ALOACEAEAEA
`US005217996A
`5,217,996
`United States Patent 15
`{11] Patent Number:
`Jun, 8, 1993
`Ksander
`[45} Date of Patent:
`
`[54]. BIARYL SUBSTITUTED 4-AMINO-BUTYRIC
`ACID AMIDES
`
`(75]
`
`Inventor: Gary Ksander, Milford, N.J.
`
`[73] Assignee: Ciba-Geigy Corporation, Ardsley,
`N.Y.
`
`[21] Appl. No.: 824,132
`
`[22] Filed:
`
`Jan. 22, 1992
`
`Int. CLS CO7C 229/34; A6IK 31/235
`[51]
`[82] U.S. Ch. cee ceeceeeseeteneseseenee §14/533; 514/563;
`546/335; 549/452; 558/267; 558/275; 560/41;
`562/450
`[58] Field of Search ..........0.0.0. 560/41, 39; 562/450;
`549/77, 452, 496; 546/335; 558/267, 275;
`514/533, 563
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`ABSTRACT
`[57]
`The invention relates to biaryl substituted 4-amino-
`butyric acid derivatives of formula I
`
`R2
`T
`XOC™CHCHCHNHCTA(CH)m—COX!
`R}
`CHp-biary!
`
`@
`
`wherein COX and COX’ independently represent car-
`boxyl or carboxyl derivatized in form of a pharmaceuti-
`cally acceptable ester or amide; R1 represents hydrogen,
`lower alkyl, C3~C7-cycloalkyl-lower alkyl, aryl-lower
`alkyl, biaryl-lower alkyl, lower alkoxy, aryl-lower alk-
`oxy, aryloxy, N-lower alkylamino, N,N-di-lower alkyl-
`amino, N-aryl-lower alkylamino, N,N-di-aryl-lower
`alkylamino, N-arylamino, N,N-diarylamino, lower al-
`kanoylamino, aryl-lower alkanoylamino or aroylamino;
`R2 represents hydrogen, hydroxy, lower alkoxy, lower
`alkyl, aryl-lower alkyl, C3-C7-cycloalkyl-lower alkyl,
`amino-lower alkyl, hydroxy-lower alkyl,
`lower al-
`kylthio-lower alkyl,
`lower alkoxy-lower alkyl, aryl-
`loweralkyithio-loweralkyl or aryl-lower alkoxy-lower
`alkyl; biaryl represents pheny! substituted by carbocy-
`clic or heterocyclic aryl; A represents a direct bond,
`lower alkylene, phenylene or cyclohexylene; m repre-
`sents 1 or zero, provided that m represents 1 when A is
`a direct bond; or pharmaceutically acceptable salts
`thereof; pharmaceutical compositions comprising said
`compounds; methods for the preparation of said com-
`pounds and for the preparation of intermediates; and
`methods of treating disorders in mammals which are
`responsive to the inhibition of neutral endopeptidases
`by administration of said compounds to mammals in
`need of such treatment.
`
`11 Claims, No Drawings
`
`4,513,009 4/1985 Roqueset al. ...
`4,721,726
`1/1988 Berger...
`4,939,261
`7/1990 Ksander...
`w 562/444
`
`5,021,430 6/1991 Ksander.......
`w» 514/332
`5,096,925
`3/1991 Ksander .......sssesereeenes 514/522
`
`514/464
`
`« 514/513
`
`:
`
`FOREIGN PATENT DOCUMENTS
`
`38758 10/1981 European Pat. Off.
`836934
`1/1985 South Africa .
`2207351
`6/1989 United Kingdom .
`
`.
`
`OTHER PUBLICATIONS
`
`C. M. Ksanderet al., J. Med. Chem. 32, 2519 (1989).
`
`Primary Examiner—José G. Dees
`Assistant Examiner—B. Frazier
`Attorney, Agent, or Firm—Norbert Gruenfeld
`
`BIOCON PHARMALTD (IPR2020-01263) Ex. 1009, p. 001
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1009, p. 001
`
`

`

`1
`
`5,217,996
`
`BIARYL SUBSTITUTED 4-AMINO-BUTYRIC ACID
`AMIDES
`
`SUMMARYOF THE INVENTION
`
`Endogenousatria] natriuretic peptides (ANP), also
`called atrial natriuretic factors (ANF) have diuretic,
`natriuretic and vasorelaxant functions in mammals. The
`natural ANF peptides are metabolically inactivated, in
`particular by a degrading enzyme which has been rec-
`ognized to correspond to the enzymeneutral endopepti-
`dase (NEP) EC 3.4. 24.11, also responsible for e.g. the
`metabolic inactivation of enkephalins.
`The aim of the present invention is to provide novel
`biaryl substituted 4-amino-butyric acid amide deriva-
`tives described below which are useful as neutral endo-
`peptidase (NEP) inhibitors, e.g. as inhibitors of the
`ANF-degrading enzyme in mammals, so as to prolong
`and potentiate the diuretic, natriuretic and vasodilator
`properties of ANF in mammals,by inhibiting the degra-
`dation thereof to less active metabolites. The com-
`poundsof the invention are thus particularly useful for
`the treatment of conditions and disorders responsive to
`the inhibition of neutral endopeptidase EC 3.4. 24.11,
`particularly cardiovascular disorders, such as hyperten-
`sion, renal insufficiency including edemaandsalt reten-
`tion, pulmonary edema and congestive heartfailure. By
`virtue of their inhibition of neutral endopeptidase, the
`compoundsofthe invention mayalso be useful for the
`treatment of pain, depression and certain psychotic
`conditions. Other potential indications includethetreat-
`ment of angina, premenstrual syndrome, Meniere’s dis-
`ease, hyperaldosteronism, hypercalciuria, ascites, glau-
`coma, asthma, inflammations and gastrointestinal disor-
`ders such as diarrhea,
`irritable bowel syndrome and
`gastric hyperacidity.
`The present
`invention relates to biaryl substituted
`4-amino-butyric acid derivatives of formula I
`
`|
`XOC—CH™CH2—CHNHC—A(CH)COX’
`R)
`CH2-biary]
`
`R2
`
`@
`
`wherein COX and COX’ independently represent car-
`boxy! or carboxy] derivatized in form of a pharmaceuti-
`cally acceptable ester or amide; R; represents hydrogen,
`lower alkyl, C3-C7-cycloalkyl-lower alkyl, aryl-lower
`alkyl, biaryl-lower alkyl, lower alkoxy, aryl-lower alk-
`oxy, aryloxy, N-lower alkylamino, N,N-di-lower alkyl-
`amino, N-aryl-lower alkylamino, N,N-di-aryl-lower
`alkylamino, N-arylamino, N,N-diarylamino, lower al-
`kanoylamino,aryl-lower alkanoylamino or aroylamino;
`R2 represents hydrogen, hydroxy, lower alkoxy, lower
`alkyl, aryl-lower alkyl, C3-C7-cycloalkyl-lower alkyl,
`amino-lower alkyl, hydroxy-lower alkyl,
`lower al-
`kylthio-lower alkyl,
`lower alkoxy-lower alkyl, aryl-
`loweralkylithio-lower alkyl or aryl-lower alkoxy-lower
`alkyl; biaryl represents phenyl substituted by carbocy-
`clic or heterocyclic aryl; A represents a direct bond,
`lower alkylene, phenylene or cyclohexylene; m repre-
`sents 1 or zero, provided that m represents | when A is
`a direct bond; or a pharmaceutically acceptable salt
`thereof.
`Pharmaceutically acceptable ester and amide deriva-
`tives are preferably prodrug derivatives, such being
`convertible by solvolysis or under physiological condi-
`
`2
`tions to the free carboxylic acids of formula I wherein
`COX and/or COX’ represent carboxyl.
`Compounds of formula I and derivatives thereof,
`depending on the nature of substituents, possess one or
`more asymmetric carbon atoms. Theresulting diastere-
`oisomers and optical antipodes are encompassed by the
`instant invention.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`The definitions used herein, unless denoted other-
`wise, have the following meanings within the scope of
`the present invention.
`|
`The term biaryl represents phenyl substituted by
`carbocyclic aryl or heterocyclic aryl as defined herein,
`ortho, meta or para to the point of attachment of the
`phenyl ring, advantageously para; biaryl is also repre-
`sented as the —CegH4,—R3 substituent
`in formulae
`herein.
`Carbocyclic aryl preferably represents preferably
`monocyclic carbocyclic aryl or optionally substituted
`naphthyl.
`Monocyclic carbocyclic aryl represents optionally
`substituted phenyl, being preferably phenyl or phenyl
`substituted by one to three substituents, such being
`advantageously lower alkyl, hydroxy, lower alkoxy,
`lower alkanoyloxy, halogen, cyano,
`trifluoromethyl,
`lower alkanoylamino or lower alkoxycarbonyl. Mono-
`cyclic carbocyclic aryl particularly preferably repre-
`sents phenylor pheny]substituted by loweralkyl, lower
`alkoxy, hydroxy, halogen, cyanoortrifluoromethyl.
`Optionally substituted naphthyl represents 1- or 2-
`naphthyl or 1- or 2-naphthyl preferably substituted by
`loweralkyl, lower alkoxy or halogen.
`Heterocyclic aryl represents preferably monocyclic
`heterocyclic aryl such as optionally substituted thienyl,
`indolyl,
`imidazolyl, furanyl, pyridyl, pyrrolyl or N-
`lower alkylpyrrolyl.
`Optionally substituted furanyl represents 2- or 3-fura-
`ny! or 2- or 3-furanyl preferably substituted by lower
`alkyl.
`Optionally substituted pyridyl represents 2-, 3- or
`4-pyridylor 2-, 3- or 4-pyridyl preferably substituted by
`loweralkyl, halogen or cyano.
`Optionally substituted thieny! represents 2- or 3-thie-
`nyl or 2- or 3-thienyl preferably substituted by lower
`alkyl.
`Optionally substituted indolyl represents preferably
`2- or 3-indolyl or 2- or 3-indoly! preferably substituted
`by loweralkyl, lower alkoxy or halogen.
`Optionally substituted imidazolyl is preferably 1- or
`2-imidazolyl or 1- or 2-imidazoly] preferably substituted
`by loweralkyl.
`Aryl as in aryl-lower alkyl, aryl-lower alkoxy, aryl-
`oxy, N-arylamino, N,N-diarylamino, aryl-lower alkoxy-
`carbonyl or aryl-lower alkanoylamino is preferably
`pheny!] or phenyl substituted by one or two of lower
`alkyl, lower alkoxy, hydroxy, lower alkanoyloxy, halo-
`gen, trifluoromethyl, cyano, lower alkanoylamino or
`lower alkoxycarbonyl.
`.
`The term “lower” referred to herein in connection
`with organic radicals of compounds respectively de-
`fines such with up to andincluding 7, preferably up and
`including 4 and advantageously. one or two carbon
`atoms. Such maybe straight chain or branched.
`A loweralkyl group preferably contains 1-4 carbon
`atomsandrepresents e.g. ethyl, n- or iso-propyl, n-, iso-,
`sec.- or tert.-butyl or advantageously methyl.
`
`20
`
`45
`
`50
`
`55
`
`60
`
`65
`
`BIOCON PHARMALTD (IPR2020-01263) Ex. 1009, p. 002
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1009, p. 002
`
`

`

`5,217,996
`
`15
`
`20
`
`25
`
`30
`
`3
`4
`A lower alkoxy group preferably contains 1-4 carbon
`Aroylamino is preferably benzoylamino or ben-
`atomsand represents for example methoxy, n-propoxy,
`zoylamino substituted on the benzene ring by lower
`isopropoxy, n-, iso-, sec.- or tert.-butoxy or advanta-
`alkyl, lower alkoxy, halogen ortrifluoromethyl.
`Carboxyl esterified in form of a pharmaceutically
`geously ethoxy.
`Aryl-lower alkyl is advantageously benzyl or phen-
`acceptable ester, represents advantageously a prodrug
`ethyl optionally substituted by one or two of lower
`ester that may be convertible by solvolysis or under
`alkyl, lower alkoxy, hydroxy, lower alkanoyloxy, halo-
`physiological conditions to the free carboxylic acid,
`such being preferably C}-C2o-alkoxycarbonyl, advanta-
`gen ortrifluoromethyl.
`geously lower alkoxycarbonyl;
`(amino, acylamino,
`Aryl-lower alkoxy represents advantageously e.g.
`benzyloxy, benzyloxy substituted by lower alkyl, lower
`mono-or di-lower alkylamino)-lower alkoxycarbonyl;
`alkoxy, lower alkanoyloxy, halogen ortrifluoromethyl,
`carboxy-lower alkoxycarbonyl,
`e.g.
`alpha-carboxy-
`lower alkoxycarbonyl;
`lower alkoxycarbonyl-lower
`or pyridylmethoxy.
`alkoxycarbonyj,e.g. alpha-lower alkoxycarbonyl-lower
`Aryloxy preferably represents phenoxy or phenoxy
`substituted by lower alkyl,
`lower alkoxy,
`lower al-
`alkoxycarbony!; a-(di-lower alkylamino, amino, mono-
`lower alkylamino, morpholino, piperidino, pyrrolidino,
`kanoyloxy, halogen ortrifluoromethyl.
`l-lower
`alkylpiperazino)-carbonyl-lower
`alkoxycar-
`N-arylamino and N,N-diarylamino represent advan-
`tageously N-phenylamino or N,N-diphenylamino op-
`bonyl; aryl-lower alkoxycarbonyl, preferably option-
`tionally substituted in the phenyl moiety or phenyl moi-
`ally (halo,
`lower alkyl or lower alkoxy)-substituted
`eties by lower alkyl, lower alkoxy, hydroxy,
`lower
`benzyloxycarbonyl, or pyridylmethoxycarbony]; 1-(hy-
`alkanoyloxy, halogen or trifluoromethyl.
`droxy, lower alkanoyloxy or lower alkoxy)-lower alk-
`The term C3-Cy-cycloalkyl represents a saturated
`oxycarbonyl, e.g. pivaloyloxymethoxycarbony};
`(hy-
`droxy, lower alkanoyloxy or lower alkoxy)-loweral-
`cyclic hydrocarbon radical which contains 3 to 7 and
`preferably 5 to 7 ring carbon andis, most preferably,
`koxymethoxycarbonyl;_bicycloalkoxycarbonyl-lower
`alkoxycarbonyl, e.g. bicyclo[2,2,1]-heptyloxycarbonyl-
`cyclopentyl or cyclohexyl.
`The term cycloalkyl-lower alkyl represents prefera-
`lower alkoxycarbonyl, especially bicyclo-{2,2,1]-hep-
`tyloxycarbonylmethoxycarbony] such as bornyloxycar-
`bly 1- or 2-(cyclopentyl or cyclohexylethyl, 1-, 2- or
`bonylmethoxycarbonyl; 1-(ower alkoxycarbonyloxy)-
`3-(cyclopentyl or cyclohexyl)propyl, or 1-, 2-, 3- or
`lower
`alkoxycarbonyl;
`5-indanyloxycarbonyl;
`3-
`4-(cyclopentyl or cyclohexyl)-butyl.
`Amino-lower alkyl
`represents preferably amino-
`phthalidoxycarbony! and (lower alkyl, lower alkoxy or
`(ethyl, propyl! or butyl), particularly omega-amino-
`halo)-substituted 3-phthalidoxycarbonyl; polyhydroxy-
`lower alkoxycarbonyl or protected polyhydroxy-lower
`(ethyl, propy! or butyl).
`alkoxycarbonyl
`in which polyhydroxy-lower alkoxy
`A N-lower alkylamino group preferably contains 1-4
`carbon atomsin the loweralkyl portion and represents,
`and protected polyhydroxy-lower alkoxy represent
`preferably dihydroxypropyloxy or trihydroxybutyloxy
`for example, N-n-propyl-amino, N-iso-propylamino,
`N-n-butylamino, N-tert.-butylamino
`and
`advanta-
`wherein hydroxy groups are free or one or more, as
`appropriate, are protected in form ofesters, e.g. a lower
`geously N-methylamino or N-ethylamino.
`alkanoyl] or a benzoyl ester, in form of ethers, e.g. a
`A N,N-di-lower alkylamino group preferably con-
`tains 1-4 carbon atomsin each lower alky] portion and
`lower alkyl] or benzyl ether, or,
`in case two vicinal
`represents, for example, N,N-dimethylamino, N-meth-
`hydroxy groupsare involved,in the form ofacetals or
`ketals, e.g. a lower alkylidene, a benzylidene or a 5- or
`yl-N-ethylamino
`and
`advantageously
`N,N-die-
`thylamino.
`6-membered cycloalkylidene derivative.
`Hydroxy-loweralkyl is for example 2-hydroxyethy]
`Protected polyhydroxy-lower alkoxycarbonyl ad-
`vantageously represents (2,2-dimethyl-1,3-dioxolan-4-
`and preferably hydroxymethyl.
`Lower alkylthio as in lower alkylthio-lower alkyl
`yl)-methoxycarbonyl.
`represents advantageously C\-Cy,-alkylthio and prefera-
`Acyl as in acyloxy or acylamino represents prefera-
`bly lower alkanoyl, carbocyclic aryl-lower alkanoyl,
`bly methylthio or ethylthio.
`Loweralkylene represents branchedorstraight chain
`aroyl, lower alkoxycarbonyl or aryl-lower alkoxycar-
`alkylene of 1 to 7 carbon atoms, advantageously straight
`bonyl, advantageously lower alkanoyl. Lower alkoxy-
`chain (or linear) alkylene, such as methylene, ethylene,
`carbony!for acyl is preferably t-butoxycarbony]l (abbre-
`viated t-BOC). Aryl-lower alkoxycarbonyl for acyl is
`propylene, butylene, pentylene or hexylene and most
`preferably benzyloxycarbony] (abbreviated CBZ).
`preferably straight chain C)-Cy4-alkylene.
`Phenylene represents preferably 1,3 or 1,4-phenylene,
`Carboxy-lower alkoxycarbonyl represents advanta-
`advantageously 1,4-phenylene.
`geously e.g. 1-carboxyethoxycarbony]l.
`Cyclohexylene represents preferably 1,4-cyclohexy-
`Lower alkoxycarbonyl-lower alkoxycarbonyl repre-
`lene.
`sents advantageously e.g. 1-(ethoxycarbonyl)ethoxycar-
`bonyl.
`Halogen (halo) preferably represents
`chioro, but may also be bromooriodo.
`al-
`alkoxycarbonyl, mono-lower
`Amino-lower
`Lower alkanoyloxy advantageously contains 2 to 5
`kylamino-lower alkoxycarbonyl, di-dower)alkylamino-
`lower alkoxycarbonyl advantageously represent e.g.
`carbon atomsandis preferably acetoxy, pivaloyloxy or
`aminoethoxycarbonyl, ethylaminoethoxycarbonyl, die-
`propionyloxy.
`thylaminoethoxycarbonyl.
`Loweralkanoylamino advantageously contains 2 to 5
`Loweralkylidene is preferably isopropylidene.
`carbon atoms and is preferably acetylamino or pro-
`Cycloalkylidene is preferably cyclohexylidene.
`pionylamino.
`A lower alkoxycarbonyl! group preferably contains 1
`Carboxyl esterified in form of a pharmaceutically
`to 4 carbon atomsin the alkoxy portion and represents,
`acceptable prodrug ester
`represents most advanta-
`for example, methoxycarbonyl, n-propoxycarbony]l,
`geously (C-Cg-alkoxycarbony], phenyloxycarbonyl,
`benzyloxycarbonyl optionally substituted on phenyl by
`iso-propoxycarbonyl or advantageously ethoxycarbo-
`lower alkyl,
`lower alkoxy, halo or trifluoromethyl,
`nyl.
`
`fluoro or
`
`40
`
`45
`
`50
`
`35
`
`65
`
`BIOCON PHARMALTD (IPR2020-01263) Ex. 1009, p. 003
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1009, p. 003
`
`

`

`5,217,996
`
`Ro
`Oo
`H
`H
`|
`i
`Y
`i
`XOC—C—CH)—C—NH—C—A—(CH)COX’
`
`(r')
`
`Ry
`
`CHp-biary!
`
`wherein COX, COX’, Ri, Ro, A, biaryl and m have the
`meanings as defined herein above for compounds of
`formula I. The compoundsof formulae Ia, Ib, Ic, Id, Ie
`and If given below are present as well, preferably in the
`enantiomeric form depicted in formula I’.
`Illustrative thereof, in the above compoundsoffor-
`mula I wherein R; is lower alkyl, the carbon atom car-
`rying said substituent is assigned the (R)-configuration;
`and the carbon atom carrying the biarylmethy] substitu-
`ent is assigned the (S)-configuration.
`Moreparticularly, the present invention is concerned
`with and hasfor its object the compoundsof formula Ia
`
`5.
`pivaloyloxymethoxycarbonyl, 1-(C2-Cy4-alkanoyloxy)-
`ethoxycarbony]l,
`(2,2-dimethy]-1,3-dioxolan-4-yl)-
`methoxycarbonyl, 5-indanyloxycarbonyl, 3-phthalidox-
`ycarbonyl,
`bornyloxycarbonylmethoxycarbonyl,
` 1-
`(C\-C4-alkoxycarbonyloxy)-ethoxycarbonyl
`or
`3-
`pyridylmethoxycarbony].
`Carboxyl derivatized in the form of a pharmaceuti-
`cally acceptable amide represents preferably carbamoy]
`or N-substituted carbamoyl, advantageously [lower
`alkylamino, arylamino, di-lower alkylamino, morpho-
`lino, N-lower alkylpiperazino, pyrrolidino, piperidino,
`perhydroazepino, (amino or acylamino)-lower alkyl-
`amino or aryl-lower alkylamino}]-carbony]l.
`Pharmaceutically acceptable salts are either pharma-
`ceutically acceptable acid addition salts for any basic
`compoundsofthe invention or salts derived from phar-
`maceutically acceptable bases for any acidic com-
`poundsof the invention.
`Pharmaceutically acceptable salts of basic com-
`poundsof the invention are acid addition salts, which
`are preferably such of therapeutically acceptable inor-
`ganic or organic acids, such as strong mineralacids, for
`example hydrohalic, e.g. hydrochloric or hydro-bromic
`acid, sulfuric, phosphoric or nitric acid; aliphatic or
`aromatic carboxylic or sulfonic acids, e.g.
`formic,
`acetic, propionic, succinic, glycollic, lactic, malic, tar-
`taric, gluconic,citric, maleic, fumaric, pyruvic, pheny]-
`acetic, benzoic, 4-aminobenzoic, anthranilic, 4-hydrox-
`wherein COOR and COOR' independently represent
`ybenzoic, salicylic, 4-aminosalicylic, pamoic, nicotinic,
`carboxyl or carboxyl derivatized in form of a pharma-
`ceutically acceptable ester; Ri represents hydrogen,
`methanesulfonic,
`ethanesulfonic,
`hydroxyethanesul-
`lower alkyl, lower alkoxy, N-lower alkylamino, lower
`fonic, 1,2-ethanedisulfonic acid, benzenesulfonic, p-tol-
`alkanoylamino, aryl-lower alkyl, aryl-lower alkoxy,
`uenesulfonic, naphthalenesulfonic, sulfanilic, cyclohex-
`aryloxy, N-arylamino or aroylamino wherein ary] in
`ylsulfamic acid, or ascorbic acid.
`each case represents phenyl optionally substituted by
`Pharmaceutically acceptable salts of the acidic com-
`lower alkyl, lower alkoxy, halogen, hydroxy, cyano,
`pounds of the invention, e.g. those having a free car-
`acyloxy ortrifluoromethyl, or aryl represents thieny] or
`boxy! group are salts formed with pharmaceutically
`furanyl optionally substituted by lower alkyl; R2 repre-
`acceptable bases, e.g. alkali metal salts (e.g. sodium,
`sents hydrogen, hydroxy,
`lower alkyl or aryl-lower
`potassium salts), alkaline earth metal salts (e.g. magne-
`alkyl wherein ary! independently has the meaning given
`sium, calcium salts), ammonium salts, mono-, di- or
`above under Rj; R3 represents phenyl, or phenyl substi-
`tri-lower (alkyl or hydroxyalkyl)-ammoniumsalts(e.g. -
`tuted by lower alkyl, lower alkoxy, halogen, cyano,
`ethanolammonium, diethanolammonium,triethanolam-
`acyloxy ortrifluoromethyl; or R3 represents thienyl or
`45
`monium, tromethaminesalts).
`furany! optionally substituted by loweralkyl; A repre-
`The compounds of the invention, of formula I and
`sents a direct bond, lower alkylene, 1,4-phenylene or
`derivatives thereof may contain several asymmetric
`1,4-cyclohexylene; m represents 1 or zero provided that
`carbon atoms, depending on the nature of the substitu-
`m represents 1 whenAis a direct bond; or a pharmaceu-
`50
`ents. Thus the compoundsofthe invention exist in the
`tically acceptable salt thereof.
`form of geometric isomers, racemates, diastereoisomers,
`Advantageously, R3 is located in the para position.
`Particularly preferred embodiments of the invention
`pure enantiomers or mixtures thereof, all of which are
`as described aboverelate to:
`within the scope of the invention.
`a) compounds wherein R3is phenyl or pheny! substi-
`For example, the compounds of formula I exist in
`_
`tuted by lower alkyl, lower alkoxy, halogen, cy-
`isomeric forms, e.g. wherein the asymmetric carbon
`ano, acyloxy ortrifluoromethyl;
`atom on the butyryl chain bearing the Ry and/orbiary!-
`b) compounds wherein A is lower alkylene, m repre-
`methyl groups mayeither exist in the S or R configura-
`sents 1 or zero, and R2 represents hydrogen, lower
`tion. The compounds of the invention, e.g.
`those of
`alkyl, hydroxy or lower alkoxy.
`formula I having said two asymmetric centers exist as
`c) compounds wherein Ry represents hydrogen,
`two different racemic diastereoisomeric forms which
`lower alkyl,
`lower alkoxy or aryl-lower alkyl
`may be called erythro and threo depending ontherela-
`wherein ary! represents phenyl optionally substi-
`tive orientation of the Ri and biarylmethy] substituents
`tuted by one or two of loweralkyl, lower alkoxy,
`of the chain. Each of the two racemates consists of the
`halogen, hydroxy, cyano, acyloxy or trifluoro-
`optically active enantiomers (or antipodes) having
`methyl; most preferably compounds wherein Rj
`(S,S), (R,R), (R,S) or (S,R) configurations, respectively.
`represents lower alkoxy or loweralkyl.
`Preferred is the threo racemic form and particularly
`A particular embodiment of the invention relates to
`the enantiomeric form depicted in formula I’
`compoundsof formula Ib
`
`R2
`(Ia)
`ROOC—CH—CH)—cH—NH—b~a—(tn,—coor’
`Ry
`
`CH
`
`R3
`
`20
`
`25
`
`35
`
`40
`
`55
`
`60
`
`65
`
`BIOCON PHARMALTD (IPR2020-01263) Ex. 1009, p. 004
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1009, p. 004
`
`

`

`7
`
`5,217,996
`
`Oo
`
`R2
`
`(Ib)
`
`ROOC—CH=CH?—-CH™~NH=C—A—(CH)n,—COOR’
`
`2
`4
`il
`HOOC—CH—CHaCH—NH—C=(CH2)—COOH
`R;
`
`(Id)
`
`.
`D
`“HKG Ry
`™()()
`
`wherein Rjis loweralkyl; n is an integer 1 through 4; or
`a pharmaceutically acceptable mono-or di-ester deriva-
`tive thereof in which one or two of the acidic hydroxy
`groups of the carboxyl functional groupsare esterified
`in form of a mono- or di-pharmaceutically acceptable
`ester; or a pharmaceutically acceptable salt thereof; or
`an optical] antipode thereof.
`Preferred are said compoundsof formula Id wherein
`Ry is methyl! and n is 2; and mono-ordi-esters thereof.
`Asdiscussed before, the butyric acid compounds of
`e.g. formula Id exist
`in two distinct diastereomeric
`forms which maybe called erythro and threo. Preferred
`are e.g. the compoundsof formulaId as the threo diaste-
`Teomer (racemate), more particularly as the enantio-
`meric form having the R-configuration at C-atom 2 and
`the S-configuration at C-atom 4 and wherein the buty-
`Tyl portion is as depicted in formula Id’
`
`(id’)
`
`H Y
`
`Qnnariin
`
`owTt
`HOOC—C—CH)—C—NH—C—(CH;),;—COOH
`i
`C
`Ri
`
`wherein R) and n are as defined under formula Id; or a
`pharmaceutical acceptable mono-or diester derivative
`thereof; or a pharmaceutical acceptable salt thereof.
`Particularly preferred are compoundsof formula Ie
`
`wherein COOR and COOR' independently represent
`carboxyl or carboxyl derivatized in form of a pharma-
`ceutically acceptable ester; Ry is hydrogen, loweralkyl,
`lower alkoxy or aryl-lower alkyl wherein aryl repre-
`sents phenyl optionally substituted by lower alkyl,
`lower alkoxy, halogen, hydroxy, cyano, acyloxy or
`trifluoromethyl; R2 represents hydrogen, hydroxy or
`lower alkoxy; R4 and Rs independently represent hy-
`drogen, lower alkyl, hydroxy, lower alkoxy, halogen,
`cyanoortrifluoromethyl; A represents lower alkylene;
`m represents I or zero; or a pharmaceutical acceptable
`salt thereof.
`Particularly preferred are compoundsof formula Ic
`
`20
`
`°
`li
`ROOC—CH—CH2—CHTNH—C—(CHa)COOR’
`Ri
`|
`
`25
`
`(ic)
`
`Ry
`
`wherein COOR and COOR’ independently represent
`carboxyl or carboxyl derivatized in form of a pharma-
`ceutically acceptable ester; Ry is lower alkyl or lower
`alkoxy; R4 represents hydrogen,
`lower alkyl,
`lower
`alkoxy, halogen, or trifluoromethyl; n represents an
`integer | through 6; or a pharmaceutical acceptable salt
`thereof.
`Preferred are compounds of formula Ic wherein
`COORand COOR’independently represent carboxyl,
`C)-C29-alkoxycarbonyl, (carbocyclic or heterocyclic
`aryl)-lower alkoxycarbonyl, (di-lower alkylamino, N-—
`lower alkyipiperazino, morpholino, pyrrolidino, piperi-
`dino or perhydrazepino)-C2 to Cg4-alkoxycarbonyl,
`dihydroxypropyloxycarbonyl protected in form of a
`ketal, 5-indanyloxycarbonyl!, 3-phthalidoxycarbonyl,
`bicycloalkoxycarbonyl-lower
`alkoxycarbonyl,
` a-
`(ower alkoxycarbonyl or di-lower alkylaminocar-
`bonyl)-lower
`alkoxycarbonyl,
`1-(lower
`alkoxycar-
`bonyloxy)-lower
`alkoxycarbonyl
`or
`1-(lower
`al-
`kanoyloxy)-lower alkoxycarbonyl; or a pharmaceuti-
`cally acceptable salt thereof.
`Particularly preferred are said compoundsof formula
`Ic wherein COOR and COOR’independently represent
`carboxyl, C}-C4-alkoxycarbonyl,
`3-pyridylmethox-
`ycarbonyl, benzyloxycarbony] optionally substituted on
`phenyl by loweralkyl, lower alkoxy, halo ortrifluoro-
`methyl, 5-indanyloxycarbonyl, 1-(C2-Cs-alkanoyloxy)-
`ethoxycarbony], 3-phthalidoxycarbonyl, (2,2’-dimethy]-
`1,3-dioxolan-4-y])-methoxycarbonyl,
`bornyloxycar-
`bonylmethoxycarbonyl, 1-(C\-C4-alkoxycarbonyloxy)-
`ethoxycarbonyl; or a pharmaceutically acceptable salt
`thereof.
`A preferred embodiment of the invention relates to
`compoundsof formula Id
`
`|“()(\CH3
`
`45
`
`4
`2
`li
`ROOC™CHCH2—CH—NH—~C—(CH32)2—COOR’
`
`
`(le)
`
`50
`
`65
`
`wherein COOR and COOR’independently represent
`carboxyl or carboxy] esterified in form of a pharmaceu-
`tical acceptable prodrug ester; or a pharmaceutically
`acceptable salt thereof.
`Particularly preferred embodiments of the invention
`as. described aboverelateto:
`(a) compounds of the above formula Ie wherein R
`and R’ independently represent hydrogen, C)-C4-
`alkyl, benzyl optionally substituted on phenyl by
`loweralkyl, lower alkoxy, halo ortrifluoromethyl,
`pivaloyloxymethyl,
`1-(C2-C4-alkanoyloxy)-ethyl,
`(2,2-dimethyl-1,3-dioxolan-4-yl)-methyl, 5-indanyl,
`3-phthalidyl, bornyloxycarbonylmethyl, 1-(C-C4-
`alkoxycarbonyloxy)-ethyl or 3-pyridylmethy]; or a
`pharmaceutically acceptablesalt thereof;
`(b) compounds of the above formula Ie wherein
`COOR'is carboxyl; and COOR represents car-
`boxyl or carboxyl derivatized in form of a pharma-
`
`BIOCON PHARMALTD (IPR2020-01263) Ex. 1009, p. 005
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1009, p. 005
`
`

`

`9
`ceutically acceptable ester; or a pharmaceutically
`acceptablesalt thereof;
`(c) compounds of the above formula Ie having the
`R-configuration at C-atom 2 and the S-configura-
`tion at C-atom 4;
`(d) the compound according to the above formula Ie
`wherein COORis ethoxycarbonyl and COOR’ is
`carboxy], namely being 4-[N-(3-carboxy-1-oxo-
`propyl)amino}-4-(p-phenylphenylmethyl)-2-
`methylbutanoic acid ethyl ester,
`the (2R,4S)an-
`tipode thereof or a pharmaceutical acceptable salt
`thereof.
`The novel compoundsofthe invention are pharmaco-
`logically potent neutral endopeptidase enzyme inhibi-
`tors which inhibit e.g. the degradation of atrial natri-
`uretic factors (ANF) in mammals. They thus potentiate
`the diuretic and natriuretic effect of exogenous or en-
`dogenous ANF in mammals.
`The compoundsofthe invention are thus particularly
`useful in mammalsas diuretic, natriuretic (saluretic) and
`antihypertensive agents for the treatmentof e.g. hyper-
`tension, congestive heart failure and edema.
`As neutral endopeptidase inhibitors, the compounds
`are also e.g. enkephalinase inhibitors so as to inhibit the
`degradation of endogenous enkephalins and may thus
`also be useful for the treatment of pain in mammals.
`The above-cited properties are demonstrable in vitro
`and in vivo tests, using advantageously mammals,e.g.
`mice, rats, dogs, monkeysorisolated organs, tissues and
`preparations thereof. Said compounds can be applied in
`vitro in the form of solutions, e.g. preferably aqueous
`solutions, and in vivo either enterally, parenterally,
`advantageously intravenously, e.g. aS a suspension or in
`aqueous solution. The dosage in vitro may range be-
`tween about 10-4 molar and 10-9 molar concentra-
`tions. The dosage in vivo may range depending on the
`route of administration, between about 0.01 and 50
`mg/kg, advantageously between about 1.0 and 25
`mg/kg.
`Theanalgesic activity can be determined by measur-
`ing the potentiation of the analgesic effects of enkepha-
`lin and derivatives thereof, and by classical analgesic
`tests, such as the phenyl-p-benzoquinone induced writ-.
`ing test [J. Pharmacol. Exp. Therap. 125, 237 (1959)]
`and the hot plate test in the mouse [J. Pharmacol. Exp.
`Therap. 107, 385 (1953).
`The antihypertensive activity can be determined in
`the spontaneously hypertensiverat, Goldblatt rat or
`Goldblatt dog by direct measurementof blood pressure.
`Advantageously, the effect is measured in the DOCA-
`salt hypertensive rat and/or renal hypertensive rat or
`dog model.
`The diuretic (saluretic) activity can be determined in
`standard diuretic screens, e.g. as described in “New
`Antihypertensive Drugs”, Spectrum Publications, 1976,
`pages 307-321, or by measuring the potentiation of
`atrial natriuretic factor-induced natriuresis and diuresis
`in the rat.
`The potentiation of ANF can also be determined by
`measuring the increase in ANF plasmalevel achieved.
`The in vitro inhibition of neutral endopeptidase
`(NEP) 3.4.24.11 can be determined as follows:
`Neutral endopeptidase 3.4.24.11 activity is deter-
`mined by the hydrolysis of the substrate glutaryl-Ala-
`Ala-Phe-2-naphthylamide (GAAP) using a modified
`procedure of Orlowski and Wilk (1981). The incubation
`mixture (total volume 125 1) contains 4.2 pg of protein
`(rat kidney cortex membranes prepared by method of
`
`20
`
`35
`
`40
`
`45
`
`30
`
`60
`
`65
`
`5,217,996
`
`.
`10
`Maedaetal, 1983), 50 mMtris buffer, pH 7.4 at 25° C.,
`500 pM substrate (final concentration), and leucine
`aminopeptidase M (2.5 yg). The mixture is incubated
`for 10 minutesat 25° C. and 100 ploffast garnet (250 pg
`fast garnet/m! of 10% Tween 20 in 1M sodium acetate,
`pH 4.2) is added. Enzymeactivity is measured spectro-
`photometrically at 540 nm. One unit of NEP 24.11 ac-
`tivity is defined as 1 nmol of 2-naphthylamine released
`per minute at 25° C. at pH 7.4. ICso values are deter-
`mined,i.e. the concentration of test compound required
`for 50% inhibition of the release of 2-naphthylamine.
`Neutral endopeptidase activity is also determined
`using ANFas a substrate. A trial natriuretic factor de-
`grading activity is determined by measuring the disap-
`pearance of rat-ANF (r-ANF) using a 3 minute reverse
`phase-HPLCseparation. An aliquot of the enzymein 50
`mM Tris HCI buffer, pH 7.4, is preincubated at 37° C.
`for 2 minutes and the reaction is initiated by the addition
`of 4 nmol of r-ANF in a total volume of 50 pl. The
`reaction is terminated after 4 minutes with the addition
`of 30 pl of 0.27% trifluoroacetic acid (TFA). Forty
`microliters of the mixture is injected into a reverse
`phase-HPLC and analyzed using a C4 cartridge in a 3
`minute, isocratic separation. Twenty-three percent of
`buffer B (0.1% TFA in 80% acetonitrile) is used. Buffer
`A is 0.1% TFA in water. Oneunit of activity is defined
`as the hydrolysis of 1 nmol of r-ANF per minute at 37°
`C. at pH 7.4. ICso values are determined,i.e. the concen-
`tration of test compound required for 50% inhibition of
`the hydrolysis of ANF.
`The test compoundis dissolved in dimethy] sulfoxide
`or 0.25M sodium bicarbonate solution, and the solution
`is diluted with pH 7.4 buffer to the desired concentra-
`tion.
`In vitro testing is most appropriate for the free car-
`boxylic acids of the invention.
`/
`The effect of the compoundsofthe invention on rat
`plasma ANFconcentration can be determined as fol-
`lows:
`,
`Male Sprague-Dawley rats (275-390 g) are anesthe-
`tized with ketamine (150 mg/kg)/acepromazine (10%)
`and instrumented with catheters in the femoral artery
`and vein to obtain blood samples and infuse ANF,re-
`spectively. The rats are tethered with a swivel system
`and are allowed to recover for 24 hours before being
`studied in the conscious, unrestrained state.
`In this assay, plasma ANFlevels are determined in
`the presence and absence of NEPinhibition. On the day
`of study, all rats are infused continuously with ANF at
`450 ng/kg/min.i.v. for the entire 5 hours of the experi-
`ment. Sixty minutes after beginning the infusion, blood
`samples for baseline ANF measurements are obtained
`(time 0) and the rats are then randomly divided into
`groupstreated with the test compound or vehicle. Ad-
`ditional blood samples are taken 30, 60, 120, 180 and 240
`minutes after administration of the test compound.
`Plasma concentrations are determined by a specific
`radioimmunoassay. The plasmais diluted (x 12.5, x25
`and X50) in buffer containing: 50 mM Tris (pH 6.8), 154
`mM NaCi, 0.3% bovine serum albumin, 0.01% EDTA.
`One hundred microliters of standards [TANF (99-126)]
`or samples are added fo 100 pl of rabbit anti-rANF
`serum an