.3 g, 133 mmol), potassium t-butoxide (95 g, 850 mmol), water (2.0 mL), and tetrahydrofuran (1.0 Heat to reflux. After 1 hour, cool to ambient temperature and partition between water and ethyl acetate. Separate the organic layer and extract with water and saturated aqueous sodium chloride. Dry the organic layer over MgSO 4 filter, and evaporate invacuo to give the title compound.

EXAMPLE 13 2 -Diethylaminoethylamino)phenyl1-1,2-diphenyl-2chloro-ethylene hydrochloride salt (CH2)2.NH ~15 5 .o

*HCI

.:Combine (E and Z)-l-[4-N-acetyl-N-(2ydiethylaminoethylamino)phenyl-l,,2-diphenyl-2-chloroethylene (53.9 g, 133 mmol) and diethyl ether. Cool to about -5 0 C. Add hydrochloric acid (gas, 10.1 g, 280 mmol) over about 5 minutes while maintaining the temperature at about 5-15 0 C to give a solid. Collect the solid by filtration, rinse with diethyl ether, and dry. Combine the solid and acetone (375 mL). Heat to reflux. After hours, cool to ambient temperature and collect the solid by filtration, rinse with acetone. Combine the solid with tetrahydrofuran (450 mL) and heat to reflux. After 18 hours, cool to ambient temperature and collect the solid by filtration, rinse with tetrahydrofuran. Again combine the solid with tetrahydrofuran (400 mL) and heat at reflux.

After 18 hours, cool to ambient temperature and collect the -34solid by filtration, rinse with tetrahydrofuran, and dry in vacuo to give the title compound (98.9% E-isomer).

Throughout the description and claims of the specification the word "comprise" and variations of the word, such as "comprising" and "comprises" is not intended to exclude other additives, components, integers or steps.

a a a EXAMPLE 14 (E)-l-(4-N-(2-Diethylaminoethylamino)phenyl}-l,2-diphenl-2chloro-ethylene citric acid salt

N-(CH

2 2 CH2CO2H

C(OH)CO

2

H

CH2C02H Combine (E)-l-(4-N-acetyl-N-(2- 15 diethylaminoethylamino)phenyl]-1,2-diphenyl-2-chloroethylene hydrochloride salt (27.6 g, 58 mmol) and ethyl acetate (350 mL). Carefully add with stirring a saturated aqueous solution of sodium bicarbonate (200 mL). After minutes, separate the organic layer and extract with a 20 saturated aqueous solution of sodium chloride. Combine the aqueous layers and extract with ethyl acetate. Dry the combined organic layers over MgSO 4 filter, and evaporate in v acuo to give a residue. Combine the residue and acetone (300 mL), filter, and add a solution of citric acid (11.1 g, 57.8 mmol) in acetone (70 mL). Stir for 5 hours to give a solid. Collect the solid by filtration, rinse with acetone, and dry in vacuo to give the title compound. Elemental Analysis calculated for C 26

H

28 C1N 2

C

6

H

8 0 7 C, 64.37; H, 6.25; N, 4.69. Found: C, 64.05; H,6.19; N, 4.59.

EXAMPLE Measurement of the Prevention of Bone Loss Female, Sprague-Dawley rats weighing 200 to 225 grams each (70-75 days of age) are obtained from Harlan Sprague Dawley, Inc. (Indianapolis, IN). For each compound, studies are carried out on four groups of animals as follows: Group 1 consists of 5 to 20 ovariectomized rats treated with test -36compound at typical doses of 0.001 to 10 mg/kg/day; Group 2 consists of 5 to 20 ovariectomized rats treated with vehicle; Group 3 consists of 5 to 20 sham ovariectomized rats (incision made but ovaries not removed) treated with test compound at typical doses of 0.001 to 10 mg/kg/day; Group 4 consists of 5 to 20 sham ovariectomized rats treated with vehicle. All rats are housed individually in suspended wire cages and provided standard rodent laboratory pellets (Purina #5001) and deionized water adlibitum.

The test compound is homogenized in distilled water containing lecithin (10 mg/ml), sodium methylparaben (1.05 mg/ml) and sodium propylparaben (0.23 mg/ml). Compound and vehicle are administered daily for 40 consecutive days by 15 gavage in a volume of 5 ml/kg. Treatment is initiated on S. i the day following ovariectomy or sham ovariectomy. Twentyfour hours after the final administration of test compound (or vehicle) the rats are sacrificed, the hind limbs removed, dissected free of the bulk of soft tissue and then 20 placed in 10% buffered formalin until further processed.

Complete ovariectomy is verified at the time of sacrifice and rats with fragments of ovarian tissue-are excluded.

After a minimum of one week in formalin, the femurs and tibias are carefully dissected free of each other and remaining soft tissue. The femurs are then air dried for a minimum of 48 hours and radiographed in a Faxitron X-ray cabinet (Hewlett-Packard, McMinnville OR) using Kodak XK-1 film exposed for 9 seconds at a tube voltage of 60 kV.

Images of radiographs are captured using a CCD-72 solidstate video camera (Dage-MTI Inc., Michigan City, IN) equipped with a 50 mm Nikon lens and a +1 or combined and +1 close-up lenses depending on the desired magnification. The captured images are digitized with a CFG framegrabber (Imaging Technology Inc., Bedford MA) and analyzed using Optimas image analysis software (Bioscan Inc., Edmonds WA).

-37- Bone density gray value of radiographic images) is measured at the distal femur. Cancellous bone loss is measured at the proximal tibia. Decreased gray value variance reflects the loss of fine structure of bone present in radiographs.

Image analysis measurements are made as follows: For determination of the gray-values of femurs, images are precisely orientated on the monitor using screen templates and analysis region (2.30 mm x 3.34 mm; 11250 pixels) is superimposed over the digitized images at the distal femur at approximately 1.2 mm proximal to the border of the medial and lateral condyles. The gray value of each picture 15 element within the analysis region is then measured and the mean gray value calculated by Optimas image analysis *software (BioScan Inc; Edmondes, Washington). To minimize 0 the effect of random noise on gray value measurements, two images of the analysis region are captured and averaged.

20 The averaged image is then treated with a 3 x 3 smoothing filter to yield the final image for gray value measurements.

In addition to mean gray value, gray value variance is also determined at the distal femur using the same analysis region used for determining mean gray value.

To control for variability in gray value measurements potentially arising from the radiographic and' imaging procedures, gray values are converted to Density Units using standard curves prepared from gray value measurements collected from a 10-step aluminum wedge radiographed along with the specimens on each film.

After removal from the formalin, tibias are placed in labeled tissue cassettes, decalcified, imbedded in parraffin, cut in 5 micron sagittal sections which are stained with aniline blue (a differential stain for bone).

-38- Trabecular bone loss is measured directly from stained sections of the proximal tibia.

For measuring trabecular bone area, tibia section images are precisely orientated on a moniter screen template and a analysis region (1.17 mm x 1.54 mm; 18240 pixels) centered over the image between the cortico-medullary margins and positioned 1.0 mm from the growth plate at the closest point. Optimas then determines the area of both the analysis region and the trabecular bone within the region (in mm 2 and calculated the percent of the region occupied by trabecular bone.

*o Statistical comparisons are done using Data Desk 15 Professional software (Odesta Corp., Northbrook

IL).

Comparisons between groups are made using the two-tailed t test for independent means with a pooled estimate of variance. Values for the right and left femurs are averaged for each animal and the combined values used to calculate descriptive statistics and for t tests. Statistical significance is assigned at P 0.05 and P 0.01.

EXAMPLE 16 Measurement of Serum Osteocalcin 25 For each compound, studies are carried out on four groups a animals as described in Example 65. Anesthetize each animal with CO2, collect blood by cardiocentesis into clot-activating serum separator tubes. Collect the serum by centrifugation and stored at Serum osteocalcin levels are measured by radioimmunoassay using the method recommended by Biomedical Technologies, Inc. from whom the required reagents are purchased. Serum is diluted 1:20 and either 100 ul of diluted sample or 100 ul rat osteocalcin standard (BT-412) is added to 100 ul of rat osteocalcin primary antibody (goat anti-rat osteocalcin, BT-413), 100 ul normal goat nonimmune -39serum (NIS) and RIA buffer [122.5 mM NaC1, 25mM Na 4 EDTA, mM NaH 2

PO

4 pH 7.4, 0.1% Tween 20 and 0.1% bovine serumalbumin (RIA grade)] to give a total volume of 500 ul.

Incubate at 4'C overnight on an orbital platform shaker (120 rpm). The second day, *[l 2 5 I]-rat osteocalcin (BT-411R) is added inl00 il (approximately 20,000 cpm) to compete with bound osteocalcin. Tubes are vortexed, then incubate overnight at 4°C on an orbital shaker. On day three, 1 ml of precipitating second antibody donkey anti-goat IgG (BT-414) in 0.1 M sodium phosphate buffer, pH 7.4, containing 2.5% polyethylene glycol and 0.05% NaN 3 is then added. The samples are mixed and incubated for 2 h at 4°C on an orbital shaker, followed by sedimentation by centrifugation at 1500 x g for 15 min at 4'C. The 15 supernatant is carefully discarded and the pellets washed twice by centrifugation with 500 ul distilled water.

Supernatant is discarded and the radioactivity associated with the pellet quantitated (2 min) using an LKB gamma counter. Osteocalcin standards run in parallel are used for determination of standard osteocalcin values. Statistical analysis for osteocalcin measurements is carried out using Instat software to do a two-sided T-test.

t An embodiment of the present invention provides a method for the treatment of a patient afflicted with bone tissue loss or osteoporosis comprising the administration thereto of a effective antiosteoporosis amount of a compound of Formula III, or IV.

The terms "bone tissue loss" as used herein refers to a disease or condition in which bone mass or density is decreased by the loss of both mineral and protein matrix components resulting in bone fragility.

The term "osteoporosis" as used herein refers to a [-disease or condition in which bone tissue loss is responsible for bone fragility resulting in one or more 15 bone fractures.

As used herein, "an effective antiosteoporosis amount" of a compound of Formula III, or IV refers to an amount which is effective, upon single or multiple dose administration to the patient, in preventing or decreasing the rate of bone tissue loss in the patient beyond that expected in the absence of such treatment.

Identification of patients in need of treatment for 25 bone tissue loss or osteoporosis is well within the ability and knowledge of one skilled in the art. The methods for identification of patients which are at risk of developing bone tissue loss or osteoporosis are known and appreciated in the medical arts, such as family history of the development of bone tissue loss or osteoporosis and the presence of risk factors associated with the development of bone tissue loss or osteoporosis. A clinician skilled in the art can readily identify, by the use of clinical tests, physical examination and medical/family history, those patients who are at risk of developing bone tissue loss or osteoporosis and thus readily determine if an individual is -41a patient in need of prophylactic treatment for bone tissue loss or osteoporosis.

An effective antiosteoporosis amount can be readily determined by the attending diagnostician, as one skilled in the art, by the use of known techniques and by observing results obtained under analogous circumstances. In determining the effective antiosteoporosis amount or dose a number of factors are considered by the attending diagnostician, including, but not limited to: the species of mammal; its size, age, and general health; the degree of or involvement or the severity of the disease; the response of the individual patient; the particular compound administered; the mode of administration; the i 15 bioavailability characteristics of the preparation administered; the dose regimen selected; the use of concomitant medication; and other relevant circumstances.

An effective antiosteoporosis amount of a compound of Formula III, or IV is expected to vary from about 0.001 milligram per kilogram of body weight per day (mg/kg/day) to about 100 mg/kg/day.

S.

In a further embodiment, the present invention provides 25 for a method of increasing bone mass and preventing bone tissue loss or osteoporosis in a patient, comprising administering an effective antiosteoporosis amount of a compound of the Formula III, or IV.

As used herein, the term "patient" refers to a warmblooded animal, such as a mouse, a rat, a hamster, a rabbit, or a human, which is afflicted with bone tissue loss or osteoporosis or is at risk of developing bone tissue loss or osteoporosis or who is in need of treatment for bone tissue loss or osteoporosis.

-42- In effecting treatment of a patient afflicted with the disease states described above or in effecting prophylactic treatment of a patient who may be afflicted with the disease states as described above, a compound of Formula III, or IV can be administered in any form or mode which makes the compound bioavailable in effective amounts, including oral and parenteral routes. For example, compounds of Formula III, or IV can be administered orally, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, rectally, and the like. Oral administration is generally preferred. One skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the compound selected the 15 disease state to be treated, the stage of the disease, and other relevant circumstances.

The compounds of Formula III, or IV can be administered alone or in the form of a pharmaceutical composition in combination with pharmaceutically acceptable carriers or excipients, the proportion and nature of which are determined by the solubility and chemical properties of the compound selected, the chosen route of administration, and standard pharmaceutical practice. The compounds of the invention, while effective themselves, may be formulated and administered in the form of their pharmaceutically acceptable acid addition salts for purposes of stability, convenience of crystallization, increased solubility and the like.

In another embodiment, the present invention provides compositions comprising a compound of Formula

III,

or IV in admixture or otherwise in association with one or more inert carriers. These compositions are useful, for example, as assay standards, as convenient means of making bulk shipments, or as pharmaceutical compositions. An assayable amount of a compound of Formula III, or IV -43is an amount which is readily measurable by standard assay procedures and techniques as are well known and appreciated by those skilled in the art. Assayable amounts of a compound of Formula III, or IV will generally vary from about 0.001% to about 75% of the composition by weight. Inert carriers can be any material which does not degrade or otherwise covalently react with a compound of Formula III, or IV. Examples of suitable inert carriers are water; aqueous buffers, such as those which are generally useful in High Performance Liquid Chromatography (HPLC) analysis; organic solvents, such as acetonitrile, ethyl acetate, hexane and the like; and pharmaceutically acceptable carriers or excipients.

15 More particularly, the present invention provides pharmaceutical compositions comprising an effective antiosteoporosis amount of a compound of Formula I, II, III, or IV in admixture or otherwise in association with one or more pharmaceutically acceptable carriers or excipients.

The pharmaceutical compositions are prepared in a manner well known in the pharmaceutical art. The carrier or excipient may be a solid, semi-solid, or liquid material 25 which can serve as a vehicle or medium for the active ingredient. Suitable carriers or excipients are well known in the art. The pharmaceutical composition may be adapted for oral or parenteral use and may be administered to the patient in the form of tablets, capsules, suppositories, solution, suspensions, or the like.

The compounds of the present invention may be administered orally, for example, with an inert diluent or with an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the compounds may be incorporated with excipients and used in the form of -44tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gums and the like. These preparations should contain at least 4% of the compound of the invention, the active ingredient, but may be varied depending upon the particular form and may conveniently be between to about 70% of the weight of the unit. The amount of the compound present in compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations according to the present invention are prepared so that an oral dosage unit form contains between 5.0-300 milligrams of a compound of the invention.

The tablets, pills, capsules, troches and the like may also contain one or more of the following adjuvants: 15 binders such as microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch or lactose, disintegrating agents such as alginic acid, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; and sweetening agents such as sucrose or saccharin may be .added or a flavoring agent such as peppermint, methyl salicylate or orange flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of "a the above type, a liquid carrier such as polyethylene 25 glycol or a fatty oil. Other dosage unit forms may contain other various materials which modify the physical form of the dosage unit, for example, as coatings. Thus, tablets or pills may be coated with sugar, shellac, or other enteric coating agents. A syrup may contain, in addition to the present compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

Materials used in preparing these various compositions should be pharmaceutically pure and non-toxic in the amounts used.

For the purpose of parenteral therapeutic administration, the compounds of the present invention may I be incorporated into a solution or suspension. These preparations should contain at least 0.1% of a compound of the invention, but may be varied to be between 0.1 and about 50% of the weight thereof. The amount of the inventive compound present in such compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations according to the present invention are prepared so that a parenteral dosage unit contains between 0.1 to 100 milligrams of the compound of the invention.

The solutions or suspensions may also include the one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic Sacid or sodium bisulfite; chelating agents such as ethylene diaminetetraacetic acid; buffers such as acetates, citrates 20 or phosphates and agents for the adjustment of tonicity a.

such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampules, disposable syringes or multiple dose vials made of glass or plastic.

a As with any group of structurally related compounds which possesses a particular generic utility, certain groups and configurations are preferred for compounds of Formulas I and II in their end-use application.

With respect to the substituent X, compounds of Formula III, and IV wherein X is chloro are generally preferred.

With respect to the substituent A, compounds of Formula III, and IV wherein A is diethylamino are generally preferred.

Claims (32)

1. A method of treating bone tissue loss or osteoporosis in a patient, comprising administering an effective antiosteoporosis amount of a compound of the Formula S S S *.SS S wherein A is a radical of the formula R N-_ R 1 G wherein R and R, are each independently hydrogen or C,-C4 alkyl; and G is HN, H 3 CN, CH 2 or O; w is an integer from 2 to 3; R 2 is hydrogen, C,-C4 alkyl, C1-C4 alkoxy, halogen, or hydroxy; R 3 is hydrogen, C,-C 4 alkyl, C1-C4 alkoxy, halogen, hydroxy, or -Y(CH 2 )zA 1 in which A, is a radical of the formula N- wherein Y is -HN-; /G 1 D- C IIMNWORDUENNWMSpECNK(Al967DrIVWC -47- R 4 and R 5 are each independently hydrogen or C1-C4 alkyl; G is HN, H 3 CN, CH 2 or 0; and z is an integer from 2 to 3; X is chloro or bromo; pharmaceutically acceptable salt thereof. A method according to claim 1 wherein w is 2. A method according to claim 1 wherein A is the radical or a

2.

3. R N- S S S *5 S~ wherein R and R, are each independently hydrogen or C,-C 4 alkyl.

4. A method according to claim 3 wherein R and R, are each independently C,-C4 alkyl.

5. A method according to claim 4 wherein R and R, are each ethyl.

6. A method according to claim 1 wherein R 3 is hydrogen.

7. A method according to claim 1 wherein X is chloro.

8. A method according to claim 1 wherein the compound is diethylaminoethylamino)phenyl]-1,2-diphenyl-2-chloro-ethylene.

9. A method of treating bone tissue loss or osteoporosis in a patient, comprising administering an effective antiosteoporosis amount of a compound of the Formula C:1WINWORDJENNYMISPECNKI419670V.DOC -48- wherein A is a radical of the formula R 1 N-G R wherein R and R, are each independently hydrogen or Cl-C4 alkyl; and G is HN, H 3 CN, CH 2 or O; w is an integer from 2 to 3; R 2 is hydrogen, C1-C4 alkyl, C1-C4 alkoxy, halogen, or hydroxy;

10 R 3 is hydrogen, C 1 -C 4 alkyl, C1-C4 alkoxy, halogen, hydroxy, or -Y(CH 2 )zA 1 in which A, is a radical of the formula N- R wherein 15 Yis-HN-; R 4 and R 5 are each independently hydrogen or C1-C 4 alkyl; G, is HN, H 3 CN, CH 2 or 0; and z is an integer from 2 to 3; X is chloro or bromo; or a pharmaceutically acceptable salt thereof. A method according to claim 9 wherein w is 2.

11. A method according to claim 9 wherein A is the radical R N- Ri wherein R and R, are each independently hydrogen or C,-C4 alkyl. C:\WINNORDUEN~NfU'ECNIO1 88701WDoC -49-

12. A method according to claim 11 wherein R and R, are each independently C 1 -C 4 alkyl.

13. A method according to claim 12 wherein R and R, are each ethyl.

14. A method according to claim 9 wherein R 3 is hydrogen.

15. A method according to claim 9 wherein X is chloro.

16. A method according to claim 9 wherein the compound, is diethylaminoethylamino)phenyl]-1,2-diphenyl-2-chloro-ethylene.

17. Use of a compound of the formula A-(CH 2 H R3 *w wherewherein R2 is hydrogen, Cl-C4 alkyl, CI-C4 alkoxy, halogen, or hydroxy; A is a radical of the formula N- G wherein R and R 1 are each independently hydrogen or C-C 4 alkyl; and G is HN, H 3 CN, CH 2 or O; w is an integer from 2 to 3; R 2 is hydrogen, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, halogen, or hydroxy; R 3 is hydrogen, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, halogen, hydroxy, or -Y(CH 2 )zA 1 in which A 1 is a radical of the formula C:kWNWORJENNYSPECNKIQ67DN.DOC R4 N G wherein Y is -HN-; R 4 and R 5 are each independently hydrogen or C 1 -C 4 alkyl; G, is HN, H 3 CN, CH 2 or 0; and z is an integer from 2 to 3; X is chloro or bromo; or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of bone tissue loss or osteoporosis. 10

18. A use according to claim 17 wherein w is 2.

19. A use according to claim 17 wherein A is the radical R N S.2 15 wherein R and R, are each independently hydrogen or C,-C 4 alkyl.

20. A use according to claim 17 wherein R and R, are each independently C,-C 4 alkyl.

21. A use according to claim 17 wherein R and R, are each ethyl.

22. A use according to claim 17 wherein R 3 is hydrogen.

23. A use according to claim 17 wherein X is chloro.

24. A use according to claim 17 wherein the compound is diethylaminoethylamino)phenyl]-1, 2 -diphenyl-2-chloro-ethylene.

Use of a compound of the formula C :IMNnV~fMNNYNYMSECNKlgno)NV.C -51- wherein A is a radical of the formula R N- G 5 RI wherein R and R, are each independently hydrogen or C1-C4 alkyl; and G is HN, H 3 CN, CH 2 or O; w is an integer from 2 to 3; R 2 is hydrogen, C1-C4 alkyl, C,-C 4 alkoxy, halogen, or hydroxy; R is hydrogen, C,-C4 alkyl, C-C4 alkoxy, halogen, hydroxy, or -Y(CH2)zAl in which A, is a radical of the formula ~R4 N- G- wherein Y is -HN-; R 4 and R 5 are each independently hydrogen or C,-C4 alkyl; G, is HN, H 3 CN, CH 2 or 0; and z is an integer from 2 to 3; X is chloro or bromo; or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of bone tissue loss or osteoporosis.

26. A use according to claim 25 wherein w is 2. C:%VANWORDJENYMSPECWIX419OWW.DOC -52-

27. A use according to claim 25 wherein A is the radical R N- RI wherein R and R 1 are each independently hydrogen or C 1 -C 4 alkyl.

28. A use according to claim 25 wherein R and R, are each independently C,-C 4 alkyl.

29. A use according to claim 25 wherein R and R, are each ethyl.

30. A use according to claim 25 wherein R 3 is hydrogen. 10

31. A use according to claim 25 wherein X is chloro. 0i

32. A use according to claim 25 wherein the compound is diethylaminoethylamino)phenyl]-1,2-diphenyl-2-chloro-ethylene. DATED 15 February, 1999 PHILLIPS ORMONDE FITZPATRICK Attorneys For: HOECHST MARION ROUSSEL, INC. o o *kolee C: VVINWOROVENYM\SPECWIIlgGTDW.DOC