`Bowler et al.
`
`{19]
`
`[54] STEROID DERIVATIVES
`
`[11] Patent Number:
`
`4,659,516
`
`[45] Date of Patent:
`
`Apr. 21, 1987
`
`[75]
`
`Inventors:
`
`Jean Bowler, Sandbach; Brian S.
`Tait, Macclesfield, both of United
`Kingdom
`
`[73] Assignee:
`
`Imperial Chemical Industries PLC,
`London, England
`
`
`
`[21] Appl. No.: 656,466
`
`[22] Filed:
`
`Oct. 1, 1984
`
`Foreign Application Priority Data
`[30]
`Oct. 12, 1983 [GB] United Kingdom ................. 8327256
`
`
`Int. Cl.4 ..................... .. C07J 00/00; A61K 31/56
`[51]
`[52] US. Cl. . . . . .
`. . . . . . . . . . . . . . . . . . .. 260/3975
`[58] Field of Search .................... .. 260/397.5; 514/182
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`3,679,716
`4,011,314
`
`7/1972 Anner et a1.
`3/1977 Petzoldtet al.
`
`260/3975
`
`.... ..
`260/3975
`
`wherein the double bond(s) between carbon atoms 6
`and 7 and/or carbon atoms 8 and 9 are optional;
`wherein the aromatic ring A may optionally bear one or
`two halogen or alkyl substituents; wherein R3 is hydro-
`gen, alkyl, or acyl; wherein R16 is hydrogen, alkyl or
`hydroxy; wherein either R17 is hydroxy or acyloxy and
`R27 is hydrogen, alkyl, alkenyl or alkynyl, or R17 and
`R27 together form 0x0 (:0); wherein R18 is alkyl;
`wherein A is alkylene, alkenylene or alkynylene option-
`ally fluorinated and optionally interrupted by —-0——,
`-—S—-, —SO—, —SOz—, —CO—, —NR—, —NR-
`CO—, —CONR—, —COO—, —-OCO— or phenylene,
`wherein R is hydrogen or alkyl; wherein R1 is hydro—
`gen, alkyl, alkenyl, cycloalkyl, halogenoalkyl, carboxy-
`alkyl, alkoxycarbonylalkyl, aryl, arylalkyl, or dialkyl-
`aminoalkyl, or R1 is joined to R2 as defined below; and
`wherein X is —-CONR2—, —CSNR2——, —NR12CO—,
`———NR12CS—, —NR12CONR2—,
`
`Primaiy Examiner—Leonard Schenkman
`Assistant Examiner—Joseph A. Lipovsky
`Attorney, Agent, or Firm—Cushman, Darby & Cushman
`
`NR22
`ll
`—NR'2—c—NR3—.
`
`ABSTRACT
`[57]
`A steroid derivative of the formula:
`
`ST—A—X—R‘
`
`—SO;;_NR2~— or —CO-—; or, when R1 is not hydrogen,
`is ——O—, —NR2—, —(NO)R2—, -——(PO)R2——, ——-NR1'
`2COO—; —NR12502—, —S—, —SO— or —SOz—;
`wherein R2 is hydrogen or alkyl or R1 and R2 together
`form alkylene or halogenoalkylene; wherein R12 is hy-
`drogen or alkyl and wherein R22 is hydrogen, cyano or
`.nitro; or a salt thereof when appropriate.
`
`wherein ST is a 7a-linked steroid nucleus of the general
`formula:
`
`8 Claims, No Drawings
`
`InnoPharma Exhibit 1016.0001
`
`
`
`1
`
`STEROID DERIVATIVES
`
`4,659,516
`
`This invention relates to new steroid derivatives
`which possess antioestrogenic activity.
`Various oestradiol derivatives are known which bear
`a carboxyalkyl substituent at the 7a-position. These
`have been used, when bound via the carboxy group to
`polyacrylamide resin or to agarose, for the purification
`of oestrogen receptors (Journal of Biological Chemis-
`try, 1978, 253, 8221); and, when conjugated with bovine
`serum albumin, for the preparation of antigens (United
`Kingdom Specification No. 1,478,356).
`We have now found that certain 7a-substituted deriv-
`atives of oestradiol and related steroids possess potent
`antioestrogenic activity.
`According to the invention there is provided a ste-
`roid derivative of the formula:
`
`ST—A—X—R ‘
`
`wherein ST is a 7a-1inked steroid nucleus of the general
`formula:
`
`
`
`wherein the dotted lines between carbon atoms 6 and 7,
`and carbon atoms 8 and 9, of the steroid nucleus
`indicate that there is an optional double bond be-
`tween carbon atoms 6 and 7, or that there are two
`optional double bonds between carbon atoms 6 and 7
`and carbon atoms 8 and 9;
`wherein the aromatic ring A may optionally bear one or
`two halogen or alkyl substituents;
`wherein R3 is hydrogen or alkyl, alkanoyl, alkoxycar-
`bonyl, carboxyalkanoyl or aroyl each of up to 10
`carbon atoms;
`wherein R16 is hydrogen, alkyl of up to 6 carbon atoms
`which is preferably in the B-configuration, or hy-
`droxy which is preferably in the a-configuration;
`wherein either R17 (in the B-configuration) is hydroxy
`or alkanoyloxy, carboxyalkanoyloxy or aroyloxy
`each of up to 10 carbon atoms; and R27 (in the a-con-
`figuration) is hydrogen or alkyl, alkenyl or alkynyl
`each of up to 6 carbon atoms;
`or R17 and R27 together form 0x0 (:0);
`wherein R13 is alkyl of up to 6 carbon atoms;
`wherein A is straight- or branched-chain alkylene, alke-
`nylene or alkynylene each of from 3 to 14 carbon
`atoms, which may have one or more hydrogen atoms
`replaced by fluorine atoms, or has the formula
`
`—A1—Y—A“—
`
`wherein A1 and A11 are each alkylene or alkenylene,
`optionally flourinated, having together a total of 2 to
`13 carbon atoms and Y is -——O—, —S—, —SO-—,
`—SOz—, ——CO—— or —-NR— wherein R is hydrogen
`or alkyl of up to 3 carbon atoms;
`or A1 is alkylene or alkenylene, optionally fluorinated,
`and A“ is a direct link or alkylene or alkenylene,
`optionally fluorinated, such that A1 and All together
`
`5
`
`10
`
`15
`
`20
`
`25
`
`3O
`
`35
`
`45
`
`50
`
`55
`
`65
`
`2
`have a total of 1 to 12 carbon atoms, and Y is —NR-
`CO—, —CONR—, —COO-—, —OCO— or pheny-
`lene wherein R has the meaning stated above;
`wherein R1 is hydrogen, or alkyl, alkenyl, cycloalkyl,
`halogenoalkyl, carboxyalkyl, alkoxycarbonylalkyl,
`aryl or arylalkyl each of up to 10 carbon atoms, or
`dialkylaminoalkyl wherein each alkyl is of up to 6
`carbon atoms, or R1 is joined to R2 as defined below;
`and wherein X is —CONR2—, —CSNR2—-,
`-—N-
`R—CO—, ——NR-—CS—, ~NR—C0NR2—,
`
`NR22
`II
`-NR‘2—C—NR2—,
`
`—SOzNR2— or -—CO—;
`or, when R1 is not hydrogen, is ———O——, —NR2—, ——(-
`NO)R2—, —(PO)R2-—, —NR-—COO—, —N-
`R—SOz—, —-S—, -SO~ or ~—SOz—;
`wherein R2 is hydrogen or alkyl of up to 6 carbon
`atoms, or R1 and R2 together form alkylene or haloge-
`noalkylene such that, with the adjacent nitrogen
`atom, they form a heterocyclic ring of 5 to 7 ring '
`atoms, one of which atoms may be a second heterocy-
`clic atom selected from oxygen, sulphur and nitro-
`gen;
`
`wherein R12 is hydrogen or alkyl of up to 6 carbon
`atoms;
`
`and wherein R22 is hydrogen, cyano or nitro;
`or a salt thereof when appropriate.
`A suitable value for the halogen or alkyl substituent
`in ring A is, for example, fluoro, chloro, bromo, iodo,
`methyl or ethyl.
`A suitable value for R3 when it is alkyl, alkanoyl,
`alkoxycarbonyl, carboxyalkanoyl or aroyl is, for exam-
`ple, methyl, ethyl, acetyl, propionyl, butyryl, pivalyl,
`decanoyl, isopropoxycarbonyl, succinyl or benzoyl. R3
`is preferably hydrogen or alkanoyl or alkoxycarbonyl
`each of up to 5 carbon atoms.
`~
`A suitable value for R16 when it is alkyl is, for exam-
`ple, methyl or ethyl. R16 is preferably hydrogen.
`A suitable value for R17 when it is alkanoyloxy, car-
`boxyalkanoyloxy or aroyloxy is, for example, acetoxy,
`propionyloxy, succinyloxy or benzoyloxy. R17 is prefer-
`ably hydroxy.
`A suitable value for R27 when it is alkyl, alkenyl or
`alkynyl is, for example, ethyl vinyl or ethynyl. R27 is
`preferably hydrogen.
`A suitable value for R18 is methyl or ethyl, especially
`methyl.
`The group ST— is preferably oestra-1,3,5(10)-triene-
`3,17,8-diol, 3-hydroxyoestra-l,3,5(10)-trien-l7-one or
`17a-ethynyloestra~1,3,5(10)-triene-3,l7B-diol,
`all
`of
`which bear the —A—X-—-—R1 substituent in the 7a—posi-
`tion, or a 3-alkanoyl ester thereof.
`One preferred value for the group —A— is a straight-
`chain alkylene group of the formula
`
`—(CH2)n‘
`
`wherein n is an integer of from 3 to 14, especially from
`7 to 11, which may have one of the hydrogen atoms
`replaced by fluorine, for example to provide the group
`-(CH2)8CHFCH2—. A may also be a branched-chain
`alkylene
`group,
`for
`example
`the
`group
`—(CH2)5CH(CH3)——, or a straight-chain alkenylene
`group, for example of the formula
`
`InnoPharma Exhibit 1016.0002
`
`
`
`3
`
`—(CH2)2CH=CH(CH2)m—
`
`4,659,516
`
`4
`wherein —A— is ——(CH2),,—, wherein n is an integer
`from 3 to 14, especially from 7 to 11, or —A— is
`
`wherein m is an integer from 0 to 10, especially from 3
`to 7.
`
`A second preferred value for the group A is a group
`of the formula
`
`--A‘—Y——-A“—
`
`wherein A1 is straight-chain alkylene or alkenylene each
`of 2 to 9 carbon atoms, especially alkylene of 4 to 6
`carbon atoms, —Y— is phenylene (ortho, meta— or,
`especially, para-) and A“ is a direct link, ethylene or
`vinylene, especially ethylene.
`A suitable value for R1 when it is alkyl, alkenyl or
`cycloalkyl is, for example, methyl, ethyl, n-propyl, iso-
`propyl, n-butyl,
`isobutyl,
`t-butyl, n-pentyl,
`t-pentyl,
`2,2-dimethylpropyl,
`l-methylbutyl, 2-methylbutyl, 3-
`methylbutyl, n-hexyl, 1,1-dimethylbutyl, 1,3-dimethyl-
`butyl, n-heptyl, n-nonyl, n—decyl, n-undecyl, allyl, cy-
`clopentyl or cyclohexyl.
`A suitable value for R1 when it is aryl or arylalkyl is,
`for example, phenyl, 2-ethy1pheny1, p-fluorophenyl,
`p-chlorophenyl, m-chlorophenyl, p-cyanophenyl, p-
`methoxyphenyl, benzyl, a-methylbenzyl, p-chloroben-
`zyl, p-fluorophenethyl or p-chlorophenethyl.
`A suitable value for R1 when it is halogenoalkyl,
`carboxyalkyl, alkoxycarbonylalkyl or dialkylaminoal-
`kyl is, for example, 2-chloro-2,2-difluoroethyl, 2,2,2-tri-
`fluoroethyl, 2,2,3,3,3-pentafluoropropyl, 3-chloropro-
`pyl, 2,2-difluorobutyl, 4,4,4-trifluorobutyl, 1H,1H-hep-
`tafluorobutyl, 4,4,5,5,5-pentafluoropentyl, 4,4,5,5,6,6,6-
`heptafluorohexyl,
`lH,1H—tridecafluoroheptyl, S-car-
`boxypentyl,
`5-methoxycarbonylpentyl
`or
`3-dime-
`thylaminopropyl.
`A suitable value for the heterocyclic ring —NR1R2 is,
`for
`example,
`pyrrolidino,
`piperidino,
`4-methyl-
`piperidino, 4~ethy1piperidino, 3-methylpiperidino, 3,3-
`dimethylpiperidino, 4-chloropiperidino, morpholino or
`4-methylpiperazino.
`A suitable value for R2 or R12 when it is alkyl is, for
`example, methyl, ethyl or n-butyl.
`One appropriate salt is an acid-addition salt of a ste-
`roid derivative which possesses an amino function, for
`example a compound wherein Y is —NR—, X is
`—NR2—- or R1 is dialkylaminoalkyl. A suitable acid-
`addition salt is, for example, a hydrochloride, hydrobro-
`mide, acetate, citrate, oxalate or tartrate.
`Another appropriate salt is a base-addition salt of a
`steroid derivative which possesses a carboxy function,
`for example a compound wherein R1 is carboxyalkyl. A
`suitable base-addition salt is, for example, a sodium,
`potassium, ammonium or cyclohexylarnine salt.
`A preferred steroid derivative of the invention has
`the formula:
`
` .
`
`'A—X—R‘
`
`wherein R17 is hydroxy and R27 is hydrogen or ethynyl,
`or R17 and R27 together form oxo;
`
`"(CH2)m
`
`(CH2)p"
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`55
`
`wherein m is an integer from 2 to 9, especially from 4 to
`6, and p is 0 to 2, especially 0 or 2; wherein R1 is alkyl,
`fluoroalkyl or cycloalkyl each of up to 10 carbon
`atoms, or phenyl, chlorophenyl or benzyl, or is linked
`to R2 as stated below;
`wherein X is —CONR2—, ~—NR12C0—, ——S—,
`—S0— or —SOz-—, wherein R2 is hydrogen or alkyl
`of up to 3 carbon atoms or together with R1 forms
`alkylene of 5 or 6 carbon atoms, and wherein R12 is
`hydrogen or alkyl of up to 3 carbon atoms.
`A particularly preferred steroid derivative of the
`invention has the last-mentioned formula wherein the
`number of carbon atoms in the two groups A and R1
`adds up to between 12 and 16, inclusive, especially 14 if
`neither Rl nor A contains a phenyl or phenylene group,
`and 16 if there is a phenylene group in ——A— or a
`phenyl group in R1.
`Specific steroid dervatives of the invention are here-
`inafter described in the Examples. Of these, particularly
`preferred compounds are:
`N-n-butyl-N-methy1-, N-2,2,3,3,4,4,4—heptafluorobutyl—
`N-methyl- and N,N-(3-methylpentamethylene)-ll-
`(3, 17B-dihydroxyoestra—1,3,5(lO)-trien-7a-yl)undeca-
`mide;
`N—n-butyl- and N-2,2,3,3,4,4,4-heptafluorobutyl-3-p-[4-
`(3, 17B-dihydroxyoestra-1,3,5(10)-trien-7a-yl)butyl]-
`phenylpropionamide;
`7a-(10-p—chloro—
`7a—(10-p-chlorophenylthiodecyl)-,
`phenylsulphinyldecyl)-,
`7a—[9-(4,4,5,5,5-penta—
`fluoropentylsulphinyl)nonyl]-,
`7a-[lO-(4,4,4—tri-
`fluorobutylsulphinyl)-decyl]-
`and
`7a—[10-(p&
`chlorobenzylsulphonyl)decyl]oestra-l,3,5(10)-triene-
`3,17B-diol; and
`7a-(9-n-heptylsulphinylnonyl)oestra-1,3,5(10)-triene-
`3,17B-diol.
`A preferred process for the manufacture of a steroid
`derivative of the invention wherein X has the formula
`——-CONR2—, —CSNR2— or —SOzNR2— comprises
`the
`reaction of
`a
`compound of
`the
`formula
`STl—A—Zl, wherein A has the meaning stated above,
`wherein ST1 either has the same meaning as stated
`above for ST, or is an equivalent 7a-linked steroid nu-
`cleus which bears one or more protecting groups for
`functional derivatives, and wherein 21 is an activated
`group derived from a carboxylic,
`thiocarboxylic or
`sulphonic acid, with an amine of the formula HNR‘RZ,
`wherein R1 and R2 have the meanings stated above,
`whereafter any protecting group in ST1 is removed by
`conventional means.
`A suitable activated group 21 is, for example, a mixed
`anhydride, for example an anhydride formed by reac-
`tion of the acid with a chloroformate such as isobutyl
`chloroformate.
`
`65
`
`A suitable protecting group in ST1 is, for example, an
`alkyl or aralkyl ether, for example the methyl or benzyl
`ether, of the 3-hydroxy function, or a tetrahydropyra-
`nyl ether of the 17B-hydroxy function.
`A preferred process for the manufacture of a steroid
`derivative of the invention wherein X has the formula
`
`InnoPharma Exhibit 1016.0003
`
`
`
`5
`—CO— comprises the reaction of an acid of the for-
`mula STl—A—COOH, wherein ST1 and A have the
`meanings stated above, with an organometallic com-
`pound of the formula Rl—M, wherein R1 has the mean-
`ing stated above and M is a metal group, for example the 5
`lithium group, whereafter any protecting group in ST1
`is removed by conventional means.
`A preferred process for the manufacture of a steroid
`derivative of the invention wherein X has the formula
`7 —S—, ——O—, —NR2— or —(P0)R2—— comprises the
`reaction of a compound of the formula STl—A—ZZ,
`wherein ST1 and A have the meanings stated above and
`wherein Zzis a displaceable group, with a compound of
`the formula RISH, RIOH, HNR1R2 or RleP—C6H5
`wherein R1 and R2 have the meanings stated above, 15
`whereafter any protecting group in ST1 is removed by
`conventional means, and whereafter a phosphonium salt
`is hydrolysed to the phosphinyl compound.
`A suitable value for Z2 is, for example, a halogen
`atom or a sulphonyloxy group, for example the me- 20
`thanesulphonyloxy or toluene-p-sulphonyloxy group.
`A preferred process for the manufacture of a steroid
`derivative of the invention wherein X has the formula
`—NR12CO—, ——NR12CS—, ——NR12CONR2—,
`
`10
`
`25
`
`NR22
`II
`-—NR12-c—NR2—,
`
`—-NR12COO— or —NR12802— comprises the reac- 30
`tion of a compound of the formula STl—A—NHRIZ,
`wherein ST, A and R12 have the meanings stated
`above, with an acylating agent derived from an acid of
`the formula RICOOH, RICSOH, RIOCOOH or RISO-
`20H; or, for the manufacture of a urea, with an isocya- 35
`nate of the formula RINCO; or, for the manufacture of
`a guanidine, with a cyanamide of
`the formula
`RlNRZ—CN, whereafter any protecting group in ST1 is
`removed by conventional means.
`A suitable acylating agent is, for example, an acyl 40
`chloride or acyl anhydride.
`The starting materials for use in all the abovemen-
`tioned processes may be obtained by reacting a steroid
`derivative of the formula
`
`45
`
`50
`
`wherein R16 and R18 have the meanings stated above
`and wherein R37 is an acyl group, for example the acetyl
`group, with a compound of the formula
`
`f‘“
`Br—AZ—an—O-SIi-C(CH3)3
`CH3
`
`55
`
`60
`
`wherein A2 either has the same meaning as stated above
`for A, or wherein ~A2—CH2— has the same meaning 65
`as stated above for A; separating the isomers at the
`7-position of the steroid nucleus to provide the 7a-iso-
`mer; hydrolysing off the dimethyl-t-butylsilyl protect-
`
`4,659,516
`
`6
`
`ing group; and converting the steroidal part of the mo-
`lecular to the required structure by conventional reac-
`tions. The intermediate product obtained, which has the
`formula:
`
`srl—ALcr-rzon
`
`wherein STI has the meaning stated above, may be
`oxidised to the corresponding carboxylic acid of the
`formula STl—AZ—COOH which provides the start-
`ing material for the first or second process of the
`invention described above;
`or it may be converted into a compound of the formula
`STl—A2—CHZZ2 by reaction with a halogenating
`agent or a sulphonylating agent to provide the start-
`ing material for the third process of the invention
`described above.
`
`The starting material for the fourth process of the
`invention described above may be obtained by using the
`third process of the invention described above except
`that an amine of the formula RIZNH2 is used in place of
`an amine of the formula HNR‘RZ.
`The intermediate of the formula STl—AZ—CHzOH
`may be oxidised to an aldehyde of the formula
`STl—AZ—CHO which may then be used, by reaction
`with
`an
`appropriately-substituted
`hydrocarbyltri-
`phenylphosphonium salt or hydrocarbyltriethylphos-
`phonate, to prepare a starting material wherein ——A— is
`alkenylene.
`'
`An alternative process for the manufacture of a ste-
`roid derivative of the invention wherein —A-—— is alke-
`nylene of the formula —A3—CH=CH—A4— com-
`prises the reaction of a compound of the formula:
`
`srl—A3CH0
`
`wherein ST1 and A3 have the meanings stated above,
`with a triphenylphosphonium salt of the formula:
`
`Rlx—A4—CH2—P+(Ph)3Q-
`
`wherein R1, X and A4 have the meanings stated above
`and wherein Q— is an anion, for example the bromide
`ion.
`
`The reaction may be carried out in solution in di-
`methyl sulphoxide in the presence of dimsyl sodium.
`The steroidal aldehyde starting material when
`—A3— is -—A2—— as defined above may be obtained by
`oxidation of the corresponding alcohol as described
`above. The steroidal aldehyde starting material wherein
`—A3— is a direct link may be obtained from the 3-keto-
`A4’6-inital steroidal starting material described above by
`reaction with cyanide to give the 3-keto-A4-7a-cyano
`compound, aromatisation, suitable protection and then
`reduction of the cyano group to the formyl group.
`The phosphonium starting material may be obtained
`by reaction of triphenylphosphine with a bromide of the
`formula
`
`Rl—X—AL—cnznr.
`
`A steroid derivative of the invention wherein ST is a
`l7B-hydroxy-steroid derivative may be converted by
`conventional reactions into the corresponding l7—keto
`steroid derivative, and thence to the corresponding
`l7B-hydroxy-l7a-hydrocarbyl steroid derivative (that
`is, a steroid derivative of the invention wherein R27 is
`alkyl, alkenyl or alkynyl). Similarly, a steroid derivative
`
`InnoPharma Exhibit 1016.0004
`
`
`
`4,659,516
`
`8
`and contain conventional excipients. Thus a tablet could
`contain diluents, for example mannitol or maize starch,
`disintegrating agents, for example alginic acid, binding
`agents, for example methyl-cellulose, and lubricating
`agents, for example magnesium stearate.
`The composition may contain, in addition to the ste-
`roid derivative of the invention, one or more antiandro—
`genic agents or antiprogestational agents.
`A composition for oral administration may conve-
`niently contain from 5 to 500 mg. of a steroid derivative
`of the invention.
`The invention is illustrated but not limited by the
`following Examples:_
`EXAMPLE 1
`
`N-Methylmorpholine (0.24 ml.) and isobutyl chloro-
`formate (0.288 ml.) were successively added to a stirred
`solution
`of
`1 1-(17B-acetoxy-3-benzoyloxyoestra-
`1,3,5(10)-trien-7a-yl)undecanoic acid (1.0 g.) in methy-
`lene chloride (17 ml.) which was cooled to -—10° C.,
`and after 30 minutes n-butylamine (0.29 ml.) was added
`and the mixture was stirred at laboratory temperature
`for 15 minutes. Saturated aqueous sodium bicarbonate
`solution (20 ml.) was added and the mixture was ex-
`tracted four times with methylene chloride (50 ml. each
`time). The combined extracts were washed with water
`(10 m1.), dried and evaporated to dryness. There was
`thus obtained as residue 1l-(l7B-acetoxy-3-benzoyloxy-
`N-n-butyloestra-l,3,5(10)-trien-7a-yl)undecanamide as
`an oil.
`
`Aqueous N-sodium hydroxide solution (8 ml.) was
`added to a stirred solution of the above amide (1.06 g.)
`in a mixture of methanol (16 ml.) and tetrahydrofuran (8
`ml.) and the mixture was stirred at laboratory tempera-
`ture for 18 hours, neutralised with aqueous N-hydro—
`chloric acid and the organic solvents were removed by
`evaporation. Water (40 ml.) was added and the mixture
`was extracted four times with methylene chloride (60
`ml. each time). The combined extracts were washed
`with water (10 ml.), dried and evaporated to dryness
`and the residue was purified by chromatography on a
`silica gel (Merck Kieselgel 60) column using a 13:7 v/v
`mixture of ethyl acetate and toluene as eluant. There
`was thus obtained N-n-butyl-l1-(3,17B-dihydroxyoes-
`tra-1,3,5(10)trien-7a-yl)undecanamide as an oil which
`was characterised by the following data:
`
`
`3.7
`3.28
`
`0.90
`
`quartet
`
`triplet
`
`1
`2
`
`3
`
`7
`of the invention wherein R3 and/or R17 are other than
`hydrogen may be obtained from the corresponding
`compounds wherein R3 and/or R17 are hydrogen by
`conventional etherification or esterification processes,
`and these may also be used in reverse to prepare the
`corresponding hydroxy compounds.
`A steroid derivative of the invention wherein A is
`alkenylene may be hydrogenated to provide the corre-
`sponding compound wherein A is alkylene.
`A steroid derivative of the invention wherein —X—
`is —CH2NR2- or —NR2CH2— may be obtained by
`the reduction, for example with borane, of the corre-
`sponding compound wherein —X—— is —CONR2— or
`—-NR2CO—.
`A steroid derivative of the invention wherein —X—-
`is —CSNH——- or —NHCS—- may be obtained by the
`reaction of the corresponding compound wherein X is
`——CONH— or —NHCO— with 2,4-bis-(4-methoxy-
`phenyl)-1,3-dithia-2,4-diphosphetane-2,4-disulphide.
`A steroid derivative of the invention wherein X is
`—(NO)R2, ——SO—— or —502— may be obtained by'the
`oxidation of the corresponding compound wherein X is
`—NR2— or —S-—.‘ The conditions for the oxidation
`will be chosen to provide the desired product; for exam-
`ple-aqueous sodium metaperiodate will oxidise the Sul-
`phur group to sulphinyl, and m-chloroperbenzoic acid
`in chloroform solution will oxidise the sulphur group to
`sulphonyl or the amine to its oxide.
`As stated above, a steroid derivative of the invention
`possesses antioestrogenic activity. This may be demon-
`strated by its effect
`in antagonising the increase in
`weight of the uterus of an immature female rat pro-
`duced by administering oestradiol benzoate to said rat.
`Thus, when a steroid derivative of the invention and
`oestradiol benzoate are co-administered for 3 days to
`such a rat, a smaller increase in uterine weight is pro-
`duced than the substantial increase which would be
`produced by the administration of oestradiol benzoate
`without the steroid derivative of the invention.
`In particular, a preferred steroid derivative of the
`invention produces an antioestrogenic effect at a dose
`which produces no partial agonist effect, unlike the
`known antioestrogens
`tamoxifen and clomiphene.
`When a preferred steroid is coadministered with oes-
`tradiol benzoate to a rat as described above, no increase
`in uterine weight whatsoever is observed at a suitable
`dose.
`
`A compound with the above pharmacological prop-
`erties is of value in the treatment of the same conditions
`in which tamoxifen is beneficial, in particular, in the
`treatment of anovulatory infertility and in the treatment
`of breast tumors. It is also of value in the treatment of
`menstrual disorders.
`When used to produce an anti-oestrogenic effect in
`warm-blooded animals, a typical daily dose is from 0.1
`to 25 mg/kg. administered orally or by injection. In
`man this is equivalent to an oral dose of from 5 to 1250
`mg./day. A steroid derivative of the invention is most
`conveniently administered to man in the form of a phar-
`maceutical composition.
`According to a further feature of the invention, there
`is provided a pharmaceutical composition comprising a
`steroid derivative of the invention together with a phar-
`maceutically acceptable diluent or carrier.
`The composition may be in a form suitable for oral or
`parenteral administration. A tablet or capsule is a partic-
`ularly convenient form for oral administration and such
`a composition may be made by conventional methods
`
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`
`Proton maggetic resonance sEtrum gin CDCIQZ
`Shift (5) Type of peak No of protons
`Assignment
`7. 16
`multiplet
`l
`aromatic
`6.65
`”
`2
`protons at
`positions
`1, 2 and 4
`position 17
`—CH2-— adjacent
`to -CO—
`—CH3 in
`n-butyl
`0.78 position 18 singlet 3
`
`
`
`
`
`
`
`Mass Spectrum
`
`M+ = 511.4039 (C33H53O3N requires 511.4024).
`M—H20 =493.
`M—(CHzCONHC4H9) = 397.
`
`InnoPharma Exhibit 1016.0005
`
`
`
`9
`
`4,659,516
`
`10
`stirred at laboratory temperature for 70 minutes, neu-
`tralised with aqueous N-hydrochloric acid and the
`methanol was removed by evaporation. The residue
`was extracted four times with ethyl acetate (60 ml. each
`time) and the combined extracts were washed with
`water (20 m1.), dried and evaporated to dryness. The
`residue was purified by chromatography on a silica gel
`column using a 7:3 v/v mixture of toluene and ethyl
`acetate as eluant.
`
`Thin layer chromatography (silica gel plates using a 7.3
`v/v mixture of ethyl acetate and toluene)
`RF: 0.3.
`The
`l 1-(17B-acetoxy—3-benzoyloxyoestra- l,3,5(10)—
`trien-7a-yl)-undecanoic acid used as starting material
`was obtained as follows:
`
`A solution of dirnethyl-t-butylsilyl chloride (37.3 g.)
`in tetrahydrofuran (40 ml.) was added to a solution of
`11-bromoundecanol (50.18 g.) and imidazole (28.95 g.)
`in tetrahydrofuran (120 m1.) and the mixture was kept at
`laboratory temperature for 1.75 hours, diluted with
`diethyl ether (300 m1.) and filtered. The filtrate was
`evaporated to dryness and the residue purified by chro-
`matography on silica gel using a 4:1 v/v mixture of
`petroleum ether (b.p. 60°—80° C.) and toluene as eluant.
`A solution of the 11-(dimethyl-t-butylsi1yloxy)unde-
`cyl bromide thus obtained (73.1 g.) in tetrahydrofuran
`(200 ml.) was added during 2 hours to a stirred suspen-
`sion of magnesium tumings (4.8 g.) in tetrahydrofuran
`(20 m1.) under normal conditions for preparation of a
`Grignard reagent, and the mixture was heated under
`reflux for 2 hours, diluted with tetrahydrofuran (100
`m1.) and cooled to —30° C. Cuprous iodide (19.05 g.,
`dried at 100° C. immediately before use) was added, the
`mixture was vigorously stirred for 10 minutes and a
`solution of 6-dehydro-lQ-nortestosterone acetate (15.48
`g.) in tetrahydrofuran (50 ml.) was added. The mixture
`was stirred for 40 minutes, acetic acid (12 ml.) was
`added. and the mixture was evaporated to dryness.
`Water (150 ml.) was added to the residue, and the mix-
`ture was extracted four times with diethyl ether (300 ml.
`each time). The combined extracts were washed with
`water (50 m1.), dried and evaporated to dryness, and the
`residue was purified by chromatography on a silica gel
`column using a 24:1 v/v mixture of toluene and ethyl
`acetate as eluant.
`
`A mixture of l7B-acetoxy-7a-[l1-(dimethyl-t-buty1-
`silyloxy)undecy1]oestr-4—ene-3-one thus obtained (11.2
`g.), acetic acid (62 ml.), water (31 m1.) and tetrahydrofu-
`ran _(56 ml.) was stirred at 50° C. for 2.75 hours and was
`then evaporated to dryness. A solution of the residue in
`pyridine (56 ml.) and acetic anhydride (28 ml.) was kept
`at laboratory temperature for 18 hours, cooled to 0° C.,
`water (10 ml.) was added and the mixture was stirred
`for 45 minutes and then evaporated to dryness. The
`residue was dissolved in diethyl ether (400 ml.) and the
`solution was washed with saturated aqueous sodium
`bicarbonate solution (20 m1.) and then with water (20
`ml.), dried and evaporated to dryness.
`A solution of the 17B-acetoxy-7a-(1l-acetoxyun-
`decyl)oestr-4—ene-3-one thus obtained (8.98 g.) in aceto-
`nitrile (50 ml.) was added rapidly to a vigorously stirred
`suspension of cupric bromide (7.75 g.) and lithium bro-
`mide (1.52 g.) in acetonitrile (120 ml.) which was heated
`under reflux under an atmosphere of argon, and the
`mixture was stirred and heated for 30 minutes and then
`cooled. Saturated aqueous sodium bicarbonate solution
`(200 ml.) was added and the mixture was extracted four
`times with ethyl acetate (200 ml. each time). The com-
`bined extracts were washed with water (50 m1.), dried
`and evaporated to dryness, and the residue was purified
`by chromatography on a silica gel column using a 9:1
`v/v mixture of toluene and ethyl acetate as eluant.
`Aqueous N-sodium hydroxide solution (8 ml.) was
`added to a stirred solution of the 17B-acetoxy—7a-(1 1-
`acetoxyundecyl)oestra-l,3,5(10)-trien-3-ol
`thus
`ob-
`tained (2.8 g.) in methanol (54 ml.) and the mixture was
`
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`
`Aqueous N-sodium hydroxide solution (6 ml.) and '
`benzoyl chloride (0.93 ml.) were added to a stirred
`solution of the 17B-acetoxy-7a—(1 1-hydroxy-undecy1)o- ,
`estra-l,3,5(10)-trien-3-ol thus obtained (1.94 g.) in ace-
`tone (20 ml.) which was cooled to 0° C., and the mixture
`was stirred for 20 minutes and then poured into a mix-
`ture of ice-water (200 ml.) and saturated aqueous so-
`dium bicarbonate solution (50 ml.). The mixture was
`extracted four times with diethyl ether (120 ml. each
`time) and the combined extracts were washed twice
`with saturated aqueous sodium bicarbonate solution (15
`ml. each time) and then with water (20 m1.), dried and
`_evaporated to dryness. The residue was purified by
`chromatography on a silica gel column using a 7:3 v/v
`mixture of toluene and ethyl acetate as eluant.
`Jones’s reagent (8N-chromic acid solution, 2.3 ml.)
`was added to a solution of the 17 B-acetoxy-3-ben-
`zoyloxy-7a-(1 1-hydroxyundecyl)oestra-1,3,5(10)-triene
`thus obtained (2.17 g.) in acetone (37 ml.) which was
`cooled to 0" C. After 15 minutes isopropanol (0.5 ml.)
`was added and the mixture was evaporated to dryness.
`Water (40 ml.) was added and the mixture was extracted
`three times with methylene chloride (60 ml. each time).
`The combined extracts were washed twice with water
`(10 ml. each time), dried and evaporated to dryness, and
`the residue was purified by chromatography on a silica
`gel column using a 7:3 v/v mixture of toluene and ethyl
`acetate as eluant. There was thus obtained 11-(17B-
`acetoxy-3-benzoyloxyoestra-l,3,5(10)-trien-7a-yl)un-
`decanoic acid;
`
`EXAMPLE 2
`
`The process described in Example 1 was repeated
`using the appropriate amine in place of n-butylamine.
`There were thus obtained the compounds described in
`the following table, all of which were oils the structures
`of which were continued by proton magnetic reso-
`nance and mass spectroscopy:
`
`OH
`
`
`
`" (CH2).oc0NR1R2
`
`HO
`
`R 1
`R2
`
`H
`H
`‘
`ethyl
`H
`n-propyl
`H
`isopropyl
`H
`isobutyl
`H
`t—butyl
`H
`3-methylbutyl
`H
`l-methylbutyl
`H
`2-methylbutyl
`H
`2,2—dimethylpropyl
`H
`n-hexyl
`H
`
`+
`+
`
`+
`
`+
`
`InnoPharma Exhibit 1016.0006
`
`
`
`11
`
` ~continued
`OH
`
`4,659,516
`
`
`
`" (CH2)1oCONR1R2
`
`HO
`
`+
`
`+
`
`
`R1
`R2
`1,1-dimethylbutyl
`H
`+
`1,3-dimethylbutyl
`H
`cyclohexyl
`H
`2,2,2-trifluoroethyl
`H
`2,2,3,3,4,4,4-heptafluorobutyl
`H
`2.2-diiluorobutyl
`H
`3-chloropropyl
`H
`phenyl
`H
`4—methoxyphenyl
`H
`4—chlorophenyl
`H
`4—cyanophenyl
`H
`2-ethylphenyl
`H
`benzyl
`H
`l-phenylethyl
`H
`5-carboxypentyl
`H
`3-dimethylaminopropyl
`H
`n-butyl
`methyl
`2,2-dimethylpropyl
`methyl
`2—methylbutyl
`methyl
`n-hexyl
`methyl
`2,2,3,3,3-pentafluoropropyl
`methyl
`2,2-difluorobutyl
`methyl
`-
`4,4,4—triiluorobutyl
`methyl
`2,2,3,3,4,4,4~heptafluorobutyl
`methyl
`benzyl
`methyl
`n-butyl
`ethyl
`n-butyl
`n-butyl
`2,2,2-trifluoroethyl
`n-butyl
`"(CH2)5’
`
`"‘
`
`+
`
`“(CH2)2—Il‘1'—(CH2)2—
`CH3
`
`—- CH CH CH —
`(
`2)2 I
`(
`2)2
`CH3
`
`—CH2(|3H(CH2)3“
`CH3
`
`“(CH2)2CHC1(CH2)2"
`
`-(CH2)2<|3H(CH2)2“
`C2H5
`
`'
`
`+
`
`+
`
`r3
`—(CH2)3$—CH2-
`CH3
`
`'A solution of ammonia in tetrahydrofuran was used as starting material.
`“Methyl 6-aminohexanoate was used as starting material, the methyl ester being
`hydrolysed during the second stage of the process.
`
`In some cases (indicated + in the above table) the
`undecanoic acid used as starting material was the 3-
`hydroxy-
`rather
`than the 3-benzoyloxy-compound,
`which was prepared by a shortened route as follows:
`The
`17B-acetoxy-7a-(1 1-acetoxyundecyl)oestr—
`4—ene-3-one, prepared as described in the 5th paragraph
`of Example 1, was hydrolysed to the corresponding
`11-hydroxyundecy1 compound as described in'the 7th
`paragraph of Example 1, and this product was purified
`by chromatography on a silica gel column using a 3:2
`v/v mixture of toluene and ethyl acetate as eluant. It
`
`5
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`12
`was then oxidised to the corresponding undecanoic acid
`as described in the 9th paragraph of Example 1, and this
`product was purified by chromatography on a silica gel
`column using a 19:1 v/v mixture of methylene chloride
`and methanol as eluant. The undecanoic acid was
`aromatised as described in the 6th paragraph of Exam-
`ple 1, except that the pH of the reaction mixture was
`adjusted to 3 before extraction into ethyl acetate. The
`product was purified by chromatography on a silica gel
`column using a 3:1 v/v mixture of diethyl ether and
`petroleum ether (b.p. 60°-80° C.) as eluant. There was
`thus obtained, as an oil, 11-(17B-acetoxy-3-hydroxyoes-
`tra—1,3,5(10)-trien-7a-yl)un