JP3090957B2 - Inhibition of matrix metalloproteases by acetylene-containing compounds - Google Patents

Description

Description: FIELD OF THE INVENTION The present invention relates to enzyme inhibitors useful for inhibiting matrix metalloproteases, and more particularly to novel biarylacetylene-containing compounds or derivatives thereof.

2. Related Art Matrix metalloproteases (also known as matrix metalloendo-proteinases or MMPs) are a type of zinc endoproteinase, which are interstitial collagenases (also known as MMP-1), stromelysin (Stromelysin) (proteoglycanase, transin
n) or also known as MMP-3), gelatinase A (also known as 72 kDa-gelatinase or MMP-2) and gelatinase B (also known as 95 kDa-gelatinase or MMP-9). Including, but not limited to. The MMP is a TIMP (Tis
It is secreted from various cells, including fibroblasts and chondrocytes, together with a natural proteinaceous inhibitor known as sue Inhibitor of Metallo Proteinase.

All of the above MMPs are capable of destroying various connective tissue components of articular cartilage or basement membrane. Each MMP is secreted as an inactive proenzyme, which must be degraded before it can exert its proteolytic activity at a later stage. In addition to the matrix breaking effect, some of the aforementioned MMPs, such as MMP-3,
For example, it has been implicated as an in vivo active for MMP-1 and MMP-9 (Ito, et al., Arch Biochem. Biophys.
267 , 211, 1988; Ogata, et al., J. Biol. Chem., 267 , 3581, 19
92). Thus, the cascade of proteolytic activity is due to excess MM
Can be induced by P-3. As a result, specific MMP-3 inhibitors will limit the activity of other MMPs that are not directly inhibited by said inhibitors.

It is also possible that MMP-3 is degraded and this inactivates other proteinases, such as endogenous inhibitors of elastase (Winyard. Et al., FEB).
S Letts. 279 , 1, 91, 1991). Thus, inhibitors of MMP-3 can affect its activity by altering the levels of endogenous inhibitors of other disrupting proteinases.

Many diseases are believed to be mediated by excessive or unwanted matrix-disrupting metalloprotease activity or by imbalance in the ratio of MMP to TIMP. These include: a) Osteoarthritis (Woessner, et al., J.
Biol. Chem., 259 (6), 3633, 1984; Phadke, et al., J. Rhe.
umatol. 10 , 852, 1983), b) Rheumatoid arthritis (Mullin
s, et al., Biochem. Biophys. Acts 695 , 117, 1983; Woolle
y, et al., Arthritis Rheum. 20 , 1231, 1977; Gravallese, e
t al., Arthritis Rheum. 34 , 1076, 1991), c) Septic arthritis (Williams, et al., Arthritis Rheum. 33 , 533, 199).
0), d) Tumor metastasis (Reich, et al., Cancer Res., 48 , 330)
7,1988 and Matricean, et al., Proc. Nat'l. Acad, Sci. USA
83 , 9413, 1986), e) Periodontal disease (Overall, et al., J. Peri)
odental Res. 22 , 81, 1987), f) corneal ulcer (Burns, et a)
l., Invest. Opthalmol. Vis. Sci. 30 , 1569, 1989), g) proteinuria (Baricos, et al., Biochem. J. 254 , 609, 1988),
h) Coronary thrombus from atherosclerotic plaque rupture (Henney, et a
l., Proc, Nat'l. Acad. Sci., USA 88 , 8154-8158, 1991),
i) Aortic aneurysm disease (Vine, et al., Clin. Sci. 81 , 233, 199)
1), j) Fertility control (Woessner, et al., Steroids 54 , 49)
1,1989), k) Dystrophobic epidermolysis bullosa (Kronberger, et al., J., Invest. Dermatol. 79 , 208, 198).
2), l) degenerative cartilage loss due to traumatic joint injury, m)
Conditions causing an inflammatory response, osteopenia mediated by MMP activity, n) temporomandibular joint disease, o) demyelination of the nervous system (Chantry, et al., J. Neurochem. 50 , 688, 1988).

The need for new treatments is particularly important in the case of arthritic diseases. Osteoarthritis (OA), Rheumatoid arthritis (RA)
And the primary damaging effect of septic arthritis is the progressive loss of articular cartilage and thus normal joint function. Non-steroidal anti-inflammatory drugs (NSAIDs) exist to control pain and swelling, but commercial drugs cannot prevent or delay the cartilage loss. The end result of the disease is a complete loss of joint function, which can only be treated by joint replacement surgery. MMP inhibitors are expected to halt or reverse the progression of cartilage loss and obviate or delay surgical intervention.

Proteases are important components at several stages in the development of metastatic cancer. In the above process, the tumor expands at the primary site due to proteolysis of the structural proteins of the basement membrane,
It metastasizes from said site, as well as defined and infiltrated at distant secondary sites. Also, tumor-induced angiogenesis is required for tumor growth and is dependent on proteolytic cell remodeling. Transfection experiments with various types of proteases indicate that matrix metalloproteases play a dominant role in the above processes for certain gelatinases A and B (MMP-2 and MMP-9, respectively). For an overview of the above areas, see Mullins, et al., Bio.
chim. Biophys. Acta 695 , 177, 1983; Ray, et al., Eur.
ir. J. 7 , 2062, 1994; Birkedal-Hansen, et al., Crit. Re.
See v. Oral Biol. Med. 4 , 197, 1993.

In addition, natural matrix metalloprotease inhibitors
Inhibition of extracellular matrix degradation by TIMP-2 (protein) prevents cancer growth (DeClerck, et al., Cancer R
es. 52, 701,1992), and TIMP-2 inhibits tumor-induced angiogenesis in experimental systems (Moses, et al., Science 248, 1408,
1990). For review, see DeClerck.e
See al., Ann. NY Acad. Sci. 732 , 222, 1994.
Furthermore, the synthetic matrix metalloproteinase inhibitor batimastat, when administered intraperitoneally, inhibits human colon tumor growth and spreads in orthotopic models in nude mice (Wang, et al., Cancer Res, 54 , 4726,199
4) and prolong the survival of mice bearing human ovarian cancer xenografts (Davies, et al., Cancer Res. 53 , 2087, 199).
3) It was proved. The use of said compounds and related compounds is described in Brown, et al., WO-9321 942A2 (931111).

Delaying metastatic cancer, promoting tumor regression, inhibiting cancer cell growth,
For delaying or preventing cartilage loss associated with osteoarthritis,
Alternatively, there are several patents and applications claiming the use of metalloprotease inhibitors for the treatment of the other diseases mentioned above (eg, Levy, et al., WO-9519).
965 A1; Beckett, et al., WO-9519956 A1; Beckett, et a
l., WO-9519957 A1; Beckett, et al., WO-9519961 A1; Br
own, et al., WO-9321942 A2; Crimmin, et al., WO-94216
25 A1; Dickens, et al., US Pat. No. 4599361; Hughes, et.
al., US Pat. No. 5190937; Broadhurst, et al., EP 574758
A1; Broadhurst, et al., EP 276436; and Myers, et al., E
P 520573 A1). The advantageous compounds of said patent have a peptide backbone with a zinc complexing group (hydroxamic acid, thiol, carboxylic acid or phosphonic acid) at one end and various side chains, both of which are naturally occurring. Amino acids and those with new functional groups. These small peptides often have poor absorption and exhibit low oral bioavailability. They also tend to undergo rapid proteolytic metabolism and therefore have a short half-life. For example,
Batimastat, a compound described in Brown, et al., WO-9321942 A2, can only be administered intraperitoneally.

Certain 3-biphenoylpropanoic acids and 4-biaryloylbutanoic acids are described in the literature as anti-inflammatory, anti-platelet aggregating,
It has been described as an anti-inflammatory, antiproliferative, antilipidemic, antirheumatic, analgesic and cholesterol lowering agent. None of the above examples describe MMP inhibition as a mechanism for the therapeutic effects claimed herein. Certain related compounds have also been used as intermediates in the manufacture of liquid crystals.

In particular, Tomcufcik, et al., US Patent 3,874,701 claims certain substituted benzoylpropionic acids for the treatment of inflammation and pain. The compounds include 3-biphenoylpropanoic acid (also known as fenbufen) described below.

Child, et al., J. Pharm. Sci., 66 , 466, 1977, describe the structure-activity relationships of some analogs of fenbufen. These include compounds with substituted biphenyl ring systems or propanoic acid moieties with phenyl, halogen,
Included are compounds substituted with hydroxyl or methyl or several compounds in which the carboxylic acid or carbonyl functionality has been converted to various derivatives. Compounds containing 4'-substituted biphenyl and substituted propanoic acid moieties in combination in one molecule are not described. Phenyl-substituted compounds shown below (compounds XL IX and L XXVII)
And methyl-substituted compounds (Compound XL VII) are described as being inactive.

Kameo, et al., Chem. Pharm. Bull. 36 , 2050, 1988 and JP
Patent 62133825 A2 describes certain substituted 3-biphenoylpropionic acid derivatives and analogs thereof, including: Various compounds with other substituents on the propionic acid moiety have been described, but they do not contain a biphenyl residue.

In the formula, X, H, 4'-Br, 4' -Cl, 4'-CH 3,
Or 2'-Br.

Cousse, et al., Eur. J. Med. Chem., 22 , 45, 1987 describe the following methyl and methylene substituted 3-biphenoyl-propanoic acids and -propenonic acids. Carbonyl is
Corresponding compounds substituted by CH ₂ OH or CH ₂ are also described.

In the formula, X, H, Cl, Br, is CH _3, O, F or NH _2.

Nichl, et al., DE Patent 1957750 also describe some of the aforementioned methylene-substituted biphenoylpropanoic acids.

El-Hashash, et al., Revue Roum. Chim. 23 , 1581, 1978
Describes products derived from β-aroyl-acrylic acid epoxides, including the following biphenyl compounds. Compounds substituted with a biphenyl moiety are not described.

Kitamura, et al., JP Patent 60209539, describes certain biphenyl compounds used as intermediates for the production of liquid crystals, including: Biphenyl is unsubstituted in the above intermediate.

Wherein R ¹ is alkyl of 1 to 10 carbons.

Thyes, et al., DE Patent 2854475, uses the following compounds as intermediates. Biphenyl groups are unsubstituted.

Sammour, et al., Egypt J. Chem. 15 , 311, 1972 and Couqu
elet, et al., Bull. Soc. Chim. Fr. 9 , 3196, 1971, describe certain dialkylamino substituted biphenoyl propanoic acids, including: The biphenyl group is not substituted in any case.

Wherein R ¹ , R ² are alkyl, benzyl, H or, together with nitrogen, morpholinyl.

Others disclose a series of biphenyl-containing carboxylic acids represented by the compounds described below, which inhibit neutral endopeptidase (NEP24.11), a membrane-bound zinc metalloprotease (Stanton, et al., Bioor
g.Med.Chem.Lett. 4 , 539, 1994; Lombaert, et al., Bioor
g.Med.Chem.Lett. 4 , 2715, 1994; Lombaert, et al., Bioor
g. Med. Chem. Lett. 5 , 145, 1995; Lombaert, et al., Bioor.
g. Med. Chem. Lett. 5 , 151, 1995).

N-carboxyalkyl derivatives having biphenylethylglycine represented by the compounds described below are stromelysin-1 (MMP-3), 72 kDA gelatinase (MM
P-2) and collagenase have been reported (Durette, et al., WO-9529689).

Effective, with improved bioavailability and biostability over prior art peptide-based compounds, and that can be optimized for use against specific target MMPs It would be desirable to obtain a suitable MMP inhibitor. These compounds are the subject of the present application.

The development of efficacious MMP inhibitors has been identified for diseases mediated by the presence or excess of MMP activity, including osteoarthritis, rheumatoid arthritis, septic arthritis, tumor metastasis, periodontal disease, corneal ulcers and proteinuria. Provide new treatments. Thiol (Beszant, et al., J. Med. Chem. 36 , 4030, 1993), hydroxamic acid (Wahl, et al., Bioorg. Med. Chem. Lett. 5 , 34)
9,1995; Conway, et al., J. Exp., Med. 182 , 449, 1995; Porte
r, et al., Bioorg.Med.Chem, Lett. 4 , 2741, 1994; Tomczu
k, et al., Bioorg. Med. Chem. Lett. 5 , 343, 1995;
no, et al., Bioorg. Med. Chem. Lett. 5 , 1415, 1995), phosphorus-based acids (Bird, et al., J. Med. Chem. 37 , 158, 1994; Mor).
phy, et al., Bioorg. Med. Chem. Lett. 4 , 2747, 1994; Korty
lewicz, et al., J. Med. Chem. 33 , 263, 1990), carboxylic acid (Chapman, et al., J. Med. Chem. 36 , 4293, 1993; Brown, et.
al., J. Med.Chem. 37 , 674, 1994; Morphy, et al., Bioorg.Me.
d. Chem. Lett. 4 , 2747, 1994; Stack, et al., Arch. Bioche.
m. Biophys. 287 , 240, 1991; Ye, et al., J. Med. Chem. 37 , 20
6,1994; Grobelny, et al., Biochemistry 24 6145, 1985; Mo
okhtiar, et al., Biochemistry 27 , 4299, 1988),
Some MMP inhibitors have been described in the literature. However, such inhibitors generally have a peptide backbone,
And thus usually exhibits low oral bioactivity due to poor absorption and a short half-life due to rapid proteolysis. Thus, there remains a need for improved MMP inhibitors.

SUMMARY OF THE INVENTION The present invention relates to compounds having matrix metalloprotease inhibitory activity. The compounds are useful for inhibiting the inhibition of matrix metalloproteases and, thus, the symptoms that contribute to the symptoms of MMP. Accordingly, the present invention provides a medicament and a method for treating the above-mentioned symptoms.

The compounds described include: (a) osteoarthritis, rheumatoid arthritis, septic arthritis,
Reduces the effects of periodontal disease, corneal ulcers, proteinuria, aortic aneurysm disease, malnutrition-type epidermolysis bullosa, conditions leading to inflammatory response, osteopenia mediated by MMP activity, temporomandibular joint disease, and demyelinating diseases of the nervous system (B) inhibition of degenerative cartilage loss due to tumor metastasis or traumatic joint damage; (c) reduction of coronary artery thrombus resulting from atherosclerotic plaque rupture; A matrix metalloprotease-inhibiting amount of is administered to a mammal.

The compounds of the present invention are also useful academic research tools for studying the function and mechanism of action of matrix metalloproteases in two systems, in vivo and in vitro. Since the compounds of the present invention can be used for denaturing MMP action due to their MMP inhibitory action, researchers should observe the effect of reduced MMP activity in biological experimental systems upon study. Can be.

The present invention has a matrix metalloproteinase inhibitory activity, and it relates to compounds represented by the general formula _{T x A-B-D-} E-G (L).

Among the above general formula (L), T _x A represents a substituted or unsubstituted or an aromatic 6-membered ring or N, O or heteroaromatic having atomic 1-2 selected independently from the group of S Represents a 5- to 6-membered ring. T represents a substituted acetylene component.

In the general formula (L), B represents an aromatic 6-membered ring or a heteroaromatic 5- to 6-membered ring having 1 to 2 atoms independently selected from the group of N, O or S. . This is referred to as ring B or B units. When N is used together with S or O in ring B, at least one heteroatom
Separated by carbon atoms.

In the general formula (L), D is Represents

In the general formula (L), E represents a chain of n carbon atoms having R ⁶ m substituents, wherein the R ⁶ groups are independently substituents or spiro or non-spiro rings Is composed. This ring may be formed in two ways;
a) two groups R ⁶ are bonded, and together with the chain atom to which the two groups R ⁶ are bonded and any intervening chain atoms, constitute a 3- to 7-membered ring; b) one group R is linked to this one group R ⁶ to form a chain, together with the chain atom to which the group R ⁶ is bonded and any intervening chain atoms;
It constitutes a 3- to 7-membered ring. The number of carbon atoms in the chain, n
Is 2 or 3, and the number m of the substituents R ⁶ is an integer of 1 to 3. The number of carbon in the entire group R ⁶ is at least two.

Each group R ⁶ is an alkyl, alkenyl, alkynyl, heteroaryl, non-aromatic cyclic group and combinations thereof optionally substituted with one or more heteroatoms as described more fully below. It is. General formula (L)
Wherein E is a substituted monocyclic or bicyclic component optionally substituted with one or more heteroatoms.

In general formula (L), G is, -PO ₃ H ₂ -M, Wherein M represents —CO ₂ H, —CON (R ¹¹ ) ₂ or —CO ₂ R ¹² , and R ¹³ represents an acyclic, naturally occurring amino acid 19 in any side chain. Represent.

The most advantageous compounds of the present invention are [Wherein, R ¹⁵ represents HOCH ₂ , MeOCH ₂ , (n-Pr) ₂ NCH ₂ , CH ₃
CO ₂ C ₂ , EtOCO ₂ CH ₂ , HO (CH ₂ ) ₂ , CH ₃ CO ₂ (CH ₂ ) ₂ , HO
_{2 C (CH 2) 2,} OHC (CH 2) 3, HO (CH 2) Ph, 3-HO-Ph
And PhCH are selected from ₂ the group consisting of OCH _2; R ¹⁶ is ].

Pharmaceutically acceptable salts of the compounds are also included within the scope of the invention.

Among the most similar related compounds of the prior art, the biphenyl moiety of the molecule is unsubstituted and the propanoic or butanoic acid moiety is either unsubstituted or has one methyl or phenyl group. ing. The presence of a larger phenyl group has been reported to render prior art compounds inactive as anti-inflammatory analgesics. For example, Child, et al., JP
See harm.Sci. 66,466 (1977). In contrast, compounds with potent MMP inhibitory activity have large size substituents on the propane or butane portion of the molecule.
Also advantageously, the biphenyl moiety of the best MMP inhibitor is
Even when the propane or butane moieties are optimally substituted, they still have a substituent at the 4 'position, and the unsubstituted biphenyl compounds of the present invention have sufficient Have activity.

The above description merely summarizes certain aspects of the present invention and is not intended to limit the invention in any way and should not be construed as limiting the invention.
Accordingly, all patents and other references cited herein are incorporated by reference in their entirety.

More particularly the description of preferred embodiments, the compounds of the present invention, a matrix metalloprotease inhibitory activity and the general _{formula: T x A-B-D} -E-G (L) wherein, T _x A is [Wherein, R ¹ represents H or alkyl having 1 to 3 carbons]
Represents a substituted or unsubstituted aromatic or heteroaromatic component selected from the group consisting of:

Throughout this specification, for the chemical structure shown,
An open bond indicates where the structure is attached to another group. For example, (Where R ⁵⁰ is Is a structural formula It is.

In the above structural formula, the aromatic ring is advantageously called ring A or A unit, T represents a substituent and is called T group or T unit. T is a substituted acetylene component, and x is 1.

Ring B of the general formula (L) is a substituted or unsubstituted aromatic or heteroaromatic ring in which all substituents render the molecule incompatible with the active site of the target enzyme. A group that disrupts the relative structure of Ring A and Ring B that is not or detrimental. The substituent may be a component such as lower alkyl, lower alkoxy, CN, NO ₂ , a halogen atom, etc., but is not limited to the above groups.

In the general formula (L), B is Wherein R ¹ represents an aromatic or heteroaromatic ring selected from the group consisting of: Said rings are referred to as ring B or B units.

In the general formula (L), D is Represents the component of

In the general formula (L), E represents a chain of n carbon atoms having m substituents R ⁶ called groups R ⁶ or R ⁶ units. Group R
⁶ is independently a substituent or constitutes a spiro ring or a non-spiro ring. This ring may be formed in two ways: a) two groups R ⁶ are bonded is, and two radicals R ⁶ are chain atoms and any intervening chain attached Together with the atoms to form a 3- to 7-membered ring, or b) one group R is linked to this one group R ⁶ to form a chain, and the chain atom to which the group R ⁶ is bonded Together with any intervening chain atoms to form a 3- to 7-membered ring. The number n of carbon atoms in the chain is 2 or 3, and the number m of substituents R ⁶ is m.
Is an integer of 1 to 3. The number of carbon in the entire group R ⁶ is at least two.

Each group R ⁶ is independently selected from the group consisting of the substituents listed below as items 1) to 14).

1) alkyl having 1 to 10 carbon atoms, provided that when the A unit is phenyl, the B unit is phenylene and m is 1
Where n is 2 and the alkyl group is located on the carbon α to the D unit and x is 1 or 2) 2) aryl having 6 to 10 carbons, wherein A When the unit is phenyl, the B unit is phenylene, the aryl group is phenyl, n is 2, m is 1 or 2, and x is 1 or 2; 9 and at least 1 N, O or S
4) arylalkyl having 6 to 10 carbon atoms and alkyl portion having 1 to 8 carbon atoms; 5) heteroaryl moiety having 4 to 9 carbon atoms and at least 1 carbon atom; A heteroarylalkyl comprising an N, O or S heteroatom, wherein the alkyl moiety has 1 to 8 carbons; 6) an alkenyl having 2 to 10 carbons; 7) an alkenyl having an aryl moiety having 6 to 10 carbons. 8) a heteroaryl moiety consisting of 4 to 9 carbons and at least one N, O or S heteroatom, wherein the alkenyl moiety has 2 to 5 carbons; Arylalkenyl; 9) alkynyl of 2 to 10 carbons; 10) arylyl having an aryl moiety having 6 to 10 carbons and an alkynyl moiety having 2 to 5 carbons 11) heteroarylalkynyl wherein the heteroaryl moiety consists of 4 to 9 carbons and at least one N, O or S heteroatom and the alkynyl moiety has 2 to 5 carbons; 12) t is 0 or 1 And R ⁷ is an integer of And aryl I of aryl-containing R ⁷ radical is selected from the group consisting of heteroaryl ingredient commensurate consisting heteroatom 4-9 carbons and at least one N, O or S - (CH _{2) t} R ⁷ In the group R ⁷ , Y represents O or S; R ¹ is the same as defined above, and u is 0, 1 or 2, provided that R ⁷ is And when the A unit is phenyl, the B unit is phenylene, m is 1, n is 2, and t is 0, and x is 1 or 2.

13) v is an integer of 1 to 4, and Z is -S-, -S (O)
—, —SO ₂ — or —O—, wherein R ⁸ is alkyl of 1 to 12 carbons, aryl of 6 to 10 carbons, 4 to 9 carbons and at least one heterocyclic of N, O or S. A heteroaryl consisting of atoms; an arylalkyl having an aryl moiety having 6 to 12 carbons and an alkyl moiety having 1 to 4 carbons;
The aryl moiety has 6 to 12 carbons and at least one N,
Having an O or S heteroatom, wherein the alkyl moiety is
Heteroarylalkyl having four; R ⁹ carbon 2-6
Alkyl, aryl of 6 to 10 carbons, heteroaryl of 4 to 9 carbons and at least one heteroatom of N, O or S and arylalkyl in which the aryl part has 6 to 10 carbons, or Heteroaryl consisting of 4 to 9 carbons and at least one heteroatom of N, O or S, wherein the alkyl moiety has 1 to 4 carbons,
However, when R ⁸ is -C (O) R ⁹ , Z is -S-
Or -O-; when Z is -O-, R ⁸ represents-
(C _q H _2q O) _r R ⁵ ; when the A unit is phenyl, the B unit is phenylene, m is 1 and n is 2
And v is 0 and x is 1 or 2-(CH ₂ ) _v
ZR ^8, and 14) w is an integer from 1 to 3, R ¹⁰ represents one to two alkyl carbon ^{_{- (CH 2) w SiR 10}} 3.

Further, any aryl or heteroaryl moiety of the group T or R ^6, _{optionally, - (CH 2) y C} (R 11) (R 12) O
_{H, - (CH 2) y} OR 11, - (CH 2) y SR 11, - (CH 2) y S
^{(O) R 11, - (} CH 2) y S (O) 2 R 11, - (CH 2) y SO 2 N
_{^{(R 11) 2, - (}} CH 2) y N (R 11) 2, - (CH 2) y N
(R ¹¹ ) COR ¹² , —OC (R ¹¹ ) ₂ O— [in this case, two oxygen atoms are bonded to the aryl ring], — (CH ₂ ) _y CO
R ¹¹ , − (CH ₂ ) _y CON (R ¹¹ ) ₂ , − (CH ₂ ) _y CO ₂ R ¹¹ , −
(CH ₂ ) _y OCOR ¹¹ , -halogen atom, -CHO, -CF ₃ , -NO
₂ , -CN and -R ¹² [wherein y is 0 to 4, R ¹¹
Represents H or an alkyl having 1 to 4 carbons, and R ¹² represents an alkyl having 1 to 4 carbons.] May have up to two substituents selected from the group consisting of

In general formula (L), G is, -PO ₃ H ₂ -M, Wherein M represents —CO ₂ H, —CON (R ¹¹ ) ₂ or —CO ₂ R ¹² , and R ¹³ represents an acyclic, naturally occurring amino acid 19 in any side chain. Represent.

Also, pharmaceutically acceptable salts of the compounds contained in formula (L) are within the scope of the invention.

The G unit is most preferably attached to the E unit at a carbon of β to the D unit, and is preferably a carboxylic acid group.

As used herein, the term "alkyl" shall refer to linear, branched, cyclic, and polycyclic materials. The term "haloalkyl" is for example-
(CH _{2) 2} Cl, is that the partially or totally alkyl groups halogenated, such as -CF ₃ and -C ₆ F _13.

In one embodiment, the invention relates to compounds of general formula (L), wherein at least one A, B unit and R ^{6 comprises} a heteroaromatic ring. Preferred compounds having a heteroaromatic ring are those in which the heteroaryl group has NR ¹ when the ring is O, S or 5-membered, and has N when the ring is 6-membered. It is a heteroaryl having 4 to 9 carbon atoms consisting of a 6-membered heteroaromatic ring. Particularly preferred compounds having a heteroaromatic ring are those in which at least one of the A and B units consists of a thiophene ring. When the A unit is thiophene, advantageously 2
At the 5-position and one substitution value T at the 5-position
have. When the B unit is thiophene, 2
It is linked to the D unit and the A unit via the positions 5 and 5, respectively.

In the general formula (L), rings A and B are preferably phenyl and phenylene, respectively, and ring A is preferably at least one ring furthest from the position of ring A attached to ring B. , The D units are preferably carbonyl groups and the G units are preferably carboxyl groups.

In another embodiment, the present invention provides that, in E units, n
Is 2 and m is 1. Thus, the compound has two carbon atoms between the D and G units and has one substituent on the two carbon chains.

In another embodiment, the present invention relates to a method wherein the ring A is a substituted or unsubstituted phenyl group, the ring B is p-phenylene, and the aryl moiety of any R ⁶ component having an aryl is
It relates to compounds of the general formula (L) having only carbon in the ring. Therefore, the compound does not have a heteroaromatic ring.

In another embodiment, the present invention is directed to compounds of general formula (L) wherein m is 1 and R ⁶ is independently a substituent. The compound has only a single substituent R ⁶ on the E unit, but the substituent is not included in the ring. Preferred compounds in said subset are of the formula: [Wherein x is 1 and the substituent T is located at the 4-position of ring A with respect to the bonding position between rings A and B]. Para substituent T of the subset is further preferably the following _{group: MeOCH 2 C≡C -, (n} -Pr) 2 NCH 2 C≡C-, CH 3 C
O ₂ CH ₂ C≡C-, EtOCO ₂ CH ₂ C≡C-, HOCH ₂ C≡C-, HO
(CH ₂ ) ₂ C≡C-, CH ₃ CO ₂ (CH ₂ ) ₂ C≡C-, HO ₂ C (C
H ₂ ) ₂ C≡C-, OHC (CH ₂ ) ₃ C≡C-, HO (CH ₂ ) ₄ C≡C
-, PhC≡C-, 3-HO-PhC≡C- and PhCH ₂ OCH ₂ C≡C
-An acetylene-containing component selected from

Another compound of the general formula (L) wherein R ⁶ is — (CH ₂ ) _t R ⁷ has t as an integer from 1 to 5. R ⁶ is-(CH
₂ ) Preferred compounds of the general formula (L) wherein _v ZR ⁸ are 1-4
As an integer and Z as —S— or —O—. Formula R ⁶ is alkyl additional compounds of (L) has a four or more carbons in the alkyl, and R ⁶ is arylalkyl is an alkyl moiety of the arylalkyl It has 2-3 carbons.

In another embodiment, the present invention relates to the present invention, wherein the number m of substituents on the E unit is 2 or 3, and when m is 2, the two groups R ⁶ are independently substituents Or together form a spiro ring, or one group R ⁶ is independently a substituent, another forms a spiro ring, and m is 3
Wherein the two groups R ⁶ are independently substituents;
And one group R ⁶ comprises a ring or two groups R ⁶
Is a ring, and one group R ⁶ is a compound of the general formula (L) in which one group is independently a substituent or three groups R ⁶ form a ring. Thus, in said subset, the E unit is di- or tri-substituted, and when di-substituted, any ring formed by two or two groups R ⁶ is a spiro ring. Yes, when trisubstituted, the group R ⁶ includes compounds that may form a spiro or non-spiro ring.

In another embodiment, the present invention relates to any of the embodiments, wherein the number m of substituents on the E unit is 1 or 2, and when m is 1, the group R ⁶
Constitutes a non-spiro ring, and when m is 2,
The general formula (L) in which two groups R ⁶ together form a non-spiro ring, or one group R ⁶ is independently a substituent and the other R ⁶ forms a non-spiro ring )). Therefore, in the above subset, the E unit is a substituent
It has one or two R ₂ , and at least one of the substituents is contained in a non-spiro ring.

More specifically, a representative compound of general formula (L) wherein one or more of the substituents R ⁶ is essential for the formation of a non-spiro ring is represented by the following structural formula: Wherein a is 0, 1 or 2; b is 0 or 1
C is 0 or 1; d is 0 or 1; c +
d is 0 or 1; e is 1-5; f is 1-4
G is 3-5; h is 2-4; i is 0
J is 0 to 3; k is 0 to 2;
The total number of R ⁶ is 0, 1 or 2; U is O, S or NR
Represents ¹ ; z is 1 or 2; each R ¹⁴ is independently alkyl of 1 to 9 carbons;
An arylalkyl having 7 and an aryl moiety having 6 to 10 carbons; an alkenyl having 2 to 9 carbons; an alkenyl moiety having 2 to 4 carbons and an aryl moiety having 6 to 10 carbons.
Aryl substituted alkenyl having 2 to 9 carbons
Number alkynyl; having alkynyl portions of two to four carbon atoms, an alkynyl aryl moiety is an aryl substituted with 6-10 _{^{carbon; -COR 2; -CO 2 R 3}} ; -CON (R 2) 2; −
(CH ₂ ) _t R ^{7 wherein} t is 0 or an integer of 1 to 4; and-(CH ₂ ) _v ZR ^{8 wherein} v is 0 or 1 to 3
Z represents -S- or -O-, and R ¹ , R ⁷
And R ⁸ are selected from the group consisting of], as defined above].

Preferred compounds of the general formula (L) in which one or more of the substituents R ₆ are essential for the formation of a non-spiro ring are of the following structural formula: [Wherein a, b, c, d, (c + d), e, g, i,
k, the total number of radicals R ⁶ , U and R ¹⁴ are the same as defined above].

The substituent T preferably has the general formula: R ³⁰ (CH ₂ ) _n 'C≡C-, wherein n' is 1 to 4 and R ³⁰ is HO-, MeO-,
_{(N-Pr) 2 N-,} CH 3 CO 2 -, CH 3 CH 2 OCO 2 -HO 2 C-, OHC
-, Ph-, 3-HO- Ph- and PhCH ₂ O-is selected from the group consisting of with the proviso that when R ³⁰ is Ph or 3-HO-Ph is, n 'is 0 ] The acetylene-containing component which has this.

Most advantageously, T is MeOCH ₂ C≡C—, (n-Pr) ₂ NC
H ₂ C≡C-, CH ₃ CO ₂ CH ₂ C≡C-, EtOCO ₂ CH ₂ C≡C-, HOC
H ₂ C≡C-, HO (CH ₂ ) ₂ C≡C-, CH ₃ CO ₂ (CH ₂ ) ₂ C≡C
-, HO ₂ C (CH ₂ ) ₂ C≡C-, OHC (CH ₂ ) ₃ C≡C-, HO (C
H ₂ ) ₄ C≡C-, PhC≡C-, 3-HO-PhC≡C- and PhCH
₂ OCH ₂ C≡C-.

The sign x defining the number of substituents T is preferably 1 or 2, most preferably 1, and when x is 1, T
Is preferably in the 4-position of ring A.

Ring A is preferably a phenyl or thiophene ring, most preferably phenyl.

Ring B is preferably a 1,4-phenylene ring or a 2,5-thiophene ring, most preferably a 1,4-phenylene ring.

 The D unit is most preferably a carbonyl group.

In group E, R ⁶ is preferably 1) an arylalkyl in which the aryl part has 6 to 10 carbons and the alkyl part has 1 to 8 carbons; 2) t is an integer of 0 or 1 to 5 R ⁷ is an imidoyl group having an aromatic group; — (CH ₂ ) R ⁷ ; or 3) v is 0 or an integer of 1 to 4, Z is S or O, and R ⁸ is carbon — (CH ₂ ) _v ZR ⁸ wherein 6-10 aryl or aryl moieties have 6-12 carbons and the alkyl moiety is arylalkyl having 1-4 carbons.

The group R ⁶ is most preferably the following, in which all aromatic moieties are advantageously substituted: 1) the aryl part is phenyl and the alkyl part is 1 to 4 carbon atoms. 2) t is an integer of 1 to 3, and R ⁷ is N- (1,2-naphthalene-dicarboximidoyl), N- (2,3-naphthalene-dicarboximidoyl) or -(CH ₂ ) _t R ⁷ , which is N- (1,8-naphthalene-dicarboximidoyl) or N-phthalimidoyl, 3) v is an integer of 1 to 3, Z is S and R ⁸ Is phenyl- (CH ₂ ) _v ZR ⁸ .

Further advantageous compounds of the general formula (L) have the following structural formula: Has an R ⁶ unit represented by

One of skill in the art will appreciate that many of the compounds of the present invention exist in enantiomeric or diastereomeric forms, and that such stereoisomers generally exhibit different activities in biological systems. It is clear that. The present invention includes all possible stereoisomers having inhibitory activity against MMPs, as well as mixtures of stereoisomers having at least one inhibitory activity, regardless of the name of the stereoisomer.

The most advantageous compounds of the present invention are listed in the following list: I) R / S 4 '-(3-hydroxy-1-propynyl) -γ-oxo-α- (3-phenylpropyl)-
[1,1'-biphenyl] -4-butanoic acid, II) S-4 '-(3-hydroxy-1-propynyl)
-Γ-oxo-α- (3-phenylpropyl)-[1,
1'-biphenyl] -4-butanoic acid, III) R-4 '-(3-hydroxy-1-propynyl) -γ-oxo-α- (3-phenylpropyl)-
[1,1′-biphenyl] -4-butanoic acid, IV) 4 ′-(3-methoxy-1-propynyl) -γ-
Oxo-α- (3-phenylpropyl)-[1,1′-biphenyl] -4-butanoic acid, V) γ-oxo-α- (3-phenylpropyl)-
4 '-(3-propyl-1-hexynyl)-[1,1'-
Biphenyl] -4-butanoic acid, VI) 4 '-[3- (acetyloxy) -1-propynyl] -γ-oxo-α- (3-phenylpropyl)-
[1,1′-biphenyl] -4-butanoic acid, VII) 4 ′-“3-{(ethoxycarbonyl) oxy} -1-propynyl] -γ-oxo-α- (3-phenylpropyl)-[1 , 1'-biphenyl] -4-butanoic acid, VIII) 4 '-(4-hydroxy-1-butynyl) -γ
-Oxo-α- (3-phenylpropyl)-[1,1′-
Biphenyl] -4-butanoic acid, IX) 4 '-[3- (acetyloxy) -1-propynyl] -γ-oxo-α- (3-phenylpropyl)-
[1,1′-biphenyl] -4-butanoic acid, X) 4 ′-(4-carboxy-1-butynyl) -γ-
Oxo-α- (3-phenylpropyl)-[1,1′-biphenyl] -4-butanoic acid, XI) γ-oxo-4 ′-(5-oxo-1-pentynyl) -α- (3-phenylpropy )-[1,1'-biphenyl] -4-butanoic acid, XII) 4 '-(6-hydroxy-1-hexynyl) -γ
-Oxo-α- (3-phenylpropyl)-[1,1′-
Biphenyl] -4-butanoic acid, XIII) γ-oxo-4 '-(phenylethynyl) -α
-(3-phenylpropyl)-[1,1'-biphenyl]
-4-butanoic acid and XIV) 4 '-[(3-hydroxyphenyl) ethynyl]-[gamma] -oxo- [alpha]-(3-phenylpropyl)-
[1,1′-biphenyl] -4-butanoic acid, XV) 1,3-dihydro-1,3-dioxo-α- [2-oxo-2- [4 ′-｛3- (phenylmethoxy) -1 -Propynyl}-[1,1'-biphenyl] -4-yl] ethyl] -2H-isoindole-2-butanoic acid and XVI) 1,3-dihydro-α- [2- [4- (hydroxyethynyl)- [1,1'-biphenyl] -4-yl] -2
-Oxoethyl] -1,3-dioxo-2H-isoindole-2-butanoic acid.

General Production Methods: The compounds of the present invention can be easily produced using known chemical reactions and methods. Nevertheless, the following general method of preparation is presented to assist reading in synthesizing inhibitors with the more detailed examples presented in the experimental section below, which provides working examples. Keep it. All variable groups of the method are the same as described in the general description, unless otherwise defined below. The variable sign n is defined independently in each case. When a variable group as described by a code (eg R ⁹ ) is used more than once in the structural formulas described, each of the above groups has the definition of the above code. May independently vary within the range.

General Methods A-The compounds of the invention in which the A and B rings are substituted phenyl and phenylene, respectively, are preferably prepared in an aprotic solvent in the presence of a Lewis acid catalyst such as aluminum trichloride, e.g. Free of substituted biphenyls M II and activated acyl-containing intermediates such as succinic or glutaric anhydride derivatives M III or acid chlorides M IV in 1,1,2,2-tetrachloroethane Manufactured by use of the Del Crafts reaction. The well-known Friedel-Crafts reaction involves many alternative solvents and Berlin
er.Org.React., 5,229,1949 and Heaney, Comp.Org.Synt
h., 2,733, 1991.

The anhydride M III is mono- or polysubstituted in an asymmetric manner, and the crude product MI-A often exists as a mixture of isomers by anhydride attack from either of the two carbonyls. ing. The resulting isomer is
Degraded to pure form by crystallization or chromatography using standard methods known to those skilled in the art.

If these are not available commercially, MM III is succinic anhydride, aldehyde or ketone Sutobbu condensation of dialkyl succinate used (resulting in R ⁶ in the side chain), subsequent catalytic hydrogenation, in order to diacid It can be prepared by hydrolysis of a half ester intermediate and subsequent conversion to anhydride M acid by reaction with acetyl chloride or acetic anhydride. Also, in the half ester, the simple is converted to the acid chloride MIV by treatment with thionyl chloride or oxalyl chloride. For an overview of Stob condensation, including a list of suitable solvents and chlorides, see Johnson and Daub, Org.
See t., 6,1,1951.

The method comprises the steps of M III (R ⁶ is H, isobutyl and H,
n-pentyl) as well as Wo
lanin. et al., US Patent No. 4771038.

Method A Method A is particularly useful for the preparation of cyclic compounds such as MI-A-3, wherein the two R ⁶ groups are linked to a methylene chain to form a 3-7 membered ring. is there. The small ring (3- to 5-membered) anhydride is the compound of the invention MI in cis form.
It is readily available only as the cis isomer giving -A-3-. Then use a base such as DBU in THF
The trans-form compound MI-A4 is produced by the treatment of MI-A3. Substituted four-membered starting material anhydrides, such as M III-A-1, are described below, as described below.
It is formed by a photochemical 2-2 reaction. This method is based on R ¹⁴
Is particularly useful for the preparation of compounds such as acetoxy or acetoxymethylene. Next Friedel
After the Kraftz reaction, the acetate can be removed by basic hydrolysis and the 2- (trimethylsilyl)
The carboxyl was protected by conversion to the ethyl ester. The resulting intermediate, wherein R ¹⁴ is CH ₂ OH, can be converted to compounds of the invention having another R ¹⁴ group by using the methods described in General Method G.

Also, the Friedel-Crafts method states that the double bond is a C-2 of a succinoyl chain (e.g., from maleic anhydride or 1-cyclopentene-1,2-dicarboxylic anhydride).
Or where two R ⁶ groups are present together on one chain carbon to form an exo-methylene (＝ CH ₂ ) group. As in the case of the use of itaconic anhydride as starting material to obtain
It is also useful when a double bond is found in the side chain. The subsequent use of said compounds is described in Method D column.

General Method B-Compound MI can also be prepared by monoalkylating dialkyl maleate MVI with an alkyl halide to form intermediate M VII, followed by intermediate M
Halomethyl biphenyl ketone M VI to give IX
It can also be prepared by a reaction cascade involving alkylation with II. Next, the compound of formula MIX is hydrolyzed using an aqueous base, heated to decarboxylate the maleic acid intermediate, and MI-B-2
(Method B-1). By using one equivalent of an aqueous base, the ester MI having R ¹² as alkyl
-B-2 is obtained, and an acidic compound (R ¹² is H) is obtained by using 2 equivalents or more of the base. Optionally, no heating is used and the diacid or acid ester MI-
B-1 is obtained.

Also, the diester intermediate M IX can be heated at 110 ° C. for about 24 hours in a sealed tube with a strong acid such as concentrated hydrochloric acid in acetic acid to give MI-B-1 (R ¹² Is). Also, the reaction of MVI with MVIII can be performed before the reaction with the alkyl halide to give the same MIX (Method B-2).

Also, the diester intermediate M XIX where R ¹² is allyl is
Upon exposure to a Pd catalyst in the presence of pyrrolidine, MI-B-2
(R ¹² is H) (Dezei
l, Tetrahedron Lett. 28, 4371, 1990).

Intermediate M VII is formed from biphenyl M II in a Friedel-Crafts reaction using a haloacetyl halide, such as bromoacetyl bromide or chloroacetyl chloride. Alternatively, biphenyl can be reacted with acetyl chloride or acetic anhydride, and the resulting product can be halogenated, for example, using bromine, to form an intermediate M VIII (X is
Br).

Method B has the advantage that when Method A produces a mixture, it produces a single part of the isomer. Method B is particularly useful when Method A is used and the side chain R ⁶ has an aromatic or heteroaromatic ring that can participate in intramolecular acylation reactions to produce by-products. This method is also very useful when the group R ⁶ adjacent to the carboxyl of the final compound has a heteroatom such as oxygen, sulfur or nitrogen or a more complex functional group such as an imide ring.

Method B If R ⁶ has a selected functional group Z, maleate M VII can be prepared by alkylation of a commercially available unsubstituted maleate with prenyl or allyl halide, and the product Ozonolysis is carried out in a reducing operation, and a desired group Z is subjected to Mitsunobu reaction (Mits
unobu, Synthesis 1, 1981). Also, the intermediate alcohol can be exposed to alkylation conditions to obtain the maleate M VII having the desired group Z.

General Method C—It is particularly advantageous to use chiral HPLC to resolve enantiomers of a racemic product mixture (eg, Arit. Et al., Chem. Int. Ed. Engl. 12, 30, 1991).
reference). The compounds of the present invention can be prepared as pure enantiomers by using a chiral auxiliary step. For example, Evans. Aldrichimica Acta, 15 (2), 23,
See 1982 and other similar references known to those skilled in the art.

General Method D - R ⁶ is an alkyl - or aryl - or heteroaryl - or acyl - or heteroaryl carbonyl - compounds where thiomethylene, patent WO 90/057
Prepared in a similar manner as described in 19. When the thus substituted itaconic anhydride MXVI (n = 1) is reacted under Friedel-Crafts conditions, the acid MI-D-
1, which can be separated from the minor isomer MI-D-5 by chromatography or crystallization. MI-D-5 can also be obtained by reacting compound MI-D-4 according to the invention (according to any of methods A to C) with formaldehyde in the presence of a base.

The compound MI-D-1 or MI-D-5 is then converted to a catalyst such as potassium carbonate, ethyldiisobutylamine, tetrabutylammonium fluoride or an azobisisobutyronitrile (AIBN) free radical initiator. Is reacted with a mercapto derivative M XVII or XM VIII in a solvent such as diethylformamide or tetrahydrofuran to give the compounds MI-D-2, MI-D-3, M
ID-6 or MI-D-7 is obtained.

Method D General Methods E-Biaryl compounds such as those of the present invention are prepared by combining an aryl or heteroaryl metal compound whose metal is zinc, tin, magnesium, lithium, boron, silicon, copper, cadmium, etc. with an aryl or heteroaryl halide or trifide. It can also be prepared by a cross-coupling reaction with Suzuki or Stille, such as a rate (trifluoromethane-sulfonate). In the following reaction formula, either Met or X is a metal, and the other is a halide or triflate (OTf). Pd (com)
Is a soluble complex of palladium such as tetrakis (triphenylphosphine) -palladium (O) or bis- (triphenylphosphine) -palladium (III) chloride. These methods are well known to those skilled in the art. For example, Suzuki, Pure Appl. Chem. 63,213 (199
4); Suzuki, Pure Appl. Chem. 63, 419, 1991; and Fariana
And Roth, “Metal-Organic Chemistry” Volume 5 (Chap
ter 1), see 1994.

The starting material M XXIII (B = 1,4-phenylene) is readily formed using methods similar to methods A, B, C or D, but using halogenated benzene rather than biphenyl as the starting material Is done. If desired, the material wherein X is a halogen atom can be converted to a bromo intermediate, such as treatment of the bromo intermediate with hexamethylditin and palladium tetrakistriphenylphosphine in toluene under reflux to form the trimethyltin intermediate. The reaction can be converted to a substance wherein X is a metal by reactions well known to those skilled in the art. The starting material M XXII (B = heteroaryl) is most advantageously prepared by Method C, but more readily available heteroaryl is used than the biphenyl starting material. Intermediate MXXII is commercially available or is readily prepared from commercially available materials by methods well known to those skilled in the art.

T, x, A, B, E and G are the same Me as in the structural formula (L)
t = metal and X = halide and triflate or Met = halide or triflate and X = metal The general method described above involves various Friedel-Crafts reactions such as methods A, B, C or D Useful for the preparation of compounds that yield mixtures in biaryl acylation patterns. In addition, the method E includes a method in which the aryl group A or B has one or more heteroatoms, such as a compound having a thiophene, furan, pyridine, pyrrole, oxazole, thiazole, pyrimidine, or pyrazine ring instead of phenyl. Particularly useful for the preparation of (heteroaryl).

General Method F—The group R ^{6 of} Method F is prepared by the following intermediate M XXV
Wherein, when taken together to form a 4-7 membered carbocyclic ring, the double bond is treated with 2 equivalents of a strong base such as lithium diisopropylamide or lithium hexamethylsilylamide; Continue with structure M XXVI
The conjugation with a ketone group can be removed by acid termination to obtain a compound having the following formula: Next, the reaction of M XXVI with a mercapto derivative using a method similar to the general method D gives the cyclic compound MI-F-1 or MI-F-2.

Method F General Methods G Compounds of the present invention wherein two groups R ⁶ are linked to form a substituted 5-membered ring are most advantageously prepared by Method G. In this method, the acid C II (R = 0H)
Has been produced using the method described in Tetrahedron 37, Suppl., 411, 1981. The acid is protected as an ester by use of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and methods well known to those skilled in the art [eg, R = benzyl (Bn) or 2- ( Trimethylsilyl) ethyl (TMSE)]. The substituted bromobiphenyl C III has been converted to its Grignard reagent by treatment with magnesium and reacts with CI to give the alcohol C VI. Alcohol CVI is removed by base treatment of the mesylate by using conditions well known to those skilled in the art to produce olefin CVII. Also, C III is
First, bromide metallization with n-butyllithium at low temperature (−78 ° C.) followed by treatment with chlorotrimethyltin is converted to a trimethyltin intermediate, CI
Enol triflate (C II) by reaction with 2- [N, N-bis (trifluoromethylsulfonyl) amino] -5-chloropyridine in the presence of a strong aprotic base
Is converted to Next, tin and enol triflate intermediate is coupled in the presence of a Pd ⁰ catalyst. Cu I and AsPh ₃ directly give intermediate C VII. Ozone decomposition of C VII (treatment with methyl sulfide) gives aldehyde C
VIII occurs. Also, following the treatment with OsO _4, converts the C VII to C VIII by HIO _4.

Method G The conversion of the key intermediate C VIII to the intended compounds of the invention is carried out in several ways, depending on whether the side-chain sensitive groups Z are identical. Reaction of CVIII with Wittig reagent and subsequent hydrogenation yields products where Z is alkyl or arylalkyl. Aldehyde C VIII using a reducing agent such as lithium tris [(3-ethyl-3-pentyl) oxy] aluminum hydride (LTEPA).
The selective reduction of affords the alcohol C IX. The alcohol was converted to phenyl ether or various heteroatom-substituted derivatives, which were used to generate the side chain Z according to Mitsunobu conditions well known to those skilled in the art (see Mitsunobu, Synthesis, 1, 1981). . Also, the alcohol of CIX was converted to a leaving group or bromide such as tosylate (CX) by conditions well known to those skilled in the art, and then the leaving group was replaced by a suitable nucleophilic group. Some examples of this type of reaction are described in Norma, et al., J. Med.
37,2552,1994. Direct acylation of alcohol C IX results in compounds of the present invention where Z is O acyl, and reaction of said alcohol with various alkyl halides in the presence of a base gives alkyl ether. In each case, the final step depends on the stability of R and Z, but in all cases 2- (trimethylsilyl) by removal of benzyl by chlorine hydrolysis or treatment with tetrabutylammonium fluoride. Removal of the acid blocking group R to generate an acid (R = H) by using conditions well known to those skilled in the art, such as the removal of ethyl.

General Methods H-The amides of the acids of the compounds of this invention can be prepared from the acids by treatment with a primary or secondary amine and a coupling agent such as dicyclohexylcarbodiimide in a suitable solvent, such as dichloromethane or dimethylformamide. Can be manufactured. These reactions are well known to those skilled in the art. The amine component may be simply alkyl or arylalkyl substituted, or may be an amino acid derivative in which the carboxyl is blocked and the amino group is free.

General Methods Compounds of the present invention wherein I- (T _x ) is alkynyl or substituted alkynyl are prepared by general method I (Austin, J. Org. Chem. 46, 2280, 1981). Intermediate M
X is prepared by methods A, B, C, D or G by starting with commercially available M III (R ¹ = Br). Cu
(I) / MX and substituted acetylene M in the presence of paradate
Reaction with XI yields compound MI-I-1 of the present invention. In certain cases, R ³ is a trialkylsilyl,
Alcohol may be blocked. In such cases, the silyl group can be removed by treatment with an acid such as trifluoroacetic acid or an HF-pyridine reagent.

Method I Suitable pharmaceutically acceptable salts of the compounds of the present invention include addition salts formed with organic or inorganic bases.
The salt-forming ions derived from these bases may be metal ions, for example aluminum ions, alkali metal ions such as sodium or potassium, alkaline earth metal ions such as calcium or magnesium or amine salt ions. Some are known for this purpose. Ammonium salts, dibenzylamine and
Arylalkylamines such as N, N-dibenzylethylenediamine, methylamine, t-butylamine, lower alkylamines such as procaine, lower alkylpiperidines such as N-ethylpiperidine, cycloalkyls such as cyclohexylamine or dicyclohexylamine Examples include amines, 1-adamantylamine, benzathine, or salts derived from amino acids such as arginine, lysine and the like. Physiologically acceptable salts such as sodium or potassium salts and amino acid salts can be advantageously used in medicine as described below.

These and other salts, which are not necessarily physiologically tolerable, are useful for isolating or purifying the acceptable products for the purposes described below. For example, (+)-in a suitable solvent
The use of commercially available enantiomerically pure amines, such as cinchonine, often results in classical resolution.
A simple enantiomeric salt crystal of a compound of the present invention can be produced, leaving the opposite enantiomer in solution, in a method called l resolution). Since one enantiomer of the described compounds of the present invention generally has a substantially greater physiological effect than its enantiomer, this active isomer exists in purified crystalline or liquid phase. The salts are prepared by reacting the compound of the invention in the acid form with an equivalent amount of the base by providing the desired basic ion in the medium in which the salt precipitates or in the aqueous medium and subsequent lyophilization. By conventional neutralization techniques, the free acid form is obtained from the salt using, for example, potassium bisulfate, hydrochloric acid, and the like.

The compound of the present invention may be a matrix metalloprotease
Inhibits MMP-3, MMP-9 and MMP-2 and to a lesser extent MMP-3
-1 and thus are effective in treating or preventing the diseases described in the state of the art.
Compounds of the invention are likely to inhibit these other MMPs to a different extent, as the other unlisted MMPs share a high degree of homology with those described, especially at the site of the catalyst. . Variations in substituents on the biaryl portion of the molecule and the propanoic or butanoic acid linkage portion of the claimed compounds have been shown to affect significant inhibition of the described MMPs. Thus, this general class of compounds `` modulates '' by choosing particular substituents to enhance the inhibition of particular MMPs associated with particular pathologies, while leaving unrelated MMPs to be less effective be able to.

Methods for treating conditions caused by matrix metalloproteases can be performed on mammals, including humans, that exhibit these conditions.

The inhibitors of the present invention are intended for use in animal and human applications. For these purposes, the inhibitors may contain one or more pharmaceutically acceptable carriers, diluents, fillers, binders and other active ingredients, depending on the intended mode of administration and dosage form. Used in pharmaceutical formulations containing excipients.

Administration of the inhibitor may be in any suitable manner known to those skilled in the art. Examples of suitable parenteral administration include intravenous, intraarticular, subcutaneous and intramuscular methods. Intravenous administration can be used to obtain a sharp control of the maximum plasma concentration of the drug. Improved half-life and targeting of the drug to the joint cavity is aided by incorporating the drug into liposomes. By incorporating a ligand outside the liposome that binds to synovial fluid-specific macromolecules, it is possible to increase the selectivity of liposome targeting to the joint cavity. Selective intramuscular, intra-articular or subcutaneous depot injection with or without encapsulation of the drug in degradable microspheres, for example composed of poly (DL-lactide-coglycolide), can be used to prolong the maintenance of the drug. Used to obtain the release of For the improved convenience of the dosing form,
Park seal system available from pharmacies (Percuseal
System) can be used. Improved convenience and patient compliance are achieved through the use of injector pens (eg, Novopin or Q pens) or needleless jet injectors (eg, from Bioject, Mediject or Becton Dickinson). Other precisely controlled releases such as prolonged zero order or pulsed release are achieved by releasing the drug through the cannula into the synovial space as needed using an infusion pump. Examples include ALZ such as ALZET osmotic pump
Includes subcutaneous infusion osmotic pump from Company A.

Nasal release involves the incorporation of the drug in a carrier (less than 200 μm) of bioadhesive particles, such as a carrier consisting of cellulose, polyacrylate or polycarbophil, combined with a suitable absorption enhancer such as a phospholipid or acylcarnitine. Is achieved by Available systems include Dan
Includes those developed by Biosys and Scios Nova.

A striking property of the compounds of the invention compared to the various peptide compounds described in the state of the art here is the outstanding oral activity of the compounds of the invention. Some compounds are
It showed oral biocompatibility in various animal models of up to 90-98%. Oral release is achieved by incorporating the drug into tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsifiers or suspensions. Oral release is achieved by incorporating the drug into an enteric-coated capsule formed to release the drug into the colon with low digestive protease activity. Examples include ORZ-CT / Osmet ^TM and PULS from ALZA.
Includes the INCAP ^™ system and the Scherer Drug Delivery System, respectively. Other systems use azo-crosslinked polymers that are degraded by colon-specific bacterial azoreductase or pH-sensitive polyacrylate polymers that are activated by increasing pH in the colon. The system described above is used in conjunction with a wide range of available absorption enhancers.

Rectal release is accomplished by incorporating the drug into a suppository.

The above-mentioned preparations can be prepared by adding a compound of the present invention to various therapeutically inert, inorganic or organic carriers well known to those skilled in the art. Examples include lactose,
Examples include, but are not limited to, corn starch or derivatives thereof, talc, vegetable oils, waxes, fats, polyols such as polyethylene glycol, water, saccharose, alcohol, glycerin and the like. Various preservatives may be required to help the stability of the formulation or to increase the biocompatibility of the active ingredient, or to produce a formulation having an acceptable flavor or aroma for oral administration. , Emulsifier, dispersant, flavoring agent, wetting agent,
Antioxidants, softeners, coloring agents, stabilizers, salts, buffers and the like are added.

The amount of pharmaceutical composition used will depend on the recipient and the condition being treated. The required amount is determined without undue experimentation by methods well known to those skilled in the art. By determining the amount of target enzyme that must be inhibited to selectively treat the disease, the required amount is calculated.

The matrix metalloprotease inhibitors of the present invention are useful not only for the treatment of the above physiological conditions, but also for activities such as metalloprotease purification and for testing matrix metalloprotease activity. This activity test may be performed in vitro using natural or synthetic enzyme preparations or in vivo, eg, where abnormal disruptive enzyme concentrations are naturally found (use of genetically mutated or transgenic animals). Or using animal models caused by administration of foreign agents or by surgery that impairs joint stability.

EXAMPLES The following examples are for illustrative purposes only and are not intended to, or in any way, be construed as limiting, the invention.

General Methods: All reactions were performed in flame-dried or oven-dried glassware under a positive pressure of argon and were magnetically stirred unless otherwise noted. Sensitive liquids and solutions were transferred via syringe or cannula and introduced into the reaction vessel through a rubber septum. The reaction product solution was concentrated using a Buchi evaporator unless otherwise noted.

Materials: Diethyl ether and tetrahydrofuran, usually
Commercial grade reagents and solvents were used without further purification except distilling from benzophenone ketyl under argon and methylene chloride from calcium hydride under argon. Many specialty organic or organometallic starting materials and reagents are available from Aldrich. 1001 West Saint Pa.
Obtained from ul Avenue, Milwaukee, WI 53233. Solvents were often obtained from EM Science as distributed by VWR Scientific.

Chromatography: Analytical thin-layer chromatography (TLC), Whatman Step
Recoated glass backed silica gel 60A F-254 25
Performed on a 0 μm plate. Visualization of the spot
Performed by one of the following techniques: (a) UV irradiation,
(B) exposure to iodine vapor, (c) phos in ethanol
Immersing the plate in a 10% solution of homolybdic acid and
Subsequent heating and (d) ethanol containing 0.5% concentrated sulfuric acid
Into a 3% solution of p-anisaldehyde in ethanol
Plate immersion and subsequent heating.

 Column chromatography was performed using 230-400 mesh EM
 Science Performed using silica gel.

 Analytical high performance liquid chromatography (HPLC) at 288 nm
4.6 × 250mm Microsorb monitored at Depending on the column,
Performed in 1 ml and semi-preparative HPLC was monitored at 288 nm at 21.4 ×
250mm Microsorb Depending on the column, perform at 24 ml per minute
Was.

Apparatus: Melting point (mp) is measured by Thomas Hoover melting point apparatus (Th
omas-Hoover melting point apparatus) and were not calibrated.

Proton ( ¹ H) nuclear magnetic resonance (NMR) spectra were measured using a GN-OMEGA300 (300 MHz) spectrometer manufactured by General Electric Company, and the carbon 13 ( ¹³ C) N
The MR spectrum is converted to General Electric GN-OM
It measured using the EGA300 (75MHz) spectrometer. Most of the compounds synthesized in the following experiments were analyzed by NMR, the spectra being in each case consistent with the intended structure.

Mass spectrum (MS) data was obtained on a Kratos Concert 1-H spectrometer with the latest version of liquid cesium secondary ion (LCIMS), fast atom bombardment (FAB). Most of the synthetic sta compounds were analyzed by mass spectroscopy in the following experiments, and the spectra were
The spectra were in each case consistent with the intended structure.

General description: For a multi-step method, successive steps are indicated by numbers.

Example 1-Preparation of Compound I Step 1 A dry two-necked three-necked round bottom flask was equipped with a stir bar, an addition funnel for pressure equalization, an inlet for argon, and a thermometer. The flask was charged with a suspension of sodium hydride (8.4 g of 95% NaH; -0.33 mol) in anhydrous THF (700 ml) and cooled in an ice-water bath. Diethyl malonate (48.54 g, 0.30 mol) was added dropwise from the addition funnel over 25 minutes. Stirring was continued for 1.5 hours before 1-bromo-3-phenylpropane (47 ml, ~ 61 g, ~ 0.30 mol) was added via addition funnel over 10 minutes. An addition funnel rinse (THF, 2 × 10 ml) was added to the reaction mixture and stirring was continued for 30 minutes. The addition funnel and thermometer were replaced with a reflux condenser and stopper and the reaction was heated at reflux for 19 hours. The mixture was cooled to room temperature and then cooled in an ice-water bath. Distilled water (400 ml) was added slowly with stirring. The layers were separated and the aqueous phase was chloroform (10
0 ml). Combined organics with 10% HCl
(250 ml) and the separated aqueous phase was back-extracted with chloroform (100 ml). Combine organics with saturated N
After washing with aHCO ₃ (250 ml), the separated aqueous phase was back-extracted with chloroform (100 ml). Dry the organics (Na ₂
SO ₄ ) and when concentrated yields a yellow oil,
This was purified by distillation under reduced pressure (0.4 Torr) on a Vigreux column. The fraction boiling at 124-138 ° C. was the clear desired product (57.21 g, 0.206 mol; 68% yield). TLC (50% hexane-dichloromethane): _Rf = 0.32.

Step 21 A one-necked round-bottomed flask was equipped with a rubber septum and an inlet for argon. In this flask,
A solution of commercially available 4-bromobiphenyl (50.00 g, 0.215 mol) in dichloromethane (100 ml) was charged. Bromoacetyl bromide (21.0 ml, 48.7 g,
0.230 mol) was added by syringe and the solution was cooled to 0 ° C. in an ice-water bath, while AlCl ₃ (34.3 g, 0.
258 mol) was added dropwise. Gas evolved from the opaque olive green reaction mixture. The reaction mixture was carefully poured into cold saturated aqueous NaHCO ₃ at room temperature for 24 h. The resulting mixture is extracted with three 200 ml portions of ethyl acetate, the combined organic phases are dried over Na ₂ SO ₄ and concentrated to give the desired product as a yellow solid in quantitative yield. TC (30% dichloromethane-
Hexane), _Rf = 0.30.

Step 3 A dry, two-necked, three-necked round bottom flask was equipped with a magnetic stir bar, an inlet for argon, and an addition funnel for pressure equalization. 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 in an ice-water bath, while sodium hydride (5.40 g of 95% NaH, 0.214 mol) was slowly added in small portions. The reaction mixture was stirred at 0 ° C. for 1 hour and the product of Step 2 (80.0 g, 0.2%) in anhydrous THF (300 ml).
15 mol) was added via addition funnel in about 20 minutes. The dark orange reaction mixture was stirred at room temperature under argon for 3 hours. The reaction vessel was cooled in an ice-water bath, while distilled water (150 ml) was carefully added. The aqueous phase is extracted with three 300 ml portions of ethyl acetate, the combined organic phases are dried over MgSO ₄ and concentrated to yield 124 g of a dark orange oil. This material was used in the following work without purification.

The orange oil was dissolved in 400 ml of 1: 1 THF: methanol and added to aqueous NaOH (4N, 500 ml, 2.00 mol). The reaction mixture was stirred at room temperature for 24 hours,
Stir for 48 hours and then at room temperature for 24 hours. Most of the MeOH was removed in vacuo and the residue was extracted with a 200 ml volume of 1: 1 ethyl acetate: hexane and a 200 ml volume of hexane. Acidify the aqueous phase with HCl and add 200 ml of ethyl acetate
The extraction was carried out twice with each time and three times with 100 ml each. The combined organic phases are dried over MgSO ₄ and concentrated to give a quantitative yield of the diacid. TLC (10% methanol-chloroform and 1% acetic acid): _Rf = 0.45.

Step 4 The crude diacid from step 3 was dissolved in 1,4-dioxane (500 ml) and heated to reflux for 24 hours. The solvent was removed under vacuum and the residue in an amount of 10 g was chromatographed on silica gel (gradient elution with 10-50% ethyl acetate-hexane containing 1% acetic acid) to give a yellow solid bed. 0.840 g of the desired product
(10%), MP174 ° C is obtained.

Step 5-In a 15 ml single-necked round bottom flask equipped with a rubber septum and a needle inlet for argon, add diethylamine 2.
6 ml, product of step 4 (0.300 g, 0.667 mM), propargyl alcohol (1.0 ml, 0.96 g, 17 mM), copper (I) iodide
(0.0220 g, 0.115 mM) and trans-dichlorobis (triphenylphosphine) paradate (0.110 g, 0.157 m
M). The resulting mixture was stirred at room temperature for 4 days. The reaction mixture is concentrated (residue 290 mg) and a part (90 mg) of the residue is purified by column chromatography on 50 g of silica gel (20% ethyl acetate-
Hexane and 0.5% acetic acid), the coupling product as a white solid (0.035 g, 40%), MP130 ° C.

Examples 2 and 3-Preparation of compounds II and III_ThreeEtOH 5% in CN, H_TwoO4.75
% And Chiralcel AD with HOAc 0.095% On the column
And produced by the chiral separation of Example 1 at a flow rate of 20 ml / min.
Was.

Example 2: First Chiralcel AD column;¹H NMR (300MHz, C
DCl_Three) Δ8.02 (d, J = 8.4Hz, 2H), 7.67 (d, J = 8.7Hz, 2
H), 7.58 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 7.
17 ~ 7.33 (m, 5H), 4.54 (s, 2H), 3.46 (dd, J = 8.1,16.8
Hz, 1H), 3.14 to 3.02 (m, 2H), 2.65 (t, J = 7.2Hz, 2H),
1.64 to 1.84 (m, 4H).

Example 3: Second Chiralcel AD column;¹H NMR (300MHz, C
DCl_Three) Δ8.02 (d, J = 8.4Hz, 2H), 7.67 (d, J = 8.7Hz, 2
H), 7.58 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 7.
17 ~ 7.33 (m, 5H), 4.54 (s, 2H), 3.46 (dd, J = 8.1,16.8
Hz, 1H), 3.14 to 3.02 (m, 2H), 2.65 (t, J = 7.2Hz, 2H),
1.64 to 1.84 (m, 4H).

Example 6-Preparation of compound VI A single port with a rubber septum and a needle inlet for argon
In a 10 ml round bottom flask, 0.5 ml of pyridine, from Example 1 (0.0070 g, 0.014 mM) and acetic anhydride (0.020 ml, 22 mg,
0.21 mM). The reaction mixture was stirred at room temperature for 2 hours, then 30 ml of 1N HCl was added. The resulting mixture is
Extracted three times with 30 ml portions of ethyl acetate, the combined organic phases were dried over MgSO ₄ and concentrated. HPLC (2.5%
Of ethyl acetate-dichloromethane and 0.01% trifluoroacetic acid) to give 3 mg (38%) of Example 6, MP1
37 ° C. was obtained.

Example 7-Preparation of compound VII A single bite with a rubber septum and a needle inlet for argon
In a 15 ml round bottom flask, 2 ml of triethylamine, 2 m of THF
l, Compound I (0.0570 g, 0.134 mM) and ethyl chloroformate (0.032 ml, 36 mg, 0.34 mM) were charged. The reaction mixture was stirred at room temperature for 16 hours, then 50 ml of 1N HCl was added. The resulting mixture was washed with 3 portions of 50 ml of ethyl acetate.
Extracted once, the combined organic phases were dried over MgSO ₄ and concentrated. Column chromatography on silica gel 10g (40% ethyl acetate hexane and 0.5% HOAc)
And the following purification by HPLC (1.5% ethyl acetate-dichloromethane and 0.01% trifluoroacetic acid)
1 mg (1.5%) was obtained. MS (FAB-LSIMS) 499 [M
+ H] ⁺ .

Example 11-Preparation of compound XI A single bite with a rubber septum and a needle inlet for argon.
In a 25 ml round bottom flask, 1 ml of CH ₂ Cl ₂ , from Example 12 (0.012
g, 0.026 mM) and Dess. et al., J. Org. Chem. 48, 4156, 1
Dess-Martin reagent manufactured by Dess-Marti
n reagent) (16 mg, 0.038 mM). The resulting mixture was stirred at 0 ° C. for 30 minutes, diluted with 30 ml of ethyl acetate and washed twice with 20 ml of 1N HCl. MgS organic layer
Dried with O ₄ and concentrated. Purification by HPLC (1.
5% ethyl acetate-dichloromethane and 0.01% trifluoroacetic acid) gave 1 mg (9%) of Example 11. ¹ H
NMR (300 MHz, CDCl ₃ ) δ 89.70 (t, J = 1.3 Hz, 1H), 8.0
5 (d, J = 8.4 Hz, 2H), 7.70 (d, J = 8.7 Hz, 2H), 7.65 (d, J
= 8.4Hz, 2H), 7.44 (d, J = 8.4Hz, 2H), 7.15 ~ 7.35 (m, 5
H), 3.46 (dd, J = 8.1, 16.8 Hz, 1H), 3.14 to 3.02 (m, 2
H), 2.67 (t, J = 7.2 Hz, 2H), 2.48 (t, J = 7.5 Hz, 2H), 2.
41 (dt, J = 1.3 Hz and 6.3 Hz, 2H), 1.96 (m, 2H), 1.64 ~
1.84 (m, 4H).

Example 1, Example 2, Example 6, Example 7, and Example 11
The following series of biphenyl-containing products were used to produce:

Example 15-Preparation of Compound XV Step 1 A solution of sodium hydride (4.35 g, 181 mM) in freshly distilled THF (100 ml) was cooled to 0 ° C. and via a dropping funnel for 40 minutes, using commercially available diallyl malonate ( 35.0 g, 190 mM). 30 at room temperature
After stirring for minutes, N-(-2-bromoethyl) phthalimide (43.9 g, 247 mM) was added in one portion to the solution,
The mixture was heated to reflux. After 48 hours, the solution was cooled to 0 ° C., quenched with 2N HCl, and concentrated to about 20% of its original volume. The concentrate was diluted with ethyl acetate (300 ml) and washed successively with a saturated aqueous solution of K ₂ CO ₃ and NaCl. The organic layer was dried over MgSO _4,
Filter and concentrate under reduced pressure. Purification by flash column chromatography (gradient elution with 5-25% ethyl acetate-hexane) gave diallyl 2-phthalimidoethylmalonate (41.2%) as a colorless oil.
g, 64%). 1H NMR (300 MHz, CDCl ₃ ) δ7.82
(M, 2H), 7.72 (m, 2H), 5.85 (m, 2H), 5.30 (m, 2H), 5.
22 (m, 2H), 4.60 (m, 4H), 3.80 (t, J = 6.6Hz, 2H), 3.46
(T, J = 7.2 Hz, 1H), 2.30 (dd, J = 13.8, 6.9 Hz, 2H).

Step 2 Cool a solution of the product of Step 1 (5.20 g, 14.6 mM) in freshly distilled THF (100 ml) to 0 ° C, while slowly adding NaH (385 mg, 16.1 mM). did.
After 40 minutes, the reaction mixture was warmed to room temperature and
Of step 2 (4.55 g, 14.6 mM) was added in small portions and the mixture was stirred for 24 hours. The reaction mixture was cooled to 0 ° C., quenched slowly with 2N HCl (300 ml) and extracted with dichloromethane once with 150 ml and twice with 100 ml each time. The combined organic phases were dried over MgSO _4, and filtered, and concentrated,
This gives 6.5 g (71%) of the desired product, which is obtained in step 3
Used without purification. TLC (30% ethyl acetate-hexane): _Rf = 0.4.

Step 3 A solution of the product of Step 2 (6.50 g, 10.4 mM) in 1,4-dioxane (also 100) is cooled to 0 ° C., while tetrakis (triphenylphosphine) palladium (0.180 g, 146 mM) ) And pyrrolidine (2.40 ml, 29.2 mM)
Was added continuously. After stirring at 0 ° C. for 2 hours and at room temperature for 4 hours, the reaction mixture was poured into 2N HCl (100 ml). The resulting mixture was extracted four times with 100 ml portions of dichloromethane, and the combined organic phases were dried over MgSO ₄ and concentrated to give the diacid as a yellow solid (9.70 ml).
g). A 3.8 g sample of the material was dissolved in 1,4-dioxane (150 ml) and heated at reflux for 1 hour. After cooling to room temperature, the solution is concentrated and the residue is chromatographed on silica gel (300 g) (gradient with 5% to 15% methanol-dichloromethane) to give the desired acid (0.300 g). But this is
Further purification by recrystallization gave 0.170 g of the desired product as a white crystalline solid (59% of the total yield from step 2).
%), MP209-210 ° C.

Step 4 A one-necked 100 ml round bottom flask equipped with a rubber septum and a needle inlet for argon and containing 2 ml of THF was charged with NaH (435 mg, 17.2 mM) and cooled to 0 ° C.
But on the other hand, propargyl alcohol (1.0 ml,
0.963 g, 17.2 mM) was added via syringe over about 5 minutes. The resulting mixture was stirred at 0 ° C. for 10 minutes and at room temperature for 30 minutes. Benzyl bromide (1.8 ml, 2.59 g, 15.1 mM) was added and the reaction mixture was stirred at room temperature for 36 hours, and pentane (150 mM
l) and washed with 100 ml of brine. The solvent was distilled off and the residue (3.5 g of a yellow oil) was used directly in step 5. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.36-7.31 (m, 5
H), 4.61 (s, 2H), 4.17 (d, J = 2.4, 2H), 2.47 (t, J = 2.
4Hz, 1H).

Step 5 The method of Step 5 of Example 1 was used to prepare that of Example 15 using the product of Step 4 and the product of Step 3 as starting materials. MP 151 ° C.

Example 16-Preparation of Formula XVI Step 1 A one-necked 100 ml round bottom flask equipped with a rubber septum and a needle inlet for argon was charged with propargyl alcohol (1.0 ml, 0.963 g, 17.2 mM) and ether (20 ml), cooled, and cooled to 0 ° C. On the other hand, NaH (435 mg,
17.2 mM) was added slowly. The resulting mixture was stirred at room temperature for 1 hour and t-butyldimethylsilyl chloride (2.60
g, 17.2 mM). The reaction mixture was stirred at room temperature for 6 hours, poured into hexane (150 ml) and 1N
And washed with HCl. The organic phase was dried over MgSO ₄ and concentrated to give 2.88 g of a yellow oil, which was used in step 2 without purification. ¹ H NMR (300 MHz, CDCl ₃ )
δ 4.29 (d, J = 2.1 Hz, 2H), 2.37 (t, J = 2.1 Hz, 1H), 0.89
(S, 9H), 0.11 (s, 3H).

Step 2 The method of Step 2 of Example 1 was used to produce the desired acetyl biphenyl using commercially available 4-iodobiphenyl and acetyl chloride. TL
C (10% ethyl acetate-hexane): _Rf = 0.3.

Step 3 The method of Step 5 of Example 1 was used to produce the desired biphenylacetyl using the product of Step 1 and the product of Step 2. TLC (10% ethyl acetate-hexane): _Rf = 0.4.

Step 4 Into a one-necked 50 ml round bottom flask equipped with a rubber septum and a needle inlet for argon, 5 ml of THF, the product of Step 3 (1.06 g, 2.94 mM) is charged and cooled to −78 ° C. On the other hand, potassium hexamethyldisilazide (61
7 mg, 2.94 mM) was added dropwise via syringe. The reaction mixture was stirred at -78 ° C for 30 minutes, trimethylsilyl chloride (0.374ml, 0.320g, 2.94mM) was added dropwise via syringe and the resulting mixture was stirred at -78 ° C for 3 hours.
The reaction mixture was warmed to 0 ° C. in 1 hour,
Bromosuccinimide (0.540 g, 2.94 mM 9) was added and the mixture was warmed to room temperature and stirred for 16 hours.
The reaction mixture was poured into an aqueous solution of saturated NH ₄ Cl in a volume of 100 ml and extracted with three 50 ml portions of dichloromethane. The combined organic phases were dried with MgSO ₄ and concentrated. Column chromatography on 200 g of silica gel (gradient elution with 0-5% ethyl acetate-hexane) gave 0.264 g (20%) of bromomethyl ketone. TLC (10% ethyl acetate-hexane): _Rf = 0.5.

Step 5 The method of Example 15, steps 2-3, was used to produce the desired biphenylphthalimide using the product of step 4. TLC (50% ethyl acetate and 1% acetic acid):
_Rf = 0.3.

Filling neck, round bottom flask 50ml equipped with a step 6 needle inlet for rubber septum and argon, CH ₂ Cl ₂ 10ml, the product from Step 5 (0.040 g, 0.067) and HF- pyridine 2ml did. The resulting mixture was stirred for 10 min at room temperature, diluted with water in an amount of 75 ml, and extracted with CH ₂ Cl ₂ in an amount of 75 ml. The organic phase was dried over MgSO _4, and concentrated. By chromatography on 5 g of silica gel or by column chromatography (25% ethyl acetate-hexane and 1% HOAc), 6 mg of Example 16 (19%
%). MP145 ° C.

Example 17 Biological Evaluation of Compounds of the Invention P218 Fluorescence Quenching Evaluation for MMP Inhibition: The P218 fluorescence quenching evaluation (Microfluorometric Profiling Assay) was performed for substances in cuvettes and for various matrix metalloproteases (Microfluorometric Profiling Assay). MMP), Knight, et al., FEBS Lett. 296, 2
It is a modification of the one first described by 63,1992.
This evaluation was performed for each compound of the present invention and three MMs.
Performed with P, MMP-3, MMP-9 and MMP-2, analyzed simultaneously, 96-well microtiter plate and
Adapted as follows on a Hamilton AT workstation.

P218 fluorescent substrate: P218 is 4-acetyl-7- at the N-terminal position
It is a synthetic substrate containing a methoxycoumarin (MCA) group and a 3- [2,4-dinitrophenyl] -L-2,3-diaminopropyl (DPA) group inside. This is a variant of the peptide reported by Knight (1992) and is used as a substrate for matrix metalloproteases. P218
When the peptide is cleaved (the putative cleavage site for the Ala-Leu bond), the fluorescence of the MCA group is excited at 328 nm on a fluorimeter and 39.
Emitted at 3 nm and detected. P218 is currently BA only at Bayer
Manufactured as CHEM. P218 has the following structure: H-MCA-Pro-Lys-Pro-Leu-Ala-Leu-DPA-Ala
-Arg-NH2 (MW1332.2) Recombinant human CHO stromerbusin (MMP-3) Recombinant human CHOPro-MMP-3: human CHO prostromerbusin-257 (pro-MMP-3) is available from Housely et al., J. Biol. Che
m.2568.4481 (1993) and expressed and purified.

Activation of Pro-MMP-3: Tris 5 mM at pH 7.5, CaCl ₂ 5 m
M, NaCl 25 mM and 1.72 μM (100 μg / ml) Pro-MMP-3 in 0.005% Brij-35 (MMP-3 buffer) were added to TPS at 25 ° C.
K (N-tosyl- (L) -phenylalanine chloromethyl ketone) trypsin (1: 100 w / pro-MMP-3)
Activated by culturing for 30 minutes in w). The reaction was stopped by the addition of soybean trypsin inhibitor (SBTI: 5: 1 w / w, relative to trypsin concentration). This activation protocol resulted in the formation of a 45 kDa active MMP-3, which still contained the C-terminal portion of the enzyme.

Production of Human Recombinant Progelatinase A (MMP-2): Recombinant Human Pro-MMP-2: Human Progelatinase A
(Pro-MMP-2) was prepared according to Fridman et al., J. Biol. Chem. 267 15398.
(1992) using a vaccine expression system.

Activation of Pro-MMP-2: 252 mg / ml Pro-MMP-2 at 1: 5
And 50 μg / ml in 25 mM Tris, 7.5 mM CaCl ₂ , 150 mM NaCl, and 0.005% Brij-35 (MMP-2 activation buffer) at pH 7.5.
to a final concentration of ml. Mercury p-aminophenylacetate (APMA) was prepared at 10 mM (3.5 mg / ml) in 0.05 NaOH. 0.5m
APMA solution was added at 1/20 of the reaction volume for a final APMA concentration of M, and the enzyme was incubated at 37 ° C for 30 minutes. Activated MMP-2 (1
(5 ml) was dialyzed twice against MMP-2 activation buffer 2 (dialysis membrane was 1% against 0.1% BSA solution in MMP-2 activation buffer).
Minutes prior to treatment, then washed extensively with H ₂ O). The enzyme was concentrated in a centricon concentrator (the concentrator was pretreated for 1 minute with a solution consisting of 0.1% BSA in MMP-2 activation buffer, then H ₂
O, further washed with MMP-2 activation buffer), diluted again and then re-concentrated twice. The enzyme was diluted to 7.5 ml (0.5 times the original volume) with MMP-2 activation buffer.

Production of human recombinant progelatinase B (MMP-9): Recombination of human Pro-MMP-9: Wilhelm et al., J. Biol. Chem.
264 17213 (1989) Human progelatinase B (pro-MMP-9) derived from U937 cDNA was expressed in a sufficiently long form using a baculovirus expression system. Chem. 260 2493 (198
Purified using the method already described by 4).

Activation of Pro-MMP-9: Tris 50 mM, pH 7.4, CaCl ₂ 10m
Active by culturing 20 μg / ml of Pro-MMP-2 in 150 mM M, NaCl and 0.005% Brij-35 (MMP-9 activation buffer) with 0.5 mM mercury aminophenylacetate (APMA) at 37 ° C. for 3.5 hours It has become. The enzyme is dialyzed against the same buffer and AP
MA was removed.

Equipment: Hamilton Microlab AT Plus: Hamilton MicroLab AT P
MMP characterization was performed automatically with lus. Program Hamilton to (1) automatically dilute serially 11 possible inhibitors from a 2.5 mM stock in 100% DMSO, (2) partition the substrate, and then transfer the inhibitor to 96 well cytofluo Distribute to plate, and (3) mix and add single enzyme to plate to initiate reaction. The program is subsequently started at the point of substrate addition, the diluted inhibitor is remixed, and the plate is automatically made for each additional enzyme by initiating the reaction by addition of the enzyme. In this way, use the same inhibitor dilution for all
MMP analysis was performed.

Following Millipore cytofluoro II. Culture, the plates were read on a cytofluoro II fluorometric plate reader, excited at 340 nm, emitted at 395 nm, and read the gain set at 80.

Buffer: Micro Fluorescence Analysis Reaction Buffer (MRB): Dilute test compounds, enzymes and P218 substrate dilutions for microfluorescence analysis with Ca
Cl ₂ 10 mM, NaCl 150 mM, 0.005% Brij-35 and 1% DMS
Manufactured in microfluorescence reaction buffer containing 2- (N-morpholino) ethanesulfonic acid (MES) at pH 6.5 with O.

Method: MMP microfluorometric characterization: Analyze final substrate concentration 6
Varying drug concentrations were performed with μM P218 and approximately 5-8 nM MMP. Program Hamilton device, 2.5m sequentially
Dilution from the M stock (100% DMSO) to 11 compounds yielded a 10-fold final compound concentration during the evaluation. First, the microfluorimetric reaction buffer (MRB) from the instrument is placed in a 1 ml Marsh dilution tube.
Discharged to 6 tube shelves. Next, 20 μl of inhibitor (2.5 mM) was removed from the sample shelves of the device and mixed with buffer in row A of the Marsh shelf to yield a 50 mM drug concentrate. The inhibitors were then serially diluted to 10 mM, 5 mM, 1 mM, 2 mM, 0.5 and 0.1 mM. Position 1 on the sample shelf contains only DMSO for the enzyme-only well under evaluation, but this does not result in an inhibitor in column 1 rows AH. The instrument then dispenses 107 μl of P218 substrate (8.2 mM in MRB) into a single 96-well cytofluoromicrotitration plate. Remix the device and add Ma to the corresponding row in the microtiter plate.
Load 14.5 μl of diluted compounds of AG in rsh shelf (row H represents state-of-the-art row, releasing 39.5 μl of MRB instead of drug or enzyme). The reaction is initiated by adding 25 μl of the appropriate enzyme (5.86 times the final enzyme concentration) to each well except for row H, state of the art from the BSA-treated reaction reservoir (enzyme reservoir at room temperature for 1 hour). 1% in Tris 50 mM, pH 7.5, containing 150 mM NaCl
Pre-treated with BSA, followed by washing with H ₂ O and drying at room temperature).

After adding the enzyme and mixing, surround the plate and
Incubate for hours. The additional enzymes are tested in the same way by dispersing the P218 substrate in the microtiter plate and starting the Hamilton program, remixing, and dispersing the drug from the same Marsh shelf to the microtiter plate. Second to be inspected
The (or third etc.) MMPs are mixed and dispersed from the reaction shelves into the microtiter plate before surrounding and culturing. Reassemble with any additional MMPs that check this.

IC50 determination during microfluorometric assay: Copy the data generated with cytofluoro II from the exported "CSV" file to the master Excel extension sheet. Data from a variety of different MMPs (one 96-well plate for the MMP) is calculated simultaneously. The% inhibition was determined by comparing the amount of hydrolysis (fluorescent units generated over 25 minutes of hydrolysis) of the enzyme-only well in column 1 with the compound-containing well for each drug concentration.
Following background removal, the percent inhibition is calculated as [(control value-treated value) / (control value)] × 100.

Inhibition rates for drugs 5, 1, 0.5, 0.1, 0.02, 0.005 and 0.0
A determination was made for an inhibitor concentration of 01 mM. To obtain IC ₅₀ values, it was used a linear regression analysis of the percentage inhibition against inhibitor concentration log.

Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification or practice of the invention described herein. It is pointed out that the specification and examples are to be considered only as illustrative, and that the actual scope and spirit of the invention is indicated by the appended claims.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61P 1/02 A61P 1/02 9/00 9/00 15/00 15/00 17/00 17/00 25/00 25/00 27/02 27/02 29/00 29/00 35/00 35/00 43/00 111 43/00 111 C07C 59/90 C07C 59/90 59/92 59/92 69/145 69/145 69/96 69 / 96 Z 229/34 229/34 C07D 209/48 C07D 209/48 Z (56) Reference JP-A-61-65842 (JP, A) WO 96/15096 (WO, A1) WO 97/43237 ( WO, A1) West German Patent Application Publication 2804293 (DE, A1) Pharm. Sci. , 66 [4] (1977), pp. 466-476 (58) Fields investigated (Int. Cl. ⁷ , DB name) C07C 59/84 A61K 31/19 A61K 31/195 A61K 31/215 A61K 31/40 A61P 1 / 02 A61P 9/00 A61P 15/00 A61P 17/00 A61P 25/00 A61P 27/02 A61P 29/00 A61P 35/00 A61P 43/00 111 C07C 59/90 C07C 59/92 C07C 69/145 C07C 69 / 96 C07C 229/34 C07D 209/48 CAPLUS (STN) REGISTRY (STN) WPIDS (STN)

Claims (5)

(57) [Claims] (1) a general formula: [Wherein, R ¹⁵ represents HOCH ₂ , (n-Pr) ₂ NCH ₂ , CH ₃ CO ₂ CH ₂ ,
EtOCO ₂ CH ₂ , HO (CH ₂ ) ₂ , CH ₃ CO ₂ (CH ₂ ) ₂ , HO ₂ C (C
_{H 2) 2, OHC (CH} 2) 3, HO (CH 2) 4, 3-HO-Ph and P
h ¹⁶ is selected from the group consisting of hCH ₂ OCH ₂ and R ¹⁶ is And a pharmaceutically acceptable salt thereof. 2. A composition having a matrix metalloprotease inhibitory activity, comprising the compound of claim 1 and a pharmaceutically acceptable carrier. (A) osteoarthritis, rheumatoid arthritis, septic arthritis, periodontal disease, corneal ulcer, proteinuria, aortic aneurysm disease, dystrophic epidermolysis bullosa, inflammatory reaction, MM
Alleviation of the effects of osteopenia, temporomandibular joint disease, demyelinating diseases of the nervous system mediated by P activity; (b) suppression of degenerative cartilage loss due to tumor metastasis or traumatic joint injury; (c) atherosclerotic plaque rupture The composition having an inhibitory action on matrix metaprotease according to claim 2, which is used for reducing a coronary thrombosis derived from: (d) a conception control effect. 4. The composition according to claim 2, which is used for alleviating osteoarthritis. 5. The composition according to claim 2, which is used for inhibiting tumor metastasis.