United States Patent
`Dukes
`[45] Date of Patent:
`Feb. 2, 1993
`
`[:9]
`
`[11] Patent Number:
`
`5,183,814
`
`lllllllllllllllll|||l|||l||||||||||l||||||lllllllllllllllllllllllllllllllll
`US005l83814A
`
`[54] SELECI‘IVE OESTROGEN THERAPY FOR
`PERIMENOPAUSAL OR
`POSTMENOPAUSAL CONDITIONS
`
`[75]
`
`Inventor: Michael Dukes, Wilmslow, United
`Kingdom
`
`[73] Assignee:
`
`Imperial Chemical Industries PLC,
`London, England
`
`[21] App]. No.: 362,043
`
`[22] Filed:
`
`Jun. 6, 1989
`
`[30]
`
`Foreign Application Priority Data
`
`4,826,831
`4,894,373
`
`5/1989 Plunkett et al. ................... .. 514/170
`1/1990 Young ............................ .. 514/239.2
`
`OTHER PUBLICATIONS
`
`-
`Volker et al, Maturitas, 1o:157—159 (1988).
`Ottosson et al, Gynecol. obstet. Invest., 18:140—146;
`296-302 (1984).
`Chemical Abstracts, vol. 109, No. 3, Jul. 18, 1988, p. 73,
`Abstract No. 17l99p.‘
`
`Primary Exam1'ner——Frederick E. Waddell
`Assistant Examiner—Raymond J. Henley, III
`Attorney, Agent, or F1'rm—Cushman, Darby & Cushman
`
`Jun. 6, 1988 [GB] United Kingdom ................. 8813353
`
`[57]
`
`ABSTRACT
`
`'[51]
`
`Int. c1.s ................. .. A61K 31/55; A61K 31/165;
`A61K 31/10
`[52] us. Cl. .................................. .. 514/171; 514/170;
`514/182; 514/622; 514/708; 514/709; 514/710;
`514/712; 514/713
`[58] Field of Search ............. .. 514/170, 171, 182,622,
`514/708. 709, 710, 712, 713
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`4,659,516 4/1987 Bowler et al.
`4,732,912
`3/1988 Pilgrim et al.
`
`................. .. 260/397.5
`...................... 514/510
`
`The invention relates to a therapeutic product compris-
`ing an oestrogen and a pure antioestrogen for simulta-
`neous, sequential or separate use in selective oestrogen
`therapy of perimenopausal or postmenopausal condi-
`tions; to a process for the manufacture of said product
`and to a pharmaceutical composition containing said
`product. The invention also relates to a pharmaceutical
`composition comprising an oestrogen and a pure antio-
`estrogen and to a process for the manufacture of said
`composition.
`
`4 Claims, No Drawings
`
`|nnoPharma Exhibit 1047.0001
`
`

`

`1
`
`5,183,814
`
`SELECTIVE OESTROGEN THERAPY FOR
`PERIMENOPAUSAL OR POSTMENOPAUSAL
`CONDITIONS
`
`10
`
`15
`
`20
`
`25
`
`This invention relates to a therapeutic product for use
`in a new method of medical treatment and, more partic-
`ularly, it relates to a product comprising an oestrogen
`and a pure antioestrogen for use in a new method for the
`treatment or prophylaxis of perimenopausal or post-
`menopausal conditions, particularly perimenopausal or
`postmenopausal osteoporosis. The invention also relates
`to a pharmaceutical composition comprising an oestro-
`gen and a pure antioestrogen and to the use thereof in
`the manufacture of a new medicament for use in the
`treatment or prophylaxis of perimenopausal or post-
`menopausal conditions.
`When a female animal, particularly a human female,
`enters the perimenopausal stage the animal’s ovaries
`begin to secrete less of the female sex hormones, partic-
`ularly oestradiol. Symptoms in women at this stage
`include the following: vasomotor disturbances (hot
`flushes), urogenital atrophy (particularly affecting the
`vagina and distal urethra), psychosomatic complaints,
`changes in lipid metabolism and osteoporosis. The rate
`of decline of ovarian function and the severity of the
`above-mentioned symptoms are highly variable be-
`tween individual women but in a substantial number of
`individuals the symptoms are sufficiently severe that
`treatment is required. Oestrogen replacement therapy 30
`has been used in women and it is generally recognised
`to be effective in combatting the typical perimenopausal
`and post-menopausal symptoms (British Medical Jour-
`nal,
`l987, 295, 914; American Journal of Obstet. and
`Gynecol., 1987, 156, 1298 and 1347). However oestrogen 35
`replacement therapy can also cause uterine hyperplasia,
`irregular vaginal menstruation and, in a small propor-
`tion of women, endometrial cancer (American Journal of
`Obstet. and Gynecol., 1987,’ 156, 1313).
`To combat the continuous unopposed stimulation of 40
`oestrogen-responsive tissues an oestrogen and a proges-
`togen are normally co-administered for part of each
`treatment period thereby causing regular vaginal men-
`struation. (American Journal of Obstet. and Gynecol.,
`1987, 156, 1304). However the continuation of men-
`strual periods is unattractive to many postmenopausal
`women and, in addition, progestogens can cause side
`effects, for example oedema, premenstrual irritability
`and breast tenderness.
`
`45
`
`Alternative therapies are therefore required.
`It has recently been shown that compounds demon-
`strating a mixture of oestrogenic and antioestrogenic
`properties in warm-blooded animals, including humans,
`may be of use in the treatment of postmenopausal condi-
`tions (European Patent Specification No. 0178862).
`Particular compounds stated to have such activity in-
`clude clomiphene and tamoxifen.‘ Comprehensive re-
`views of the clinical usage of these compounds are
`available. for example a review of clomiphene by Clark
`et al. in Pharmacology and Therapeutics, 1982, Volume
`15, pages 467 to 519, and a review of tamoxifen by Furr
`et al. in Pharmacology and Therapeutics, 1984, Volume
`25, pages 127-205.
`It has also recently been shown that a treatment re-
`gime comprising the dosing of a small amount of an
`oestrogen, for example oestrone sulphate or natural
`conjugated oestrogens, followed by the closing of an
`antioestrogen, for example tamoxifen or clomiphene led
`
`50
`
`55
`
`65
`
`2
`to the partial inhibition of the maximum oestrogen-
`induced stimulation of uterine endometrial tissue (A.
`Kauppila et al., Gynecol. abstet. Invest, 1988, 25, 58 and
`Arch. Gynecol., 1983, 234, 49).
`It has now been found that administration of an oes-
`trogen and a pure antioestrogen, whether simulta-
`neously, sequentially or separately, results in the oestro-
`gen being selectively effective in some oestrogen-
`responsive tissues, for example bone, and being selec-
`tively opposed in other oestrogen-responsive tissues, for
`example the endometrium of the uterus, and this is the
`basis of the present invention.
`_
`A pure antioestrogen is a compound which possesses
`antioestrogenic activity and no oestrogenic activity.
`This may be demonstrated in rats by the effect of the
`compound in antagonising the increase in weight of the
`uterus of an immature female rat produced by adminis-
`tering oestradiol benzoate to said rat. Thus, when each
`of a pure antioestrogen and oestradiol benzoate are
`administered for 3 days to such a rat, a smaller increase
`in uterine weight is produced than the substantial in-
`crease which would be produced by the administration
`of oestradiol benzoate alone. Unlike the known antioes-
`
`trogens tamoxifen and clomiphene, when a pure antio-
`estrogen is administered alone to a rat no increase in
`uterine weight whatsoever is observed.
`It is disclosed in European Patent Specification No.
`138504 that certain preferred steroidal antioestrogens
`are pure antioestrogens. It is also disclosed in European
`Patent No. 124369 that certain preferred non-steroidal
`antioestrogens are pure antioestrogens.
`According to the present invention there is provided
`a product comprising an oestrogen and a pure antioes-
`trogen for simultaneous, sequential or separate use in
`selective oestrogen therapy of perimenopausal or post-
`menopausal conditions.
`.
`In a particular product of the invention the oestrogen
`component of a product of the invention is oestradiol,
`ethinyloestradiol, oestriol, oestrone, natural conjugated
`oestrogens, piperazine oestrone sulphate, mestranol,
`chlorotrianisene, dienoestrol, stilboestrol or hexoestrol
`or a pharmaceutically-acceptable ester thereof.
`A pharmaceutically-acceptable ester of the oestrogen
`component of a product of the invention is, for example,
`an alkyl or aryl ester each of up to.12 carbon atoms. It
`will be appreciated that an ester of a steroidal oestrogen
`may be formed at the 3-position, the 17-position or at
`both of these positions. It will also be appreciated that
`an ester may be formed at one or both of the phenolic
`groups in some non-steroidal oestrogens, for example
`stilboestrol and hexoestrol. A suitable alkyl ester of up
`to 12 carbon atoms is, for example, an acetate, propio-
`nate, butyrate, valerate, hexanoate, heptanoate, octane-
`ate, cyclopentylpropionate, nonanoate, decanoate, un-
`decanoate or dodecanoate. A suitable aryl ester of up to
`12 carbon atoms is, for example, a benzoate, toluate or
`naphthoate. A preferred ‘pharmaceutical]y-acceptable
`ester of the oestrogen component of a product of the
`invention includes, for example, oestroadiol benzoate,
`oestradiol cyclopentylpropionate, oestradiol dipropio-
`nate, oestradiol heptanoate, oestradiol undecanoate,
`oestradiol valerate and stilboestrol dipropionate.
`In a further particular product of the invention the
`pure antioestrogen is
`N-n-butyl-N-methyl-, N-lH,lH-heptafluorobutyl-N-
`methyl-or N,N-(3-methylpentamethylene)-1l-(3,17B-
`dihydroxyoestra-1,3,5(10)trien-7a-yl)undecanamide;
`
`|nnoPharma Exhibit 1047.0002
`
`

`

`5,183,814
`
`3
`or N-ll-I,1H-heptafluorobutyl-3-p-[4-
`N-n-butyl-
`(3,l 7B-dihydroxyoestra-1,3,5(lO)-triene-7a-yl)butyl]-
`phenylpropionamide;
`7a-(10-p-chloro-
`7a-(l0-p-chlorophenylthiodecyl)-,
`phenylsulphinyldecyl)-,
`7a-[9-(4,4,5,5,5-penta-
`fluoropentylsulphinyl)nonyl]-,
`7a-[10-(4,4,4-tri-
`fluorobutylsulphinyl)decyl]-or 7a-[10-(p-chlorobenzyl-
`sulphinyl)decyl]-oestra-1,3,5(10)triene-3,175-diol; or
`7a-(9-n-heptylsulphinylnonyl)oestra-l,3,5(l0)-triene-
`3,l7B-diol.
`In a further particular product of the invention the
`pure antioestrogen is a compound of the formula:
`
`NU—A—x—R'
`
`l0
`
`15
`
`20
`
`25
`
`30
`
`35
`
`50
`
`60
`
`65
`
`wherein NU is 6-hydroxy-2-p-hydroxyphenylnapth-
`l-yl and A is —(CH2)1o—, —(CH2)].1— or —(CH2)5-
`(l,4-phenylene)-(CH2)2—;
`or NU is 1,2,3,4-tetrahydro-6-hydroxy-2-p-hydroxy-
`phenylnaphth-l-yl (either the 1RS,2RS or 1RS,2SR
`isomer), or
`l,2,3,4-tetrahydro-6-hydroxy-2-p-hydro»
`yphenyl-2-methylnapth-1-yl
`(either
`the 1RS,2RS or
`lRS,2SR isomer), and A is -—(CHz);o—, —(CH2)n—
`or —(CH2)4-(1,4-phenylene)-(CI-l2)2—;
`or NU is (1RS,2RS)-5-hydroxy—2-p-hydroxyphenyl-
`indan-l-yl or
`(lRS,2RS)-5-hydroxy-2-p-hydroxyphe-
`nyl-2-methylindan-l-yl and A is —(CH2)1o—-—, —(CH2)-
`11-
`or
`-—(CH2)4-(1,4-phenylene)-(CH2)2—;
`and
`wherein XR1 is —CONR‘R3 wherein R2 is hydrogen or
`methyl and R‘ is n-butyl, 1H,1H-heptafluorobutyl, n-
`pentyl or n-hexyl, or XRI
`is —SR‘, —S0R1 or
`—-SOzRl wherein R1 is n-pentyl, n-hexyl, 4,4,5,5,5-pen-
`tafluoropentyl
`or
`1H,lH,2H,2H,3H,3H-hepta-
`fluorohexyl.
`In a further particular product of the invention the
`pure antioestrogen is
`N-n-butyl-, N-n-butyl-N-methyl-, N-n-pentyl. N-
`(1H, 1 H-heptafluorobutyl)-or
`N-(1H,1H-hepta-
`fluorobutyl)-N-methyl-3-p-[5-(6-hydroxy-2-p-hydroxy-
`phenylnaphth-1-yl)pentyl]phenylpropionamide;
`N-methyl-N-(1H,1H-heptafluorobutyl)-p-[4-
`[(1RS,2RS)-6-hydroxy-2-p-hydroxphenyl-2-methyl-
`1,2,3,4-tetrahydronaphth-1-yl]-butyl]pheny1propiona-
`mide;
`(lRS,2RS)-1-[4-[p-(2—n-hexylthioethyl)phenyl]-
`butyl]-2-p-hydroxyphenyl-l,2,3,4-tetrahydronaphth-6-
`ol or the corresponding 4,4,5,5,5-pentafluoropentylthio
`derivative, or the corresponding hexylsulphinyl, hexyl-
`sulphonyl or pentafluoropentylsulphinyl derivatives;
`2-p-hydroxyphenyl-1-[5-[p-(2-n-hexylthioethyl)-
`phenyl]pentyl]naphth-6-ol or the corresponding hexyl-
`sulphinyl derivative; or (1RS,2RS)-l-[4[p-(2-n-hexylthi-
`oethyl)phentyl]butyl]-2-p-hydroxyphenyl-2-methyl-
`l,2,3,4-tetrahydronaphth-6-ol or
`the corresponding
`4,4,5,5,5-pentafluoropentylthio derivative, or the corre-
`sponding hexylsulphinyl or pentafluoropentylsulphinyl
`derivative, or the corresponding (lRS,2SR) isomers of 55
`both the hexylthio and hexylsulphinyl derivatives.
`A preferred product of the invention comprises an
`oestrogen and a pure antioestrogen for use as stated
`above wherein the oestrogen is oestradiol or ethinyloes-
`tradiol, or a pharmaceutical]y-acceptable ester thereof,
`and the pure antioestrogen is 7a-[9-(4,4,5,5,5-penta-
`fluoropentylsulphinyl)nonyl]oestra-l,3,5(10)-triene-
`3,1713-diol or (1RS,2RS)-2-p-hydroxyphenyl-2-methyl-
`1-[9-(4,4,5,5,5-pentafluoropentylsulphiny1)nonyl]-
`l,2,3,4-tetrahydronaphth-6-ol.
`A particularly preferred product of the invention
`comprises an oestrogen and a pure antioestrogen for use
`as stated above wherein the oestrogen is oestradiol,
`
`45
`
`4
`oestradiol benzoate, oestradiol valerate or oestradiol
`undecanoate and the pure antioestrogen is 7a-[9-
`(4,4,5,5,5-pentafluoropentylsulphinyl)nonyl]oestra-
`1,3,5(10)-triene-3,17B-diol.
`According to a further feature of the invention there
`is provided a process for the manufacture of a product
`comprising an oestrogen and a pure antioestrogen for
`simultaneous, sequential or separate use in selective
`oestrogen therapy of perimenopausal or postmeno-
`pausal conditions, which process comprises bringing
`together said oestrogen and said pure antioestrogen.
`In a further feature of the invention there is provided
`a process for the manufacture of a product comprising
`an oestrogen and a pure antioestrogen for simultaneous
`use in selective oestrogen therapy of perimenopausal or
`postmenopausal conditions, which process comprises
`bringing into admixture said oestrogen and said pure
`antioestrogen.
`A product of the invention may be administered to a
`warm-blooded animal, including a human, in the form
`of a pharmaceutical composition. Thus according to a
`further feature of the present invention there is pro-
`vided a pharmaceutical composition which comprises
`the product of the invention together with a phar-
`maceutically-acceptable diluent or carrier.
`As mentioned above a product of the invention is
`useful
`for
`selective
`oestrogen
`therapy
`of
`perimenopausal or postmenopausal conditions. It will
`be understood that there is no absolute requirement that
`the oestrogen and pure antioestrogen components of the
`product of the invention must be dosed simultaneously.
`Sequential or separate use of these components may
`also provide selective oestrogen therapy and such use is
`to be understood to fall within the definition of a prod-
`uct of the invention. Thus it will be appreciated that a
`pharmaceutical composition according to the present
`invention includes a composition comprising an oestro-
`gen, a pure antioestrogen and a pharmaceutically-
`acceptable diluent or carrier. Such a composition con-
`veniently provides the product of the invention for
`simultaneous use in selective oestrogen therapy of
`perimenopausal or postmenopausal conditions. A phar-
`maceutical composition according to the present inven-
`tion also includes separate compositions comprising a
`first composition comprising an oestrogen and a phar-
`maceutically-acceptable diluent or carrier, and a second
`composition comprising a pure antioestrogen and a
`pharmaceutically-acceptable diluent or carrier. Such a
`composition conveniently provides the product of the
`invention for sequential or separate use in selective
`oestrogen therapy of perimenopausal or postmeno-
`pausal conditions.
`The compositions of the invention may be in a form
`suitable for oral use (for example as tablets, capsules,
`aqueous or oily suspensions, emulsions or dispersible
`powders or granules), for topical use (for example as
`creams, ointments, gels, or aqueous or oily solutions or
`suspensions; for example for use within a transdermal
`patch), for parenteral administration (for example as a
`sterile aqueous or oily solution or suspension for intra-
`venous, subcutaneous,
`intramuscular or intravascular
`dosing), or as a suppository for rectal dosing or as a
`pessary for vaginal dosing.
`The compositions of the invention may be obtained
`by conventional procedures using conventional phar-
`maceutical excipients, well known in the art.
`
`|nnoPharma Exhibit 1047.0003
`
`

`

`5,183,814
`
`5
`Suitable pharmaceutically acceptable excipients for a
`tablet formulation include, for example, inert diluents
`such as lactose, sodium carbonate, calcium phosphate
`or calcium carbonate, granulating and disintegrating
`agents such as corn starch or alginic acid; binding
`agents such as gelatin or starch; lubricating agents such
`as magnesium stearate, stearic acid or talc; preservative
`agents such as ethyl or propyl p-hydroxybenzoate, and
`anti-oxidants, such as ascorbic acid. Tablet formulations
`may be uncoated or coated either to modify their disin-
`tegration and the subsequent absorption of the active
`ingredient within the gastrointestinal tract, or to im-
`prove their stability and/or appearance, in either case
`using conventional coating agents and procedures well
`known in the art.
`
`l0
`
`15
`
`6
`rally-occurring gums such as gum acacia or gum traga-
`canth, naturally-occurring phosphatides such as leci-
`thin, esters or partial esters derived from fatty acids and
`hexitol anhydrides (for example sorbitan monooleate)
`and condensation products of the said partial esters with
`ethylene oxide such as polyoxyethylene sorbitan mono-
`oleate. The emulsions may also contain sweetening,
`flavouring and preservative agents.
`The pharmaceutical compositions may also be in the
`form of sterile injectable aqueous or oily suspensions,
`which may be formulated according to known proce-
`dures using one or more of the appropriate dispersing or
`wetting agents and suspending agents which have been
`mentioned above. A sterile injectable preparation may
`also be a sterile injectable solution or suspension in a
`non-toxic parenterally-acceptable diluent or solvent, for
`example a solution in 1,3-butanediol, in a vegetable oil
`(such as arachis oil, castor oil or coconut oil) or in a
`mineral oil (such as liquid paraffin).
`Conveniently the subcutaneous or intramuscular in-
`jection of an aqueous suspension or an oily solution or
`suspension of a pharmaceutical composition of the in-
`vention provides a depot of the active ingredients at the
`injection site from which those ingredients may leach
`out over a period of time to provide the sustained re-
`lease thereof.
`
`20
`
`25
`
`Compositions for oral use may be in the form of hard
`gelatin capsules in which the active ingredient is mixed
`with an inert solid diluent, for example, calcium carbon-
`ate, calcium phosphate or kaolin, or as soft gelatin cap-
`sules in which the active ingredient is mixed with water
`or an oil such as peanut oil, liquid paraffin or olive oil.
`Aqueous suspensions generally contain the active
`ingredient in finely powdered form together with one
`or more suspending agents, such as sodium carboxy-
`methylcellulose, methylcellulose, hydroxypropylme-
`thylcellulose, sodium alginate, polyvinyl-pyrrolidone,
`gum tragacanth and gum acacia; dispersing or wetting
`agents such as lecithin or condensation products of an
`alkylene oxide with fatty acids (for example polyox-
`ethylene stearate), or condensation products of ethylene
`oxide with long chain aliphatic alcohols, for example
`heptadecaethyleneoxycetanol, or condensation prod-
`ucts of ethylene oxide with partial esters derived from
`fatty acids and a hexitol such as polyoxyethylene sorbi-
`tol monooleate, or condensation products of ethylene
`oxide with partial esters derived from fatty acids and
`hexitol anhydrides, for example polyethylene sorbitan
`monooleate. The aqueous suspensions may also contain
`one or more preservatives (such as ethyl or propyl
`p-hydroxybenzoate, anti-oxidants (such as ascorbic
`acid), colouring agents,
`flavouring agents, and/or
`sweetening agents (such as sucrose, saccharine or aspar-
`tame).
`Oily suspensions may be formulated by suspending
`the active ingredient in a vegetable oil (such as arachis
`oil, castor oil, sesame oil or coconut oil) or in a mineral
`oil (such as liquid paraffin). The oily suspensions may
`also contain a thickening agent such as beeswax, hard
`paraffin or cetyl alcohol. Sweetening agents, such as
`those set out above, and flavouring agents may be
`added to provide a palatable oral preparation. These
`compositions may be preserved by the addition of an
`anti-oxidant such as ascorbic acid.
`Dispersible powders and granules suitable for prepa-
`ration of an aqueous suspension by the addition of water
`generally contain the active ingredient together with a
`dispersing or wetting agent, suspending agent and one
`or more preservatives. Suitable dispersing or wetting
`agents and suspending agents are exemplified by those
`already mentioned above. Additional excipients, such as l
`sweetening, flavouring and colouring agents, may also
`be present.
`The pharmaceutical compositions of the invention
`may also be in the form of oil-in-water emulsions. The
`oily phase may be a vegetable oil, such as castor oil,
`soya bean oil or arachis oil, or a mineral oil, such as, for
`example, liquid paraffin or a mixture of any of these.
`Suitable emulsifying agents may be, for example, natu-
`
`30
`
`35
`
`45
`
`50
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`55
`
`65
`
`Suppository formulations may be prepared by mixing
`the active ingredient with a suitable non-irritating ex-
`cipient which is solid at ordinary temperatures but liq-
`uid at the rectal temperature and will therefore melt in
`the rectum to release the drug. Suitable excipients in-
`clude, for example, cocoa butter and polyethylene gly-
`cols.
`
`Topical formulations, such as creams, ointments, gels
`and aqueous or oily solutions or suspensions, may gen-
`erally be obtained by formulating an active ingredient
`with a conventional, topically acceptable, vehicle or
`diluent using conventional procedure well known in the
`art.
`
`According to a further feature of the invention there
`is provided a process for the manufacture of a pharma-
`ceutical composition as defined above which comprises
`bringing into admixture a product as defined above
`together with a pharmaceutical]y-acceptable diluent or
`carrier.
`
`The invention also provides a method of selective
`oestrogen therapy of perimenopausal or postmeno-
`pausal conditions which comprises administering simul-
`taneously, sequentially or separately to a warm-blooded
`animal an effective amount of a product as defined
`above. The invention also provides the use of a product
`as defined above for the manufacture of a new medica-
`ment for use simultaneously, sequentially or separately
`in selective oestrogen therapy of perimenopausal or
`postmenopausal conditions.
`It will be appreciated that the definition of the prod-
`uct of the invention and the pharmaceutical composi-
`tion of the invention includes only those products or
`compositions which are useful in a new method for the
`treatment or prophylaxis of perimenopausal or post-
`menopausal condition. Pharmaceutical compositions
`comprising an oestrogen and a pure antioestrogen, to-
`gether with a pharmaceutical]y-acceptable diluent or
`carrier, are novel. In European Patent Sepcifications
`Nos. 138504 and 124369 it is disclosed that the antioes-
`trogenic activity of the compounds disclosed therein
`may be demonstrated by the co-administration of a test
`compound and oestradiol benzoate to an immature fe-
`
`|nnoPharma Exhibit 1047.0004
`
`

`

`5,183,814
`
`7
`male rat. Antioestrogenic activity is demonstrated by
`antagonism of the increase in weight of the uterus of the
`rat which is produced when oestradiol benzoate alone is
`administered to said rat. It is to be noted that, during
`those tests, the oestradiol benzoate was given by subcu-
`taneous injection whereas the test compound was given
`separately either orally or subcutaneously.
`According to a further aspect of the invention there is
`provided a pharmaceutical composition comprising an
`oestrogen and a pure antioestrogen together with a
`pharmaceutically-acceptable diluent or carrier.
`The pharmaceutical compositions of this feature of
`the invention may be obtained by conventional proce-
`dures using conventional pharmaceutical excipients
`well known in the are such as, for example, those dis-
`closed above.
`
`This aspect of the invention also provides a process
`for the manufacture of a pharmaceutical composition as
`defined immediately above which comprises bringing
`into admixture an oestrogen and a pure antioestrogen
`together with a pharmaceutical]y-acceptable diluent or
`carrier.
`
`This aspect of the invention also provides a method
`of selective oestrogen therapy of perimenopausal or
`postmenopausal conditions which comprises adminis-
`tering to a warm-blooded animal an effective amount of
`a pharmaceutical composition as defined immediately
`above. The invention also provides the use of a pharma-
`ceutical composition as defined immediately above for
`the manufacture of a new medicament for use in selec-
`
`tive oestrogen therapy of perimenopausal or postmeno-
`pausal conditions.
`As stated above a product of the invention is of use in
`selective oestrogen therapy of perimenopausal or post-
`menopausal conditions. Selective oestrogen therapy
`may be demonstrated using the standard procedure set
`out below:
`
`a) an in vivo assay measuring the antioestrogenic
`activity of a compound and any oestrogenic activity
`possessed by that compound. This may be demonstrated
`in rats by the effect of the compound in antagonising the
`increase in weight of the uterus of an immature female
`rat produced by administering oestradiol benzoate to
`said rat. Thus, when each of a pure antioestrogen and
`oestradiol benzoate are administered for 3 days to such
`a rat, a smaller increase in uterine weight is produced
`than the substantial increase which would be produced
`by the administration of oestradiol benzoate without the
`pure antioestrogen. Unlike the known antioestrogens
`tamoxifen and clomiphene, when a pure antioestrogen is
`administered alone to a rat no increase in uterine weight
`whatsoever is observed.
`The oestrogenic activity of a compound may be dem-
`onstrated in rats by the effect of the compound when it
`is administered alone to said rat on the uterine weight of
`the animal.
`
`b) An in vivo assay in mature rats measuring the
`antioestrogenic activity of a compound by the effect of
`the compound when closed during a test period of 28
`days in antagonising the protective effect on the ani-
`mals’ bone density of their endogenous oestrogens. The
`bone density of a group of ovariectomised rats in which
`endogenous oestrogen levels are much reduced serves
`as a control for the effect expected to be produced by a
`fully effective antioestrogen.
`The antioestrogenic activity of the compound in ma-
`ture rats can also be measured in the same assay by
`measuring the effect of the compound in antagonising
`
`8
`the effect of the animals’ endogenous oestrogens which
`serve to increase the weight of their uteri.
`A comparison of the potencies of the antioestrogenic
`effects of a compound as measured by its effects on the
`animals’ bone density and uterine weights allows the
`selectivity of the antioestrogenic effects'of the com-
`pound to be measured.
`Although the pharmacological properties of a prod-
`uct of the invention vary with the structures of the
`oestrogenic and antioestrogenic components and with
`the route of administration, in general a product of the
`invention comprises:
`(i) an oestrogen which possesses oestrogenic activity in
`the above test (a) at doses in the range, for example,
`0.002-2.0 mg/kg orally or in the range, for example,
`0.0001—O.1 mg/kg subcutaneously;
`(ii) a pure antioestrogen which possesses antioestro-
`genic activity in the above tests (a) and (b) at doses in
`the range, for example, in test (a): ED5o0.05—5 mg/kg
`orally or ED5o 0.01—1.0 mg/kg subcutaneously;
`in
`test
`(b):
`antiuterotrophic
`effect:
`ED5o<20
`mg/kg/day orally, <2 mg/kg/day subcutaneously
`or intramuscularly and <10 mg/kg/injection when
`closed as an intramuscular depot injection; reduction
`in bone density: ED5o>2O mg/kg/day orally, >5
`mg/kg/day subcutaneously or intramuscularly and
`>10 mg/kg/injection when dosed as an intramuscu-
`lar depot injection.
`A product of the invention is thereby seen to be sur-
`prisingly selective as the activity of the pure antioestro-
`gen component is expressed to a high degree within
`uterine tissue but to a lesser degree on bone.
`The size of the dose, for therapeutic or prophylatic
`purposes, of a product of the invention as defined above
`will naturally vary according to the nature and severity
`of the conditions presented, the age and menopausal
`state of the animal and the route of administration.
`In general the minimum quantity of the oestrogenic
`component of a product of the invention as defined
`above will be chosen so as to provide a beneficial effect
`with regard to the nature and severity of the conditions
`presented. The quantity of the pure antioestrogenic
`component is then chosen to antagonise to a substantial
`degree the effect of the oestrogenic component on the
`uterine tissue. Methods of evaluating the condition of
`uterine tissue are well known to the man skilled in the
`art, for example, by examination of a specimen of endo-
`metrial
`tissue taken by, for example, suction or, for
`example, by way of a biopsy.
`So far as the oestrogenic component of a product of
`the invention as defined above is concerned the size of
`the dose and routes of administration conventionally
`utilised in oestrogen replacement therapy may be used.
`Thus, for example, a tablet containing, for example, 0.5
`to 2 mg of oestradiol, oestradiol benzoate, natural con-
`jugated oestrogens or oestradiol valerate may be admin-
`istered daily. Alternatively a tablet containing 10 to 100
`pg of ethinyloestradiol may be administered daily. Al-
`ternatively the oestrogenic component may be adminis-
`tered by, for example, intramuscular injection utilising,
`for example, 1 to 10 mg of oestradiol benzoate dissolved
`in an oil such as ethyl oleate; for example, transdermal
`means utilising, for example, 10-100 pg of oestradiol
`contained within a transdermal patch; or, for example,
`vaginal application utilising, for example, daily applica-
`tion of 0.5 to 2 mg of natural conjugated oestrogens
`contained within 0._5 to 5 ml of a cream.
`
`10
`
`15
`
`20
`
`25
`
`30
`
`_
`
`35
`
`45
`
`50
`
`55
`
`60
`
`65
`
`|nnoPharma Exhibit 1047.0005
`
`

`

`5,183,814
`
`10
`weighed and their volumes were determined using Ar-
`chimedes Principle. The femurs were then burned and
`the residual ash was weighed. From these data, gross
`femur density and bone mineral density were calculated
`as follows:
`
`Gross Femur Density=Femur Weight/Femur Volume
`Bone Mineral Density=Femur Ash Weight/Femur
`Volume
`.
`.
`The results shown below in Tables I and II demon-
`strate that at a dose of 2 mg/kg/day subcutaneously the
`test compound selectively inhibits the action of the
`animals’ endogenous oestrogen on their uteri (90%
`inhibition of uterine weight) whereas there was no sig-
`nificant inhibition of either bone mineral density or of
`gross femur density.
`
`TABLE I
`
`9
`So far as the antioestrogenic component of a product
`of the invention as defined above is concerned the size
`of the dose is chosen such that the effect of the oestro-
`genic component on uterine tissue is antagonised to a
`substantial degree whereas the beneficial effect of the
`oestrogenic component on bone is substantially unop-
`posed. Thus, for example, the antioestrogenic compo-
`nent may be formulated in like manner to the oestro-
`genie component, for example as a tablet, an oily solu-
`tion suitable for intramuscular injection, within a trans-
`dermal patch, or within a cream suitable for vaginal
`application. The daily administration of one or more
`tablets containing conveniently 50 mg to 5 g, and pref-
`erably 50 mg to 500 mg, of a pure antioestrogen may be
`used. Preferably the pure antioestrogen may be adminis-
`tered‘ by the periodic intramuscular injection of, for
`example, an aqueous suspension or an oily solution or
`suspension containing 50 mg to 5 g of the pure antioes-
`trogen. Preferably an oily solution, for example a solu-
`tion containing arachis or castor oil, an alcohol such as
`benzyl alcohol and 50 mg to 500 mg of the pure antioes-
`trogen is employed. Such an injection provides a depot
`of the pure antioestrogen which thereafter leaches out
`from the injection site to provide a selective antioestro-
`genic effect for a period of, for example, one to six
`weeks.
`
`As mentioned above a product of the invention is
`useful
`for
`selective
`oestrogen
`therapy
`of
`perimenopausal or postmenopausal conditions. As pre-
`viously mentioned perimenopausal and postmenopausal
`conditions include, for example, vasomotor disturb-
`ances (hot
`flushes), urogenital atrophy (particularly
`affecting the vagina and the distal urethra), psychoso-
`matic complaints, changes in the lipid metabolism and
`oesteoporosis. The selective antioestrogenic effect of
`the pure antioestrogenic component of a product of the
`invention, as demonstrated by a greater antioestrogenic
`effect on the uterus of a rat than on the bone of the rat,
`allows the beneficial effect of the oestrogenic compo-
`nent of the product of the invention to be selectively
`applied to the bone and prevents the detrimental effect
`of an unopposed oestrogenic effect on the uterus. The
`utero-selective effect of the pure antioestrogenic com-
`ponent o