WO1997043245A1 - Inhibition de metalloproteases matricielles par des composes contenant de l'acetylene - Google Patents

Inhibition de metalloproteases matricielles par des composes contenant de l'acetylene Download PDF

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WO1997043245A1
WO1997043245A1 PCT/US1997/007921 US9707921W WO9743245A1 WO 1997043245 A1 WO1997043245 A1 WO 1997043245A1 US 9707921 W US9707921 W US 9707921W WO 9743245 A1 WO9743245 A1 WO 9743245A1
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compounds
mmp
group
carbons
acid
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PCT/US1997/007921
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Brian R. Dixon
Jinshan Chen
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Bayer Corporation
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Priority to CA002253796A priority Critical patent/CA2253796C/fr
Priority to BR9709077A priority patent/BR9709077A/pt
Priority to AU29386/97A priority patent/AU710759B2/en
Priority to EP97923622A priority patent/EP0912496A1/fr
Priority to JP09540980A priority patent/JP3090957B2/ja
Publication of WO1997043245A1 publication Critical patent/WO1997043245A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/44—Iso-indoles; Hydrogenated iso-indoles
    • C07D209/48—Iso-indoles; Hydrogenated iso-indoles with oxygen atoms in positions 1 and 3, e.g. phthalimide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/34—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C59/00—Compounds having carboxyl groups bound to acyclic carbon atoms and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
    • C07C59/40—Unsaturated compounds
    • C07C59/76—Unsaturated compounds containing keto groups
    • C07C59/90—Unsaturated compounds containing keto groups containing singly bound oxygen-containing groups

Definitions

  • the matrix metalloproteases are a famiK of zmc endoproteinases which include, but are not limited to. interstitial collagenase (a.k.a. MMP-1 ).
  • stromelysin a.k.a. proteoglvcanase, transin, or MMP-3
  • geiatmase A a.k.a .
  • MMP-2 72kDa-gelat ⁇ nase or MMP-2
  • geiatmase B a.k.a. 95kDa-gelat ⁇ nase or MMP-9.
  • MMPs are secreted by a variety of cells including fibroblasts and chondrocytes, along with natural proteinaceous inhibitors known as TIMPs (Iissue Inhibitor of MetalloProtemase).
  • MMPs are capable of destroying a variety of connective tissue components of 5 articular cartilage or basement membranes. Each MMP is secreted as an inactive proenzyme which must be cleaved m a subsequent step before it is able to exert its own proteolytic activity. In addition to the matrix destroying effect, certain of these MMPs such as MMP-3 have been implemented as the tn vivo activator for other MMPs such as MMP-1 and MMP-9 (Ito, et al., Arch
  • MMP-3 can cleave and thereby inactivate the endogenous inhibitors of other proteinases such as elastase ( inyard, et al., FEBS Letts. 222. 1. 91, 1991). Inhibitors of MMP-3 could thus influence the activity of other destructive protemases by modifyin ⁇ he of their endogenous inhibitors.
  • a number of diseases are thought to be mediated by excess or undesired mam -destroying eulloprotease activity or by an imbalance in the ratio of the MMPs to the TLMPs
  • These include a) osteoa ⁇ h ⁇ tis (Woessner, et al., J. Biol.Chem. 259(6). 3633, 1984, Phadke, et al., J Rheumatol
  • OA osteoarthritis
  • RA rheumatoid arthritis
  • septic arthritis the progressive loss of articular cartilage and thereby normal joint function.
  • No marketed pharmaceutical agent is able to prevent or slow this cartilage loss, although nonsteroidal anti- inflammatory drugs (NSAlDs) have been given to con ⁇ ol pain and swelling.
  • NSAlDs nonsteroidal anti- inflammatory drugs
  • MMP inhibitors are expected to halt or reverse the progression of cartilage loss and obviate or delay surgical inter, ention Proteases are critical elements at several stages in the progression of metastatic cancer.
  • WO-9421625 Al Dickens, et al.. U.S. Pat. No. 4,599,361 ; Hughes, et al., U.S. Pat. No. 5.190.937, Broadhurst, et al.. EP 574758 Al ; Broadhurst, et al., EP 276436; and Myers, et al, EP 520573 Al
  • the preferred compounds of these patents have peptide backbones with a zinc complexmg group (hydroxamic acid, thiol. carboxylic acid or phosphinic acid) at one end and a va ⁇ ety of sidechains. both those found in the natural ammo acids as well as those with more novel functional groups.
  • Such small peptides are often poorly absorbed, exhibiting low oral bioavailability. They are also subject to rapid proteolytic metabolism, thus having short half lives.
  • batimastat the compound desc ⁇ bed in Brown, et al., WO-9321942 A2, can only be given mtra pe ⁇ toneally.
  • 3-biphenoylpropanoic and 4-biaryioylbutano ⁇ c acids are described in the literature as ami- inflammatory, anti-platelet aggregation, anti-phlogistic, anti-proliferative, hypolipidemic, antirheumaric, analgesic, and hypocholesteroiemic agents. In none of these examples is a reference made to MMP inhibition as a mechanism for the claimed therapeutic effect.
  • Certain related compounds are also used as intermediates in the preparation of liquid crystals. Specifically. Tomcufcik, et al., US patent 3.784.701 claims certain substituted benzoylpropionic acids to treat inflammation and pain. These compounds include 3-biphenoylpropanoic acid (a.k.a. fenbufen) shown below.
  • Child, et al.. J. Phar . Sci. 6 >, 466, 1977 describes structure-activity relationships of several analogs of fenbufen. These include several compounds in which the biphenyl ring system is substituted or the propanoic acid portion is substituted with phenyl, halogen, hydroxyl or methyl, or the carboxylic acid or carbonyl functions are converted to a variety of de ⁇ vatives. No compounds are described which contain a 4'-substituted biphenyl and a substituted propanoic acid portion combined in one molecule. The phenyl (compounds XLEX and LXXVU) and methyl (compound XL VII) substituted compounds shown below were described as inactive.
  • Nichl, et al. DE patent 1957750 also desc ⁇ bes certain of the above methylene substituted biphenoylpropanoic acids.
  • R' is an alkyl of 1 - 10 carbons.
  • Thyes. et al.. DE patent 2854475 uses the following compound as an intermediate.
  • the biphenyl group is not substituted.
  • R', R 2 alkyl, benzyl, H, or, together with the nitrogen, morpholinyl.
  • Others have disclosed a se ⁇ es of biphenyl-containing carboxylic acids, illustrated by the compound shown below, which inhibit neural endopeptidase (NEP 24.1 1 ).
  • a membrane-bound zmc metalloprotease Stimon. et al., Bioorg. Med. Chem. Lett. 4, 539, 1994; Lombaert. et al., Bioorg. Med. Chem. Lett. 4. 2715. 1994: Lombaert, et al.. Bioorg. Med. Chem. Lett. 5_. 145. 1995: Lombaert. et al.. Bioorg. Med. Chem. Lett, £, 151. 1995).
  • N-carboxyalkyl derivatives containing a biphenylethylglycine are inhibitors of stromelysin- 1 (MMP-3), 72 kDA geiatmase (MMP-2) and collagenase (Durette, et al.. WO-9529689).
  • MMP inhibitors which possess improved bioavailability and biological stability relative to the peptide-based compounds of the prior art, and hich can be optimized for use against particular target MMPs.
  • Such compounds are the sub j ect of the present application.
  • MMP inhibitors would afford new therapies for diseases mediated b> the presence of. or an excess of MMP activity, including osteoarth ⁇ tis.
  • inhibitors of MMPs have been descnbed in the literature, including th ols (Beszant. et al . J Med Chem 3_£, 4030, 1993).
  • hydroxamic acids (Wahl. et al. Bioorg. Med. Chem. Lett. . 349. 1995. Conway. et al. J. Exp Med. ! ⁇ 2. 9. 1995; Porter, et al..
  • This invention provides compounds having matrix metalloprotease inhibitory activity. These compounds are useful for inhibiting mat ⁇ x metalloproteases and, therefore, combating conditions to which MMP's cont ⁇ bute. Accordingly, the present invention also provides pharmaceu t ical compositions and methods for treating such conditions.
  • the compounds described relate to a method of treating a mammal comp ⁇ sing admuuste ⁇ ng to the mammal a mat ⁇ x metalloprotease inhibiting amount of a compound according to the invention sufficient to: ⁇ a) alleviate the effects of osteoarth ⁇ tis, rheumatoid arthntis. septic arth ⁇ tis. pe ⁇ odontal disease, comeal ulceration. proteinu ⁇ a, aneurysmal aortic disease, dystrophobic epidermolysis. bullosa, conditions leading to inflammatory responses, osteopenias mediated by MMP activity, tempero mandibular joint disease, demyelating diseases of the nervous system;
  • the compounds of the present invention are also useful scientific research tools for studying functions and mechanisms of action of mat ⁇ x metalloproteases in both in vtvo and m vitro systems.
  • the present compounds can be used to modulate MMP action, thereby allowing the researcher to observe the effects of reduced MMP activity in the expe ⁇ mental biological system under study.
  • This invention relates to compounds having matrix metalloprotease inhibitory activity and the generalized formula:
  • T,A represents a substituted or unsubstitute ⁇ aromatic 6-membered nng or heteroaromatic 5 - 6 membered nng containing 1 - 2 atoms independently selected from the group of N, 0, or S T represents a substituted acetylemc moiety
  • B represents an aromatic 6-membered nng or a 5 heteroaromatic 5 - 6 membered nng contaimng I - 2 atoms independently selected from the group
  • D represents
  • E represents a chain of n carbon atoms bea ⁇ ng m substituents R 6 in which the R ⁇ groups are independent substituents. or constitute spiro or ⁇ onspiro ⁇ ngs. Rings may be formed in two ways: a) two groups R 6 are joined, and taken together with the cham atom(s) to w hich the two R 6 group(s) are attached, and any mtervemng chain atoms, constitute a 3 - 7 i membered nng, or b) one group R 6 is joined to the chain on which this one group R 6 resides, and taken together with the cham atom(s) to which the R 6 group is attached, and any mtervemng chain atoms, constitutes a 3 - 7 membered ring.
  • the number n of carbon atoms in the cham is 2 or 3
  • the number m of R 6 subsutuents is an integer of 1 - 3.
  • Each group R* is alkyl, alkenyl, alkynyl, heteroaryl, non-aromatic cyclic, and combinations thereof optionally substituted with one or more heteroatoms as described more fully below.
  • E is a substituted mono- or bicyclic moiety optionally substituted with one or more heteroatoms.
  • G represents -PO,H : -M
  • M represents -CO : H. -CON(R") : or -CO : R' ⁇ and R 13 represents any of the side chains of the 19 noncyciic naturally occurring ammo acids.
  • R' 5 is selected from the group comprising: HOCH 2 , MeOCH 2 , (n-Pr) : NCH : , CH 3 CO : CH,, EtOCO : CH 2 ⁇ HO(CH,) 2 , CH,CO : (CH 2 ) 2 , HO 2 C(CH 2 ) 2 , OHC(CH 2 ) 3 , HO(CH,) # Ph, 3-HO-Ph. and PhCH,OCH,; and R' 6 is selected from the group comprising: HOCH 2 , MeOCH 2 , (n-Pr) : NCH : , CH 3 CO : CH,, EtOCO : CH 2 ⁇ HO(CH,) 2 , CH,CO : (CH 2 ) 2 , HO 2 C(CH 2 ) 2 , OHC(CH 2 ) 3 , HO(CH,) # Ph, 3-HO-Ph. and PhCH,OCH,; and R' 6 is
  • the biphenyl portion of the molecule is unsubstituted, and the propanoic or butanoic acid portion is either unsubstituted or has a single methyl or phenyl group. Presence of the larger phenyl group has been reported to cause prior art compounds to be inactive as anti-inflammatory analgesic agents. See, for example, Child, et al., J.
  • the compounds of the present invention are matenals having mat ⁇ x metalloprotease inhibitory activity and the generalized formula:
  • R 1 represents H or alkyl of 1 - 3 carbons.
  • an open bond indicates the point at which the structure joins to another group.
  • the aromatic ring is referred to as the A ring or A unit
  • T represents a substituent group, referred to as a T group or T unit.
  • T is a substituted acetylenic moiety and x is 1.
  • the B nng of generalized formula (L) is a substituted or unsubstituted aromatic or heteroaromatic ring, in which any substituents are groups which do not cause the moiecule to fail to fit the active site of the target enzyme, or disrupt the relative conformations of the A and B rings, such that they would be detrimental.
  • substituents may be moieties such as lower alkyl, lower alkoxy, CN, NO 2 , halogen, etc., but are not to be limited to such groups.
  • B represents an aromatic or heteroaromatic ring selected from the group consisting of: in which R 1 is defined as above. These rings are referred to as the B ring or B unit.
  • D represents the moieties:
  • E represents a chain of n carbon atoms bearing m substituents R 6 . referred to as R 6 groups or R 4 units.
  • the R 6 gr° u P s are independent substituents, or constitute spiro or nonspiro rings. Rings may be formed in two ways: a) two groups R 6 are joined, and taken together with the chain atom(s) to which the two R* group(s) are attached, and any intervening chain atoms, constitute a 3 - 7 membered ring, or b) one group R 4 is joined to the chain on which this one group R 6 resides, and taken together with the chain atom(s) to which the R 6 group is attached, and
  • any mtervemng chain atoms constitutes a 3 - 7 membered nng.
  • the chain is 2 or 3. and the number m of R 6 substituents is an integer of 1 - 3.
  • the number of carbons in the totality of R 6 groups is at least two.
  • Each group R* is independently selected from the group consisting of the substituents listed below as items 1 ) - 14).
  • alkyl of 1 - 10 carbons provided that if the A unit is phenyl, the B unit is phenylene.
  • m is 1.
  • n is 2.
  • the alkyl group is located on the alpha carbon relative to the D unit, then x is 1 or 2.
  • phenylene the aryl group is phenyl.
  • n is 2. and m is 1 or 2, then x is 1 or 2;
  • arylalkyl in which the aryl portion contains 6 - 10 carbons and the alkyl portion contains 1 - 8 carbons;
  • heteroaryl-alkyl in which the heteroaryl portion comp ⁇ ses 4 - 9 carbons and at least one N. O. or S heteroatom, and the alkyl portion contains 1 - 8 carbons;
  • portion contains 2 - 5 carbons
  • heteroaryl-alkenyl in which the heteroaryl portion comprises 4 - 9 carbons and at least one N, O, or S heteroatom and the alkenyl portion contains 2 -5 carbons;
  • aryl-alkynyi in which the aryl portion contains 6 - 10 carbons and the alkynyl portion contains 2 - 5 carbons;
  • heteroaryl-alkynyl in which the heteroaryl portion comprises 4 - 9 carbons and at least one N, 0, or S heteroatom and the alkynyi portion contains 2 - 5 carbons:
  • R 7 in which t is 0 or an integer of 1 - 5 and R' is selected from the group consisting of:
  • R 7 group comprises 4 - 9 carbons and at least one N, 0, or S heteroatom.
  • Y represents O or S;
  • R 1 , R 2 , and R 3 are as defined above, and u is 0, 1, or 2 provided that when R 7 is
  • heteroarylalkyi in which the aryl portion contains 6 to 12 carbons and at least one N. O, or S heteroatom and the alkyl portion contains 1 to 4 carbons
  • -C(0)R' in which the R 9 represents alkyl of 2 to 6 carbons, aryl of 6 to 10 carbons, heteroaryl comp ⁇ sing 4 to
  • arylalkyl in which the aryl portion contains 6 to 10 carbons or is a heteroaryl comprising 4 to 9 carbons and at least one N, O, or S heteroatom, and the alkyl portion contains 1 to 4 carbons with the provisos that when
  • R' is -C(O)R' .
  • Z is -S- or -O-; when Z is -0-, R 1 may also be - ⁇ C q H ⁇ O) ⁇ 5 in which q, r, and R 5 are as defined above; and when the A unit is phenyl, the B unit is phenylene.
  • m is 1 , n is
  • aryl or heteroaryl portions of any of the T or R* groups optionally may bear up o two substituents selected from the group consisting of - ⁇ CH j ) y C(R")(R 12 )OH, -(CH 2 ) y OR n ,
  • G represents -PO ; H : -M,
  • M represents -CO : H. -CON(R") 2 . or -CO,R 12 . and R 13 represents any of the side chains of the 1 noncyclic naturally occurring amino acids.
  • the G unit is most preferably attached to the E unit at the carbon ⁇ to the D unit and is preferably a carboxyiic acid group.
  • alkyl means straight, branched, cyclic, and polycyclic materials.
  • haloalkyl means partially or fully halogenated alkyl groups such as - ⁇ CH : ) : C1. -CF, and -C 6 F I3 for example.
  • the invention relates to compounds of generalized formula (L) in which at least one of the units A, B. and R* comprises a heteroaromatic ring.
  • Preferred heteroaromatic ring-containing compounds are those in which the heteroaryl groups are heteroaryl of 4 - 9 carbons comprising a 5 - 6 membered heteroaromatic ring containing O, S, or NR l when the ring is 5-membered, and N when said ring is 6-membered.
  • Particularly preferred heteroaromatic ring-containing compounds are those in which at least one of the A and B units comprises a thiophene ring.
  • a unit is thiophene, it is preferably connected to B unit at position 2 and carries one substituent group T on position 5.
  • B u t is thiophene. it is preferably connected through positions 2 and 5 to D and A units respectively.
  • the A and B rings are preferably phenyl and phenylene. respectively, the A nng preferably bears at least one substituent group T preferably located on the position furthest from the position of the A ring which is connected to the B nng, the D unit is preferably a carbonyl group, and the G unit is preferably a carboxyl group.
  • the invention relates to compounds of generalized formula (L). in the E unit of which n is 2 and m is 1. These compounds thus possess two carbon atoms between the
  • the invention relates to compounds of generalized formula
  • the invention relates to compounds of generalized formula (L) in which m is 1 and R 6 is an independent substituent.
  • L generalized formula (L) in which m is 1 and R 6 is an independent substituent.
  • R 6 is -(CH 2 ) t R 7 have t as an integer of 1-5
  • the invention relates to compounds of generalized formula (L) in which the number of substituents m on the E unit is 2 or 3; and when m is 2, both groups R* are independent substituents, or together constitute a spiro ring, or one group R* is an independent substituent and the other constitutes a spiro nng; and when m is 3, two groups R 6 are independent substituents and one group R 6 constitutes a ring, or two groups R 6 constitute a ring and one group
  • R" is an independent substituent, or three groups R 6 are independent substiments.
  • This subset therefore contains compounds in which the E unit is di- or tri- substituted, and in the disubstituted case any rmgs formed by one or both R 6 groups are spiro rings, and in the trisubstituted case, the R 6 groups may form either spiro or nonspiro ⁇ ngs.
  • the invention relates to compounds of generalized formula
  • ⁇ therefore contains compounds in which the E umt cames one or two substituents R°. and at least one of these substituents is involved in a nonspiro nng.
  • each group R 14 is independently selected from the group consisting of:
  • alkyl of 1 - 9 carbons arylalkyl m which the alkyl portion contains 1 - 7 carbons and the aryl portion contains 6 - 10 carbons; alkenyl of 2 - 9 carbons; aryl -substituted alkenyl in which the alkenvl portion contains 2 - 4 carbons and the aryl portion contains 6 • 10 carbons; alkynyl of 2 - 9 carbons: aryl -substituted alkynvl in which the alkynyl portion contains 2 - 4 carbons and the aryl portion contains 6 - 10 carbons; aryl of 6 - 10 carbons; -COR 2 ; -CO 2 R 3 ; -CON(R 2 ),.
  • R 7 in which t is 0 or an integer of 1 - 4: and -(CH : XZR' in which v is 0 or an integer of 1 to 3. and Z represents -S- or -0-.
  • R'. R 7 . and R* have been defined above.
  • Preferred compounds of generalized formula (L) in which one or more of the substituent groups R 6 are involved in the formation of nonspiro ⁇ ngs have E units of the following structures
  • the substiment group T is preferably an acetylene contaimng moiety with the general formula:
  • the subsc ⁇ pt x. which defines the number of T substiments. is preferably 1 or 2, most preferably 1. and when x is I the T is preferably on the 4- position of ring A.
  • the A nng is preferably a phenyl or thiophene nng, most preferably phenyl.
  • the B ring is preferably a 1 ,4-phenyiene or 2.5-thiophene nng, most preferably 1.4-phenylene.
  • the D unit is most preferably a carbonyl group.
  • R* is preferably:
  • arylalkyl wherein the aryl portion contains 6 - 10 carbons and the alkyl portion contains 1 - 8 carbons;
  • aryl portion contains 6 to 12 carbons and the alkyl portion contains 1 to 4 carbons.
  • the group R* is most preferably the following, wherein, any aromatic moiety is preferably substimted: 1 ) arylalkyl wherein the aryl portion is phenyl and the alkyl portion contains 1 - 4 carbons;
  • variable groups of these methods are as described in the generic description if they are not specifically defined below.
  • the variable subscript n is independently defined for each method.
  • the raw product MI-A often exists as a mixture of isomers via attack of the anhydride from either of the two carbonyls.
  • the resultant isomers can be separated into pure forms by crystallization or chromatography using standard methods known to those skilled in the art.
  • the succinic anhyd ⁇ des Mill can be prepared via a Stobbe Condensation of a dialkyl succinate with an aldehyde or ketone (resulting in side chain R 6 ), followed by catalytic hydrogenation. hydrolysis of a hemiester intermediate to a diacid.
  • Method A is especially useful for the preparation of cyclic compounds such as MI-A-3, in which two Ro groups are connected in a methylene chain to form a 3-7 member ring.
  • Small ring (3-5 member) anhydrides are readily available only as cis isomers which yield cis invention compounds MI-A-3
  • the trans compounds Ml-A-4 are then prepared by treatment of MI-A-3 with a base such as DBL ' in THF.
  • the substimted four member ring starting material anhydrides such as Mffi-A-l are formed in a photochemical 2-2 reaction as shown below This method is especially useful for the preparation of compounds in which R 14 is acetoxy or acetoxymethylene.
  • the compounds MI can be prepared via a reaction sequence involving mono-aikylation of a dialkyl malonate MVI with an alkyl halide to form intermediate MVII, followed by alkylation with a halomethyl biphenyl ketone MVffi to yield intermediate MIX.
  • Compounds of structure MIX are then hydrolyzed with aqueous base and heated
  • esters MI-B-2 with R ⁇ : as alkyl are obtained, and using more than
  • the diester intermediate MIX can be heated with a strong acids such as
  • Intermediates MVII are formed from biphenyls MH in a Friedel-Craf reaction with haloaceryl halides such as bromoacetyl bromide or chloroacetyl chloride.
  • Method B has the advantage of yielding single regio isomers when Method A yields mixtures.
  • Method B is especially useful when the side chains R 6 contain aromatic or heteroaromatic rings that may participate in intramolecular acyiation reactions to give side products if Method A were to be used. This method is also very useful when the R 6 group adjacent to the carboxyl of the
  • final compound contains heteroatoms such as oxygen, sulfur, or nitrogen, or more complex functions such as imide rings.
  • malonate MVII can be prepared by alkylating a commercially available unsubsttmted malonate with prenyl or allyl halide, subject this product to ozonalysis with reductive work-up, and the desired z group can be coupled via a Mitsunobu reaction (Mitsunobu, Synthesis 1, 1981).
  • the intermediate alcohol can be subjected to alkylation conditions to provide malonate MVII containing the desired Z group.
  • n General Method E - Biaryl compounds such as those of this application may also be prepared by Suzuki or Stille cross-coupling reactions of aryl or heteroaryl metallic compounds in which the metal is zinc. tin. magnesium, lithium, boron, silicon, copper, cadmium or the iike with an aryl or heteroaryl halide or t ⁇ flate (trifluoromethane-sulfonate) or the like. In the equation below either Met or X is the metal and the other is the halide or triflate (OTf).
  • Pd(com) is a soluble complex of palladium such as tetrakis(t ⁇ phenylphosphine)-palladium(O) or bis- (triphenylphosphine)- palladiumdu) chloride.
  • palladium such as tetrakis(t ⁇ phenylphosphine)-palladium(O) or bis- (triphenylphosphine)- palladiumdu) chloride.
  • the materials in which X is halo can be converted to those in which X is metal by reactions well known to those skilled in the art, such as treatment of a bromo intermediate with hexamethyiditin and palladium tetrakistriphenyiphosphine in toluene at reflux to yield the t ⁇ methyltin intermediate.
  • the intermediates MXXtl are either commercial or easily prepared from commercial materials by methods well known to those skilled in the art.
  • Method E is also especially useful for the preparation of products in which the aryl groups.
  • a or B. contain one or more heteroatoms (heteroaryls) such as those compounds that contain thiophene, furan, pyridine, pyrrole, o azole. thiazole, pyrimidine or pyrazine rings or the like instead of phenyls.
  • V Conversion of key intermediate CVHI to the targeted patent compounds is accomplished m several ways dependmg on the identity of side chain function Z.
  • Reaction of CVITJ with W ig reagents followed by hydrogenation yields products in which Z is alkyl and or arylalkyl
  • Selective reduction of aldehyde CVHJ with a reducing agent such as lithium tns [(3-ethyl- 3pentyl ) oxy]alummum hyd ⁇ de (LTEPA) yields alcohol CK.
  • the alcohol is converted to phenyl ethers or a vanety of heteroatom substimted de ⁇ vatives used to generate sidecham Z via the
  • CEX is converted to a leaving group such as tosylate (CX) or bromide by conditions well known to those skilled in the art and then the leaving group is displaced by an appropriate nucleophile.
  • Amides of the acids of the invention compounds can be prepared from the acids by treatment in an appropnate solvent such as dichloromethane or dimethylformamide with a primary or secondary amine and a coupling agent such as dicyclohexylcarbodiimide. These reactions are well known to those skilled in the art.
  • the amme component can be simple alkyl or arylalkyl substimted or can be amino acid de ⁇ vatives in which the carboxyl is blocked and the amino group is free.
  • Suitable pharmaceutically acceptable salts of the compounds of the present invention include addition salts formed with organic or inorganic bases.
  • the salt forming ion de ⁇ ved from such bases can be metal ions, e.g., aluminum, alkali metal ions, such as sodium or potassium, alkaline earth metal ions such as calcium or magnesium, or an amme salt ion, of which a number are known for this purpose.
  • Examples include ammonium salts, arylalkylamines such as dibenzyiamine and .V ⁇ ' -dibenzylethylenediamine, lower alkylamines such as methylamme, -butylamine, procame, lower alkylpiperidines such as N-ethylpiperidine, cycloalkylamines such as cyclohexylamine or dicyclohexyiamine, 1-adamantylam ⁇ ne, benzathine, or salts derived from amino acids like argmine, lysine or the like.
  • the physiologically acceptable salts such as the sodium or potassium salts and the amino acid salts can be used medicinally as described below and are preferred.
  • the salts are produced by reacting the acid form of the invention compound with an equivalent of the base supplying the desired basic ion in a medium which the salt precipitates or in aqueous medium and then Ivophihzing
  • the free acid form can be obtained from the salt by conventional neutralization techniques, e.g.. with potassium bisulfate, hydrochlo ⁇ c acid, etc.
  • the compounds of the present invention have been found to inhibit the mat ⁇ x metalloproteases MMP-3. MMP-9 and MMP-2, and to a lesser extent MMP-1 , and are therefore useful for treating or preventing the conditions referred to in the background section.
  • MMPs not listed above share a high degree of homology with those listed above, especially in the catalytic site, it is deemed that compounds of the invention should also inhibit such other MMPs to varying degrees. Varying the substiments on the biaryl portions of the molecules, as well as those of the propanoic or butanoic acid chains of the claimed compounds, has been demonstrated to affect the relative inhibition of the listed MMPs.
  • compounds of this general class can be lk tuned" by selecting specific substiments such that inhibition of specific MMP(s) associated with specific pathological conditions can be enhanced while leaving non-invoived MMPs less affected.
  • SUBSTITUTE SHEET (RULE 26 ⁇ ) The method of treating matrix metalloprotease-mediated conditions may be practiced in mammals, including humans, that exhibit such conditions.
  • inhibitors of the present invention are contemplated for use in veterinary and human applications. For such purposes, they will be employed in pharmaceutical compositions containing active ingredient(s) plus one or more pharmaceutically acceptable earners, diluents, fillers, binders, and other excipients. depending on the administration mode and dosage form contemplated.
  • Administration of the inhibitors may be by any suitable mode known to those skilled in the art.
  • suitable parenteral administration include intravenous, intraarticular, subcutaneous and intramuscular routes.
  • Intravenous administration can be used to obtain acute regulation of peak plasma concentrations of the drug.
  • Improved half-life and targeting of the d g to the joint cavities may be aided by entrapment of the drug in liposomes. It may be possible to improve the selectivity of liposomal targeting to the joint cavities by incorporation of ligands into the outside of the liposomes that bind to synovial-specific macromolecules.
  • intramuscular, intraarticular or subcutaneous depot injection with or without encapsulation of the dmg into degradable microspheres e.g., comprising poiy(DL-lactide-co-glycolide) may be used to obtain prolonged sustained drug release.
  • degradable microspheres e.g., comprising poiy(DL-lactide-co-glycolide)
  • an ⁇ .p. implanted reservoir and septum such as the Percuseal system available from Pharmacia.
  • injector pens e.g. the Novo Pin or Q-pen
  • needle-free jet injectors e.g. from Bioject, Mediject or Becton Dickinson
  • Prolonged zero-order or other precisely controlled release such as pulsatile release can also be achieved as needed using implantable pumps with delivery of the drug through a cannula
  • Examples include the subcutaneously implanted osmotic pumps available from ALZA, such as the ALZET osmotic pump.
  • Nasal delivery may be achieved by incorporation of the drug into bioadhesive paniculate earners ( ⁇ 200 -m) such as those comp ⁇ smg cellulose, polyacrylate or polycarbophil. in conjunction with suitable absorption enhancers such as phospholipids or acylcamitines.
  • bioadhesive paniculate earners ⁇ 200 -m
  • suitable absorption enhancers such as phospholipids or acylcamitines.
  • Oral delivery may be achieved by incorporation of the drug into tablets, coated tablets, dragees, hard and soft gelatine capsules. solutions, emulsions or suspensions. Oral delivery may also be achieved by incorporation of the dmg into enteric coated capsules designed to release the dmg into the colon where digestive protease activity is low. Examples include the OROS-CT/OsmetTM and PULSENCAPTM systems from ALZA and Scherer Drug Delivery Systems respectively. Other systems use azo-crosslinked polymers that are degraded by colon specific bacterial azoreductases, or pH sensitive polyacrylate polymers that are activated by the rise in pH at the colon. The above systems may be used in conjunction with a wide range of available absorption enhancers.
  • Rectal delivery may be achieved by incorporation of the dmg into suppositories.
  • the compounds of this invention can be manufactured into the above listed formulations by the addition of various therapeu ⁇ caliy inert, inorganic or organic carriers well known to those skilled in the art. Examples of these include, but are not limited to, lactose, com starch or derivatives thereof, talc, vegetable oils, waxes, fats, polyols such as polyethylene glycol, water, saccharose. alcohols, glycerin and the like. Various preservatives, emulsifiers, dispersants.
  • flavorants we ⁇ mg agents, antioxida ts, sweeteners, colorants, stabilizers, salts, buffers and the like are also added, as required to assist in the stabilization of the formulation or to assist in increasing bioavailability of the active ⁇ ngredient(s) or to yield a formulation of acceptable flavor or odor in the case of oral dosing.
  • the amount of the pha ⁇ naceutical composition to be employed will depend on the recipient and the condition being treated. The requisite amount may be determined without undue experimentation by protocols known to those skilled in the art. Alternatively, the requisite amount may be calculated, based on a determinanon of the amount of target enzyme which must be inhibited m order to treat the condition.
  • the atnx metalloprotease inhibitors of the invention are useful not only for treatment of the physiological conditions discussed above, but are also useful in such activities as purification of metalloproteases and testing for matrix metalloprotease activity.
  • activity testing can be both in vitro using natural or synthetic enzyme preparations or in vivo using, for example, animal models in which abnormal destructive enzyme levels are found spontaneously (use of genetically mutated or transgenic animals) or are induced by administration of exogenous agents or by surgery which disrupts joint stability.
  • Step I A dry 2-L, three-necked, round-bonomed flask was equipped with a stir bar. a pressure equalizing addition funnel, an argon inlet and a thermometer. The flask was charged with a suspension of sodium hydride (8.4 g of 95% NaH; -0.33 mol) in dry THF (700 mL) and was cooled with an ice water bath. Diethyl malonate (48.54 g, 0.30 mol) was added dropwise from the addition funnel over 25 min. Stirring was continued for 1.5 h before adding l-bromo-3-phenylpropane (47 mL. -61 g, -0.30 mol) over 10 min via the addition funnel.
  • l-bromo-3-phenylpropane 47 mL. -61 g, -0.30 mol
  • Step 2 A 1 -L, one-necked, round bottom flask was equipped with a mbber septum and an argon
  • Step 3 A dry 2-L, three-necked, round-bottomed flask was equipped with a magnetic stir bar, an argon inlet, and a pressure equalizing addition funnel. The flask was charged with a solution of the product of step 1 (63.0 g, 0.227 mol) in THF (500 mL). The reaction vessel was cooled with an ice
  • the reacuon mixture was stirred for 1 h at 0 °C, and a solution of the product of step 2 (80.0 g, 0.215 mol) in dry THF (300 mL) was added via addition funnel over ca. 20 nun.
  • the deep orange reaction mixture as stirred at room temperature under argon for 3 h.
  • the reaction vessel was cooled in an ice water bath while distilled water (150 mL) was added cautiously.
  • the aqueous phase was extracted with three 300 mL portions of ethyl acetate, the combined orgamc phases were dried over
  • Step 4 The unpurified diacid from step 3 was dissolved in 1,4-dioxane (500 mL) and heated to reflux for 24 h. The solvent was removed m vacuo, and a 10 g portion of the residue chromatographed on siiica gel (gradient elution with 10-50% ethyl acetate-hexanes containing 1% acetic acid) to afford 0.840 g (10%) of the desired product as a yellow solid. MP 174 °C.
  • Step 5 A one-necked, 15-mL. round-bonomed flask equipped with a rubber septum and an argon needle inlet was charged with 2.6 mL of diethyiamine, the product of step 4 (0.300 g, 0.667 mmol).
  • propargyl alcohol 1.0 mL. 0.96 g, 17 mmol.
  • copper (I) iodide 0.0220 g, 0.1 15 mmol
  • :r ⁇ /w-dichlorobis(triphenylphosphine)palladate 0.1 10 g, 0.157 mmol
  • reaction mixture was concentrated (290 mg residue) and pan of the residue (90 mg) was purified via column chromatography on 50 g of silica gel (20% ethyl acetate-hexanes with 0.5% acetic acid) afforded the coupling product as a white solid (0.035 g, 40%) of coupling product as a white solid.
  • Example 2 and Example 3 were prepared by chiral separation of Example 1 on a Chiralcel AD * column (2 cm x 25 cm) using 5% EtOH, 4.75% H,O and 0.095% HOAc in CH 3 CN, flow rate
  • Example 6 Preparation of Compound VI
  • Example 1 (0.0070 g, 0.014 mmol), and acetic anhyd ⁇ de (0.020 mL. 22 mg, 0.21 mmol). The reaction mixture was stirred for 2 h at room temperature, and then added to 30 mL of IN HC1. The resulting mixture was extracted with three 30 mL portions of ethyl aceute, the combined orgamc phases were d ⁇ ed over MgSO 4 and concentrated. Purification via HPLC (2.5% ethyl acetate-dichloromethane with 0.01% trifluoroacetic acid) afforded 3 mg (38%) of Example 6. MP 137 °C.
  • Example 7 Preparation of Compound VII
  • t ⁇ ethylamme 2 mi of THF
  • compound I 0.0570 g, 0.134 mmol
  • ethyl chlorofo ⁇ nate 0.032 mL. 36 mg, 0.34 mmol
  • the reaction mixture was stirred for 16 h at room temperature and then added to 50 mL of IN HC1.
  • the resulting mixture was extracted with three 50 mL portions of ethyl acetate, the combined organic phases were dried over MgS0 4 and concentrated.
  • Example II Example 2
  • Example 7 Example 1 1
  • Step 1 A solution of sodium hydride (4.35 g, 181 mmol) in freshly distilled THF (100 mL) was cooled to 0 °C and treated with commercially available diallyl malonate (35.0 g, 190 mmol) over 40 mm via a dropping funnel. After stirring at room temperature for 30 min, ⁇ -(2-bromoethyl)phthaiimide (43.9 g, 247 mmol) was added to the solution in one portion and the mixture was heated to reflux. After 48 h the solution was cooled to 0 °C, quenched with 2N HC1 and concentrated to about 20% of its original volume.
  • the concentrate was diluted with ethyl acetate (300 mL) and washed successively with saturated aqueous solutions of : C0 3 and NaCl.
  • the organic layer was dried over MgSO 4 , filtered and concentrated under reduced pressure.
  • Purification by flash column chromatography gradient elution with 5-25% ethyl acetate-hexanes) afforded diallyl 2-phthalimidoethylmalonate (41.2 g, 64%) as a colorless oil.
  • Step 2 A solution of the product of step 1 (5.20 g, 14.6 mmol) in freshly distilled THF (100 mL ) was cooled to 0 °C. while NaH (385 mg, 16.1 mmol) was slowly added.
  • Step 3 A solution of the product of step 2 (6.50 g, 10.4 mmol) in 1,4-dioxane (100 mL) was cooled to 0 C. while tet ⁇ akis(triphenyIphosphine)palladium (0.180 g, 146 mmol) and pyrrolidine (2.40 mL, 29.2 mmol) were added sequentially. After stimng for 2 h at 0 'C and 4 h at room temperature, the reaction mixture was poured into 2N HCl ( 100 mL).
  • Step 4 A one-necked, 100-mL. round-bonomed flask equipped with a rubber septum and an argon needle inlet containing 2 mi of THF was charged with NaH (435 mg, 17.2 mmol) and cooled to 0 ⁇ C while propargyl alcohol (1.0 mL, 0.963 g, 17.2 mmol) was added via syringe over ca. 5 min. The resulting mixture was stirred at 0 °C for 10 min and at room temperature for 30 min.
  • Step 5 The procedure of Example 1 , step 5 was used to prepare Example 15 using the product from
  • step 4 and the product of step 3 as starting materials.
  • MP 151 * C Example 16 - Preparation of Compound XVI t-BuM ⁇ iSiO.
  • Step 1 A one-necked, 100-mL, round-bonomed flask equipped with a mbber septum and an argon needle miet was charged with propargyl alcohol (1.0 mL. 0.963 g, 17.2 mmol). ether (20 ml), and cooled to 0 °C while NaH (435 mg, 17.2 mmol) was added siowly. The resulting mixture was stirred at room temperature for 1 h. and /-butyldimethylsilyl chloride (2.60 g, 17.2 mmol) was added.
  • Step 2 The procedure of Example 1, step 2 was used to prepare the desired acetyl biphenyl using commercially available 4-iodobiphenyl and acetyl chloride.
  • Step 3 The procedure of Example I, step 5 was used to prepare the desired biphenyl acetylene using
  • Step 4 one-necked. 50-mL. round-bottomed flask equipped with a mbber septum and an argon needle inlet was charged with 5 mL of THF. the product of step 3 (1.06 g, 2.94 mmol), and cooled to -78 °C while potassium hexamethyldisiiazide (617 mg. 2.94 mmol) was added dropwise via syringe. The reaction mixture was stirred at -78 °C for 30 min. t ⁇ methylsilyl chlonde (0.374 mL.
  • Step 6 A one-necked. 50-mL. round-bottomed flask equipped with a rubber septum and an argon
  • the P218 quenched fluorescence assay (Micro fluorometric Profiling Assay) is a modificauon of that originally described by Knight, et al.. FEBS Lett. 296. 263, 1992 for a related substance and a variety of matrix metalloproteinases (MMPs) in cuvettes.
  • MMPs matrix metalloproteinases
  • P218 Fluorogcnic Substrate is a synthetic substrate contaimng a 4-acetyl-7- methoxycouma ⁇ n ( MCA) group in the N-termmal position and a 3-[2. 4-dimtrophenyl]-L-2.3- diaminopropionyl (DPA) group internally. This is a modification of a peptide reported by Knight ( 1992) that was used as a substrate for mat ⁇ x metalloprotemases. Once the P218 peptide is cleaved (putative clip site at the Ala-Leu bond), the fluorescence of the MCA group can be detected on a fluorometer with excitation at 328 nm and emission at 393 nm. P218 is currently being produced BACHEM exclusively for Bayer. P218 has the structure:
  • H-MCA-Pro-Lys-Pro-Leu- l/ ⁇ - eii-DPA-Ala-Arg-NH2 (MW 1332.2)
  • Recombinant Human CHO Stromelvsin (MMP-31 Recombinant Human CHO Pro-.vfX(P-3 Human CHO pro-stromelys ⁇ n-257 (pro-MMP-3) was expressed and punfied as described by Housley, et al., J. Biol. Chem. 2 , 4481. 1993.
  • Pro-MMP-2 Human pro-gelatinase A (pro-MMP-2) was prepared using a vaccinia expression system according to the method of Fridman, et al., J. Biol. Chem. 267. 15398. 1992.
  • Activation of Pro-MMP-2 Pro-MMP-2 at 252 mg mL was diluted 1:5 to a final concentration of 50 ⁇ g mL solution in 25 mM Tris at pH 7.5, 5 mM CaCX 150 mM NaCl. and
  • -.Aminophenyimercuric acetate (.APMA) was prepared in 10 mM (3.5 mgmL) in 0.05 NaOH. The APMA solution was added at 1 20 the reaction volume for a final AMPA concentration of 0.5 mM. and the enzyme was incubated at 37 °C for 30 mm.
  • Activated MMP-2 (15 mL) was dialyzed twice vs. 2 L of MMP-2 activation buffer (dialysis membranes were pre-treated with a solution consisting of 0.1% BSA in MMP-2 activation buffer for 1 min. followed by extensive H : O washing). The enzyme was concentrated on Centricon concentrators (concentrators were also pre-treated a solution consisting of 0.1 % BSA in MMP-2 activation buffer for 1 mm., followed by washing with H,0. then MMP-2 activation buffer) with re- dilution followed by re-concentration repeated twice. The enzyme was diluted to 7.5 mL (0.5 times the original volume) with MMP-2 activation buffer.
  • Recombinant Human Pro-MMP-9 Human pro-gelatinase B (pro-MMP-9) derived from L ' 937 cDNA as described by Wtlhelm, et al. J. Biol. Chem.2&L 17213, 1989 was expressed as the full-length form using a baculovirus protein expression system. The pro-enzyme was purified using methods previously described by Hibbs, et al. J. Biol. Chem.26JL 2493, 1984.
  • Pro-MMP-9 activation buffer 50 mM Tris at pH 7.4, lOmM CaCU, 150 mM NaCL and 0.005% Brij-35 (MMP-9 activation buffer) was activated by incubation with 0.5
  • APMA acetate
  • the MMP-Profiling Assay is performed robotically on a
  • the Hamilton is programmed to (1) senaily dilute up to 1 1 potential inhibitors automatically from a 2.5 mM stock in 100% DMSO, (2) distnbute substrate followed bv inhibitor to a 96 well Cytofiuor plate; and (3) add a single enzyme to the plate with mixing to sta ⁇ the reacuon.
  • Microfluoromemc Reacnon Buffer Dilution of test compounds, enzymes, and P 18 substrate for the microfluoromemc assay were made m microfluoromet ⁇ c reaction buffer consisting of 50 mM 2-(N-mo ⁇ phol ⁇ no)ethanesulfon ⁇ c acid (MES) at pH 6 5 with 10 mM CaCl,, 150 mM NaCl, 0.005% B ⁇ j-35 and 1% DMSO et ⁇ Q s:
  • MES 2-(N-mo ⁇ phol ⁇ no)ethanesulfon ⁇ c acid
  • MMP Mtcrofluorome c Profiling Assay The assay is done with a final substrate concentration of 6 ⁇ M P218 and approximately 5 to .8 nM MMP with va ⁇ able dmg concentrations.
  • the Hamilton is programmed to se ⁇ ally dilute up to 11 compounds from a 2.5 mM stock (100%
  • DMSO DMSO
  • the instrument delivers va ⁇ ous amounts of microfluoroment ⁇ c reacuon buffer (MRB) to a 96 tube rack of 1 ml Marsh dilution tubes.
  • MRB microfluoroment ⁇ c reacuon buffer
  • the instrument picks up 20 ⁇ l of inhibitor (2.5 mM) from the sample rack and mixes it with a buffer in row A of the Marsh rack, resulting in a 50 ⁇ M drug concentration.
  • the inhibitors are then se ⁇ ally diluted to 10, 5, 1, .2, .05 and .01 ⁇ M.
  • Position 1 on the sample rack contains only DMSO for the "enzyme-only" wells in the assay, which results in no inhibitor in column 1 , rows A through H.
  • the instrument then distributes 107 ⁇ l of P218 substrate (8.2 ⁇ M in
  • the instrument re-mixes and loads 14.5 ⁇ l of diluted compound from rows A to G in the Marsh rack to corresponding rows in die microtiter plate.
  • the enzyme reservoir is pretreated with 1% BSA in 50 mM Tris, pH 7.5 containing 150 mM NaCl for 1 hour at room temp., followed by extensive HjO washing and drying at room temp.).

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Abstract

L'invention concerne des composés inhibiteurs de métalloprotéases matricielles, leurs compositions pharmaceutiques et une méthode de traitement de maladies utilisant ces composés. Les composés de l'invention ont la formule générale (I) dans laquelle R15 est choisi dans le groupe comprenant: HOCH¿2?, MeOCH2, (n-Pr)2NCH2, CH3CO2CH2, EtOCO2CH2, HO(CH2)2, CH3CO2(CH2)2, HO2C(CH2)2, OHC(CH2)3, HO(CH2)4, Ph, 3-HO-Ph, et PhCH2OCH2; et R?16¿ représente (a) ou (b). Ces composés sont utiles pour inhiber des métalloprotéases matricielles et par conséquent combattre des états auxquels les métalloendo-protéinases matricielles MMP contribuent, tels que, l'arthrose, la polyartrite rhumatoïde, l'arthrite septique, les maladies périodontiques, l'ulcération de la cornée, la protéinurie, les maladies aortiques anévrismales, l'épidermolyse distrophobique, les lésions buleuses, des états conduisant à des réactions inflammatoires, les ostéopénies induites par l'activité de MMP, des maladies des articulations temporomendibulaires, les maladies démiliénisantes du système nerveux, la métastase de tumeurs ou la perte de cartilage dégénérative suivant des lésions articulaires traumatiques, et la thrombose coronaire due à une rupture des plaques athéroscléreuses. La présente invention concerne également des compositions pharmaceutiques et des méthodes de traitement de ces états.
PCT/US1997/007921 1996-05-15 1997-05-12 Inhibition de metalloproteases matricielles par des composes contenant de l'acetylene WO1997043245A1 (fr)

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CA002253796A CA2253796C (fr) 1996-05-15 1997-05-12 Inhibition de metalloproteases matricielles par des composes contenant de l'acetylene
BR9709077A BR9709077A (pt) 1996-05-15 1997-05-12 Compostos inibindo metaloproteinase de matriz composição tendo atividade inibitória de metaloprotease de matriz processos para inibir atividade de metaloprotease de matriz e para tratar um mamífero
AU29386/97A AU710759B2 (en) 1996-05-15 1997-05-12 Inhibition of matrix metalloproteases by acetylene containing compounds
EP97923622A EP0912496A1 (fr) 1996-05-15 1997-05-12 Inhibition de metalloproteases matricielles par des composes contenant de l'acetylene
JP09540980A JP3090957B2 (ja) 1996-05-15 1997-05-12 アセチレン含有化合物によるマトリックスメタロプロテアーゼの阻害

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WO2000044711A1 (fr) * 1999-01-27 2000-08-03 American Cyanamid Company INHIBITEURS DES TACE A BASE D'ACIDES HYDROXAMIQUES DES ACIDES PHOSPHINIQUE ET $G(b)-SULFONAMIDO ACETYLENIQUE
EP1031349A1 (fr) * 1999-02-25 2000-08-30 Bayer Aktiengesellschaft Utilisation d'acides 4-biarylbutyriques et 5-biarylpentanoiques substitués pour traiter les maladies cérébrales
US6288063B1 (en) 1998-05-27 2001-09-11 Bayer Corporation Substituted 4-biarylbutyric and 5-biarylpentanoic acid derivatives as matrix metalloprotease inhibitors
US6326516B1 (en) 1999-01-27 2001-12-04 American Cyanamid Company Acetylenic β-sulfonamido and phosphinic acid amide hydroxamic acid TACE inhibitors
US7141607B1 (en) 2000-03-10 2006-11-28 Insite Vision Incorporated Methods and compositions for treating and inhibiting retinal neovascularization
US8765814B2 (en) 2010-07-08 2014-07-01 Kaken Pharmaceutical Co., Ltd. N-hydroxyformamide derivative and medicament containing same
WO2015150362A2 (fr) 2014-04-03 2015-10-08 Bayer Pharma Aktiengesellschaft Dérivés chiraux d'acide cyclopentanecarboxylique à disubstitution 2,5 et leur utilisation
WO2015150350A1 (fr) 2014-04-03 2015-10-08 Bayer Pharma Aktiengesellschaft Acides cyclopentanecarboxyliques à disubstitution 2,5 pour traiter des maladies des voies respiratoires
WO2015150363A1 (fr) 2014-04-03 2015-10-08 Bayer Pharma Aktiengesellschaft Acides cyclopentanecarboxyliques à disubstitution 2,5 et leur utilisation

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2112716A1 (de) * 1971-03-17 1972-10-05 Thomae Gmbh Dr K Neue 4-(4-Biphenylyl)-4-oxobuttersaeuren,ihre Salze und ihre Ester
US3784701A (en) * 1970-09-21 1974-01-08 American Cyanamid Co Compositions containing substituted benzoylpropionic acids and method of use to treat inflammation and pain
WO1996015096A1 (fr) * 1994-11-15 1996-05-23 Bayer Corporation Acides 4-biarylbutyrique ou 5-biarylpentanoique substitues et leurs derives en tant qu'inhibiteurs de metalloproteases matrices

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3784701A (en) * 1970-09-21 1974-01-08 American Cyanamid Co Compositions containing substituted benzoylpropionic acids and method of use to treat inflammation and pain
DE2112716A1 (de) * 1971-03-17 1972-10-05 Thomae Gmbh Dr K Neue 4-(4-Biphenylyl)-4-oxobuttersaeuren,ihre Salze und ihre Ester
WO1996015096A1 (fr) * 1994-11-15 1996-05-23 Bayer Corporation Acides 4-biarylbutyrique ou 5-biarylpentanoique substitues et leurs derives en tant qu'inhibiteurs de metalloproteases matrices

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHICCARELLI, F. S. ET AL: "Disposition and metabolism of fenbufen in several laboratory animals", ARZNEIM.-FORSCH. (1980), 30(4A), 707-15 CODEN: ARZNAD;ISSN: 0004-4172, 1980, XP002043281 *
CHILD, R. G. ET AL: "A new nonsteroidal antiinflammatory analgesic:.gamma.-oxo(1,1'- biphenyl)-4-butanoic acid (Fenbufen). Chemistry and activity of analogs", ARZNEIM.-FORSCH. (1980), 30(4A), 695-702 CODEN: ARZNAD;ISSN: 0004-4172, 1980, XP002043280 *
CHILD, RALPH G. ET AL: "Fenbufen, a new anti-inflammatory analgesic: synthesis and structure-activity relations of analogs", J. PHARM. SCI. (1977), 66(4), 466-76 CODEN: JPMSAE, 1977, XP000560277 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6288063B1 (en) 1998-05-27 2001-09-11 Bayer Corporation Substituted 4-biarylbutyric and 5-biarylpentanoic acid derivatives as matrix metalloprotease inhibitors
WO2000044711A1 (fr) * 1999-01-27 2000-08-03 American Cyanamid Company INHIBITEURS DES TACE A BASE D'ACIDES HYDROXAMIQUES DES ACIDES PHOSPHINIQUE ET $G(b)-SULFONAMIDO ACETYLENIQUE
US6326516B1 (en) 1999-01-27 2001-12-04 American Cyanamid Company Acetylenic β-sulfonamido and phosphinic acid amide hydroxamic acid TACE inhibitors
EP1031349A1 (fr) * 1999-02-25 2000-08-30 Bayer Aktiengesellschaft Utilisation d'acides 4-biarylbutyriques et 5-biarylpentanoiques substitués pour traiter les maladies cérébrales
WO2000050017A2 (fr) * 1999-02-25 2000-08-31 Bayer Aktiengesellschaft Utilisation de derives substitues d'acides 4-biarylbutyrique et 5-biarylpentanoique pour le traitement des maladies cerebrales
WO2000050017A3 (fr) * 1999-02-25 2001-02-01 Bayer Ag Utilisation de derives substitues d'acides 4-biarylbutyrique et 5-biarylpentanoique pour le traitement des maladies cerebrales
US7141607B1 (en) 2000-03-10 2006-11-28 Insite Vision Incorporated Methods and compositions for treating and inhibiting retinal neovascularization
US8765814B2 (en) 2010-07-08 2014-07-01 Kaken Pharmaceutical Co., Ltd. N-hydroxyformamide derivative and medicament containing same
WO2015150362A2 (fr) 2014-04-03 2015-10-08 Bayer Pharma Aktiengesellschaft Dérivés chiraux d'acide cyclopentanecarboxylique à disubstitution 2,5 et leur utilisation
WO2015150350A1 (fr) 2014-04-03 2015-10-08 Bayer Pharma Aktiengesellschaft Acides cyclopentanecarboxyliques à disubstitution 2,5 pour traiter des maladies des voies respiratoires
WO2015150363A1 (fr) 2014-04-03 2015-10-08 Bayer Pharma Aktiengesellschaft Acides cyclopentanecarboxyliques à disubstitution 2,5 et leur utilisation

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TW381079B (en) 2000-02-01
PA8429301A1 (es) 2000-05-24
ID16910A (id) 1997-11-20
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AR007097A1 (es) 1999-10-13
JPH11511179A (ja) 1999-09-28
HN1997000088A (es) 1997-06-18
YU18697A (en) 1999-11-22
HRP970245B1 (en) 2002-06-30
JP3090957B2 (ja) 2000-09-25
CN1139570C (zh) 2004-02-25
CO5080759A1 (es) 2001-09-25
AU2938697A (en) 1997-12-05
CA2253796A1 (fr) 1997-11-20
HRP970245A2 (en) 1998-04-30
SV1997000035A (es) 1999-01-14
TNSN97084A1 (fr) 2005-03-15
CA2253796C (fr) 2003-10-28
AU710759B2 (en) 1999-09-30
CN1225623A (zh) 1999-08-11
EP0912496A1 (fr) 1999-05-06

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