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`European Patent Office
`Office européen des brevets
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`® PUbIlcat'On number:
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`0 346 014 B1
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`EUROPEAN PATENT SPECIFICATION
`
`9 6
`
`9
`
`Date of publication of patent specification: 03.68.94 @ Int. Cl.5: A61K 31/565, //(A61K3‘I/565,
`31 :165
`ED Application number: 893055612
`)
`
`@ Date of filing: 02.06.89
`
`@ Therapeutic product for the treatment of peri- or postmenopausal conditions.
`
`
`
`Priority: 06.06.88 GB 8813353
`
`Date oi publication of application:
`13.12.89 Bulletin 39/50
`
`Publication of the grant of the patent:
`03.08.94 Bulletin 94/31
`
`@ Proprietor: ZENECA LIMITED
`Imperial Chemical House, 9 Millbank
`London SW1P 3JF(GB)
`
`@ Inventor: Dukes, Michael
`54 Styal Road
`Wilmslow Cheshire, 5K9 4AQ(GB)
`
`Representative: Tait, Brian Steele et al
`ICl Group Patents Service Dept.
`PO Box 6
`Shire Park
`Bessemer Road
`
`Welwyn Garden City Herts AL71HD (GB)
`
`Designated Contracting States:
`AT BE CH DE ES FR GB GR IT LI LU NL SE
`
`References cited:
`EP-A- 0 "I24 359
`EP~A- 0 138 504
`
`CHEMICAL ABSTRACTS, vol. 109, no. 3, 18th
`July 1988, page 73, abstract no. 17199p, Co-
`lumbus, Ohio, US; N. FROEHLANDEH et al.:
`"Growth hormone and somatomedin C dur-
`
`ing post-menopausal replacement therapy
`with estrogen alone and in combination with
`an antiestrogen“, & MATURITAS 1988, 9(4).
`297-302
`
`EP0346014B1
`
`Note: Within nine months from the publication of the mention of the grant of the European patent, any person
`may give notice to the European Patent Office oi opposition to the European patent granted. Notice of opposition
`shall be filed in a written reasoned statement.
`It shall not be deemed to have been filed until the opposition fee
`has boon paid (Art. 99(1) European patent convention).
`
`Rank Xerox (UK) Business Services
`(31013091333)
`
`InnoPharma Exhibit 1055.0001
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`
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`EP 0 346 014 B1
`
`Description
`
`This invention relates to a therapeutic product for use in a new method of medical treatment and, more
`particularly,
`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 postmenopausal conditions, particularly
`perimenopausal or postmenopausal osteoporosis. The invention also relates to a pharmaceutical composi—
`tion comprising an oestrogen and a pure antioestrogen and to the use thereot in the manufacture of a new
`medicament for use in the treatment or prophylaxis of perimenopausal or postmenopausal conditions.
`When a female animal, particularly a human temale, enters the perimenopausal stage the animal‘s
`ovaries begin to secrete less of the female sex hormones, particalarly 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 between individual women but
`in a substantial number of individuals the symptoms are
`sufficiently severe that treatment is required. Oestrogen replacement therapy has been used in women and
`it
`is generally recognised to be effective in combatting the typical perimenopausal and post-menopausal
`symptoms
`1987a
`914;
`1987,1529
`1298 and 1347). However oestrogen replacement therapy can also cause uterine hyperplasia,
`irregular
`vaginal menstruation and, in a small proportion of women, endometrial cancer (American Journal of Obstet.
`and Gynecol., 1987,1§§, 1313).
`To combat the continuous unopposed stimulation of oestrogen-responsive tissues an oestrogen and a
`progestogen are normally co-administered for part of each treatment period thereby causing regular vaginal
`
`menstruation. (American Journal of Obstet. and Gynecol, 1987, 156, 1304). However the continuation of
`menstrual periods is unattractive to many postmenopausal women and, in addition, progestogens can cause
`side effects, for example oedema, premenstrual irritability and breast tenderness.
`Alternative therapies are therefore required.
`It has recently been shown that compounds demonstrating a mixture of oestrogenic and antioestrogenic
`properties in warm-blooded animals, including humans, may be of use in the treatment of postmenopausal
`conditions (European Patent Specification No..0178862). Particular compounds stated to have such activity
`include clomiphene and tamoxifen. Comprehensive reviews 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 at. in Pharmacology and Therapeutics,
`1984, Volume 25, pages 127-205.
`It has also recently been shown that a treatment regime comprising the closing to postmenopausal
`women of the oestrogen ethinyloestradiol led to an increase in serum growth hormone (GH) levels whereas
`the periodic dosing of both ethinyloestradiol and the antioestrogen tamoxifen led to a reduction of GH levels
`
`to pro-treatment levels [N. Froehlander et al., Maturitas, 1988, 9(4), 297 (Chem. Abstracts, 109, 17199p)].
`It has also recently been shown that a treatment regime comprising the dosing of a small ampunt of an
`oestrogen, for example oestrone sulphate or natural conjugated oestrogens, followed by the dosing of an
`antioestrogen, for example tamoxifen or clomiphene led to the partial inhibition of the maximum oestrogen-
`
`induced stimulation of uterine endomatrial tissue (A. Kauppila et al., Gynecol. obstet. Invest., 1988, 25, 58
`and Arch. Gynecol, 1983, Egg, 49).
`It has now been found that administration of an oestrogen and a pure antioestrogen, whether
`simultaneously, sequentially or separately,
`results in the oestrogen being selectively effective in some
`oestrogen-responsive tissues,
`for example bone, and being selectively 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 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 alone. 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.
`
`is disclosed in European Patent Specification No. 138504 that certain preferred steroidal an~
`It
`tioestrogens are pure antioestrogens.
`It
`is also disclosed in European Patent No. 124369 that certain
`preferred non-steroidal antioestrogens are pure antioestrogens.
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`According to the present invention there is provided a product comprising an oestrogen and a pure
`antioestrogen as a combined preparation for simultaneous, sequential or separate use in selective oestrogen
`therapy of perimenopausal or postmenopausal conditions, the pure antioestrogen being selected from
`fl-n-butyI-fl-methyl-, fl-iHnH-heptailuorobutyl-fl-methyl- or EEG-methylpentamethylene)-11-(3,17i9—
`dihydroxyoestra—1,3,5(10)—trien—7a-yl)undeoanamide;
`[\lvn—butyl-
`or
`Int-1H,1H-heptatluorobutyl—3-p—[4-(3,17/3—dihydroxyoestra—1 ,3,5(10)—trien—7a—yl)butyl]-
`phenylpropionamide;
`7a-(10-Q-chlorophenylthiodecyl)—, 7a-(10-p-chlorophenylsulphinyldecyl)—, 7a-[9-(4,4,5,5.5-pentailuoropen-
`tylsulphinyl)nonyl]~, 7a-[10~(4,4,4-tritluorobutylsulphinyl)decyl1- or 7a~[10~(p~ohlorobenzylsulphinyl)decyl]«
`oestra—t ,3,5(10)-triene-3,17;3-diol;
`7a-(9-n-hepty|sulphinylnonyl)oestra—1,3,5(10)-triene-3,17fi-diol; and
`a compound oi the formula:—
`
`NU-A-X-R1
`
`-(CH2)5-(1,4-
`
`-(CH2)M- or
`
`wherein NU is 6—hydroxy-Z-p—hydroxyphenylnaphth-i-yl and A is -(CH2)1o-,
`phenylene)-(CH2)2~;
`or NU is 1,2,3,4-tetrahydro—6-hydroxy-2—p-hydroxyphenylnaphth-t-yl (either 1RS,2RS or 1RS,ZSR iso-
`mer), or 1,2,3, 4-tetrahydro-6—hydroxy-Z—p—hydroxyphenyi—2—methylnaphth-1-yl
`(either
`the lRS,2RS or
`1RS,2SFl isomer), and A is ~(CH2)1a—, -(CH2)11-or —(CH2)4—(1,4-phenylene)~(CH2)g—;
`or NU is (1RS.2RS)—5-hydroxy-Z-p-hydroxyphenylindan-1-yl or (1RS,2HS)-5~hydroxy—2-p-hydroxyphenyl-2-
`methylindan-1-yl and A is ~(CH2)10—, -(CH2)H-or -(CH2)4-(1,4-phenylene)-(CH2)2*;
`and wherein XR‘ is ~CONR‘R2 wherein R2 is hydrogen or methyl and Ft‘
`is n-butyl, 1H,1H~heptailuorobutyl,
`n-pentyl or n-hexyl, or
`)(Rl
`is
`-SRl, SOFl1 or -502Rl wherein Ft1
`is n-pentyl, n-hexyl, 4,4,5,5,5-pen-
`tailuoropentyl or IH, i H,2H,2H,3H,3H-heptafluorohexyl.
`the invention is
`In a particular product of
`the invention the oestrogen component of a product of
`oestradiol, ethinyloestradiol, oestriol, oestrone, natural conjugated oestrogens, piperazine oestrone sutphate,
`mestranol, chlorotrianisene, dienoestrol, stilboestrol or hoxoestrol or a pharmaceutipally-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 stilboestrot and hexoestrol. A suitable eikyl ester of up to 12 carbon atoms is, for
`example, an acetate, propionate, butyrate, valerate, hexanoate, heptanoate, octanoate, cyclopentyl-
`propionate, nonanoate, decanoate, undecanoate or dodecanoate. A suitable aryl ester of up to 12 carbon
`atoms is, for example, a benzoate, toluate or naphthoate. A preferred pharmaceutically-aoceptable ester of
`the oestrogen component of a product of the invention includes, for example, oestradiol benzoate, oestraoliol
`cyclopentylpropionate, oestradiol dipropionate, oestradiol heptanoate, oestradiol undeoanoate, oestradiol
`vaierate and stilboestrol dipropionate.
`In a further particular product of the invention the pure antioestrogen is
`_l\_l-n-butyi-, fi-n—butylfl-methyla fl-n-pentyi, fl—(l H,1 H-heptafluorobutyl)—or fl-(l H,1 H-heptailuorobutylyfl-
`methyl43-p-[5-(6-hydroxy-2-p-hydroxyphenylnaphth-1-yl)pentyllphenylpropionamide;
`fl-methyl-fl-(l H,i H-heptafluorobutyl)-p-[4-[(1 RS,2RS)-6-hydroxy-2-p-hydroxphenyl-2-methyl-1 2,3,4-
`ietrahydronaphth-1-yl]~butyl] phenylpropionamide;
`the
`(1RS,2RS)-I-[4—[_p—(2—n—hexylthioethyl)phenyl]butyi]—2—p—hydroxyphenyl-1,2,3,4-tetrahydronaphth-6—ol or
`corresponding 4,4,5,5,5-pentatluoropentylthio derivative, or the corresponding hexylsulphinyl, hexylsulphonyl
`or pentatluoropentylsuiphinyl derivatives;
`Q—E-hydroxyphenyl—t-[5-[p-(2—n~hexylthioethyl)phenyl]pentyl1naphth-6—ol or the corresponding hexylsulphinyl
`derivative; or
`
`(1R8, 2R8)—1{Hp-(2-n-hexylthioethyl)phentyl]butyl]—2-E-hydroxypl'ienyl-2—methyl-1,2,3,4-tetrahydronaphth-6-
`0] or the corresponding 4,4,5.5,5-pentatluoropentylthio derivative, or the corresponding hexylsulphinyl or
`pentaiiuoropentylsulphinyl derivative, or the corresponding (1RS,ZSR) isomers of 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 ethinyloestradiol, or a pharmaceutically-acceptable ester
`thereof, and the pure antioestrogen is 7a—[9-(4,4,5,5,5-pentailuoropentylsulphinyi)nonyl]oestra—1,3,5(10)—
`iriene-3,t7ri-diol or (1RS,2RS)-2-p-hydroxyphenyl-Z-methyi-t-[9-(4,4,5,5,5-pentafluoropentylsulphinyl)nonyl]-
`
`InnoPharma Exhibit 1055.0003
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`EP 0 346 014 B1
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`1 ,2,3,4-tetrahydronaphth—E-cl.
`A particularly preferred product of the invention comprises an oestrogen and a pure antioestrogen for
`use as stated above wherein the oestrogen is oestradiol, oestradiol benzoate, oeslradiol valerate or
`oestradiol undecanoate and the pure antioestrogen is 7a—[9—(4,4,5,5,5-pentatluoropeniylsulphinyl)nonyl]—
`oestra-1,3,5(10)-triene—3,17fi—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 as a combined preparation for simultaneous,
`sequential or separate use in selective oestrogen therapy of perimenopausal or postmenopausal conditions,
`which process comprises bringing together said oestrogen and said pure antioestrogen.
`In a further feature ot 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
`provided a pharmaceutical composition which comprises the product of
`the invention together with a
`pharmaceutisally—acceptable diluent or carrier.
`for selective oestrogen therapy of peri-
`As mentioned above a product of the invention is useful
`menopausal or postmenopausal conditions. It will be understood that there is no absolute requirement that
`the oestrogen and pure antioesirogen 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 product of the invention. Thus it
`will be appreciated that a pharmaceutical composition according to the present
`invention includes a
`composition comprising an oestrogen, a pure antioestrogen and a pharmaceutically-accepiable diluent or
`carrier. Such a composition conveniently provides the product of the invention for simultaneous use in
`selective oestrogen therapy of perimenopausal or postmenopausal conditions. A pharmaceutical composi-
`tion according to the present invention also includes separate compositions comprising a first composition
`comprising an oestrogen and a pharmaceuticalIy—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 postmenopausal 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), tor topical use (for
`example as creams, ointmenis, 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 city solution or
`suspension for intravenous, 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
`pharmaceutical excipients, well known in the art.
`inert
`for example.
`Suitable pharmaceuticaliy acceptable excipients tor a tablet formulation include,
`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 disintegration and the subsequent absorption of the active ingredient within
`the gastrointestinal tract, or to improve their stability and/or appearance,
`in either case using conventional
`coating agents and procedures well known in the art.
`Compositions for oral use may be in the form oi hard gelatin capsules in which the active ingredient is
`mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft
`gelatin capsules in which the active ingredient
`is mixed with water or an oil such as peanut oil,
`liquid
`parailin or olive oil.
`Aqueous suspensions generally contain the active ingredient in finely powdered form together with one
`or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl—
`methylceliulose, 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
`polyoxethyiene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols, for
`example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters
`derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation
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`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,
`tiavouring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame).
`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 preparation of an aqueous suspension by the addition oi
`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 exempli-
`fied by those already mentioned above. Additional excipients, such as sweetening, ilavouring 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 arachls oil, or a mineral oil, such
`as,
`for example,
`liquid paraiiin or a mixture oi any oi
`these. Suitable emulsifying agents may be, for
`example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides
`such as lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides (tor example
`sorbitan monooleate) and condensation products of
`the said partial esters with ethylene oxide such as
`polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, tlavouring and preserva-
`live agents.
`The pharmaceutical compositions may also be in the form of sterile injectable aqueous or oily
`suspensions, which may be iormulated according to known procedures 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, tor 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 injection oi an aqueous suspension or an oily solution
`or suspension of a pharmaceutical composition of the invention provides a depot of the active ingredients at
`the injection site lrom which those ingredients may leach out over a period of time to provide the sustained
`release thereof.
`
`Suppository formulations may be prepared by mixing the active ingredient with a suitable non—irritating
`excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt
`in the rectum to release the drug. Suitable excipients include, for example, cocoa butter and polyethylene
`glycols.
`Topical formulations, such as creams, ointments, gels and aqueous or oily solutions or suspensions,
`may generally 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
`pharmaceutical composition as defined above which comprises bringing into admixture a product as defined
`above together with a pharmaceutically-acceptable diluent or carrier.
`The invention also provides the use of a product as defined above for the manufacture of a combined
`preparation for use simultaneously, sequentially or separately in selective oestrogen therapy of peri-
`menopausal or postmenopausal conditions.
`the invention and the pharmaceutical
`the product of
`It will be appreciated that
`the definition of
`composition of
`the invention includes only those products or compositions which are useful
`in a new
`method for the treatment or prophylaxis of perimenopausal or postmenopausal condition. Pharmaceutical
`compositions comprising an oestrogen and a pure antioestrogen, together with a pharmaceutically-accept—
`able diluent or carrier, are novel.
`In European Patent Sepciiica‘tions Nos. 138504 and 124369 it is disclosed
`that the antioestrogenic activity of the compounds disclosed therein may be demonstrated by the co-
`administration of a test compound and oestradiol benzoate to an immature female 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 subcutaneous injection whereas the test compound was given separately
`either orally or subcutaneously.
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`the invention there is provided a pharmaceutical composition
`According to a further aspect of
`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
`procedures using conventional pharmaceutical excipients well known in the art such as, for example, those
`disclosed above.
`
`the manufacture of a pharmaceutical
`the invention also provides a process for
`This aspect of
`composition as defined immediately ab0ve which comprises bringing into admixture an oestrogen and a
`pure antioestrogen together with a pharmaceutically-acceptable diluent or carrier.
`The invention also provides the use of a pharmaceutical composition as defined immediately above for
`the manufacture of a new medicament
`for use in selective oestrogen therapy of perimenopausal or
`postmenopausal conditions.
`As stated above a product of the invention is of use in selective oestrogen therapy of perimenopausal
`or postmenopausal conditions. Selective oestrogen therapy may be demonstrated using the standard
`procedure set out below:—
`a) an
`vivg 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 oi 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 an-
`tioestrogen is administered alone to a rat no increase in uterine weight whatsoever is observed.
`The oestrogenic activity of a compound may be demonstrated 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 i_n v_ivg 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
`animals‘ 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 mature rats can also be measured in the same
`assay by measuring the effect of the compound in antagonising the efiect of the animals' endogenous
`oestrogens which serve to increase the weight of their uteri.
`A comparison of the potencies of the antioeetrogenic 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
`compound to be measured.
`Although the pharmacological properties of a product 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, 0002-20 mg/kg orally or in the range, for example, 00001-01 mg/kg subcutaneously;
`(ii) a pure antloestrogen which possesses antioestrogenic activity in the above tests (a) and (b) at doses
`in the range, for example, in test (a): EDso 0.05-5 mg/kg orally or EDso 0.01-1.0 mg/kg subcrrtaneously;
`in test
`(b): antiuterotrophic effect:— ['30er < 20 mg/kg/day orally, < 2 mgilrg/day subcutaneously or
`intramuscularly and < 10 mg/kg/injection when closed as an intramuscular depot iniection; reduction in
`bone density:— [3050 > 20 mg/kg/day orally, > 5 mg/kg/day subcutaneously or intramuscularly and > 10
`mg/kg/iniection when dosed as an intramuscular depot injection.
`A product of the invention is thereby seen to be surprisingly selective as the activity of the pure
`antioestrogen 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 antioestrogenlc 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
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`InnoPharma Exhibit 1055.0006
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`EP 0 346 014 B1
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`specimen of endometrial 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 conjugated oestrogens or oestradiol valerate may be administered daily. Alternatively a tablet
`containing 10 to 100 ug of ethinyloestradiol may be administered daily. Alternatively the oestrogenic
`component may be administered 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 ug of oestradiol contained within a transdormal patch; or, for example, vaginal application
`utilising, for example, daily application of 0.5 to 2 mg of natural conjugated oestrogens contained within 0.5
`to 5 ml of a cream.
`
`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 oestrogenic component on uterine tissue is
`antagonised to a substantial degree whereas the beneficial effect of the oestrogenic component on bone is
`substantially unopposed. Thus, for example,
`the antioestrogenic component may be formulated in like
`manner to the oestrogenic component, for example as a tablet, an oily solution suitable for intramuscular
`injection, within a transdermal 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 preferably 50 mg
`to 500 mg, of a pure antioestrogen may be used. Preferably the pure antioestrogen may be administered
`by the periodic intramuscular injection oi,
`for example, an aqueous suspension or an oily solution or
`suspension containing 50 mg to 5 g of the pure antioestrogen. Preferably an oily solution, for example a
`solution containing arachis or castor oil, an alcohol such as benzyl alcohol and 50 mg to 500 mg of the pure
`antioestrogen is employed. Such an iniection provides a depot oi the pure antioestrogen which thereafter
`leeches out from the injection site to previde a selective antioestrogenic effect for a period of, for example,
`one to six weeks.
`
`As mentioned some a product of the invention is useful for selective oestrogen therapy of peri—
`menopausal or postmenopausal conditions. As previously mentioned perimenopausal and postmenopausal
`conditions include,
`for example, vasomotor disturbances (hot flushes), urogenital atrophy (particularly
`affecting the vagina and the distal urethra), psychosomatic complaints, changes in the lipid metabolism and
`oesteoporosis. The selective antioestrcgenic 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 component 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 utcrowselective effect of the pure antioeslrcgenic component of a product of the invention
`will allow the beneficial sheet of the oestrogenic component of a product of the invention to be applied to
`other oestrogen-responsive tissues,
`for example those causing vasomotor disturbances, pyschosomatic
`complaints and changes in lipid metabolism.
`The invention will new be illustrated in the following non—limiting Examples.
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`Example 1
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`Assay in Mature Rats Of the Selective Antloestrogenic Activity of a Pure Antioestrogen
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`The pure antioestrogen used was (1 RS,2RS)—2-p-hydroxyphenyl-2-methyl-1-[9—(4,4,5,5.5—pentailuoropen-
`tylsulphinyl)nonyl]—13,3,4-tetrahydronaphth-6-ol.
`The compound was given subcutaneously as a solution in arachis oil at doses of 2 mgfkg/day and 10
`mg/kg/day to two groups of 5 mature rats for a total of 28 days. Further groups of 5 mature rats served as
`an untreated control group. A further group of 5 mature rats was ovariectomised to serve as another control
`group. At the end of the treatment period the weights of the uteri of the test and control groups of rats were
`determined.
`In addition the femurs were dissected, weighed and their volumes were determined using
`Archimedes Principle. The femurs were then burned and the residual ash was weighed. From these data,
`gross femur density and bone mineral density were calculated