United States Patent 15
`Karjalainen et al.
`
`[11] Patent Number:
`
`[45] Date of Patent:
`
`4,544,664
`* Oct. 1, 1985
`
`[54] ANTIHYPERTENSIVE SUBSTITUTED
`IMIDAZOLE DERIVATIVES
`
`Attorney, Agent, or Firm—Armstrong, Nikaido,
`Marmelstein & Kubovecik
`
`[75]
`
`Inventors: Arto J. Karjalainen; Kauko O. A.
`Kurkela, both of Oulu, Finland
`
`[73] Assignee:
`{*] Notice:
`
`Farmos Group, Ltd., Turku, Finland
`The portion of the term ofthis patent
`subsequent to Jun. 8, 1999 has been
`disclaimed.
`
`[21] Appl. No.: 396,000
`(22] Filed:
`Jul. 7, 1982
`
`Foreign Application Priority Data
`[30]
`Jul. 10, 1981 [GB] United Kingdom ................ 8121333
`
`(Sh) Minti Cit cicscccass A61K 31/415; CO7D 233/56;
`CO7D 233/64
`F521, Ss OL, siscsccsercomancceavesesresea 514/396; 514/397;
`514/400; 548/335; 548/336; 548/342
`[58] Field of Search.............. 548/335, 336, 341, 342,
`548/345; 542/458, 400, 468; 424/273 R;
`514/397, 400, 396
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`7/1960 Zaugg et all. -..-.....cscesecreeees 548/342
`2,946,804
`
`6/1982 Karjalainen etal. ..
`«- 542/548
`4,333,947
`4/1984 Karjalainen et al. «0-00. 424/274
`4,443,466
`FOREIGN PATENT DOCUMENTS
`
`0024829
`0034474
`0034473
`
`..
`3/1981 European Pat. Off.
`..
`8/1981 European Pat. Off.
`..........
`8/1981 European Pat. Off.
`OTHER PUBLICATIONS
`
` ” 548/335
`
`Kelley, J., et al., J. Pharm. Sci., 70(3), 341-343, (1981).
`
`ABSTRACT
`[57]
`The invention provides nove] compounds of the for-
`mula:
`
`Ri
`
`Sf
`
`Cc
`
`| Ne
`
`R2
`
`.
`N
`
`N
`|H
`
`R a4
`
`1
`
`or
`
`4 I
`:
`H
`
`=
`
`®
`
`ay
`
`Rs
`Re
`
`R7
`
`R
`
`5
`
`R?
`
`wherein the various substituents are defined herein be-
`low. Processes for the preparation of these compounds
`are described, as are novel pharmaceutical compositions
`comprising at least one of the compounds oftheirsalts.
`The compounds and their non-toxic salts exhibit valu-
`able pharmacological activity and are useful
`in the
`treatment of mammals, especially as antihypertensive
`agents. Furthermore, some of the compounds have
`proved to possess antithrombotic and diuretic activity.
`Antimycotic and antifungal properties have also been
`found.
`
`Primary Examiner—Richard A. Schwartz
`
`12 Claims, No Drawings
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 1
`
`

`

`1
`
`4,544,664
`
`2
`
`ANTIHYPERTENSIVE SUBSTITUTED
`IMIDAZOLE DERIVATIVES
`DESCRIPTION
`
`5
`
`CH
`ee
`
`or
`—CH(—CH2CH2CH2CH3)—
`>C=CH—CH3, >C=CH—CH2CH3,
`
`>C—CH2,
`
`Thepresent invention relates to substituted imidazole
`derivatives and their non-toxic, pharmaceutically ac-
`CH;
`ceptable acid addition salts, and their preparation, to
`pharmaceutical compositions containing the same, and 19 ilon
`to their use.
`Jf
`The imidazole derivatives of the present invention
`have the general formula:
`
`2
`
`> C=CH—CH?2CH2CH3; Rois H, —CH3, —CH2CH3,
`—CH2CH2CH;,
`
`15
`
`@)
`
`Rs
`Re
`
`ni [ ‘eeOF 3
`
`R3
`
`<
`
`N
`
`c
`
`a
`H
`
`CHa
`
`ff
`
`—CH—CH3,
`
`20 _CH»CH2CH2CH; or OH; Rio is H, —CH;,
`—CH2CH3, —CH2CH2CH3,
`
`or
`
`Rs
`
`N en{On
`ni I
`we
`q
`"
`a
`
`R2
`
`(uy 25
`
`#0
`
`CH;
`
`4
`—CH—CH3
`
`or —CH2CH2CH2CH;; Ry};
`—CHCH2CH3,
`aoe
`—cH—cH;
`
`is H, —CH3, —CH2CH3,
`
`.
`‘
`wherein R; is H, an alkyl] of 1 to 4 carbon atoms, e.g.
`‘ or —CH2CH2CH2CH;3;nis 0 to 4; provided that.
`
`
`methyl or —CH2OH; R2is H or CH3; R3 is —CH3, 35
`hi
`is
`OH. Riis Hor
`C
`*
`
`
`
`_—CH>CHs, —CHaCHbCHs, R; isHorwhen Rg is OH, CH; and R;is
`Rs
`
`aeeee
`
`—CH}CH)CH2CH3, —CH;,CH=CH), or
`
`40 {OFR
`
`,
`then Rs, Rg and R7are notall simultaneously hydro-
`gen;
`
`Rs
`
`Re:
`
`R7
`
`.
`
`==CH2, —CH—CH3, —CH—CH2—CH3,
`
`when Rj, Rzand RgallarehydrogenandR;is
`45
`{OF
`and Rs is H or OH;or R3 and Rg together represent 59 {OF
`
`Rs
`Re
`
`R7
`
`CH
`
`Z
`=C—CH3,
`
`or =CH—CH2CH2CH;3; X is
`
`Ri Ryo
`Ro
`Rs
`|
`—CH—CH— or —C=C—;
`
`Rs, Rg, and R3, which can be the sameordifferent are
`H, —CH3, —CH2CH3, halogen, OH or —OCH:or Rs
`is hydrogen and R¢ and R7 together form an —O—CH-
`2—O—bridge between two adjacent carbon atomsin
`the phenyl group; —CHRs— is —CH2—, —-CH(CH-
`3)—, —CH(—CH2CH3)—, —CH(—CH2CH2CH3)—,
`
`55
`
`then Rs, Rs, R7 are not all simultaneously hydrogen;
`Rs and Rg are not simultaneously hydrogen; and
`Rj) and Rygare notsimultancously hydrogen.
`Because of the tautomerism in the imidazole ring the
`compoundsof the general formula I and I are 4(5)-sub-
`stituted imidazole derivatives.
`The non-toxic pharmaceutically acceptable acid addi-
`60 tion salts of these compoundsare also within the scope
`of the invention.
`The compounds of the formula (I) and (ID) form acid
`addition salts with both organic and inorganic acids.
`They can thus form many pharmaceutically usable acid
`addition salts, as, for instance, chlorides, bromides, sul-
`fates, nitrates, phosphates, sulfonates,
`formates,
`tar-
`trates, maleates, citrates, benzoates, salicylates, ascor-
`bates and thelike.
`
`65
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 2
`
`

`

`4
`3
`4-[2-(2,6-dimethylphenyl)-1-methyletheny]]-2-
`Theinvention includes within its scope pharmaceuti-
`cal compositions comprising at least some of the com-
`methylimidazole
`poundsof formula(I) or (II) or a non-toxic, pharmaceu-
`4-[2-(2,6-dimethylphenyl)-1-methyletheny]]-5-
`tically acceptable salt thereof, and a compatible phar-
`methylimidazole
`maceutically
`acceptable carrier therefor.
`3
`.
`The ravition Socriten, for example, the following aiciuentinhoki
`specific compounds of formula (1):
`5-2,
`imethylphenyl)-1-methyl-1-pentenyl-
`4-[a,a-bis(2-methylphenyl)hydroxymethy]Jimidazole
`Jimidazole
`4-[[a-(2-methylphenyl)]-2-methylbenzyl]imidazole
`4-[3-(2,6-dimethylpheny)-1-ethyl-1-propenyl]imidazote
`4-(a-phenylbenzyl)-5-methylimidazole
`10 4-[5-(2,6-dimethylpheny])-1-methyl-1-pentenyl]-5-
`4-[[a-(2,6-dimethylpheny])]-a-methyl]hydroxymethyl-
`methylimidazole
`Jimidazole
`4-[5-(2,6-dimethylpheny])-1-methylpenty]]imidazole
`4-[[a-(2,3-dimethylpheny!)]-a-methy!Jhydroxymethyl-
`4-[4-(2,6-dichlorophenyl)-|-methyl-1-butenyl]imidazole
`Jimidazole
`15 4(2-(2,6-dimethylpheny!)-I-ethylethy]]imidazole
`Pheansshinesottoinerenrensses
`4-[2-(2,6-dimethylphenyl)-2-ethylethyl]imidazole
`4-[[a-(2-methylphenyl)}-2-methylbenzyl}-5-
`4-[2-(3,4-methylenedioxyphenyl)propylimidazole
`methylimidazole
`4-[2-(2-bromo-4,5-methylenedioxypheny|)propy!-
`4-[(a-methyl)-2,6-dimethylbenzyl]imidazole
`Hraiteete
`4-[(a-methyl)-2,3-dimethylbenzyl]imidazole
`The compoundsof the present invention have been
`4-[(a-ethyl)-3-methylbenzyl]imidazole
`found to possess excellent antihypertensive activity.
`4-[(a-butyl)-2,3-dimethylbenzyl]imidazole
`Preliminary tests have shown that
`they also possess
`4-[(a-methyl)-2,3-dimethylbenzyl}-2-methylimidazole
`other valuable pharmacological properties, for exam-
`4-[(a-propyl)-2-methylbenzyllimidazole
`35 ple, antithrombotic and diuretic effect. Antimycotic and
`4-[{(a-methyl)-2-methylbenzy]]imidazole
`antifungal properties have also been found.
`Hens)ee While all of the compounds of formula (I) and (ID
`aan ay nS ee anya
`essentially satisfy the objectives of the present inven-
`4-[a-butyl-a-(2,3-dimethylphenyl)-hydroxymethy!-
`- tion, certain groups of compounds remain preferred.
`Jimidazole
`Onesuch preferred group is represented by formula (I)
`4-[a-methy]-a-(2,3-dimethylphenyl)-hydroxymethy]]-2-
`wherein Ra is hydrogen, R3 is alkyl and Rs, Re and Rz7,
`methylimidazole
`whichcan be the sameordifferent, each are hydrogen,
`4-[a-propyl-a-(2-methylpheny!)-hydroxymethyl-
`methyl, ethyl or halogen. Another preferred group of
`Jimidazole
`35 compoundsis represented by formula (II), wherein Rs,
`4-(a-methy]-2-chlorobenzyl)imidazole
`Reand R3, which can be the same ordifferent, each are
`eeee hydrogen, methyl, ethyl or halogen.
`In such com-
`imidazole
`:
`:
`:
`:
`4-[a-methyl-a-(2,5-dimethylphenyl)-hydroxymethyI-
`pounds, those in which Ry fs hydrogen or methyl Ro -
`Jimidazole
`a hydrogen or methyl, Rg or Ry, is methyl, ethyl or iso-
`4-[a,a-bis-(2,3-dimethylphenyl)hydroxymethy|-
`peapyl Bey aad Ban aoe: Hyper eA RO aay he
`Jimidazole
`mentioned. Especially the compounds wherein n is
`4-[a-(2,3-dimethylpheny!)-2,3-dimethylbenzyl-
`greater than 0 possess valuable antimycotic properties.
`Jimidazole
`Especially good antihypertensive properties have been
`4-[(a-ethyl)-2,6-dimethylbenzyl]imidazole
`4s found in compoundsof formula (II) wherein n is O and
`4-[(a-ethyl)-2,3-dimethylbenzy]]imidazole
`X is
`1-(4-imidazolyl)-1-(2,3-dimethylphenylethylene
`1-(4-imidazolyl)-1-(2,6-dimethyIphenylethylene
`1-(4-imidazolyl)-1-(2,3-dimethylphenyl)propene
`1-(4-imidazoly])-!-(2,3-dimethylphenyl)pentene
`The following specific compounds of formula (II):
`4-[2-(2,6-dimethylphenyl)-1-methylethyl]imidazole
`4-{2-(2,6-dimethylphenyl)propyllimidazole
`oeSeeenieeeipenotniienie
`wLe;0~
`iC
`n
`=
`r
`1
`1.12-02.6-dimsthylphenyl)-2-hydroayethy!imidazole
`4-(2-phenylpropyl)imidazole
`4-[2-(2,6-dimethylphenyl)- |-methyletheny!]Jimidazole
`4-[2-(2,6-dimethylphenyl)-1-propenyl]imidazole
`4-[2-(4-chlorophenyl)-|-methylpropyl]imidazole
`4-(2-methyl-4-phenyl-1-buteny])imidazole
`
`According to the feature of the invention, the com-
`pounds of formula (1) wherein Rs is OH and the com-
`poundsofformula (II) are made by a Grignard reaction,
`55:
`‘
`ar
`in which an imidazolylketone of the formula
`9
`N :z
`ni [
`
`4,544,664
`
`5
`
`20
`
`nT re
`—c=c—.
`
`50
`
`60
`
`4-[5-(2,6-dimethylphenyl)- |-methy!-1-penteny|-
`Jimidazole
`4-[3-(2,6-dimethylphenyl)-2-methy]-1-propenyl-
`Jimidazole
`4-[2-(2,6-dimethylphenyl)-1-ethylethenyl]imidazole
`4-[2-(2,3-dimethylphenyl)- 1-methyletheny]]imidazole
`4-[2-(2,6-dimethylphenyl)-!-isopropylethenylJimidazole
`
`Ra
`
`N
`|
`i
`wherein Rj, R2 and Rj are as defined before, is reacted
`with an arylalkyl magnesium halide derivative or aryl
`magnesium halide derivative of the formula:
`
`65
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 3
`
`

`

`6
`
`°I
`
`l
`C—OCH:
`
`R2
`
`N
`
`ri [N
`
`k
`
`5
`
`4,544,664
`
`Rs
`
`Re
`
`Ry
`
`(CH2)n, ~MgHal
`
`wherein Rs, Rg and R7 are as defined before, n’ is 0 to 5
`and Hal is a halogen atom to give compounds ofthe
`formula (IL)
`
`(IIT)
`
`{“
`
`wherein Ry, Ra, R3, Rs, Re, R7 and n’are as before.
`The arylalkylmagnesium halide derivative can be, for
`example, an arylalkyimagnesiumbromide derivative,
`which is prepared by reacting the corresponding ary-
`lalkylbromide derivative with magnesium. Suitable sol-
`vents for the reaction include a variety of ethers, prefer-
`ably tetrahydrofuran. The arylalkylmagnesiumhalide
`derivative is prepared in the usual way by adding the
`arylalkylmagnesiumhalide derivative in a suitable sol-
`vent, e.g. tetrahydrofuran, dropwise onto magnesium
`turnings covered by tetrahydrofuran, at
`the boiling
`point of the reaction mixture. When the magnesium
`turnings have reacted, the mixtureis cooledslightly and
`the 4-imidazole derivative is added in solid form in smal]
`
`portionsorin tetrahydrofurane solution. After the addi-
`tion, the reaction mixture is refluxed until all of the
`4-imidazole derivative has reacted. The reaction time
`varies between one and five hours.
`Anotherprocess for the preparation of compounds of
`formula (III) is a Grignard reaction in which a com-
`poundof the formula (IV)
`
`S
`
`15
`
`20
`
`25
`
`35
`
`wherein Ri and R2 are as before, is reacted in a first
`step/with a Grignard reagent of the formula
`
`Rs.
`
`Re
`
`R7
`
`(CH2),,, —MgHal
`
`wherein Rs, Rg, R7 and n’ are as before, to give a com-
`pound offormula (IV), which in a second step without
`isolation is reacted with a Grignard reagent of the for-
`mula
`
`R3MgHal
`
`wherein R3is as defined before.
`Compounds of formula (I) wherein R4 is H can be
`prepared by reduction of compounds of formula (III)
`wherein n' is 0 with hydrogen. A suitable catalystis e.g.
`palladium-on-carbon.
`Unsaturated compounds of formula (I) wherein R3
`and Ry, are —-CH2, —CH—CH3, —CH—CH2CH3,
`
`CH;
`=C—CH;
`
`or =CH—CH?CH?CH;or formula (II) wherein Ryo is
`hydrogen are prepared by dehydrating compounds of
`formula (III):
`
`45
`
`ay)
`
`50
`
`55
`
`yee
`
`b
`
`(up)
`
`Rs
`
`Ce
`
`°
`
`Rs
`
`tccuy,,{OF
`Nn
`n-~ LR2
`
`|H
`
`wherein R|-R7 and n’ are as before, is reacted with a
`compoundofthe formula
`
`R3MgHal
`
`wherein R3 is an alkyl or aryl as defined before and Hal
`is halogen. Yet another process for the preparation of
`compoundsof formula (III) is a Grignard reaction in
`which an imidazole carboxylic acid alkyl ester, prefera-
`bly the methyl ester of the formula
`
`65
`
`wherein Rj, R2, Rs, Re, R7 are as defined before, R3is
`an alkyl or aryl as defined before and n’is 0 to 5, to give
`a compound of the formula (V)
`
`Ri
`C=CH—(CH2)y
`
`R2
`
`N
`
`Ri< L
`N
`hi
`
`)
`
`Rs
`Rg or
`
`Ry
`
`
`
`Petition for Inter Partes Review of US 8,455,527
`AmnealPharmaceuticals LLC — Exhibit 1004 — Page 4
`
`

`

`a
`scontinued
`8
`
`8
`ri? |
`own
`H
`
`ll
`C—(CH2)
`
`4,544,664
`
`Rs
`
`Ro
`7”
`
`'
`i
`wherein Rj, R2, Rs, Re, R7, and n and n’ are as defined
`before; Ri; is an alkyl as defined before and Rg is an
`alkeny] as defined before.
`The dehydrationis preferably performedbyrefluxing
`in an appropriate acidic solution, e.g. concentrated hy-
`drochloric acid or heating for example with potassium-
`hydrogen sulfate.
`The compounds of formula (V) can further be re-
`duced with hydrogenin the presenceofa palladium-on-
`carbon catalyst
`to the corresponding saturated com-
`pounds of formulae (I) and (11).
`Compoundsof formula (II) wherein Ri is hydrogen
`are prepared by a Wittig reaction which comprises
`reacting an imidazole aldchyde of the formula
`
`N
`ni L
`,
`H
`
`ae
`
`“
`
`*
`
`wherein R, and R2 are as before, with an aralkylidene-
`triphenylphosphorane of the formula:
`
`Rio
`(CgHs)3P=C—(CH2)y
`
`Rs
`Ro
`Ry
`wherein Rs, Re, R7, Rio and n are as defined before, to
`give the unsaturated compoundsof formula (11), which
`in a further step can be reduced to the corresponding
`saturated compounds of formula (II) as described
`above.
`The aralkylidenetriphenylphosphoranes are prefera-
`bly prepared by reacting the corresponding aralkyltri-
`phenylphosphonium halide of the formula:
`Rs
`as
`2
`Re
`(CH2)¢—CH—P—(CeHs); Hal©
`Ry
`
`wherein Rs, Re, R7, Rio and n are as before and Halis
`halogen, with a basic reagent, preferably butyllithium.
`In the Grignard- and Wittig-syntheses described
`above, the free nitrogen atom in the imidazole starting
`material can be protected by different methods. Suitable
`protecting groupsare for example benzyl, triphenylsilyl
`or dialkoxymethane. The removal of the protecting
`group can be performedin different ways, and depends
`on the kind of protecting group used. For example, a
`dialkoxymethane group is removed by acidic hydrolysis
`and a benzyl group by sodium in liquid ammonia.
`The present invention further provides yet another
`method for preparing compounds of the invention.
`
`8
`Thus, according to this embodimentofthe invention, a
`starting material of the formula (V1) or (VID)
`
`Re
`
`Ry
`or
`
`Rs
`
`Re
`Ry
`
`CH—C—C—R3
`Lid14
`Ris
`
`Ri2 Ris
`rot
`(CH2),—X—C—C—R?
`I b
`i Ris
`
`Vv
`
`(vu)
`
`wherein R2, R3, Rs, Rg, R7 and n are as hereinbefore
`defined; wherein R12, Ri3, Rig and Ris, which can be
`the same or different, are each hydrogen, hydroxy,
`mercapto, halogen, amino, —O— alkyl of 1 to 7 carbon
`atomsor
`
`10
`
`15
`
`20
`
`25
`
`i
`-o-c—
`O—-C—R,
`wherein R is an alkyl; or wherein Riz and Rj4 can be
`combined to form a keto group, or R13 and Ris can be
`combined to form a keto group, or both R12and Ry4 and
`30 R43 and Rys can simultaneously form keto groups;
`is
`reacted with a reagent capable of converting said start-
`ing material to the corresponding imidazole of the for-
`mula:
`
`Rs
`CH
`
`R2
`
`N
`
`RX I
`N
`ti
`
`a5
`
`40
`
`45
`
`Rs or
`
`Re
`
`Rz
`
`R aa
`:
`
`N
`
`"
`H
`
`X—(CH2),
`
`|
`
`R2
`
`Rs
`
`Ro
`
`Rr
`
`50
`
`wherein Ri, R2, R3, Rs, Re, R7, X and n are defined as
`before. Reagents capable of converting the depicted
`starting material to the corresponding imdidazole in-
`55 clude NH3+CH20 (or a source of ammonia and form-
`aldehyde);
`
`Il
`|
`Se H—C—O7~—NH4*; HCONH3; Rj—C—NH2;
`Ry
`
`or Ri:CHO and NH3. Choice of an appropriate reagent
`varies with the particular starting material employed.
`When R, is hydrogenit is preferable to employ form-
`amide as the reagent in cases where,
`in place of the
`bromine atom in the aforementioned starting materials,
`there is instead a hydroxyl, amino or acetyl group. In
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 5
`
`

`

`4,544,664
`
`10
`
`¥
`
`Qo-7
`2—
`é T- ( )
`2—O—Zz
`
`whereY is the arylalkylresidue determined by the for-
`mula (I) and (II), R is an alkyl group of 1 to 7 carbon
`atoms or an ary] radical of 6 to 10 carbon atoms.
`Preferably, the hydrolysis is carried out by boiling
`the starting material, an N-acylated imidazole deriva-
`tive, in an aqueoussolution of an inorganic acid until the
`reaction is completed.
`Yet another process for the preparation of the com-
`pounds of formula (I) and (II) comprises hydrogenating
`a starting material of the formula:
`
`5
`
`10
`
`15
`
`20
`
`25
`
`40
`
`45
`
`350
`
`55
`
`9
`these instances, formamideis used in excess and acts in
`part as the solvent. Generally, the reaction is run at the
`boiling point of formamidefor a period of time ranging
`from one to five hours.
`
`Yet another process for the preparation of the com-
`pounds of formula (I) and (II) comprises reacting form-
`amide with a benzene derivative of the formula:
`
`Rs
`Rg
`
`R7
`
`R‘
`Rs
`CH—QorLO)
`
`R7
`
`wherein Rs, R6, R7, Rg, n and X are as defined herein-
`above, and Q is a radical of formula:
`
`Il
`£%;
`ee nea or —CH—C—R2
`R2
`R2
`Hal
`
`wherein R is a substituted and unsubstituted alkyl, ary-
`lalky] or aryl group, and R2, and Hal are as defined
`hereinabove. Preferably the reaction is performed by
`vigorously boiling the benzene derivative in formamide,
`the reaction time varying with the particular material
`employed.
`Reaction times typically are from 30 minutes to 8
`hours. Obviously, the formamide treatment will be fol-
`lowed by reaction with an appropriate acid (¢.g. HCl)
`when Q in thestarting material is
`
`Oo
`0
`|
`Il
`“eeeS
`Ra
`
`in order to obtain the corresponding compoundoffor-
`mula (1) and (II).
`Similarly, when a starting material wherein Q is
`
`iiwer)
`
`R2
`
`is employed, then the formamide treatmentwill be fol-
`lowed by hydrogenation,
`thus affording the desired
`compoundofformula (J) and (II).
`A further process for the preparation of the com-
`pounds of the formula (I) and (II) comprises hydroly-
`sing a corresponding N-acetylated compound of the
`formula (J) and (II)
`
`Y
`
`Ri
`
`RY
`
`! t
`
`N
`
`N
`
`Y
`
`Y
`
`N
`
`| H
`
`rs [ ° ri L or RK [
`N
`Ra
`N
`R2
`N
`77R”
`
`wherein Y is as defined before and R'is an arylor alky]
`and R”is an aryl group. The hydrogenation is conve-
`niently conducted in the presence of a suitable catalyst
`and under a hydrogen atmosphere, with stirring or
`using metallic sodium in liquid ammonia. Suitable cata-
`lysts include platinum oxide, palladium-on-carbon and
`Raney nickel. Reaction temperatures vary with the
`particular starting material employed, with typical tem-
`peratures being 25°-70° C.
`Yet another method for the preparation of the com-
`pounds of formula (I) or (ID) wherein X is
`
`Rg
`Rg
`—CH—CH—
`
`comprises reacting a N-trialkylsilylimidazole of the
`formula
`
`N (
`
`J
`N
`y-$-¥
`
`|Y
`
`wherein Y is an alkyl group, preferably methyl, with an
`arylalkylhalogenide of the formulae
`
`
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 6
`
`

`

`11
`
`Rs
`Rs
`CH—Halor R¢
`
`R7
`
`ms
`R¢
`
`R?
`
`4,544,664
`
`Rg
`Ry
`CH—CH—Hal
`
`12
`wherein Rg is an alkyl as before, in the presence of a
`Lewis acid, for example zinc (II) chloride, to give a
`compoundof the formula (XII)
`
`Rs O
`Re
`(CH2),—CH—CH—C—CH;
`
`(XID
`
`Rs
`Rg
`
`R?
`
`The compound of formula (XII) is further bromi-
`nated as before to give compoundsof the formula (VII).
`When Rg and R32are hydrogen yet another method
`for the preparation of compoundsof the formula (VII)
`can be applied. In this method a halide of the general
`formula (XIID)
`
`Rs.
`Rg
`
`Ra
`
`Ro
`(CH2),—CH—Hal
`
`(XUD
`
`is reacted with lithiated N,N-dimethylhydrazone of
`acetone followed by hydrolysis to give a compound of
`the general formula (XIV)
`
`Rs
`Rg
`
`Ry
`
`°
`Ro
`(CH2),,—CH—CH;C—CH;
`
`(XIV)
`
`which compounds are brominated as before to give
`compoundsofthe formula (VII).
`According to another method for the preparation of
`compounds of the formula (VII), compounds of the
`formula (VIII) are selectively brominated using as bro-
`minating agent for example 2-carboxyethyltriphenyl-
`phosphonium perbromide, which has the formula (XV)
`
`G
`(C6H5)3—PS—CH2CH2,C—OH Br;
`
`(XV)
`
`the preparation of com-
`Yet another method for
`poundsof the formula (VII) is possible via a directed
`aldol condensation, in which for example a compound
`of the formula (XVI)
`
`Rs
`Re
`
`Rr
`
`oO
`lt
`(CH2),—C—R 0
`
`(XVI
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`is reacted with the compound (XD)in the presence of a
`Lewis acid followed by dehydration to give a com-
`pound of the formula (XVID
`
`wherein R3, Rs, Rs, Rr, Rgand Ro are as before and Hal
`is a halogen atom,in the presence of a Lewisacid, for
`example titanium tetrachloride, aluminium chloride or
`zinc chloride. As solvent can be used for example meth-
`ylene chloride or chloroform. The reaction is prefera-
`bly carried out at room temperaturestirring the starting
`materials for 6-12 hours.
`The intermediates of formula (VI) and (VII) can be
`prepared for exampleas follows:
`An aldehyde of the formula
`
`Rs
`
`Ré
`
`(CH2)y—CHOoRa
`
`wherein Rs, Ré, R7 and n are as before, is reacted in
`alkaline or acidic conditions with a ketone, preferably
`acetone, to give a compound of the formula (VIII) via
`direct aldol condensation:
`
`Rs
`Ré
`
`(CH?),-—" CH=C—C—-CH3oRy
`
`Rs O
`
`(VIN)
`
`wherein Rgis an alkyl as defined before, which com-
`pound in a secondstepis catalytically reduced to give
`the corresponding saturated compoundof the formula:
`
`R
`
`Re
`
`(CH2),—CH2—CH=C—CH3oRa
`
`Rs
`|
`
`O
`sil
`
`(Ix)
`
`which compoundin a third step is regioselectively bro-
`minated in methanol to give compoundsof formula VII.
`Another method for the preparation of the com-
`pounds ofthe general formula (VII) is the regioselec-
`tive alkylation process of ketones in whih for example a
`halide compoundof the formula (X)
`
`EO)bc
`
`Rs
`
`Rz
`
`Ro
`I
`
`(X)
`
`is reacted with a trimethylsilylenolether derivative of
`the general formula (XD
`
`OTMS
`Rs—CH=C—CH3
`
`65
`
`(xD)
`
`Rs
`R6
`
`Rr
`
`Rig ku O
`(CH2)y-—-C=C—C—CH2R2
`
`(XVID
`
`
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 7
`
`

`

`4,544,664
`
`13
`which compound is further brominated as before to
`give a compound of the formula (VII).
`When Rj; is hydrogen, compounds of the formula
`(VIDcan be prepared from compounds ofthe formula
`(XVI), wherein these are reacted with 1-lithiated N,N-
`dimethylhydrazone of methylalkylketone of the for-
`mula (XVIII)
`
`5
`
`(XVII)
`
`10
`
`CH;
`4
`N—N—CH;
`LiCH2—C—CH2R>.
`
`14
`
`°
`ll
`C—CHoRio
`
`°
`CH=CH—C—CH2Ri0
`
`(XXILD)
`
`(XXIV)
`
`Rs
`Re
`
`R7
`
`R.
`R6
`R7
`
`In the first step this condensation gives unsaturated
`1s ketones of the formulae (XXV) and (XXVI)
`
`Here in the first step compounds of the formula
`(XIX)are achieved,
`
`Rs
`
`Re
`
`Ry
`
`to
`(CH2)p-—C=CH—C—CH2R2
`
`(XIX)
`
`20
`
`the bromination of which compounds are performed
`following the method above.
`The preparation of compoundsofthe general formula
`(VII) can be accomplished from compounds of the
`general formula (XVII) by hydrogenation of the car-
`bon-carbon bouble bond as well. The brominationin the
`second step leads to compoundsof the formula (VII).
`Alkylation of compounds of the general
`formula
`(XVID when R2 and Rio are hydrogen can be accom-
`plished, too. In this method a compoundofthe formula
`(XX)
`
`Rs.
`Re
`
`Ry
`
`Rs O
`I
`(CH2),—CH=C—C—CH;
`
`(XX)
`
`25
`
`30
`
`35
`
`40
`
`is reacted with an alkylation reagent such as dialkyllithi-
`ocuprate (XXI) which undergoes 1,4-conjugate addi-
`tion
`
`4:
`
`(R)2CuLi
`
`(XX)
`
`50
`
`to form compoundsofthe formula (XII).
`Condensation of an arylalkylketone or its vinylogue
`with 4-imidazole aldehydes of the formula (XXII)
`
`N
`
`nt 1N
`
`CHO
`
`Ra
`
`i
`
`provides further another method for the preparation of
`compoundsaccording this invention. The condensation
`is performed for example in aqueous alcohol catalyzed
`by sodium hydroxide. Arylalkylketones or their viny-
`logues have the general formulae (XXIII) and (XXIV)
`
`(XXII)
`
`55
`
`60
`
`65
`
`.
`Rg
`Ro
`
`Rs
`Ro
`Ry
`
`R2
`rey
`C—C=CH |
`
`\- Ri
`
`(XXV)
`
`}H
`
`»
`(XXVI)
`S
`f ‘ig
`CH=CH—C—C=CH | Sr,
`N|H
`
`which compounds are then hydrogenated to the end
`products according to the formulae (XXVII) and
`(XXVHI)
`
`Rs.
`Re
`Ry
`
`Rs
`Re
`Ry?
`
`2,
`
`R
`Rig
`CH3—CH—CH32,
`
`|
`
`n
`R10
`CH2CH2;CH2CH—CH?
`
`N
`
`*H
`
`2,
`
`(XXVID
`
`\—a,
`
`(XXVHD
`N
`| Sri.
`nH
`
`Yet another method for the preparation of com-
`pounds of formula (1) wherein R4 is H comprises react-
`ing a compound of the formula
`
`~O°“Ce
`nit [
`Re
`Ry
`
`R:
`
`whercin Rj, R2, Rs, Re and Rj are as before and R is an
`alkyl of 1 to 4 carbon atoms with a Grignard reagent of
`the formula
`
`R3a—CH2MgHal
`
`
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 8
`
`

`

`15
`
`4,544,664
`
`in a mixture of tetrahydrofuran and toluene with reflux-
`ing to give a compoundof the formula
`
`Ri
`
`N
`
`N
`H
`
`-
`CH
`
`R2
`
`Rs
`Re
`Ry
`
`Asstated herein above, the compoundsofthe general
`formula (1) and (11) and their non-toxic, pharmaceuti-
`cally acceptable acid addition salts have valuable phar-
`macological properties and have been found to possess
`excellent antihypertensive properties.
`Tests have shown that they also possess other phar-
`macological properties as well, for example, antithrom-
`botic activity. Furthermore, antimycotic and antifungal
`properties have been found, too.
`The processes described above for the preparation of
`compoundsof formula (II) wherein X is
`
`oH Rio
`-—Cc=c—
`
`result mainly in the trans isomer of the compound. The
`trans isomer can be converted to the cis isomer accord-
`ing to known methods,e.g. by heating it in the presence
`of an acid or byirradiating it with ultraviolet light.
`Administration of isomeric compoundsof formula (I)
`and (II), their non-toxic, pharmaceutically acceptable
`salts or mixtures thereof may be achieved parenterally,
`intravenously or orally. Typically, an effective amount
`of the derivative is combined with a suitable pharma-
`ceutical carrier. As used herein,
`the term “effective
`amount” encompasses those amounts which yield the
`desired activity without causing adverse side-effects.
`The precise amount employedin a particularsituationis
`dependent upon numerous factors such as method of
`administration, type of mammal, condition for which
`the derivative is administered, etc., and of course the
`structure of the derivative.
`The pharmaceutical carriers which are typically em-
`ployed with the derivatives of the present invention
`may besolid or liquid and are generally selected with
`the planned mannerof administration in mind. Thus,for
`example, solid carriers include lactose, sucrose, gelatin
`and agar, while liquid carriers include water, syrup,
`peanutoil and olive oil. Other suitable carriers are well-
`known to those skilled in the art of pharmaceutical
`formulations. The combination ofthe derivative and the
`carrier may be fashioned into numerous acceptable
`forms, such as tablets, capsules, suppositories, solutions,
`emulsions, and powders.
`The anti-hypertensive propertics of the imidazole
`derivatives of the present invention have been deter-
`mined by the following procedure. Sprague-Dawley
`rats of normal weight were first anesthetized with ure-
`thane. After this, the femoral artery was connected by
`way of a polyethylene tube with a blood pressure trans-
`ducer. The test substance was then injected into the
`femoral vein or given intraperitoneally and the blood
`pressure and the pulse frequency were registered with a
`recorder.
`
`16
`In a further test for anti-hypertensive properties un-
`anesthetized Wistar
`spontaneous hypertensive rats
`(SHR) were used. The test derivative was administered
`perorally by way of a tube into the stomach. The blood
`5 pressure was measured from thetail using an indirect
`bloodless method.
`The diuretic activity was studied in rats by collecting
`the urine output during 0-5 hoursafter i.p. injection of
`;
`.
`the compounds. Before the test the animals were fasting
`10 overnight and got 10 ml water p.o. immediately before
`the injection.
`The antithrombotic activity was investigated in vitro.
`The inhibiting activity of the compounds against ADP-
`and collagen-induced aggregation of thrombocytes was
`measured. In the test thrombocytes from a cow was
`used. To 1.2 ml! of plasma containing 250000 throm-
`bocytes/mm? were added 50 ul of a solution of the
`compound to be tested. After 10 min incubation either
`ADPor collagen was added. The aggregation of the
`thrombocytes was
`turbidimetrically determined at
`A= 605 n m.
`The antimicrobial activity was determined in vitro
`according to a qualitative test for antibacterial and anti-
`fungal activity, using the agar diffusion method, against
`the following standard organisms: Staphylococcus au-
`reus, Streptococcus pyogenes, Escherichia coli, Proteus
`mirabilis, Pseudomonas aeruginosus, Candida albicans
`and Aspergillus niger.
`The antifungal activity was determined in vitro
`against the following fungi: Trichophyton rubrum, Trich-
`ophyton mentagrophytis, Microsporum canis, Epidermoph-
`yton floccosum, Chrysosporum, Candida albicans, Can-
`dida guilliermondi and Saccaromyces cerevisiae. The
`fungi were cultured by plating on an agar nutrient me-
`dium. The compoundto be tested was added before the
`incubation. A measure of the efficiency of the com-
`poundtested is the radius ofthe circle, within which the
`growth of the fungi has been inhibited.
`Acute toxicity was determined by using female mice
`of NMRI-Strain with an age of about 7 months and
`weighing 30-40 g. The administration of the test com-
`pound was Lv.
`Thus
`the compound 4-[2-(2,6-dimethylphenyl)-|!-
`methyletheny!]imidazole, which has a LDs9 value of 40
`mg/kg i.v.
`in mice, was found in the blood pressure
`study with anesthetized rats of normal weight described
`aboveto cause a registrable lowering of the blood pres-
`sure at a dose of 3 pg/kg i.e. Ata dose of 10 ng/kg iv.
`the blood pressure lowering was quite clear and at a
`dose of 100-300 pg/kg iv. the reduction of the blood
`pressure was on an average 38%. The duration of the
`effect was at least 30 minutes (after which time the
`determination was interrupted). A blood pressure low-
`ering of more than 40% was obtained when 2 mg/kg of
`the compound was administered perorally. The dura-
`tion of the effect wasat least 5 h.
`The compound 4-[2-(2,6-dimethylpheny])-1-methyle-
`thyllimidazole (LDs7=40 mg/kgi.v. in mice) caused a
`blood pressure lowering of 20 percent measured 30
`minutes after the administration at a dose of 100 pg/kg
`i.v. When 10 mg/kg of the compound was given per-
`orally, a blood pressure drop of 25% was obtained.
`Duration wasat least 5 h.
`The
`compound 4-{(a-methy])-2,6-dimethylbenzy]-
`Jimidazole (LDso=150 mg/kg i.v.
`in mice) caused a
`blood pressure lowering of 30% at a dose of I-10
`mg/kgi.v. (30 min. after administration).
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`35
`
`60
`
`65
`
`Petition for Inter Partes Review of US 8,455,527
`Amneal Pharmaceuticals LLC — Exhibit 1004 — Page 9
`
`

`

`4,544,664
`
`18
`stated. The compounds which are indicated as bases are
`tested in deuteri um methanol, deu terium acetone or
`deuterium chloroform, while the values for compounds
`which are indicated as hydrochlorides were determined
`in deuterium oxide. The presented I3C-NMR-spectrum
`were determined with a Bruker WP80DS apparatus.
`T he mass-spectra were determined with a Perkin(cid:173)
`Elmer RMU-6E apparatus using direct inlet system.
`The temperature employed was the fowest temperature
`10 needed for the evaporation of the compound as base. In
`the examples the strongest and the most essential frag(cid:173)
`ment-ions from a structural viewpoint are given as m/e
`values. In parenthesis is given the intensity of the frag-
`ment-ion in relation to the main peak.
`
`.
`17
`The compound 4-((a-methyl)-2,3-dimethylbenzyl(cid:173)
`]imidazole (LDso=40 mg/kg i.v. in mice) caused a
`blood pressure drop of 55% at IO µ.g/kg i.v. (after 30
`min.). Given perorally (I mg/kg) the compound gave a
`blood pressure drop of 20%. The duration was at least 5
`5 h.
`The compound 4-[2-(2,6-dimethylphenyl)propyl(cid:173)
`]i'!'idazole, which has a LDsovalue of200 mg/kg i.v. in
`mice gave a blood pressure drop of 30% at a dose of 3
`mg/kg i.v., measured 30 min. after administration.
`The compound 4-(2-(2,6-dimethylphenyl)-2-ethyle(cid:173)
`thyl]imidazole gave a blood pressure drop of about 25%
`at a dose of 3 mg/kg i.v., measured 30 min. after admin(cid:173)
`istration.
`The compound 4-[2-(2,6-dimethylphenyl)-1-ethyle- 15
`thenyl]-imidazole which has a LDso value of 110 mg/kg
`i. v. in mice, gave a blood pressure drop of 35% at a dose
`of 3 mg/kg intraperitoneally measured 30 min. after
`administration.
`4.9 ~ (0,2 mo!) of dry magnesium turnings are cov-
`4-(a-methyl-2-methylbenzyl- 20
`The
`compound
`ered with 50 ~l of dry tetrah~drofuran. The mixture is
`)imidazole, which has a LDso value of JOO mg/kg i. v. in
`heated to bo1hng and a solution of 34 g (0.2 mol) of
`mice, gave a blood pressure drop of 20% at a dose of0.3
`2-bro~otoluene in 50 ml dry tetrahydrofuran is added
`mg/kg intraperitoneally measured 30 min. after admin-
`dropw1se at such a rate that smooth react