PCT
`
`International Bureau
`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`
`
`
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`3
`(51) I(l:1(t)€;'E1:f1t2i‘il_173/161;3t;Iit5$l551jSigifiitfigle5
`C07C 237/30, C071) 295/06, 211/14
`C071) 207/06, A6lK 31/135
`
`(11) International Publication Number:
`(43) International Publication Date:
`
`W0 94/11337
`26 May 1994 (26.05.94)
`
`(21) International Application Number:
`
`PCT/SE93/00927
`
`(22) International Filing Date:
`
`5 November 1993 (05.11.93)
`
`(74) Agents: WIDEN, Bjorn et a1.; Kabi Pharmacia AB, S-751
`82 Uppsala (SE).
`
`(30) Priority data:
`9203318-2
`
`6 November l992(06.l1.92)
`
`SE
`
`(81) Designated States: AU, CA, Fl, HU, JP, NO, US, Euro-
`pean patent (AT, BE, CH, DE, DK, ES, FR, GB, GR,
`IE, IT, LU, MC, NL, PT, SE).
`
`(71) Applicant for all designated States except US): KABI
`PHARMACIA AB [SE/ SE]; S-751 82 Uppsala (SE).
`
`Published
`With international search report.
`
`(72) Inventors; and
`(75) Inventors/Applicants (for US only) .' JOHANSSON, Rolf,
`Arne [SE/ SE]; Daggstigen 8 B, S-141 38 Huddinge (SE).
`MOSES, Pinchas [SE/ SE]; Dalvéigen 6, S-132 00 Saltsj6-
`Boo (SE). NILVERBANT, Lisbeth [SE/ SE]; Lillsj6n§iis-
`vagen 11, S-161 35 Bromma (SE). SPARF, Bengt, Ake
`[SE/SE]; Drottningstigen 6, S-142 65 Trangsund (SE).
`
`(54) Title: NOVEL 3,3-DIPHENYLPROPYLAMINES, THEIR USE AND PREPARATION
`
`(57) Abstract
`
`The invention relates to 3,3-diphenylpropylamines of formula (I), wherein R1 signifies hydrogen or methyl, R2 and R3 in-
`dependently signify hydrogen, methyl, methoxy, hydroxy, carbamoyl, sulphamoyl or halogen, and X represents a tertiary
`amino group of formula (II), wherein R4 and R5 signify non-aromatic hydrocarbyl groups, which may be the same or differ-
`ent and which together contain at least three carbon atoms, and wherein R4 and R5 may form a ring together with the amine
`nitrogen, their salts with physiologically acceptable acids and, when the compounds can be in the form of optical isomers,
`the racemic mixture and the individual enantiomers. The invention also relates to methods for their preparation, pharma-
`ceutical compositions containing the novel compounds, and the use of the compounds for preparing drugs.
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 1
`
`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 1
`
`

`
`FOR THE PURPOSES OF INFORMATION ONLY
`
`Codes used to identify States party to the PCT on the front pages of pamphlets publishing international
`applications under the PCT.
`
`AT
`AU
`BB
`BE
`BF
`BC
`BJ
`BR
`BY
`CA
`CF
`CG
`CH
`Cl
`CM
`CN
`CS
`CZ
`DE
`DK
`ES
`Fl
`FR
`GA
`
`Austria
`Australia
`_ Barbados
`Belgium
`Burkina Faso
`Bulgaria
`Benin
`Brazil
`Belarus
`Canada
`Central African Republic
`Congo
`Switzerland
`Céte d‘lvoire
`Cameroon
`China
`Czechoslovakia
`Czech Republic
`Germany
`Denmark
`Spain
`Finland
`France
`Gabon
`
`-
`
`United Kingdom
`Georgia
`Guinea
`Greece
`Hungary
`Ireland
`Italy
`Japan
`Kenya
`Kyrgystan
`Democratic People's Republic
`of Korea
`Republic of Korea
`Kazakhstan
`Liechtenstein
`Sri Lanka
`Luxembourg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`Mali
`Mongolia
`
`‘
`
`Mauritania
`Malawi
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Slovenia
`Slovakia
`Senegal
`Chad
`Togo
`Tajikistan
`Trinidad and Tobago
`Ukraine
`United States of America
`Uzbekistan
`Viet Nam
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 2
`
`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 2
`
`

`
`W0 94/ 11337
`
`PCT/SE93/00927
`
`NOVEL 3,3-DIPHENYLPROPYLAMINES, THEIR USE AND PREPARATION
`
`The present invention relates to novel
`
`therapeutically active compounds, methods for their
`
`preparation, pharmaceutical compositions containing the
`
`novel compounds, and the use of the compounds for
`
`preparing drugs.
`
`W0 89/06644 discloses 3,3-diphenylpropylamines having
`
`anticholinergic activity.
`
`In accordance with the present
`
`10
`
`invention novel therapeutically active compounds have now
`
`been found,
`
`some of which are formed as metabolites in
`
`mammals when treated with the 3,3-diphenylpropylamines
`
`disclosed in the above-mentioned WO publication. These
`
`metabolites have at least as favourable anti-cholinergic
`properties as the parent compounds and can thus be used
`
`15
`
`for the control of events mediated by acetylcholine,
`
`like
`
`urination.
`
`The novel compounds of the present invention are
`
`represented by the general formula I
`
`20
`
`25
`
`30
`
`35
`
`wherein R1 signifies hydrogen or methyl, R2 and R3
`
`independently signify hydrogen, methyl, methoxy, hydroxy,
`
`carbamoyl, sulphamoyl or halogen,
`
`and X represents a
`
`tertiary amino group of formula II
`
`/R4
`‘\R5
`
`_N
`
`II
`
`wherein R4 and R5 signify non—aromatic hydrocarbyl groups,
`
`which may be the same or different and which together
`
`contain at least three carbon atoms, preferably at least
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 3
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 3
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`

`
`W0 94/ 1 1337
`
`PCI‘/SE93/00927
`
`four carbon atoms, especially at least five carbon atoms,
`
`and wherein R4 and R5 may form a ring together with the
`
`amine nitrogen, said ring preferably having no other
`
`heteroatom than the amine nitrogen.
`
`5
`
`The compounds of formula I can form salts with
`physiologically acceptable acids, organic and inorganic,
`
`‘[5
`
`and the invention comprises the free bases as well as the
`
`salts thereof. Examples of such acid addition salts
`
`include the hydrochloride, hydrobromide, hydrogen
`
`10
`
`fumarate, and the like.
`
`When the novel compounds are in the form of optical
`
`isomers,
`
`the invention comprises the racemic mixture as
`
`well as the individual isomers as such.
`
`In the compounds of formula I, R2 is preferably
`
`15
`
`hydrogen, and R3 is preferably hydrogen or hydroxy.
`
`R2 is preferably in 3-, 4- or 5—position.
`
`R3 is preferably in 2-position with respect to the
`
`propylamine group.
`
`20
`
`The HOCH2—group is preferably in 5-position.
`Preferably, each of R4 and R5 independently signifies
`C1_8-alkyl, especially C1_6-alkyl, or adamantyl, R4 and R5
`
`together comprising at least three, preferably at least
`four carbon atoms. R4 and R5 may carry one or more hydroxy
`groups, and they may be joined to form a ring together
`
`25 with the amine nitrogen atom.
`
`Presently preferred tertiary amino groups X in
`
`formula I include the following groups a) — h):
`
`30
`
`a)
`
`—N:
`
`CH(CH )
`32,
`CH(CH3)2
`
`b)
`
`-N:
`
`CH
`
`3
`C(CH3>3
`
`,
`
`C)
`
`/CH
`-N\
`
`3
`UCH332
`
`CH CH»
`2
`3
`
`35
`
`d)
`
`CH
`CH
`3
`3
`\C{__cH
`/
`_N
`\
`C—-—CH
`/ \
`CH3
`
`CH3
`
`2
`
`2
`
`’
`
`e)
`
`CH
`CH
`3 /
`3
`\c-——— H2
`/
`—N
`CH2 ,
`\ _._ /
`CH2
`C
`CH \CH
`3
`
`3
`
`
`
`f)
`
`*
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 4
`
`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 4
`
`

`
`wo 94/11337
`
`PC1"/SE93/00927
`
`g)
`
`5
`
`CH ——CH
`/ 2
`
`2
`
`-N
`\\cH2__CH7
`-
`
`3
`
`r
`
`h’
`
`xi/
`" \
`
`CH "" H2
`2
`CH
`/ 2
`CH2-CH2
`
`Preferably, R4 and R5 are both isopropyl.
`
`A presently preferred specific compound of formula I
`
`is N,N-diisopropyl-3-(2-hydroxy-5—hydroxymethylphenyl)-3-
`
`10
`
`phenylpropylamine.
`
`The compounds of formula I may,
`
`in accordance with
`
`the present invention, be prepared by per se conventional
`
`methods, and especially by
`
`a)
`
`reducing the group R5CO in a 3,3-diphenylpropylamine
`
`15
`
`of formula III
`
`R6CO
`
`III
`
`20
`
`25
`
`A30
`
`35
`
`wherein R1 to R3 and X are as defined above, R5 is
`
`hydrogen or R70, where R7 is hydrogen,
`
`(preferably lower)
`
`alkyl, alkenyl, alkynyl or aryl
`
`(such as phenyl) and any
`
`hydroxy groups may be protected, such as by methylation or
`
`benzylation, or
`
`b)
`
`reacting a reactively esterified 3,3—diphenylpropanol
`
`of formula IV
`
`HOCH2
`
`\Q—oR1
`
`IV
`
`wherein R1 to R3 are as defined above and any hydroxy
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 5
`
`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 5
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`

`
`W0 94/1 1337
`
`PCT/SE93/00927
`
`groups may be protected, and wherein Y is a leaving group,
`
`preferably halogen or an alkyl or arylsulphonyloxy group,
`
`with an amine of formula V
`
`H - X
`
`V
`
`wherein X is as defined above, or
`
`c)
`
`reducing a 3,3—diphenylpropionamide of formula VI
`
`HOCH2
`
`QORI
`CH-CH,-CO—X
`
`V1
`
`wherein R1 to R3 and X are as defined above and any
`
`hydroxy groups may be protected, preferably using a
`
`complex metal hydride, or
`
`d)
`
`N-methylating a secondary 3,3-diphenylpropylamine of
`
`formula VII
`
`\@—OR1
`\CH-CH2-CH2-NH-Z
`Q19
`
`2
`
`VII
`
`wherein R1 to R3 and X are as defined above and any
`
`hydroxy groups may be protected, and wherein Z has the
`
`same meaning as R4 and R5 with the exception of methyl, Z
`preferably being a hydrocarbyl group comprising at least
`
`three carbon atoms,
`
`the N—methylation preferably being
`
`carried out using formaldehyde or formic acid, or
`
`10
`
`15
`
`20
`
`25
`
`30
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 6
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 6
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`

`
`W0 94/ 11337
`
`PCT/SE93/00927
`
`e)
`
`reducing a 3,3-diphenylpropenamine of formula VIIIa
`
`or a 3,3-diphenylpropylamine of formula VIIIb
`
`C=CH-CH2-X
`ER3
`
`VIIIa
`
`R2
`
`W
`
`3
`
`R
`
`VIIIb
`
`R2
`
`10 wherein R1 to R3 and X are as defined above and any
`
`hydroxy groups may be protected, and W signifies a hydroxy
`
`group or a halogen atom, preferably by means of catalytic
`
`hydrogenation,
`
`f)
`
`reacting a 3,3—diphenylpropylamine of formula IX
`
`15
`
`20
`
`HalMg
`
`I
`
`.
`\ . hm
`/C:-Z-an-:2-C.-‘.1,-X
`2}§o‘~R3
`
`R
`
`IX
`
`wherein R1 to R3 and X are as defined above, and Hal is
`
`halogen, with formaldehyde or a formaldehyde equivalent
`
`(such as s-trioxane), or
`
`25
`
`g)
`
`oxidizing the methyl group of a diphenylpropylamine
`
`of formula X
`
`CH3‘
`
`30
`
`-..
`\,.. ,._.
`/\.f‘1-\..._,-\.:‘:..4~X
`2 /:,-“\ Q3
`"
`"
`R/‘
`
`X
`
`wherein R1 to R3 and X are as defined above, and
`
`35
`
`i)
`
`when necessary splitting off hydroxy protecting
`
`groups in the compounds obtained, if desired after mono-
`
`or di—halogenation of one or both of the phenyl rings,
`
`and/or
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 7
`
`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 7
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`

`
`W0 94/ 1 1337
`
`PCHYSE93/00927
`
`ii)
`
`if desired converting the obtained bases of formula I
`
`into salts thereof with physiologically acceptable acids,
`
`or vice versa, and/or
`
`iii) if desired separating an obtained mixture of optical
`
`isomers into the individual enantiomers, and/or
`
`iv)
`
`if desired methylating an ortho—hydroxy group in an
`
`obtained compound of formula I, wherein R1 is hydrogen
`
`and/or R3 is hydroxy.
`
`The oxidation in process g) above may be performed
`
`10
`
`chemically, electrochemically or enzymatically. Chemical
`
`oxidation is advantageously performed using a metal salt
`
`or oxide like ceric ammonium nitrate, manganese oxides,
`
`chromium oxides, vanadinium oxides, cobalt acetate,
`
`aluminium oxide, bismuth molybdate or combinations
`
`15
`
`thereof. Chemical oxidation may also be effected by
`
`peracids, with or without a catalyst, or with halides.
`Electrochemical oxidation may be conducted with or without
`
`a catalyst. For enzymatical oxidation, it is preferred to
`
`use bacteria or yeast (e.g. Candida Guilliermondi, Candida
`
`20
`
`Tropicalis).
`
`The removal of hydroxy protecting groups according to
`
`i) above can e.g. be done by treatment with hydrobromic
`
`acid, borontribromide or by catalytic hydrogenation.
`
`The separation of mixtures of optical isomers,
`
`according to ii) above,
`
`into the individual enantiomers
`
`can e.g. be achieved by fractional crystallization of
`
`salts with chiral acids or by chromatographic separation
`
`on chiral columns.
`
`The starting compounds of formula III and IX may be
`
`prepared as described in the preparation example described
`
`below. The starting materials used in processes b)
`
`to e)
`
`and g) may be prepared as described in the afore-mentioned
`
`WO 89/06644 (the disclosure of which is incorporated by
`
`reference herein) with due consideration of the disclosure
`
`in the present preparation example.
`
`In accordance with the present invention,
`
`the
`
`compounds of formula I,
`
`in the form of free bases or salts
`
`with physiologically acceptable acids, can be brought into
`
`25
`
`30
`
`35
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 8
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 8
`
`

`
`W0 94/ 11337
`
`PCIYSE93/00927
`
`suitable galenic forms, such as compositions for oral use,
`
`for injection, for nasal spray administration or the like,
`
`in accordance with accepted pharmaceutical procedures.
`
`Such pharmaceutical compositions according to the
`
`invention comprise an effective amount of the compounds of
`
`formula I in association with compatible pharmaceutically
`
`acceptable carrier materials, or diluents, as is well
`
`known in the art. The carriers may be any inert material,
`
`organic or inorganic, suitable for enteral, percutaneous
`
`or parenteral administration, such as: water, gelatin, gum
`
`arabicum,
`
`lactose, microcrystalline cellulose, starch,
`
`sodium starch glycolate, calcium hydrogen phosphate,
`
`magnesium stearate,
`
`talcum, colloidal silicon dioxide, and
`
`the like. Such compositions may also contain other
`
`pharmaceutically active agents, and conventional
`
`additives,
`
`such as stabilizers, wetting agents,
`
`emulsifiers, flavouring agents,
`
`buffers,
`
`and the like.
`
`The compositions according to the invention can e.g.
`
`be made up in solid or liquid form for oral
`
`administration, such as tablets, capsules, powders,
`
`syrups,
`
`elixirs and the like,
`
`in the form of sterile
`
`solutions, suspensions or emulsions for parenteral
`
`administration,
`
`and the like.
`
`The compounds and compositions can, as mentioned
`
`above, be used for the same therapeutical indications as
`
`the compounds of the above-mentioned WO 89/06644,
`
`i.e. for
`
`the treatment of acetylcholine—mediated disorders, such as
`
`10
`
`15
`
`20
`
`25
`
`urinary incontinence. The dosage of the specific compound
`
`the mode of
`will vary depending on its potency,
`administration,
`the age and weight of the patient and the
`
`30
`
`severity of the condition to be treated. The daily dosage
`
`may, for example,
`
`range from about 0.01 mg to about 4 mg
`
`per kilo of body weight, administered singly or multiply
`
`in doses e.g.
`
`from about 0,05 mg to about 200 mg each.
`
`35
`
`The invention will be further illustrated by the
`
`following non-limiting example and pharmacological tests.
`
`Reference will be made to the accompanying drawing where
`
`the only figure (Fig. 1)
`
`shows bladder pressure inhibition
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 9
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 9
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`

`
`W0 94/1 1337
`
`PCHVSE93/00927
`
`curves for a compound of the present invention and a prior
`
`art compound, respectively.
`
`General
`
`N.M.R data were acquired on a Jeol JNM-EX 270 Fourier
`
`transform spectrometer. Spectra were recorded with
`
`tetramethylsilane (TMS)
`
`as internal standard at 30°C.
`
`Infrared spectra were recorded on a Perkin Elmer 599B
`
`instrument. Non-corrected melting points were obtained on
`
`a Koeffler apparatus. Gas chromatography was performed on
`
`10
`
`a HP 5940 instrument with a 10 m HP-1 column and the oven
`
`heated in the linear temperature gradient mode.
`
`EXAMPLE 1
`
`(+)—N,N-Diisopropyl-3-(2-hydroxy-5—hydroxymethylphenyl)-3-
`
`15
`
`phenylpropylamine (+) mandelate, and (-)-N,N—diisopropyl—
`3-(2—hydroxy—5—hydroxymethylphenyl)-3-phenylpropylamine
`
`(-) mandelate
`
`a) 6-Bromo—4-phenyl—3,4—dihydro-coumarine
`
`A solution of p-bromophenol
`
`(138 g, 0.8 mole),
`
`cinnamic acid (148 g,
`
`1.0 mole), acetic acid (200 g) and
`
`20
`
`conc. sulfuric acid was refluxed for 2 h. Volatile
`
`25
`
`30
`
`35
`
`material was distilled at reduced pressure. The residual
`
`syrup was cooled and triturated with cold water, giving a
`
`semi-crystalline mass. This was washed extensively with
`
`water, saturated sodium carbonate and finally with water
`
`again. The material was filtered through a sintered glass
`
`funnel, and then mixed with an equal weight of ethanol.
`
`The slurry was stirred at room temperature for 1 h and
`
`then filtered. The resulting product was washed briefly
`
`with ethanol and then diisopropyl ether. After drying, 135
`
`g (55.7%) of the title compound was isolated as white
`
`crystals, melting at 117°C.
`
`b) Methyl 3-(2-benzyloxy-5-bromophenyl)—3-phenyl-
`
`propanonate
`
`6—Bromo—4-phenyl-3,4-dihydro—coumarine (290 g, 0.96
`
`mole) was dissolved in a mixture of methanol
`
`(1 L) and
`
`acetone (1 L). To the above solution were added potassium
`
`carbonate (160 g,
`
`1.16 mole),
`
`a—chlorotoluene (140 g, 1.1
`
`mole) and sodium iodide (30 g, 0.47 mole), and the mixture
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 10
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 10
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`

`
`W0 94/ 1 1337
`
`PCJVSE93/00927
`
`was stirred under reflux for 3 h. The solution was
`
`concentrated by distillation, and the residue treated with
`
`water and extracted with diethyl ether. The ethereal layer
`
`was washed with water, saturated sodium carbonate solution
`
`and water, successively. The organic layer was dried over
`
`sodium sulfate, filtered
`
`and then evaporated to give 420
`
`g (zlO0%) of the title compound as a light yellow oil.
`
`c) 3- 2-benz lox -5-bromo hen l -3- hen l ro anol
`
`Methyl 3-(2—benzyloxy-5—bromophenyl)-3-
`
`phenylpropanonate (112 g, 0.26 mole) was dissolved in
`
`tetrahydrofuran (250 mL) and added dropwise under nitrogen
`
`atmosphere to a suspension of lithium aluminiumhydride
`
`(5.9 g, 0.16 mole)
`
`in tetrahydrofuran (250 mL). The
`
`mixture was stirred overnight under nitrogen atmosphere.
`
`The excess hydride was decomposed by addition of a small
`
`amount of HCl
`
`(aq,
`
`2 M). The solution was filtered on a
`
`pad of Celatom, and the solids were washed thoroughly with
`
`ether. The combined ethereal solution was washed with HCl
`
`10
`
`15
`
`(2 M), water, sodium hydroxide (2 M) and then with water
`again. The organic solution was dried over sodium sulfate,
`
`20
`
`filtered and evaporated to give 98.5 g (95%) of the title
`
`compound as a colourless oil. A small fraction of the oil
`
`was crystallized from diisopropyl ether/petroleum ether
`
`giving crystals which melted at 70°C.
`
`25
`
`d) 3-(2-benzyloxy-5-bromophenyl)—3-pheny1propyl-p-
`
`toluenesulfonate
`
`To a solution of 3-(2-benzyloxy-5-bromophenyl)-3-
`
`phenylpropanol
`
`(107 g, 0.24 mole)
`
`in dichloromethane (300
`
`mL) and pyridine (75 mL) at 0°C was added p-toluene
`
`sulfonylchloride (57 g, 0.3 mole).
`
`The solution was
`
`stirred at 0°C overnight and then evaporated at reduced
`
`pressure and at a bath temperature below 50°C. The
`
`remainder was poured onto water and then the mixture was
`
`extracted with diethyl ether. The organic layer was washed
`
`with water, HCl
`
`(2 M) and water successively, and finally
`
`dried over sodium sulfate. After filtration the ethereal
`
`30
`
`35
`
`solution was evaporated at a bath temperature of <50°C
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 11
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 11
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`
`10
`
`giving 137 g (z100%) of 3-(2—benzyloxy-5-bromophenyl)-35
`
`phenylpropyl-p-toluenesulfonate as a pale yellow oil.
`
`N N-Diiso ro 1-3- 2-benz lox -5—bromo hen 1 -3-
`
`phenylpropylamine
`
`3-(2—benzyloxy-5-bromophenyl)—3—phenylpropyl—p-
`
`toluenesulfonate (115 g, 0.2 mole) was dissolved in a
`
`mixture of acetonitrile (150 g) and diisopropylamine (202
`
`g, 2.0 mole) and the mixture was refluxed for 4 days. The
`
`solution was evaporated, and to the resulting syrup was
`
`added sodium hydroxide (2 M, 200 mL). The mixture was
`
`concentrated, cooled and then extracted with diethyl
`
`ether. The ethereal layer was extensively washed with
`
`water. The amine was extracted with excess sulfuric acid
`
`(1 M). The aqueous layer was washed with diethyl ether and
`
`then basified with sodium hydroxide (11 M). The mixture
`
`was then extracted with diethyl ether. The organic layer
`
`was washed with water, dried over sodium sulfate, filtered
`
`and then evaporated to give 78.6 g (78%) of N,N—
`
`diisopropyl—3—(2-benzyloxy-5—bromophenyl)-3-
`
`phenylpropylamine as a pale yellow oil. The 1-H N.M.R
`
`spectrum was in accordance with the above structure.
`
`f) Resolution
`
`To a solution of N,N-diisopropyl-3-(2-benzy1oxy-5-
`
`bromophenyl)-3—phenylpropylamine (255 g, 0.53 mole)
`
`in
`
`ethanol
`
`(750 g) was added L-(+)-tartaric acid (80 g, 0.53
`
`mole). When all material was dissolved, diethyl ether (90
`
`g) was added and crystallization commenced. After being
`
`stored at room temperature overnight,
`
`the formed salts
`
`were filtered off, washed with fresh ethanol-diethyl ether
`
`solution (2:1) and dried to give 98 g of white crystals
`
`melting at 156°C.
`
`[a]23= 16.3“ (c
`
`5.1, ethanol)
`
`The mother liquor from the precipitation with L-(+)—
`
`tartaric acid was evaporated. The resulting syrup was
`
`treated with sodium hydroxide (2 M) and extracted with
`
`diethyl ether. The organic phase was washed with water,
`
`dried over sodium sulfate, filtered and then evaporated,
`
`giving 170 g of free base. The base (170 g, 0.35 mole) was
`
`dissolved in ethanol
`
`(500 mL), and D-(-)-tartaric acid (53
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
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`11
`
`g, 0.53 mole) was added. When all had dissolved, diethyl
`
`ether (50 mL) was added and crystallization commenced. The
`
`crystals were filtered off and washed with fresh ethanol-
`
`diethyl ether solution giving 105 g of crystals melting at
`
`154-155°C.
`
`[a]22 = -16.4° ( c = 5.0, methanol)
`
`The mother liquor was concentrated, basified and
`
`treated as above, yielding 80 g of free base. This base
`
`was dissolved in ethanol, and treated with L-(+)-tartaric
`acid as described above, yielding additional 20 g of the
`
`dextrorotatory form of the salt.
`
`(M.p. 156°C).
`
`In an analogous manner, 20 g of the levorotatory form
`
`could be obtained.
`
`The pooled dextrorotatory form was dissolved in water
`and basified with sodium hydroxide (2 M). The mixture was
`then extracted with diethyl ether. The organic phase was
`
`washed with water, dried over sodium sulfate,
`
`filtered and
`
`finally evaporated to give the chiral amine (88 g) as a
`
`colourless oil.
`
`[a]22 = 16.3“ (c = 5.1, ethanol)
`
`10
`
`15
`
`In an analogous fashion,
`
`the levorotatory base was
`
`20
`
`obtained (90 g).
`
`[a]22 = -16.l° (c = 4.2, ethanol). The
`
`optical purity as assessed by chromatography was >99%.
`
`gl)
`
`+ -N N-Diiso ro
`
`l-3- 2-benz lox -5-carbox hen 1 -3-
`
`phenylpropylamine hydrochloride
`
`A mixture of magnesium (12.2 g, 0.5 mole), ethyl
`
`bromide (2 g), and iodine (a small crystal) in dry diethyl
`
`ether (200 mL) was warmed until the reaction started.
`
`(+)-
`
`N,N-diisopropyl-3-(2-benzyloxy—5-bromophenyl)-3-
`
`phenylpropylamine (45.6 g, 0.095 mole) and ethyl bromide
`
`(32.7 g, 0.3 mole) dissolved in dry diethyl ether (250 mL)
`
`were then added dropwise under nitrogen atmosphere. The
`
`mixture was refluxed for 1.5 h and then cooled in an
`
`acetone/dry-ice bath, whereupon powdered dry ice (zloo g)
`
`was added gently. Tetrahydrofuran was added when needed to
`
`prevent the mixture from solidification. The reaction
`
`mixture was stirred for 0.5 h when ammonium chloride (200
`
`25
`
`30
`
`35
`
`mL,
`
`20% w/w) was added. The mixture was stirred vigorously
`and then
`
`until two transparent phases were formed,
`
`filtered through a pad of Celatom. The aqueous layer was
`
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`12
`
`washed with diethyl ether and then acidified with
`
`hydrochloric acid to pH 1. The precipitated semi-
`
`crystalline gum was washed with water, and then
`
`transferred to a round bottom flask. The product was dried
`
`5
`
`toluene,
`by co-evaporation with acetone, benzene,
`diisopropyl ether and methanol, successively. The title
`compound (35.1 g, 77%) was isolated as friable shiny
`
`flakes and used without any further purification.
`
`g2)
`
`hen l -3-
`l-3- 2-benz lox -5—carbox
`'
`"
`
`
`10
`
`phenylpropylamine hydrochloride
`
`This product was isolated in 81 % yield in a
`
`corresponding way as described above from (-)-N,N-
`
`diisopropy1-3-(2-benzyloxy-5—bromophenyl)-3-
`
`phenylpropylamine.
`
`15
`
`hl)
`
`(+)-N,N—Diisopropyl—3-(2-benzyloxy—5-carbomethoxy-
`
`phenyl)—3—phenylpropylamine
`
`(+)-N,N-Diisopropyl-3-(2-benzyloxy—5-carboxyphenyl)-
`
`3-phenylpropylamine (34 g, 0.07 mole) was dissolved in
`
`methanol
`
`(300 mL) containing sulfuric acid (6 g) and
`
`20
`
`refluxed for 6 h. The solution was then cooled and
`
`concentrated. To the mixture were added ice-water and a‘
`
`slight excess of saturated sodium carbonate solution. The
`
`mixture was then extracted with diethyl ether. The organic
`phase was washed with water, dried over sodium sulfate,
`
`25
`
`filtered and evaporated, giving 30 g (93%) of crude ester.
`
`Recrystallisation from diisopropyl ether gave white
`
`crystals melting at 85-86°C. The 1-H N.M.R. spectrum was
`
`in accordance with the above structure.
`
`h2)
`
`- —N N-diiso ro l-3- 2-benz lox -5—carbomethox —
`
`30
`
`phenyl)-3—phenylpropylamine
`
`The title compound was obtained from (-)-N,N-
`
`diisopropyl-3-(2-benzyloxy-5-carboxyphenyl)-3-
`
`phenylpropylamine in a similar manner as described above
`
`for the dextro isomer in a 93 % yield.
`
`-
`
`35
`
`i1)
`
`(-)-N,N-Diisopropyl—3-(2-benzyloxy-5-hydroxymethyl-
`
`phenyl)-3—phenylpropylamine
`
`(+)—N,N-Diisopropyl—3-(2-benzyloxy—5—
`
`carbomethoxyphenyl)-3-phenylpropylamine (30 g, 0.065 mole)
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 14
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`
`dissolved in diethyl ether (250 mL) was added dropwise
`
`under nitrogen to a suspension of lithium aluminiumhydride
`
`(1.9 g, 0.05 mole)
`
`in dry diethyl ether (150 mL). The
`
`mixture was stirred overnight at room temperature, and the
`
`excess hydride was decomposed by the addition of water (z5
`
`g). The mixture was stirred for 10 min, when sodium
`
`sulfate (s) was added. After stirring for 20 minutes,
`
`the
`
`mixture was filtered and then evaporated to give 28.4 g of
`
`‘the title compound as a colourless oil.
`
`10
`
`i2)
`
`+ -N N—Diiso ro l-3- 2-benz lox —5—h drox eth l-
`
`phenyl)-3—phenylpropylamine
`
`The title compound was obtained in an analogous
`
`fashion as described above for the levo isomer from (-)-
`
`N,N-diisopropyl—3-(2—benzyloxy-5-carbomethoxyphenyl)-3-
`
`15
`
`phenylpropylamine.
`
`jl)
`
`+ -N N—Diiso ro l-3- 2—h drox -5-h drox eth 1-
`
`phenyl)-3-phenylpropylammonium (+) mandelate
`
`(+)~N,N-Diisopropyl-3-(2—benzy1oxy-5-hydroxymethyl-
`
`20
`
`25
`
`phenyl)—3—phenylpropylamine (28.2 g, 0.065 mole) was
`
`dissolved in methanol
`
`(300 g). Raney Nickel
`
`(one tea-
`
`spoon) was added and the mixture was hydrogenated at
`
`atmospheric pressure until the theoretical amount of
`
`hydrogen was consumed. The progress of the reaction was
`
`monitored by gas chromatography. The mixture was then
`
`filtered through a pad of Celatom, and the solvent was
`
`removed by evaporation at a bath temperature <50°C. The
`
`resulting oil was dissolved in diethyl ether, and the
`
`ethereal solution was washed with water, dried over sodium
`
`sulfate and evaporated giving 22.2 g of a colourless oil.
`
`30
`
`[a]22 = 16.7°
`
`(c = 4.9, ethanol).
`
`To the above oil, dissolved in 2-propanol
`
`(50 g) was
`
`added S—(+)-mandelic acid (9.6 g, 0.06 mole)
`
`in 2-propanol
`
`(50 g). Dry diethyl ether (50 g) was added, and the
`
`solution was left for several hours. The resulting heavy,
`
`35
`
`white crystals were filtered off and washed with a mixture
`
`of 2-propanol and diethyl ether (1:1 v/v) and then dried,
`
`yielding 25 g of the title compound which melted at 148°C.
`
`[a]22 = 38.3°
`
`(c = 5.1, methanol).
`
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`
`The 1-H N.M.R.
`
`spectrum was in accordance with the
`
`above structure.
`
`Chiral purity as assessed by H.P.L.C. was >99%.
`
`Elementary Anal. Theor.: C: 73.0 H: 8.0 N: 2.8
`
`0:
`
`Found:
`
`C: 72.9 H: 8.1 N: 3.0 O:
`
`16.2
`
`16.5
`
`j2)
`
`(-)—N,N—Diisopropyl—3—(2—hydroxy-5-hydroxymethyl-
`
`phenyl)—3—phenylpropylammonium (-) mandelate
`
`The title compound was obtained from (-)—N,N—
`
`diisopropyl-3-(2—benzyloxy-5-hydroxymethylphenyl)-3-
`
`10
`
`phenylpropylamine in an analogous manner to that described
`
`in jl) above.
`
`Elementary Anal.
`
`Theor.: C:
`
`73.0 H: 8.0 N: 2.8
`
`O:
`
`Found:
`
`C:
`
`73.2 H: 8.1 N: 3.0 O:
`
`16.2
`
`16.5
`
`The free base had an optical rotation of [a]22 =
`
`15
`
`-15.5“ (c = 5.0, ethanol).
`
`The l-(-)—mandelic acid salt had a m.p.
`
`of 147-148°C
`
`and an optical rotation [a]22 = -37.9“ (c = 4.7,
`
`methanol).
`
`20
`
`25
`
`30
`
`The optical purity as assessed by H.P.L.C. was >99 %.
`
`Pharmacology
`
`Pharmacological tests performed with one compound of
`
`the invention and three prior art compounds disclosed in
`
`the above mentioned WO 89/06644 will now be described. The
`
`following compounds were used:
`
`(A)
`
`(+)N,N—diisopropyl-3—(2—hydroxy—5-methylphenyl)-3-
`
`phenylpropylamine, hydrochloride (W0 89/06644):
`
`(B) N,N-diisopropyl—3-bis-(2-hydroxyphenyl)propylamine
`
`hydrochloride (WO 89/06644);
`
`(C)
`
`(+)N,N—diisopropyl-3-(5-chloro-2-hydroxyphenyl)-3-(2-
`
`hydroxyphenylpropylamine, hydrochloride (W0 89/06644);
`
`(D) N,N-diisopropyl-3-(2—hydroxy—5—hydroxymethylphenyl)-3-
`
`phenylpropylamine (-) mandelic acid salt (Example 1
`
`above).
`
`Raised index numerals in the text below refer to
`
`35
`
`literature references listed at the end of the
`
`description.
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 16
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`
`Muscarinic Receptor Binding Studies
`
`The tissue preparations and the general methods used
`
`have been described in detail elsewhere for the parotid
`
`glandl, urinary bladderz, heart3 and cerebral cortex3,
`
`respectively. Male guinea pigs (250-400 g body weight)
`
`were killed byia blow on the neck and exsanguinated. The
`
`brain was placed on ice for dissection of the cerebral
`
`cortex (grey matter only). Urinary bladders, hearts and
`
`parotid glands were dissected in a Krebs-Henseleit buffer
`
`(pH 7.4) containing 1 mM phenyl methyl sulfonyl fluoride
`
`(PMSF, a protease inhibitor). Dissected tissues were
`
`homogenized in an ice-cold sodium-potassium phosphate
`
`buffer (50 mM, pH 7.4) containing 1 mM PMSF, using a
`
`Polytron PT—1O instrument (bladder, heart, parotid) and a
`
`Potter—Elvehjem Teflon homogenizer (cortex). All
`
`homogenates were finally diluted with the ice-cold
`
`phosphate/PMSF buffer to a final protein concentration of
`
`S 0.3 mg/ml and immediately used in the receptor binding
`
`10
`
`15
`
`assays. Protein was determined by the method of Lowry et
`
`20
`
`al.
`
`(1951)4, using bovine serum albumin as the standard.
`
`The muscarinic receptor affinities of the unlabelled
`compounds A to 9 identified above were derived from.
`
`competition experiments in which the ability to inhibit
`the receptor specific binding of (-)3H-QNB (1-
`
`25
`
`quinuclidinyl[phenyl—4-3H]benzilate, 32.9 Ci/mmole) was
`
`monitored as previously described3r5. Each sample
`
`contained 10 ul of (-)3H-QNB (final concentration 2 nM),
`
`10 pl solution of test compound and 1.0 ml tissue
`
`homogenate. Triplicate samples were incubated under
`
`30
`
`35
`
`conditions of equilibrium, i.e., at 25°C for 60 minutes
`
`(urinary bladder), 80 minutes (heart and cerebral cortex)
`
`or 210 minutes (parotid gland), respectively. Non-specific
`
`binding was determined in the presence of 10 MM unlabelled
`
`atropine. Incubations were terminated by centrifugationz,
`
`and the radioactivity in the pellets was determined by
`
`liquid scintillation spectrometryz.
`
`IC50-values (concentration of unlabelled compound
`producing 50% inhibition of the receptor specific (—)3H—
`
`Petitioner Mylan Pharmaceuticals Inc. — Exhibit 1005 — Page 17
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`Petitioner Mylan Pharmaceuticals Inc. - Exhibit 1005 - Page 17
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`
`16
`
`QNB binding) were graphically determined from the
`
`experimental concentration-inhibition curves. Affinities,
`
`expressed as the dissociation constants Ki, were
`
`calculated by correcting the IC50 for the radioligand-
`
`induced parallel shift and differences in receptor
`
`concentration, using the method of Jacobs et al.
`
`(1975)5.
`
`The binding parameters for (-)3H-QNB (KD and receptor
`
`densities) used in these calculations were determined in
`
`separate series of experiments1'3. The Ki values obtained
`
`for bladder, heart, parotid and cortex, respectively, are
`
`presented in Table 1 below.
`
`Functional
`
`in vitro studies
`
`Male guinea pigs, weighing about 300 g, were killed
`
`by a blow on the neck and exsanguinated. Smooth muscle
`
`strips of the urinary bladder were dissected in a Krebs-
`
`Henseleit solution (pH 7.4). The strip preparations were
`
`vertically mounted between two hooks in thermostatically"
`
`controlled (37°C) organ baths (5 ml). one of the hooks was
`
`10
`
`15
`
`20
`
`adjustable and connected to a force transducer (FT 03,
`Grass Instruments). The Krebs-Henseleit solution was
`
`25
`
`30
`
`35
`
`continuously bubbled with carbogen gas (93.5% O2/6.5% CO2)
`
`to maintain the pH at 7.4. Isometric tension was recorded
`
`by a Grass Polygr