United States Patent
`
`[19]
`
`[11] Patent Number:
`
`6,114,341
`
`Miillcr et al.
`
`[45] Date of Patent:
`
`*Sep. 5, 2000
`
`US{JU6l l434lA
`
`......................... .. 5141399
`5,-‘I995 Miiller et al.
`............. ..
`514;'400
`5,-‘I996 Miiller et al.
`611996 M1'iller—Gliemann et al.
`11,-‘I996 Miiller ...................... ..
`111998 Gasler et al.
`
`
`
`5,420,149
`5,521,206
`5,52?,su9
`5,5?6,342
`5,705,493
`
`FOREIGN PATENT DOCUMENTS
`
`509359
`0513533 A2
`513533
`560163
`0622358 Al
`2200584
`4302956
`4309968
`
`._-‘uropean Pat. Off. .
`1011992
`ll,-‘I992 European Pal. Oil‘.
`.
`1111992 European Pal. O11‘.
`.
`9,-‘I993 European Pal. Oil‘.
`.
`ll,-‘I994 European Pal. Oil‘.
`.
`T"/1922 Germany .
`8,-‘I994 Germany .
`911994 Germany .
`OTHER PUBLICATIONS
`
`RA. Glennon unrl M. von Stradtmann, J. Heterocycl. Chem.
`vol. 12, pp. 135-138, (1975).
`C./\. (irob und 0. Weissbach, llelv. Chim. Acla 44, pp.
`1748-1753, (1961).
`AN. Kost, R.S. Sagitullin, V.I. Gorbunov und N, N, Mody—
`anov, Khim. Gelerosikl. Soedin vol. 6, 359-363, (1970);
`English translation pp. 334-337.
`
`Priirtary E.rm1rmer—Mukund J. Shah
`Assistrtrtt hbrrrrrtiner Tarrilhom "I". Ngo
`Att0rr2e_y,Ager1t‘, or F.irm—Norris, McLaughlin & Marcus,
`PA.
`
`[57]
`
`ABSTRACT
`
`The pyrimido['l,2—a]indoles according to the invention are
`prepared by reacting appropriately substituted phenylaeetic
`acid derivatives with phenylglyeinols. The pyrimido[1,2—a]
`indoles can be used as active com pounds in niedicanients, in
`particular in merlicamcnts with antiatheroselerotie activity.
`
`'18 Claims, No Drawings
`
`[54]
`
`PYRlMl]_)()[1,2-A]IND()L];",S
`
`[75]
`
`Inventors: Ulrich Miiller, Wuppertal; Peter
`Eckenberg, Erkrath; Rudi Griitzmann,
`Solingen; Hilmar Biscltofi; Dirk
`Denier‘, both of Wuppertal; Stefan
`Wohlfeil, Ililden, all of Germany;
`Stefan Lohmer, Milan, Italy; Ulrich
`Nielsch; Peter Kolkhof, both of
`Wuppertal, Germany
`
`[73] Assignee: Bayer Aktiengesellschaft, Leverkusen,
`Germany
`
`| * ] Notice:
`
`This patent issued on a continued pros-
`ecution application filed under 37' CPR
`1.53[d], and is subject to the twenty year
`patent
`term provisions of 35 U.S.C.
`154(a)(2).
`
`[21] Appl. No.: I]81'829,[|15
`
`[22]
`
`Filed:
`
`Mar. 31, 1997
`
`[30]
`
`Foreign Application Priority Data
`
`Apr. 4, 1996
`
`[DE]
`
`Germany ......................... .. 196 13 550
`
`Int. Cl.7 ........................ .. /\01N 43,554; C()7'|) 239,300
`[5]]
`[52] U.S. Cl.
`........................................... .. 5'l4,r’267; 5441252
`[58] Field of Search ............................ .. 5143267; 5441252
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`3,850,957"
`4,283,455
`5,306,820
`5,352,687
`
`1111924 White ................................. .. 26U;"309.6
`1111988 Cliffe . . . . . . . . . . .
`. . .. 514.-"220
`411994 Decker et al.
`546.3153
`10.31994 Muller el al.
`......................... .. 5l4.u"34l
`
`
`
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`
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`6,114,341
`
`1
`
`PYRIMIDO[1,2-A]INDOI.ES
`
`The present invention relates to pyrimido[l,2-a]indoles,
`to processes for
`their preparation and to their use as
`medicarnents,
`in particular as antiatherosclerotic medica-
`ments.
`
`is known that elevated blood levels of triglycerides
`It
`(hypeitriglyceridaemia)
`and
`cholesterol
`(hypercholesterolaemia) are associated with the develop-
`ment of atherosclerotic changes in vessel walls and coronary
`heart disease.
`
`10
`
`furthermore, a distinctly increased risk of
`There is,
`developing coronary heart disease when these two risk
`factors occur in combination, which is in turn associated
`with an overproduction of apolipoprotein ll-100.
`Ilence
`there is a continuing pressing need to provide effective
`medicarnents for controlling atherosclerosis and coronary
`heart disease.
`
`The present invention relates to pyrimido[l,2—a]indoles of
`the general formula (I)
`
`20
`
`
`
`(U
`
`30
`
`in which
`
`A, D, E, G, L and M are identical or dilferent and
`represent hydrogen, halogen, trilluoromethyl, carboxyl,
`hydroxyl, straight-chain or branched alkoxy or alkoxy-
`carbonyl with, in each ease, up to 6 carbon atoms or
`straight-chain or branched alkyl with up to 6 carbon
`atoms, which in turn can be substituted by hydroxyl or
`by straight-chain or branched alkoxy with up to 4
`carbon atoms,
`
`R' and R2 are identical or different and represent
`hydrogen, cycloalkyl with 3 to 8 carbon atoms or
`straight-chain or branched alkyl with up to 10 carbon
`atoms, which is optionally substituted by cycloalkyl
`with 31o 6 carbon atoms, or represent phenyl which is
`optionally substituted by halogen or trifluoromethyl, or
`R1 and R3 form, together with the carbon atom, a 4-8-
`membered cycloalkyl ring,
`
`and
`
`R3 represents phenyl which is optionally substituted up to
`3 times, identically or differently, by nitro, carboxyl,
`halogen, cyano or by straight-chain or branched alkenyl
`or alkoxycarbonyl with, in each case, up to 6 carbon
`atoms or by straight-chain or branched alkyl with up to
`6 carbon atoms, which is optionally substituted by
`hydroxyl, carboxyl or by straight-chain or branched
`alkoxy or alkoxycarbonyl with, in each case, up to 6
`carbon atoms, andfor is optionally substituted by a
`group of the formula —OR’‘ or —NR5R°,
`in which
`
`R4 is hydrogen or straight-chain or branched alkyl or
`alkenyl with, in each ease, up to 6 carbon atoms,
`R5 and R6 are identical or different and denote phenyl,
`hydrogen or straight —chain or branched alkyl with up to
`
`2
`6 carbon atoms, or denote straight-chain or branched
`acyl with up to 8 carbon atoms, which is optionally
`substituted by a group of the formula —NR7R8,
`in which
`
`R7 and R8 are identical or dillerent and denote hydrogen
`or straight-chain or branched acyl with up to 8 carbon
`atoms;
`where appropriate in an isorneric form and the salts thereof.
`The pyrimido[l,2-a]indoles according to the invention
`can also be in the fonn of their salts. Salts which may be
`generally mentioned here are those with organic or inorganic
`bases or acids.
`
`the
`Physiologically acceptable salts are preferred for
`purpose of the present invention. Physiologically acceptable
`salts of the compounds according to the invention may be
`salts of the substances according to the invention with
`mineral acids, carboxylic acids or sulphonic acids. Particu-
`larly preferred examples are salts with hydrochloric acid,
`hydrobromie acid, sulphuric acid, phosphoric acid, meth-
`anesulphonic acid, ethanesulphonic acid, toluenesulphonic
`acid, benvienesulphoriic acid, naphthalenedisulphonic acid,
`acetic acid, propionic acid, lactic acid, tartaric acid, citric
`acid, fumaric acid, maleic acid or benzoic acid.
`Physiologically acceptable salts may likewise be metal or
`ammonium salts of the compounds according to the inven-
`tion which have a free carboxyl group. Part ieularly preferred
`examples are sodium, potassium, magnesium or calcium
`salts, and ammonium salts which are derived from ammonia
`or organic amines such as, for example, ethylamine, di— or
`triethylamine, di— or triethanolamine, dicyclohexylamine,
`dimethylaminoethanol, arginine, lysine, ethylenediamine or
`2—phenylethylamine.
`The compounds according to the invention can exist in
`stereoisomeric forms which either are related as image and
`mirror image (enantiomers) or are not related as image and
`mirror image (diastereomers). The invention relates to the
`enantiomers or diastereomers or mixtures thereof in each
`case. These mixtures of eriantiorners and diastereomers can
`
`40
`
`be separated into the stereoisomerically pure components in
`a manner known per se.
`Preferred compounds of the general formula (I) are those
`in which
`
`A, D, E, G, I. and M are identical or different and
`represent hydrogen,
`fluorine, chlorine, bromine,
`trifluoromethyl, carboxyl, hydroxyl, straight-chain or
`branched alkoxy or alkoxycarbonyl with, in each case,
`up to 4 carbon atoms or straight-chain or branched
`alkyl with up to 4 carbon atoms, which can in turn be
`substituted by hydroxyl or by straight-chain or
`branched alkoxy with up to 3 carbon atoms,
`R1 and R3 are identical or different and represent
`hydrogen, eyelopropyl, cyclobutyl, cyclopentyl,
`cyclohexyl, cycloheptyl, cyclooctyl or straight-chain or
`branched alkyl with up to 8 carbon atoms, which is
`optionally substituted by eyelopropyl, cyclopentyl or
`cyclohexyl, or represent phenyl which is optionally
`substituted by fluorine, chlorine or bromine, or
`R1 and R2 form, together with the carbon atom, a 4-7-
`membered cyeloalliyl ring,
`
`and
`
`50
`
`G0
`
`R3 represents phenyl which is optionally substituted up to
`3 times, identically or ditferently, by nitro, carboxyl,
`fluorine, chlorine, bromine, cyano, by straight-chain or
`branched alkenyl or alkoxycarbonyl with, in each case,
`up to 4 carbon atoms or by straight-chain or branched
`alkyl with up to 5 carbon atoms, which is optionally
`
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`4
`where appropriate in an isomeric form and the salts thereof.
`A process for the preparation of the compounds of the
`general formula (I) according to the invention has also been
`found and is characterized in that racemic or else already
`enantiomerically pure carboxylie acids or their activated
`derivatives of the general formula (II)
`
`(“J
`
`
`
`racemic or enanliomerically pure in which
`
`A, I), E, G, I., M, R1 and R2 have the indicated meaning,
`and
`
`R9 represents hydroxyl or represents an activating radical,
`preferably chlorine,
`are amidated with phenylglyeinols of the general formula
`(III)
`
`[III]
`
`R3
`
`J\/
`
`0”
`
`_
`[I35
`
`3
`substituted by hydroxyl, carboxyl or by straight-chain
`or branched alkoxy or alkoxyearbonyl with,
`in each
`case, up to 5 carbon atoms, andfor is optionally sub-
`stituted by a group of the formula —()R" or —NR5Rfi,
`in which
`
`R4 denotes hydrogen or straight-chain or branched alkyl
`or alkenyl with, in each ease, up to 4 carbon atoms,
`R5 and R“ are identical or different and denote phenyl,
`hydrogen or straight -chain or branched alkyl with up to
`5 carbon atoms, or straight—chain or branched aeyl with
`up to 6 carbon atoms, which is optionally substituted by
`a group of the formula —NR7R“,
`in which
`
`10
`
`R7 and R* are identical or different and denote hydrogen
`or straight—chain or branched acyl with up to 6 carbon
`atoms,
`where appropriate in an isomeric form and the salts thereof.
`Particularly preferred compounds of the general formula
`(I) are those in which
`A, D, E, G, L and M are identical or different and
`represent hydrogen,
`fluorine, chlorine, bromine,
`trifluoromethyl, carboxyl, hydroxyl, straight-chain or
`branched alkoxy or alkoxycarbonyl with, in each case,
`up to 3 carbon atoms or represents straight-chain or
`branched alkyl with up to 3 carbon atoms,
`R1 and R2 are identical or ditferent and represent
`hydrogen, cyclopropyl, cyclobutyl, cyclopentyl,
`cyclohexyl, cycloheptyl, cyclooctyl or
`represent
`straight-chain or branched alkyl with up to 6 carbon 30
`
`20
`
`atoms, which is optionally substituted by cyclopentyl
`or cyclohexyl, or represent phenyl which is optionally
`substituted by fluorine, chlorine or bromine, or
`R‘ and R: form, together with the carbon atom, a 5-7-
`membered cycloalkyl ring,
`
`and
`R3 represents phenyl which is optionally substituted up to
`3 times,
`identically or diffc-I-cntly, by hydmxyl,
`trifluoromethyl,
`trifluoromethoxy, earboxyl, or by 40
`straight—chain or branched alkoxy, alkyl or alkoxyear—
`bonyl with, in each ease, up to 3 carbon atoms,
`
`35 in which
`
`R3 has the indicated meaning’
`in inert solvents, where appropriate in the presence of
`hast:-‘F iifldfor iiflt-‘1i"3T)’ 51153130993-
`The process according to the invention can be illustrated
`by way of example by the following formula diagram:
`
`Me
`
`N \
`
`/ NX Me
`
`(T(){)H
`
`(IDL
`
`IIOBT
`
`g
`
`HEN/"\/0H
`
`3 of 34
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`6,114,341
`
`-continued
`
`Me
`
`N \-
`
`/
`
`/ N
`
`Mn:
`
`(Ti 3 —‘NH
`
`/K\/on
`
`
`
`Suitable solvents for the amidation in this case are inert
`
`organic solvents which are not changed under the reaction
`conditions. These include ethers such as diethyl ether or
`tetrahydroluiran, halogenated hydrocarbons such as
`dichloromethane,
`trichloromethane,
`tetrachloromethane,
`'l,2- dichloroethanc, trichloroethane, tctrachloroethanc, 1,2-
`dichloroethylene or trichloroethylene, hydrocarbons such as
`benzene, xylene, toluene, hexane, cyclohexane, or petro-
`leum fractions, nitromethane, dimethylformamide, acetone,
`aoetonitrile or hexamethylphosphoric triamide. It is likewise
`possible to employ mixtures of
`the solvents.
`Dichloromethane, tetrahydrofuran, acetone or dimethylfor—
`mamide are particularly preferred.
`Bases which can be employed for the process according
`to the invention are, in general, inorganic or organic bases.
`These preferably include alkali metal hydroxides such as, for
`example, sodium hydroxide or potassium hydroxide, alka-
`line earth metal hydroxides such as, for example, barium
`hydroxide, alkali metal carbonates such as sodium carbonate
`or potassium carbonate, alkaline earth metal carbonates such
`as calcium carbonate or alkali metal or alkaline earth metal
`
`alcoholates such as sodium or potassium methanolate,
`sodium or potassium ethanolate or potassium tert-butoxide,
`or organic amines [trialkyl
`(C,—C,,)amines) such as
`triethylamine, or heterocycles such as 1,4-diazabicyclo
`2.2.2]octane [DABCO), 1,8—diazabicyclo[5.4.0]un(|ee—7—
`ene {DBU), pyridine, dimethylaminopyridine, methylpiperi-
`dine or morpholine. It is also possible to employ as bases
`alkali metals such as sodium and hydrides thereof such as
`sodium hydride. Sodium and potassium carbonates and
`triethylamine are preferred.
`The base is employed in an amount of from 1 mol to 5
`mol, preferably from 1 mol to 3 mol, based on 1 mol of the
`compound of the general formula (II).
`The reaction is generally carried out at a temperature in
`the range from 0° C. to 150° C., preferably from +20° C. to
`+110° C.
`The reaction can be carried out under atmospheric,
`elevated or reduced pressure (for example 0.5 to 5 bar).
`Atmospheric pressure is generally employed.
`The reaction can, where appropriate, also take a course
`via the activated stage of the acid halides which can be
`prepared from the corresponding acids by reaction with
`thionyl chloride, phosphorus trichloride, phosphorus
`pentachloride, phosphorus tribromide or oxalyl chloride.
`The bases listed above may also be employed as acid-
`binding aids for the amidation.
`Likewise suitable as ancillary substances are dehydrating
`reagents. These include, for example, carbodiimides such as
`diisopropylcarbodiimide, dicyclohexylcarbodiimideorN-(3-
`
`dirnethylaminopropyl)-N‘-ethylcarbodiimide hydrochloride
`or carbonyl compounds such as carbonyldiimidazolc or
`1,2oxazolium compounds such as 2—ethyl—5—phenyl—l,2—
`oxazolium-3'-sulphonate or propanephosphonic anhydride
`or
`isobutyl chloroformate or benzotriaxolyloxy
`(dimethylamino)phosphonium hexafluorophosphate or
`diphenylphosphoryl azide or methanesulphonyl chloride,
`where appropriate in the presence of bases such as triethy—
`lamine or N~ethylmorpholine or N—methylpiperidine or
`dicyclohexylcarbodiimide and N-hydroxysuccinimide.
`The ancillary substances are generally employed in an
`amount of from 0.5 to 3 mol, preferably from '1 to '1.5 mol,
`based on 1 mol ol‘ the appropriate carboxylic acids.
`The carboxylic acids of the general formula (II) are novel
`and can be prepared by initially preparing, by reacting
`compounds ol‘ the general formula (IV)
`
`(IVJ
`
`20
`
`30
`
`40
`
`_
`
`in which
`
`50
`
`G0
`
`R‘ and R: have the indicated meaning,
`T represents a typical leaving group such as, for example,
`chlorine, bromine, iodine,
`tosylate or mesylate, and
`preferably represents bromine,
`
`and
`
`R10 represents ((Tl—(I_,)-alkyl,
`with compounds of the general formula (V)
`
`A
`
`G
`
`N/TALL
`or \
`l
`A
`13/ X / N¢\.\a
`
`65
`
`in which
`
`A, D, E, G, L and M have the indicated meaning,
`
`4 of 34
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`
`7
`the compounds of the general formula (VI)
`
`6,114,341
`
`and
`
`8
`
`A
`r\ S
`D+k
`E}
`
`G
`NZ?/fl
`T
`X / —.\’
`N M
`
`(TH-3
`
`/ \
` C.‘(}3R1U
`T R1
`R3
`
`(VI)
`
`_
`
`m
`
`in which
`
`A, D, L], (i, l_, M, R1, R2 and Rm have the ahovemen-
`tioned meaning,
`in inert solvents, where appropriate in the presence of
`bases,
`
`and subsequently hydrolysing the esters by conventional 7”
`methods.
`'
`Enantiomerically pure acids of the formula (Ila) or (IIb}:
`
`([Ia,1
`
`A
`I4\/
`‘K
`DT
`/x
`E
`
`l
`
`G
`N/‘*0.-/}|
`/
`T
`X N'.\
`M j
`
`X
`
`CH2
`
`1‘
`\ /
`
`Cn—oo—R"
`l
`R‘
`
`(If
`
`(uh)
`
`V
`~‘
`
`30
`
`35
`
`Z represents halogen, preferably bromine,
`the enantiomerically pure menthyl esters of the general
`fonnula (Diet) or [IXb)
`
`H_;( / 1
`\cn—co3—R“ of
`ti
`
`\ ,,
`
`113:: /
`
`(IXa]
`
`[[Xb)
`
`in which
`
`R‘, R2 and R“ have the indicated meaning,
`converting the latter in a next step by a halogenation into
`the
`compounds of the general formula (Xa) or (Xb)
`
`(Na)
`
`011‘)
`
`T—CII_3 / l
`\\ '\(|?H—C0;—R“ or
`
`R:
`
`T—C[Ig / l
`\ \(|:H—(T()3—Rl1
`
`R2
`
`40
`
`in which
`
`in which R1 and R: are not hydrogen and R9 represents
`hydroxyl, are furthermore obtained by preparing, from the
`D— or L—menthyl esters of the general formula (V11)
`
`II_xC
`
`/ I
`~\ --""‘t'.TH-_3—(T()_s—R“
`
`(V11)
`
`50
`
`in which
`
`R” represents D— or L—menthyl,
`formula
`by reaction with eompounds of the general
`[VIIIa) or (Vlllb) R'—Z (VIIIa) or R2—Z (Vlllb)
`in which
`
`R1 and R2 have the indicated meaning,
`
`R1, R2, and R” have the indicated meaning, and T
`represents halogen,
`subsequently preparing, by reaction with the compounds
`oft he general formula (V), the enantiomerieally pure
`compounds of the general formula (X121) or (XIb]
`
`(X[a]
`
`
`
`50f34
`
`PENN EX. 2217
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`
`

`
`6,114,341
`
`(X[h,1
`
`10
`
`ll_xC‘ / I
`\ ‘\'\(IT;T(T()3tl5Lt
`
`R!
`
`01‘
`
`[}{IVa]
`
`(XIV-"b;
`
`/ \
`
`\ \'\'(_‘.l1-—E_‘.03tBu
`'3
`R
`
`converting the latter as described above by a halogenation
`into the enantiomerically pure compounds of the general
`formula (XVa) or (XVTJ)
`
`(XWJ
`
`(XWJ
`
`.].._/H.,(7 / I
`\\ \'\Ti‘a—c03i15u
`
`R]
`
`01'
`
`Tr
`
`Hgc / I
`\ ‘
`
`(‘.l:l—-CO3tBu
`,L,
`
`wherein R‘ and R3 have the indicated meaning and T‘
`represents halogen and converting the latter by reaction with
`the compounds of the general formula (V) into the enantio-
`merically pure esters oi‘ the general formula (X\/la] or
`(XVlb):
`
`(XV[a]
`
`(xvrhg
`
`
`
`
`10
`
`20
`
`30
`
`40
`
`50
`
` 9
`
`-continued
`
`I)—
`
`in which
`
`l_, M, R’, R2 and R“ have the indicated
`
`(3.
`A, l), E,
`meaning,
`or then converting the latter by hydrolysis into the enan-
`Iiomerically pure acids of the general formula (Ila) and
`[llb).
`It is additionally possible to prepare enantiomcrically pure
`acids of the formula (Ila) or (llb) by initially converting
`racemic carboxylic acids of the general formula (Xlla) or
`(Xllb)
`
`(xua)
`
`(xuh;
`
`II»: / I
`|—cI7H—(t(),H
`\ RI
`
`Hgc / I
`|—fi‘1I-—C03tI
`\ .,R.
`
`in which R1 and R2 have the indicated meaning,
`
`by reaction with (R)- or (S)-phenylethylamine in inert
`solvents and subsequent crystallization of the phen-
`ethylammonium salts and subsequent hydrolysis of the
`salts, into the enantiomerically pure compounds of the
`general formula (Xllla) or (Xlllb)
`
`(X[IIa,1
`
`(XIIIb)
`
`H3(T / I
`\ \§t1—c03t1
`
`R1
`
`lI3C / I
`\ ¥‘cI:H——co3t1
`
`R2
`
`in which R1 and R: have the indicated meaning,
`
`preparing from the latter in another step with isobutene, in
`inert solvents and in the presence of acids, the enan-
`tiornerically pure esters of the general formula (XlVa)
`or (XIV/b)
`
`65
`
`in which A, D, E, G, L, M, R‘ and R2 have the indicated
`meaning and in the final steps as described above, preparing
`the corresponding enantiomcrically pure acids of the for-
`mula (Ila) or (llb) and activated derivatives.
`
`6 of 34
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`CFAD V. UPENN
`
`lPR2015-01836
`
`

`
`6,114,341
`
`10
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`20
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`30
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`40
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`50
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`11
`the processes are conventional
`Solvents suitable for
`organic solvents which are not changed under the reaction
`conditions. These preferably include ethers such as diethyl
`ether, dioxane, tetrahydrofuran, glycol dimethyl ether, or
`hydrocarbons such as benzene,
`toluene, xylene, hexane,
`cyclohexane or petroleum fractions, or halogenated hydro-
`carbons such as dichloromethane,
`trichloromethane,
`tetrachloromethane, dichloroethylene, trichloroethylene or
`chlorobenzene, or ethyl acetate,
`triethylamine, pyridine,
`dimethyl sulphoxide, dimethylformamide, hexamethylphos-
`phoric triamide, acetonitrile, acetone or nitromethane. It is
`likewise possible to use mixtures of the said solvents.
`Dimethylformamide, toluene and tetrahydrofuran are pre-
`ferred.
`
`Bases which can be employed for the processes according
`to the invention are in general inorganic or organic bases.
`These preferably include alkali metal hydroxides such as, for
`example, sodium hydroxide or potassium hydroxide, alka-
`line earth metal hydroxides such as, for example, barium
`hydroxide, alkali metal carbonates and bicarbonates such as
`sodium carbonate, sodium bicarbonate or potassium
`carbonate, alkaline earth metal carbonates such as calcium
`carbonate, or alkali metal or alkaline earth metal alcoholates
`such as sodium or potassium methanolate, sodium or potas-
`sium ethanolate or potassium tert—butoXide, or organic
`amines [trialkyl(C1—C(,)-amines) such as triethylamine, or
`heterocycles such as 1.4-diazabieyclo[2.2.2]oetane
`(D/\B(T[)),
`l,8-diazabicyclo[5.4.0]undec-7'-ene (DBU),
`pyridine, dimethylaminopyridine, methylpiperidine or mor-
`pholine. lt is also possible to employ as bases alkali metals
`such as sodium or hydrides thereof such as sodium. hydride.
`Sodium bicarbonate, potassium carbonate and potassium
`tert-butoxide, DBU or D/\l3(.'() are preferred.
`Solvents suitable for the hydrolysis are water or the
`organic solvents cI.tstomary for a hydrolysis. These prefer-
`ably include alcohols such as methanol, ethanol, propanol,
`isopropanol or butanol, or ethers such as tetrahydrofuran or
`dioxane, or dimethylformamide, or dimethyl sulphoxide.
`Alcohols such as methanol, ethanol, propanol or isopropanol
`are particularly preferably used. It is likewise possible to
`employ mixtures of the said solvents.
`The hydrolysis can, where appropriate, also be carried out
`with acids such as, for example, trilluoroacetic acid, acetic
`acid, hydrochloric acid, hydrobromic acid, methanesul—
`phonic acid, sulphuric acid or perchloric acid, preferably
`with trifluoroacetic acid.
`The hydrolysis is generally carried out at a temperature in
`the range from 0° (T. to +l00° (L, preferably from +20” (T.
`to +80° C.
`The hydrolysis is generally carried out under atmospheric
`pressure. However,
`it
`is also possible to employ reduced
`pressure or elevated pressure (for example from 0.5 to 5
`bar).
`When carrying out the hydrolysis, the base is generally
`employed in an amount of from 1 to 3 mol, preferably from
`1 to 1.5 mol, based on '1 mol ofthe ester. Equimolar amounts
`of the reactants are particularly preferably used.
`The hydrolysis of tert-butyl esters is generally carried out
`with acids such as, for example, hydrochloric acid or trif-
`luoroaeetic acid, in the presence of one of the abovemen-
`tioned solvents andfor water or mixtures thereon preferably
`with dioxane or tetrahydrofuran.
`General process [A] according to the invention is gener-
`ally carried out at a temperature in the range from —30° C.
`to +'l50° C., preferably from 80° C. to 150° C.
`Suitable and preferred conditions for the individual steps
`to prepare enantiomerically pure acids are the following:
`
`12
`Compounds of the general formula (IXa) and (IXb) are
`preferably prepared in dimethylformamide and potassium
`tert-butanolale at a temperature in the range from —l0° C. to
`+10° C.
`Ilalogenation to give the compounds of the general for-
`mula (Xa) and (Xb) is carried out in chlorobenzene with
`1,3-c|ibromo—S,5—dimethylhydantoin in the presence of azo—
`bisisobutyronitrile at a temperature in the range from 0° C.
`to 110° C.
`The reaction to give the compounds of the general for-
`mula (XIa) and (XIb) takes place under a protective gas
`atmosphere in dimethylformamide and potassium tert-
`butanolate at a temperature in the range from 0° C. to 30° C.
`Ilydrolysis of the compounds of the general formula (Xla)
`and (XIb) can be carried out as described above, with the
`IIBr;"fon'nic acid system being particularly preferred. The
`hydrolysis is carried out at a temperature in the range from
`20° C. to 100° C.
`Suitable and preferred activating reagents are trilluo-
`romethanesulphonyl chloride, mesyl chloride, oxalyl chlo-
`ride and thionyl chloride. Thionyl chloride is particularly
`preferred.
`The reaction to give the compounds of the general for-
`mula (Xllla) and (Xlllb) takes place in the lirst step pref-
`erably in tetrahydrofuran and triethylamine, and in the
`second step in the waterfhydrochloric acid system. The
`reaction is carried out at a temperature in the range from 30°
`C. to 70° C.
`Concentrated sulphuric acid is particularly preferably
`employed as acid for preparing the compounds of the
`general fon'nula (XIVa) and (XI\/b) according to the inven-
`tion. The preparation is carried out with methylene chloride.
`In the further work—up step, potassium carbonate is
`employed as base. The reaction takes place at a temperature
`in the range from 0° C. to +20° C., particularly preferably at
`10° (f.
`
`The compounds of the general formula (XIVa) and
`(XIVb) are halogenated with N—bromosuccinimide in meth-
`ylene chloride in the presence of azobisisobutyronitrile.
`The base is generally employed in an amount of from 0.05
`mol to 10 mol, preferably from 1 mol to 2 mol, in each case
`based on 1 mol of the compounds of the general formulae
`(IV), (Vllla) and (Vlllb) and (Xla) and [XIb).
`The processes according to the invention are generally
`carried out under atmospheric pressure. However, it is also
`possible to carry out the princess under elevated pressure or
`under reduced pressure (for example in a range from 0.5 to
`5 bar).
`The compounds of the general formula [[11] are known
`per se.
`The compounds of the general formula (IV), (Villa) and
`(Vlllb) are known or can be prepared in analogy to known
`methods.
`
`The compounds of the general formula (V) are in some
`cases known or novel, but can then be prepared in analogy
`to published methods.
`The compounds of the general formula (VII) are novel as
`species and are prepared from the corresponding acid.
`The enantiomerically pure compounds of the general
`formula (IXa) and (IXb) are, with the exception of R‘ or
`R°=isopropyl, novel and can be prepared as described
`above.
`
`The compounds of the general formulae (Xa), and (Kb),
`(XIa), (Xlb) are novel and can be prepared as described
`above.
`
`The compounds of the general formula (XIVa) and
`(XIVb) are in some cases known or can be prepared by
`customary methods.
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`7 of 34
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`PENN EX. 2217
`CFAD V. UPENN
`
`lPR2015-01836
`
`

`
`6,114,341
`
`10
`
`13
`The enantiomerically pure compounds of the general
`formula [XVa) and (XVI:-) and (XVIa) and (XVIb) are novel
`and can be prepared as described above.
`The compounds of the general formula (VI) are novel and
`can be prepared as described above.
`The compounds of the general formula (I) according to
`the invention have a spectrum of pharmacological effects
`which was unpredictable.
`They can be used as active compounds in medicaments
`for reducing changes in vessel walls and for treating coro-
`nary heart disease, heart
`failure, brain dysfunction,
`ischacmic brain disease, stroke, disturbances of blood flow,
`microcirculation disturbances and thromboses.
`Furthermore, proliferation of smooth muscle cells plays a
`crucial part in the occlusion of vessels. The compounds 15
`according to the invention are suitable for inhibiting this
`proliferation and thus preventing atherosclerotic processes.
`The compounds according to the invention are distin-
`guished by lowering the ApoB 100—associated lipoproteins
`(Vl.l)l. and its brea.kdown products such as, for example, 20
`LDL), and ApoB 100, the triglycerides and cholesterol.
`Hence they have valuable pharmacological properties which
`are superior by comparison with the prior art.
`Surprisingly, the effect of the compounds according to the
`invention initially comprises reducing or completely inhib-
`iting the formation andfor
`the release of Apoli 100-
`associated lipoproteins from liver cells, which results in a
`lowering of the VI.l)I. plasma level. This VI.[)I. lowering
`must be associated with a lowering of the plasma levels of
`ApoB 100, I.[)I., triglycerides and cholesterol; thus a plu-
`rality of the abovemenlioned risk factors involved in
`changes in vessel walls are reduced simultaneously.
`The compounds according to the invention can therefore
`be employed for
`the prevention and treatment of
`atherosclerosis, of obesity, pancreatitis and of constipation.
`1. Inhibition of the Release of ApoB 100—associated Lipo-
`proteins
`inhibition of the release of Apoll
`to detect
`The test
`100—associated lipoproteins from liver cells took place in
`vitro with cultivated liver cells, preferably with cells of the
`human line HepG2. These cells are cultured under standard
`conditions in medium for culturing eukaryotic cells, prefer-
`ably in RPM] 1640 with 10% fetal calf serum. I[ep(i2 cells
`synthesize, and secrete into the culture supernatant, ApoB
`l{l0-associated lipoprotein particles which in principle have
`a similar structure to the VLDL and LDL particles to be
`found in the plasma.
`These particles can be detected using an immunoassay for
`human LDL. This immunoassay takes place with antibodies
`which had been induced against human I.DI. in rabble-3 under
`standard conditions. The anti-I.[)I.antibodies (rah anti-I.|)I.
`Abs) were purified by aflinity chromatography on an immu-
`nosorbent with human I.l)I.. These purified rab anti-I.|)I.
`Abs are adsorbed onto the surface of plastic. This adsorption
`expediently takes place onto the plastic surface of microtitre
`plates with 96 wells, preferably on MaxiSorp plates. If ApoB
`100—associated particles are present in the supernatant from
`llep (52 cells, thesearticles can bind to the insolubilized rab
`anti—LDL Abs, resulting in an immune complex which is
`bound to the plastic surface. Unbound proteins are removed
`by washing. The immune complex present on the plastic
`surface is detected using monoclonal antibodies which had
`been induced against human LDL, and had been purified,
`under standard conditions. These antibodies were conju-
`gated to the enzyme peroxidase. Peroxidase converts the
`colourless substrate TMB into a coloured product in the
`presence of H202. After acidification of the reaction mixture
`
`14
`with II2S(),,, the specific absorption of light at 450 nm is
`determined and is a measure of the amount of ApoB 100-
`associated particles secreted into the culture supematant by
`the HepG2 cells.
`Surprisingly, the cornpounds according to the invention
`inhibit the release of ApoB 100—associated particles. The
`IC5,, indicates the concentration of substance at which the
`absorption of light is inhibited by 50% compared with the
`control (solvent control without substance).
`
`|-;x_ ix-0_
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`1(i::o[ 134]
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`2. Determination of VLDL Secretion in Vivo in Hamsters
`The elfect of the test substances on VLDL secretion in
`vivo is investigated on hamsters. To do this, golden hamsters
`are premedicated with atropine (83 mgfkg
`and then
`anaeslheliyied with Ketavet [83 rngfkg
`and Nembutal
`(SI) mgfkg i.p.). When the animals’ reflexes have been lost,
`the jugular vein is exposed and cannulated. Subsequently,
`0.25 mlfkg of a 20% strength solution of Triton WR-1339 in
`physiological saline is administered. This detergent inhibits
`lipoprotein lipase and thus leads to an increase in the
`triglyceride level because there is no catabolism of secreted
`VI.|)I. particles. This triglyceride increase can be used as a
`measure ofthe VLDL secretion rate. Blood is taken from the
`animals by puncture of the retroorbital venous plexus before
`and one and two hours after administration ofthe detergent.
`The blood is incubated at room temperature for two hours
`and then at 4° C. overnight
`in order to complete the
`coagulation. It is then centrifuged at 10,000 g for 5 minutes.
`The triglyceride concentration in the serum obtained in this
`way is determined using a modified commercially obtain-
`able enzyme assay (Merckotest® Triglyceride No. 14354).
`100,ttl of serum are mixed with 100 pl of assay reagent in
`96-well plates and incubated at room temperature for 10
`minutes. Subsequently, the optical density is determined at
`a wavelength of 492 nm in an automatic plate reader (SLT
`Spectra). Serum samples with a triglyceride concentration
`which is too high are diluted with physiological saline. The
`triglyceride concentration present in the samples is deter-
`mined using a standard plot measured in parallel. In this
`model, test substances are administered either intravenously
`immediately before administration ofthe detergent or orally
`or subcutaneously before induction of anaesthesia.
`3. Inhibition of Intestinal Triglyceride Absorption in Vivo
`(Rats)
`Substances to be investigated for their inhibitory elfect on
`triglyceride absorption in vivo are administered orally to
`male Wistar rats with a body weight between "I 70 and 230
`g. For this purpose, the animals are divided into groups of 6
`animals 18 hour