PCT
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`wo 99/46272
`
`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`International Bureau
`
`(51) International Patent Classification 6 :
`C07F 9/572, A61K 31/675, GOlN 33/58,
`C07F 9/40, 9/6571
`
`Al
`
`(11) International Publication Number:
`
`(43) International Publication Date:
`
`16 September 1999 (16.09.99)
`
`(21) International Application Number:
`
`PCT/EP99/01617
`
`(22) International Filing Date:
`
`9 March 1999 (09.03.99)
`
`(74) Agent: VAN SOMEREN, Petronella, Francisca, Hendrika,
`Maria; Arnold & Siedsma, Sweelinckplein 1, NL-2517 GK
`The Hague (NL).
`
`(81) Designated States: AL, AM, AT, AU, AZ, BA, BB, BG, BR,
`BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GD,
`GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP,
`KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK,
`MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI,
`SK, SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZW,
`ARIPO patent (GH, GM, KE, LS, MW, SD, SL, SZ, UG,
`ZW), Eurasian patent (AM, AZ, BY, KG, KZ, MD, RU, TJ,
`TM), European patent (AT, BE, CH, CY, DE, DK, ES, Fl,
`FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OAPI patent
`(BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE,
`SN, TD, TG).
`
`Published
`With international search report.
`
`(30) Priority Data:
`98200733.8
`
`9 March 1998 (09.03.98)
`
`EP
`
`(71) Applicant (for all designated States except US): FONDA TECH
`BENELUX N.V. [BE/BE]; Joe Englishstraat 54, B-2140
`Borgerhout (BE).
`
`(72) Inventors; and
`SCHARPE, Simon,
`(75) Inventors/Applicants (for US only):
`Lodewijk [BE/BE]; Universiteitsplein 1, B-2610 Antwer(cid:173)
`pen (BE). DE MEESTER, Ingrid, Anna, Jozef [BE/BE];
`Fort 7-straat 7, B-2610 Wilrijk (BE). BELIAEV, Alexandre
`Arkadicvitch [RU/PT]; Rua Particular Engenheiro Alberto
`Martins Mesquita, 224, P-4480 Mindelo-Vila Do Conde
`(PT). LAMBEIR, Anne-Marie, Virginie, Renee [BE/BE];
`Sparrendreef 35, B-3001 Heverlee (BE). AUGUSTYNS,
`Koen, Jan, Ludovicus [BE/BE]; Heike 2, B-2322 Minder(cid:173)
`hout (BE). HAEMERS, Achiel, Jean-Marie [BE/BE]; De
`Knok 2, B-9830 Sint-Martens-Latem (BE). GOOSSENS,
`Filip, Jozef, Anny [BE/BE]; Van Bellinghenlaan 3, B-1910
`Kampenhout (BE). HENDRIKS, Dirk, Frans [BE/BE];
`Zwaluwenlaan 9, B-2360 Aartselaar (BE).
`
`(54) Title: SERINE PEPTIDASE MODULATORS
`
`(57) Abstract
`
`The present invention relates to new compounds having modulatory (inhibitory and stimulatory) activity on serine peptidases and
`proteases in general and dipeptidyl peptidase IV, prolyl oligopeptidase (PO), dipeptidyl peptidase II (DPP II), fibroblast activation protein
`a (FAPa), lysosomal Pro-X carboxypeptidase and elastase in particular. These new compounds can be used for the treatment of a variety
`of disease states in which these peptidases are involved.
`
`AstraZeneca Exhibit 2154
`Mylan v. AstraZeneca
`IPR2015-01340
`
`Page 1 of 78
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`

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`Codes used to identify States party to the PCT on the front pages of pamphlets publishing international applications under the PCT.
`
`FOR THE PURPOSES OF INFORMATION ONLY
`
`AL
`AM
`AT
`AU
`AZ
`BA
`BB
`BE
`BF
`BG
`BJ
`BR
`BY
`CA
`CF
`CG
`CH
`CI
`CM
`CN
`cu
`cz
`DE
`DK
`EE
`
`Albania
`Annenia
`Austria
`Australia
`Azerbaijan
`Bosnia and Herzegovina
`Barbados
`Belgium
`Burkina Faso
`Bulgaria
`Benin
`Brazil
`Belarus
`Canada
`Central African Republic
`Congo
`Switzerland
`Cote d'Ivoire
`Cameroon
`China
`Cuba
`Czech Republic
`Gennany
`Denmark
`Estonia
`
`ES
`FI
`FR
`GA
`GB
`GE
`GH
`GN
`GR
`HU
`IE
`IL
`IS
`IT
`JP
`KE
`KG
`KP
`
`KR
`KZ
`LC
`LI
`LK
`LR
`
`Spain
`Finland
`France
`Gabon
`United Kingdom
`Georgia
`Ghana
`Guinea
`Greece
`Hungary
`Ireland
`Israel
`Iceland
`Italy
`Japan
`Kenya
`Kyrgyzstan
`Democratic People's
`Republic of Korea
`Republic of Korea
`Kazakstan
`Saint Lucia
`Liechtenstein
`Sri Lanka
`Liberia
`
`LS
`LT
`LU
`LV
`MC
`MD
`MG
`MK
`
`ML
`MN
`MR
`MW
`MX
`NE
`NL
`NO
`NZ
`PL
`PT
`RO
`RU
`SD
`SE
`SG
`
`Lesotho
`Lithuania
`Luxembourg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`The fonner Yugoslav
`Republic of Macedonia
`Mali
`Mongolia
`Mauritania
`Malawi
`Mexico
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Singapore
`
`SI
`SK
`SN
`sz
`TD
`TG
`TJ
`TM
`TR
`TT
`UA
`UG
`us
`uz
`VN
`YU
`zw
`
`Slovenia
`Slovakia
`Senegal
`Swaziland
`Chad
`Togo
`Tajikistan
`Turkmenistan
`Turkey
`Trinidad and Tobago
`Ukraine
`Uganda
`United States of America
`Uzbekistan
`VietNam
`Yugoslavia
`Zimbabwe
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`W099/46272
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`PCT/EP99/01617
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`1
`
`SERINE PEPTIDASE MODULATORS
`
`Field of the invention
`The present invention relates to novel
`5 modulators (inhibitors and stimulators) of serine
`peptidases and proteases in general and dipeptidyl
`peptidase IV, prolyl oligopeptidase (PO), dipeptidyl
`peptidase II (DPP II), fibroblast activation protein a
`(FAPa), lysosomal Pro-X carboxypeptidase and elastase in
`10 particular. The invention further relates to the
`preparation and use of these compounds for selective
`modulation (inhibition or stimulation) of serine
`peptidases and proteases and to pharmaceutical
`preparations comprising them. The terms "peptidase" and
`15 "protease" are used interchangeably.
`
`Background of the invention
`Serine peptidasesjproteases, like granzymes,
`mast cell tryptase, elastases, trypsin-like enzymes,
`20 prolyl oligopeptidase, dipeptidyl peptidase II and
`dipeptidyl peptidase IV are involved in various processes
`that take place in the body, such as blood coagulation,
`inflammation, immune response, and control of peptide
`hormone metabolism in general. Although serine peptidases
`25 are a physiological necessity they may also constitute a
`potential health hazard in case serine peptidase activity
`in the body is not controlled.
`Serine peptidases have been described to be
`involved in various medical indications. Blood
`30 coagulation serine proteases are for example responsible
`for vascular clotting as well as cerebral and coronary
`infarction. Chymotrypsin-like enzymes and plasmin are
`involved in tumour invasion, tissue remodeling and clot
`dissociation. Pancreatitis, emphysema, rheumatoid
`35 arthritis, inflammation and adult respiratory distress
`syndrome may in some instances be caused by the
`uncontrolled proteolysis by other serine proteases such
`as elastase.
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`15
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`Serine peptidases form a large group with many
`members that are divided into clans and families. One
`member of the clan sc is dipeptidyl peptidase IV (DPP IV,
`EC 3.4.14.5), which is a highly specific exopeptidase
`5 with a serine type mechanism of protease activity,
`cleaving off dipeptides from the amino-terminus of
`peptides with proline or alanine at the penultimate
`position. In addition the slow release of dipeptides of
`the type X-Gly or x-ser is reported for some naturally
`10 occurring peptides. DPP IV is constitutively expressed on
`epithelial and endothelial cells of a variety of
`different tissues, and is also found in body fluids. In
`the hematopoietic system, DPP IV was identified as the
`leukocyte antigen CD26.
`Prolyl oligopeptidase (PO, EC 3.4.21.26) was
`discovered in the human uterus as an oxytocin-degrading
`enzyme. The enzyme shows a high specificity for proline
`residues and hydrolyses the peptide bond at its carboxyl
`side, provided the proline is not at the peptide amino-
`20 terminus. This endopeptidase has like DPPIV, a serine
`type mechanism and it is characterised by its activity on
`oligopeptides. PO cleaves specifically the Pro-Xaa bond
`in biological active peptides (substance P, ocytoxin,
`vasopressin, gonadoliberin, bradykinin, neurotensin) and
`25 it is likely to participate in the in vivo regulation of
`their actions. A role for PO in memory and other neural
`processes has been proposed (Yoshimoto T. and Ito K. in
`Handbook of proteolytic enzymes, eds. Barrett et al.,
`Academic Press, 1998, p. 272-374).
`Fibroblast activation protein a (FAPa) was
`discovered as a cell surface antigen of cultured normal
`fibroblasts. Its expression in vivo revealed to be very
`restricted on normal cells. In contrast, activated tumor
`stromal fibroblasts found in certain carcinomas express
`35 high levels of FAPa. The biological role of FAPa
`expression remains to be elucidated but speculations on
`functions in tissue remodeling and repair have been made
`(Rettig, FAPa in Barrett supra, p. 385-389).
`
`30
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`10
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`Dipeptidyl peptidase II (DPPII, EC 3.4.14.2)
`releases N-terminal dipeptides from oligopeptides, provi(cid:173)
`ded their N-termini are unsubstituted. The preferred P1
`residues are Ala and Pro. An increase in serum DPPII has
`5 been observed in cancer patients and extremely high
`levels of DPPII are present in human carcinoma cells.
`DPPII can be inhibited by the classical (unspecific)
`inhibitors of serine type peptidases (J.K.McDonald in
`Barrett, supra, p. 408-411).
`Elastases are defined by their ability to
`release soluble peptides from insoluble elastin fibers by
`a proteolytic process called elastinolysis. Elastase
`belongs to the chymotrypsin family of leucocyte serine(cid:173)
`type proteases. Human leucocyte elastase (EC 3.4.21.37)
`15 preferentially cleaves peptides with a Val in P1 but also
`peptide bonds with Ala, Ser and Cys in P1 are hydrolyzed
`and it is believed to possess an extended substrate(cid:173)
`binding site. The possible involvement of leucocyte
`elastase in inflammatory diseases, triggered the search
`20 for development of specific inhibitors. Moreover, a
`pathological role in lung emphysema, cystic fibrosis and
`adult respiratory distress syndrome has been suggested
`(J. Bieth in Barrett, supra, p. 54-60; D.Farley et al. in
`Pharmaceutical Enzymes, ed. A. Lauwers and s. Scharpe,
`25 Marcel Dekker, Inc., 1997, p. 306-326).
`Lysosomal Pro-X carboxypeptidase
`(prolylcarboxypeptidase, angiotensinase C, EC 3.4.16.2)
`cleaves c-terminal amino acids from peptides with the
`general structure X-Pro-Y, where X is either a blocking
`30 group, another protected amino acid, or a peptide, and Y
`is an aromatic or aliphatic amino acid with a free
`carboxylic group. The enzyme is recovered from the
`lysosomal fraction of different tissues. Although the
`enzyme has an acidic pH optimum for small synthetic
`35 substrates (pH 5.0), it retains 50% of its maximal
`activity at physiological pH towards larger peptide
`substrates. (Des-Arg9)-bradykinin and angiotensin II are
`possible natural substrates for lysosomal Pro-X
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`carboxypeptidase (Tan, F. and Erdos, E. in Barrett et
`al., supra, p. 405-407)
`Because of their role in various physiological
`processes it is desirable to interfere in the activity of
`5 serine peptidases. such interference can be either
`stimulation or inhibition. Various types of serine
`peptidase inhibitors have been described in for example
`EP-764 151 of the present inventors. The application
`inter alia describes compounds of the general formula z-
`10 Xaa-Y' wherein Z may or may not be present and is a
`protecting group, Xaa represents a dipeptide or an amino
`acid and Y' may be a phosphonate, such as a diphenyl
`phosphonate. Further research of the present inventors
`has revealed that the toxicity thereof is not yet
`15 acceptable and the potency/efficacy are not sufficient.
`US-5,543,396 of Powers et al. relates to
`proline phosphonate derivatives. The phosphonate may be
`substituted with one or two phenyl groups which in turn
`may be mono-, di- or trisubstituted with a halogen, c 1-c6
`20 alkyl, c 1-c6 perfluoralkyl, c 1-c6 alkoxy, N02 , CN, OH, co2H,
`amino, c 1-c6 alkyl amino, c 2-c12 dialkylamino, c 1-c6 acyl,
`and C1-c6 alkoxy-co-, c 1-c6 alkyl-s-. The present
`inventors, in the research that led to this invention,
`developed independently the same compounds. However, they
`25 found that regarding toxicity, stability and efficacy
`these compounds did not perform optimally.
`It is therefore the object of the present
`invention to provide inhibitors of serine peptidases/
`proteases that have a more optimal combination of
`30 inhibitor capacity, stability in plasma, safety,
`bioavailability, duration of action and straightforward
`synthesis. In addition, the invention has for its object
`to provide compounds that have a stimulating activity on
`serine peptidases; proteases. These two types of
`35 compounds of the invention will also be identified herein
`as "modulating compounds". More in particular, the
`invention provides compounds having such a more optimal
`combination for modulating the activity of DPP IV, PO,
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`DPP II, FAPa, lysosomal Pro-X carboxypeptidase and
`elastase.
`
`Thus, in the research that led to the present
`invention, the influence of different functional groups
`5 on the inhibitory (or stimulatory) activity of
`phosphonates was investigated. Prolylpyrrolidine diphenyl
`phosphonates were synthesized, substituted on the phenyls
`with hydroxyl, methoxy, acylamino, sulfonylamino, ureyl,
`methoxycarbonyl and alkylaminocarbonyl groups. The
`10 phenylesters were also replaced by other groups with good
`leaving group capacities such as trichloroethyl and
`trifluoroethyl. The inhibitory activity in vitro and in
`vivo on DPP IV and other serine peptidases, the stability
`and specificity of these compounds was tested.
`It was then found that compounds as claimed in
`claim 1 are very potent modulators, in particular
`inhibitors, of serine peptidasesjproteases in general and
`DPP IV, DPP II, PO, FAPa, lysosomal Pro-X carboxy(cid:173)
`peptidase and elastase in particular. The compounds as
`20 listed in claim 12 were found to be potent inhibitors of
`DPP IV and PO.
`The compounds of the invention are based on
`peptides. These peptides are constituted by either
`naturally occurring amino acids or other amino acids. The
`25 c-terminal carboxyl function is replaced by a phosphonate
`group.
`
`15
`
`The compounds of the invention are represented
`by the general formula I:
`
`30
`
`35
`
`(I)
`
`wherein
`A is
`---(R2) or H or c 1-c6 alkyl or halogenoalkyl,
`except perfluoroalkyl,
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`5
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`20
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`the phenyl group is mono-, di- or trisubstituted
`with R1 or R2;
`X is a peptide- or amino acid-derived moiety;
`A and the phenyl group substituted with R1 may
`optionally form a biphenyl diester;
`all R1. substituents and R2 substituents are each
`independently selected from the group consisting of:
`a) c1-c6 acylamino;
`b) aroylamino, optionally substituted at the o-
`10 and/or p- andjor m- position with alkyl, in particular c1-
`c6 alkyl, andjor a halogen;
`c) c1-c6 alkylsufonylamino;
`d) arylsulfonylamino, optionally substituted at
`the o- andjor p- andjor m- position with alkyl, in
`15 particular c1-c6 alkyl, andjor a halogen;
`e) a aminoacylamino wherein the a aminoacyl
`represents a side chain blocked or unblocked a-amino acid
`residue with the L, D or DL configuration at the a-carbon
`atom selected from the group consisting of:
`alanine, methionine, methionine sulfoxide,
`arginine, homoarginine, phenylalanine, aspartic
`acid, proline, hydroxyproline, asparagine,
`serine, cysteine, threonine, histidine,
`glycine, tyrosine, glutamic acid, pyroglutamic
`acid, tryptophan, glutamine, valine, norvaline,
`isoleucine, lysine, leucine, norleucine,
`thioproline, homoproline, 1,2,3,4-tetrahydro(cid:173)
`isoquinoline-3-carboxylic acid (Tic),
`2,3-dihydroindol-2-carboxylic acid,
`a-naphtylglycine, a-phenylglycine,
`4-amidinophenylglycine, 4-phenylproline,
`4-amidinophenylalanine, 0-benzyl tyrosine,
`omega-acetyl lysine, a-aminobutyric acid,
`citrulline, homocitrulline, ornithine, o(cid:173)
`methylserine, o-ethylserine, s-methylcysteine,
`S-ethylcysteine, S-benzylcysteine, homoserine,
`4-dehydroproline, penicillamine, ~-(2-
`thienyl)alanine, NH2-CH(CH2CHEt2)-COOH, a-
`
`25
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`30
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`5
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`10
`
`aminoheptanoic acid, NH2-CH(CH2-1-naphthyl)(cid:173)
`COOH, NH2-CH(CH2-2-naphthyl)-COOH, NH2-CH(C~­
`cyclohexyl)-COOH, NH2-CH[CH-(cyclohexyl) 2]-COOH,
`NH2-CH(CH2-cyclopentyl)-COOH, NH2-CH[CH-
`(cyclopentyl)2]-COOH, NH2-CH(CH2-cyclobutyl)(cid:173)
`COOH, NH2-CH[CH-(cyclobutyl) 2]-COOH, NH2-CH(CH2-
`cyclopropyl)-COOH, NH2-CH[CH-(cyclopropyl) 2](cid:173)
`COOH, 5,5,5-trifluoroleucine,
`hexafluoroleucine, (S)-azetidine-2-carboxylic
`acid, (S)-pipecolic acid, (S)-oxazolidine-4-
`carboxylic acid, (R)-thiazolidine-4-
`carboxylacid (L-thioproline), sarcosine;
`f) residue selected from the group consisting
`of 3-aminobenzoic acid, E-aminocaproic acid,
`{3- alanine;
`g) Y-NH-CO-NH-;
`h) y I 02CCH (NHCO-Y) -CH2-;
`i) Y'NHCO-;
`j) CH3-0-CO-Y 1 -NH-CO-;
`k) CH3-CH2-0-CO-Y'-NH-CO-;
`wherein Y is c1-c6 alkyl, aryl or H and Y' is
`c1-c6 alkyl,
`and pharmaceutically acceptable salts thereof.
`In a specific embodiment of the invention X is
`25 a moiety of the general formula {AA)P-aa-,
`wherein:
`p indicates that there may be 0, 1, 2, 3, 4 or
`5 residues AA, which can be the same or
`different within one molecule;
`AA and aa are selected from one of the
`following:
`a) a-amino carboxylic acids with in a position
`an optionally substituted c1-c6 alkyl or aryl or
`aralkylmoiety;
`b) alanine, methionine, methionine sulfoxide,
`arginine, homoarginine, phenylalanine, aspartic
`acid, proline, hydroxyproline, asparagine,
`serine, cysteine, threonine, histidine,
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`15
`
`glycine, tyrosine, glutamic acid, pyroglutamic
`acid, tryptophan, glutamine, valine, norvaline,
`isoleucine, lysine, leucine, norleucine,
`thioproline, homoproline, 1,2,3,4-tetrahydro-
`isoquinoline-3-carboxylic acid (Tic),
`2,3-dihydroindol-2-carboxylic acid,
`a-naphtylglycine, a-phenylglycine,
`4-amidinophenylglycine, 4-phenylproline,
`4-amidinophenylalanine, 0-benzyl tyrosine,
`omega-acetyl lysine, a-aminobutyric acid,
`citrulline, homocitrulline, ornithine, o(cid:173)
`methylserine, o-ethylserine, s-methylcysteine,
`S-ethylcysteine, S-benzylcysteine, homoserine,
`4-dehydroproline, penicillamine, P-(2-
`thienyl)alanine, NH2-CH(CH2CHEt2)-COOH, a(cid:173)
`aminoheptanoic acid, NH2-CH(CH2-1-naphthyl)(cid:173)
`COOH, NH2-CH(CH2-2-naphthyl) -COOH, NH2-CH(CH2-
`cyclohexyl)-COOH, NH2-CH[CH-(cyclohexyl) 2]-COOH,
`NH2-CH(CH2-cyclopentyl)-COOH, NH2-CH[CH(cid:173)
`(cyclopentyl)2]-COOH, NH2-CH(CH2-cyclobutyl)(cid:173)
`COOH, NH2-CH[CH-(cyclobutyl) 2]-COOH, NH2-CH(CH2-
`cyclopropyl)-COOH, NH2-CH[CH-(cyclopropyl) 2](cid:173)
`COOH, 5,5,5-trifluoroleucine,
`hexafluoroleucine, (S)-azetidine-2-carboxylic
`acid, (S)-pipecolic acid, (S)-oxazolidine-4-
`carboxylic acid, (R)-thiazolidine-4-
`carboxylacid (L-thioproline), 3-aminobenzoic
`acid, sarcosine, €-aminocaproic acid, p(cid:173)
`alanine,
`wherein the alpha amino residue may be side
`chain blocked or unblocked and has the L, D, or
`DL configuration at the alpha carbon atom;
`and pharmaceutically acceptable salts thereof.
`In an alternative embodiment of the invention X
`35 is M- (AA) -aa-
`P
`wherein:
`p, AA and aa are as defined above; and
`M is selected from:
`
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`a)
`
`b)
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`5
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`10
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`15
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`25
`
`the group consisting of optionally
`-CSNH2 , -S02NH2 ,
`substituted -CONH2 ,
`phenyl-S02-, phenyl-CH2so2-, 2-furyl(cid:173)
`acryloyl; and
`the group of protecting groups
`consisting of:
`acetyl, adamantyloxycarbonyl, benzyl(cid:173)
`oxycarbonyl, benzoyl, benzyl,
`t-butoxycarbonyl, t-butyl,
`2,4-dinitrophenyl, formyl,
`fluorenylmethoxycarbonyl,
`4-methoxybenzyl, tosyl, trifluoro(cid:173)
`acetyl, trityl, phthaloyl,
`phenylalkylcarbonyl, 2-indanylacetyl,
`2-(1,2,3,4-tetrahydronaphtyl)acetyl,
`4-(4-benzylphenoxy)alkyl;
`In a specific embodiment X represents AA-aa-,
`wherein aa is proline and AA is as defined above. In an
`alternative embodiment X represents AA-aa-, wherein AA
`20 and aa are both proline. In case aa is alanine, R1 and R2
`may further be selected from:
`1) H, halogen, N02 , CN, OH, COOH
`m) amino, c1-c6 alkylamine, C2-C12 dialkylamino,
`n) c1-c6 acyl
`o) c1-c6 alkoxy-co-
`p) c1-c6 alkyl-s-.
`In such compounds aa is alanine and preferably at least
`the AA coupled to aa is proline or phenylalanine.
`Preferred examples of the compounds are Phe-Ala-diphenyl-
`30 phosphonate or Pro-Ala-diphenylphosphonate and
`pharmaceutically acceptable salts thereof.
`The invention can be divided in three groups of
`compounds.
`In preferred compounds of this invention the
`35 phosphonate group is a diphenyl-phosphonate group
`(indicated with the symbol P(OPh)2), which is preferably
`substituted. The first group thus consists of compounds
`of the general formula II:
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`x(cid:173)
`o
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`5
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`(II)
`
`wherein the substituents R1, R2 and X are as defined
`above (group 1).
`Alternatively, the compounds are 2,2' biphenyl
`diesters of a-aminoalkyl phosphonic acid having the
`10 general formula III:
`
`(III)
`
`15
`
`20
`
`wherein the substituents R1, R2 and X are as defined
`above (group 2).
`The third group (group 3) consists of compounds
`having the general formula IV:
`---R1
`
`0
`
`0
`
`X-P
`0
`
`(IV)
`
`A
`25 wherein X and Rl are as defined above, A is H or C1-c6
`alkyl or halogenoalkyl, except perfluoroalkyl.
`Specific members of these three groups will be
`listed below.
`For inhibition of the serine protease cathepsin
`30 G, X is preferably selected from Cbz-Gly-Leu-Phe-, Z-Phe(cid:173)
`Pro-Phe, and Suc-Val-Pro-Phe-. For prolyl oligopeptidase
`X may be selected from among the following: Cbz-Gly-Gly(cid:173)
`Pro-, Cbz-Pro-Pro-, Boc-Val-Pro-Val-, MeO-Suc-Ala-Ala(cid:173)
`Ala-Val-, MeO-Suc-Ala-Ala-Pro-Val-. For DPP IV, X is as
`35 follows: Ala-Pro-, Pro-Pro-, Ala-Pip-, Phe-Pro-, Ile(cid:173)
`Pro-, Arg-Pro-, pF-Phe-Pro-, cyclohexylala-Pro-, Pro(cid:173)
`azetidine-, Phe-azetidine-, Lys-Pro-, Lys-azetidine-. X
`is selected from among Suc-Lys(Cbz)-Val-Pro-Val-, Z-Ala-
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`Ala-Ala and Boc-Val-Pro-Val- in case the enzyme to be
`inhibited is human leukocyte elastase. For Granzyme A, X
`may be selected from Cbz-(4-amidinophenylalanine)-, z(cid:173)
`Met-, 3-phenyl propanoyl-Pro-(4-aminophenylalanine)-,
`5 Cbz-Thr-(4-amidinophenylglycine)-, and Boc-D-Phe-Pro-(4-
`amidinophenylalanine)-. X may be Z-Phe-Pro-Phe-, Z-Phe-,
`suc-Val-Pro-Phe-, MeO-Suc-Ala-Ala-Pro-Phe-, or Meo-suc(cid:173)
`Ala-Ala-Ala-Phe- when the enzyme to be inhibited is
`chymotrypsine. For trypsin-like serine-type proteases, X
`10 may be selected from among Cbz-Orn-, Cbz-Lys-Ala-, Cbz(cid:173)
`Lys, Cbz-HomoLys-, Cbz-(4-amidinophenylalanine)-, Cbz-(4-
`amidinophenylglycine)-, Ph-CH2-so2-Gly-Pro-(4-amidino(cid:173)
`phenylglycine)-, 3-(2-furyl)acryloyl-(4-amidinophenyl(cid:173)
`glycine), Cbz-Lys-(4-amidinophenylglycine)-, Cbz-Lys-Ala-
`15 (4-amidinophenylglycine)-, Cbz-Thr-(4-amidinophenyl(cid:173)
`glycine)-, 3-(2-furyl)acryloyl-(4-amidinophenylalanine)-,
`Cbz-Ala-(4-amidinophenylglycine)-, Cbz-Ala-Ala-Ala-(4-
`amidinophenylglycine)-, 2-phenoxybenzoyl-Pro-(4-
`amidinophenylglycine)-, 3-phenoxybenzoyl-Pro-(4-
`20 amidinophenylglycine)-, 3-phenyl propanoyl-Pro-(4-
`amidinophenylalanine)-, 3,3-diphenyl propanoyl-Pro-(4-
`amidinophenylglycine)-. For inhibition of va protease of
`s.aureus X can be either Acetyl.Glu- or Acetyl.Asp-.
`R1 and R2 are preferably selected from the
`25 group consisting of: 3-AcNH, 4-AcNH, 4-MeS02NH, 3-H2NCONH,
`3-H2NCONH 1 4-(N-Bz-Gly-NH) 1 4-(H-Gly-NH) 1 4(H-(~)-Ala-NH) 1
`4- ( (~) -Pyr-NH) 1 4- ( (2~) -Me02CCH(NHAc) CH2) 1 4-Me02CI 4-
`(Et02CCH2NHCO) 1 4-(Me02C(CH2 ) 2NHCO), 4-CH3 (CH2 ) 2NHCO.
`Particularly preferred compounds of group 1
`30 (formula II) are the following:
`Di(3-acetamidophenyl) 1-(benzyloxycarbonyl-(S)(cid:173)
`prolyl)pyrrolidine-2(R1S)-phosphonate (lOd);
`Di(4-acetamidophenyl) 1-(benzyloxycarbonyl-(S)(cid:173)
`prolyl)pyrrolidine-2(R1S)-phosphonate (lOe);
`Di(4-methylsulfonylaminophenyl) 1-(benzyloxy-
`carbonyl-(S)-prolyl)pyrrolidine-2(R1S)-phosphonate
`( lOf) ;
`
`35 -
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`Di{3-ureylphenyl) 1-{benzyloxycarbonyl-(S)(cid:173)
`prolyl)pyrrolidine-2(R,S)-phosphonate {lOq);
`Di[4-(N-benzoylglycylamino)phenyl]-1-(benzyl(cid:173)
`oxycarbonyl-{S)-prolyl)pyrrolidine-2(R,S}-phospho-
`nate {lOh);
`Di[4-(N~benzyloxycarbonylglycylamino)phenyl]-1-
`(benzyloxycarbonyl-(S)-prolyl)pyrrolidine-2(R,S)(cid:173)
`phosphonate (lOi};
`Di[4-(N-benzyloxycarbonyl-(S)-alanylamino)phenyl]-
`1-(benzyloxycarbonyl-(S)-prolyl)pyrrolidine-2(R,S)-
`phosphonate (lOj);
`Di[4-((S)-pyroglutamylamino)phenyl]-1-(benzyloxy(cid:173)
`carbonyl-(S)-prolyl)pyrrolidine-2(R,S)-phosphonate
`(10k);
`Di{4-[-(S)-(2-methoxycarbonyl-2-acetamido)ethyl]-
`phenyl}1-(benzyloxycarbonyl-(S)-prolyl)pyrrolidine-
`2(R,S)-phosphonate (101);
`Di(4-methoxycarbonylphenyl)1-(tert-butyloxycarbonyl(cid:173)
`(S)-prolyl)pyrrolidine-2(R,S)-phosphonate (10m);
`Di{4-[(ethoxycarbonyl)methylaminocarbonyl]phenyl}
`1-(benzyloxycarbonyl-(S)-prolyl)-pyrrolidine-
`2{R,S)-phosphonate {lOn);
`Di{4-[2-(methoxycarbonyl)ethylaminocarbonyl]phenyl}
`1-(benzyloxycarbonyl-(S)-prolyl)-pyrrolidine-
`2(R,S)-phosphonate (lOo);
`Di[4-(n-propylaminocarbonyl)phenyl] 1-(benzyloxy(cid:173)
`carbonyl-{S)-prolyl)-pyrrolidine-2{R,S)-phosphonate
`(lOp);
`Di(3-acetamidophenyl) 1-({S)-prolyl)pyrrolidine-
`2(R,S)-phosphonate hydrochloride {lld);
`Di(4-acetamidophenyl) 1-({S)-prolyl)pyrrolidine-
`2{R,S)-phosphonate hydrochloride (lle);
`Di(4-methylsulfonylaminophenyl) 1-((S)-prolyl)(cid:173)
`pyrrolidine-2(R,S)-phosphonate hydrochloride (llf);
`Di(3-ureylphenyl) 1-((S)-prolyl)pyrrolidine-2(R,S)(cid:173)
`phosphonate hydrochloride (llq);
`Di[4-(N-benzoylglycylamino)phenyl]-1-((S)-pro1yl)(cid:173)
`pyrrolidine-2(R,S)-phosphonate hydrochloride (llh);
`
`5
`
`10
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`15 -
`
`20 -
`
`25
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`30
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`35 -
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`Di[4-(N-glycylamino)phenyl]-1-((S)-prolyl)(cid:173)
`pyrrolidine-2(R,S)-phosphonate trihydrochloride
`(11i);
`Di(4-(S)-alanylaminophenyl)-1-((S)-prolyl)pyrroli-
`dine-2(R,S)-phosphonate trihydrochloride (11j);
`Di(4-(S)-pyroglutamylaminophenyl)-1-((S)-prolyl)(cid:173)
`pyrrolidine-2(R,S)-phosphonate hydrochloride (11k);
`Di{4-[-(S)-(2-methoxycarbonyl-2-acetamido)ethyl](cid:173)
`phenyl} 1-((S)-prolyl)pyrrolidine-2-phosphonate
`hydrochloride (111);
`Di{4-[(ethoxycarbonyl)methylaminocarbonyl]phenyl}
`1-((S)-prolyl)pyrrolidine-2(R,S)-phosphonate (11n);
`Di{4-[2-(methoxycarbonyl)ethylaminocarbonyl]phenyl}
`1-((S)-prolyl)-pyrrolidine-2(R,S)-phosphonate
`hydrochloride (11o);
`Di[4-(n-propylaminocarbonyl)phenyl] 1-((S)-prolyl)(cid:173)
`pyrrolidine-2(R,S)-phosphonate hydrochloride (11p).
`A compound having an inhibitory activity for
`DPP IV, is for example 2,2'-Biphenyl 1-((S)-prolyl)-
`20 pyrrolidine-2(R,S)-phosphonate hydrochloride (19) or a
`pharmaceutically acceptable salt.
`Other specific compounds of group 2 (formula
`III) are the following:
`2,2'-Biphenyl 1-(benzyloxycarbonyl-(S)-prolyl)-
`pyrrolidine-2(R,S)-phosphonate (17a);
`2,2'-Biphenyl 1-(t-butyloxycarbonyl-(S)-prolyl)(cid:173)
`pyrrolidine-2(R,S)-phosphonate (17b).
`An example of a compound of group 3 (formula
`
`25
`
`IV) is:
`30 -
`2-(2'-Hydroxyphenyl)phenyl methyl 1-(S)-prolyl)-
`pyrrolidine-2(R,S)-phosphonate hydrochloride (18);
`The advantage of compounds of the invention
`over Powers et al. (supra) follows from the comparison of
`the properties of the leaving groups that are created
`35 from Powers• compounds and from the compounds of the
`invention when the inhibitors enter into a covalent
`binding with the serine in the active center of the
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`peptidases. The information comes from the Merck Index,
`12th edition, Merck & Co, Inc. USA (1996).
`Powers' leaving groups are among others 2-
`methylphenol (o-cresol), 3-methylphenol (m-cresol) and 4-
`5 methylphenol (p-cresol). The compounds belong to the
`group of cresols, which are desinfectants. These
`compounds are toxic to humans. Chronic poisoning from
`oral or percutaneous absorption may produce digestive
`disturbances, nervous disorders, vertigo, skin eruptions,
`10 jaundice, oliguria, uremia. Another leaving group in
`Powers' compounds is 4-hydroxybenzoic acid methylester
`(methyl paraben, Nipagin M) which is also a preservative
`in foods beverages and cosmetics and of which allergic
`reactions are frequently observed. 1,4-Benzenediol
`15 (hydroquinone) is a photographic developer and reducer
`and is used as an antioxidant. At very low concentrations
`there is no systemic toxicity. However, ingestion of more
`than 1 g results in nausea, vomiting, shortness of
`breath, cyanosis, convulsions and collapse. It is lethal
`20 in a dose above 5 g.
`The leaving groups of the compounds of the
`invention however, are for example 4-hydroxyacetanilide
`(paracetamol, compound lle) which is used commonly as a
`very safe analgesic/antipyretic drug. Another leaving
`25 group is 4-hydroxyhippuric acid ethyl ester (compound
`lln) of which no toxicity has been noticed, and hippuric
`acid is known to be a metabolite occurring in human
`metabolism and a normal constituent in human urine.
`From the above it follows that in comparison to
`30 Powers' compounds, the compounds of the invention do not
`present safety problems which could obstruct their
`further pharmaceutical development.
`The compounds of the invention can be used for
`the therapy of pathological states associated with
`35 excessive, impaired or unbalanced activity of said
`enzymes.
`
`Starting from the available information on the
`correlation between particular serine protease activities
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`and various disease states the skilled person will be
`able to define therapeutical utilities for the modulatory
`compounds of the invention. Hereinbelow examples of such
`disease states will be listed and support for their
`5 utility given.
`Thus, the invention relates to the compounds
`for use as a therapeutical agent. In particular, the
`invention relates to the compounds for use in the
`treatment or prophylaxis of inflammation, vascular
`10 diseases, organ specific or systemic auto-immune diseases
`(e.g. Graves' disease or multiple slerosis, inflammatory
`bowel disease), joint diseases, muscle diseases,
`neurological diseases, obesity, diseases associated with
`benign and malign cell transformation, spreading of
`15 malignant cells, conditions of glucose-intolerance,
`abnormal growth or growth retardation, rejection of
`foreign cells or tissues after transplantation,
`abnormalities in blood cell development, abnormal blood
`clotting, pain, or diseases of the central nervous
`20 system.
`
`Support for the pharmaceutical utility of the
`compounds of the invention in the above indications
`follows from the following publications, which
`demonstrate the correlation of specific serine peptidases
`25 with medical indications. Inhibition of these enzymes can
`thus be used as treatment or prophylaxis.
`The involvement of plasma prekallikrein or
`kallikrein in shock is described by w. Colman in Barrett,
`A., et al., supra, page 147-153.
`The compounds of the invention can be used for
`prevention and treatment of thrombosis and conjunction
`therapy of acute myocardial infarction by specifically
`inhibiting Factor X (Vlasuk, G.P. in New Therapeutic
`Agents in Thrombosis and Thrombolysis, Sasahara, A.,
`35 Loscalzo, J., eds (1997), pages 261-283), thrombin
`(Shafer, J.A. in New Therapeutic Agents in Thrombosis and
`Thrombolysis, (supra) pages 143-157) and Factor VII
`(factor VII-Tissue Factor) (Shafer, J.A. in New
`
`30
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`Therapeutic Agents in Thrombosis and Thrombolysis,
`(supra) pages 225-260).
`Various serine peptidases are known to be
`involved in inflammation. That this is the case is
`5 described for elastases (Bank u. et al., in Cellular
`Peptidases in Immune Function and Diseases, Ansorge, s. &
`Langner, J., Plenum Press (1997), pages 231-242, D.
`Farley et al. in Lauwers, A. and Scharpe, s. eds.,
`supra), kallikrein (Chao, J. in Barrett et al., supra,
`10 pages 97-101; Naidoo, Y. & Bhoola, K. in The kinin
`System, Framer, S.G., Academic Press (1998), pages 187-
`197); and Erdos, E.G. & Skidgel, R.A. in The Kinin
`System, supra, pages 112-141), cathepsin G (Flad, H.D. et
`al., in Cellular Peptidases in Immune Function and
`15 Diseases, supra, pages 223-230), DPP IV (Tanaka, s. et
`al., Immunopharmacology 40 (1998), 21-26), and granzymes
`(Berthou, c. et al., Pathol. Biol. 46 (1998), 617-624).
`DPP IV (Cheng, H.C. et a1. J. Biol. Chem. 273
`(1998) 24207-24215), PO (Goossens et al., Eur. J. Clin.
`20 Chem. Clin. Biochem. 34 (1996), 17-22; and Ishino, T