JP2002515428A - Protease inhibitor - Google Patents

Abstract

  (57) [Summary] The present invention relates to diacylcarbohydrazide compounds that inhibit proteases including cathepsin K and pharmaceutically acceptable salts, hydrates and solvates thereof, pharmaceutical compositions of such compounds, novel intermediates of such compounds, and Osteoporosis; gingival diseases including gingivitis and periodontitis; arthritis, more particularly osteoarthritis and rheumatoid arthritis; Paget disease; hypercalcemia of malignancy; A method of treating a disease of bone loss or cartilage or matrix destruction, comprising inhibiting the bone loss or excessive cartilage or matrix destruction by administering a compound of the invention to a patient in need of such treatment. .

Description

DETAILED DESCRIPTION OF THE INVENTION

FIELD OF THE INVENTION The present invention relates generally to diacylcarbohydrazide protease inhibitors, particularly such inhibitors of cysteine and serine proteases, more particularly compounds that inhibit cysteine proteases, and even more particularly the papain superfamily. More particularly, it relates to compounds that inhibit the cysteine proteases of the cathepsin family, and most particularly to compounds that inhibit cathepsin K. Such compounds are particularly useful in the treatment of cysteine protease related diseases, particularly those of excessive bone or cartilage loss, such as osteoporosis, periodontitis and gingivitis.

BACKGROUND OF THE INVENTION Cathepsins are a family of enzymes that are part of the papain superfamily of cysteine proteases. Cathepsins B, H, L, N and S have been described in the literature. Recently, a cathepsin K polypeptide and a cDNA encoding such a polypeptide were disclosed in U.S. Patent No. 5,501,969 (referred to there as cathepsin O). Cathepsin K has recently been expressed, purified and characterized (Bossard, MJ et al. (1996) J. Biol. Chem. 271, 12517-12524; Dra
ke, FH et al., (1996) J. Biol. Chem. 271, 12511-12516; Bromme, D. et al., (1996).
J. Biol. Chem. 271, 2126-2132).

[0003] Cathepsin K has been variously designated in the literature as cathepsin O or cathepsin O2. It is believed that the designation cathepsin K is more appropriate. Cathepsins function in the normal physiological process of proteolysis in animals, including humans, for example, in the degradation of connective tissue. However,
High levels of these enzymes in the body can cause physiological conditions that lead to disease. Thus, cathepsins include, but are not limited to, pneumocystis carinii, trypanosome cruzi (tryp
sanoma cruzi), Trypanosoma brucei (trypsanoma brucei)
brucei) and Crithidia fusiculata;
It has been implicated as a causative agent in various disease states including schistosomiasis, malaria, tumor metastasis, metachromatic cerebral white matter atrophy, muscular dystrophy, muscular atrophy and the like. See International Publication No. WO 94/04172 published March 3, 1994 and references cited therein. See also European Patent Application EP 0 603 873 A1 and references cited therein. P. gingivalis (P.
gingivallis) have been implicated in the pathogenesis of gingivitis (Potempa, J. et al. (1994) Perspectives in Drug Discovery and Desig).
n, 2, 445-458).

[0004] Cathepsin K is thought to play a causal role in diseases of excessive bone or cartilage loss. Bone consists of a protein matrix into which spindle-shaped or planar crystals of hydroxyapatite have been incorporated. Type I collagen represents the major structural protein of bone that forms about 90% of the protein matrix. The remaining 10% of the matrix consists of several non-collagenous proteins including osteocalcin, proteoglycans, osteopontin, thrombospondin, fibronectin and bone sialoprotein. The skeleton undergoes remodeling at discrete locations for a lifetime. These positions, or remodeling units, are
It then undergoes a circulation consisting of the phase of bone replacement.

[0005] Bone resorption is performed by osteoclasts, which are multinucleated cells of the hematopoietic lineage. Osteoclasts adhere to the bone surface, forming a tightly sealed zone, and then
That is, the film is rippled over a wide area on the (reabsorption) surface. This results in encapsulated intracellular compartments on the bone surface that are acidified by the proton pump on the corrugated membrane, into which osteoclasts secrete proteolytic enzymes. The low pH of the compartment dissolves the hydroxyapatite crystals on the bone surface, while proteolytic enzymes digest the protein matrix. Thus, absorption pits or pits are formed. At the end of the phase of the cycle, the osteoclasts form a new protein matrix, which then calcifies. In some disease states, such as osteoporosis and Paget's disease, the normal balance between bone resorption and formation is disrupted, and there is a net loss of bone in each cycle. Ultimately, this can lead to bone weakness and create a large fracture risk with minimal trauma.

[0006] Several published studies indicate that inhibitors of cysteine proteases are effective in inhibiting osteoclast-mediated bone resorption and have an essential role for cysteine proteases in bone resorption It revealed that. For example, Delais
se et, Biochem. J., 1980, 192 , 365 discloses a series of protease inhibitors in a mouse bone organ culture system, inhibitors of cysteine proteases (e.g., leupeptin, Z-Phe-Ala-CHN 2) is Prevents bone resorption while indicating that the serine protease inhibitor was not effective. Delaisse et al., Biochem., Biophy.
s. Res. Commun., 1984, 125, 441 reported that E-64 and leupeptin also prevented bone resorption in vivo when measuring acute changes in serum calcium in rats on a calcium deficient diet. To be effective. Lern
er et al., J. Bone Min. Res., 1992, 7, 433 disclose that cystatin, an endogenous cysteine protease inhibitor, inhibits PTH-stimulated bone resorption in mouse calvaria. Delaisse et al., Bone, 1987, 8, 305, Hill et al., J. Cell. B
iochem., 1994, 56, 118 and Everts et al., J. Cell.Physiol., 1992, 150, 221.
Other studies have also reported a correlation between inhibition of cysteine protease activity and bone resorption. Tezuka et al., J. Biol. Chem., 1994, 269, 1106, Inaoka.
Biochems Biophys. Res. Commun., 1995, 206, 89 and Shi et al., FEBS Lett.
, 1995, 357, 129 disclose that under normal conditions, cathepsin K, a cysteine protease, is abundantly expressed in osteoclasts and may be the major cysteine protease present in these cells.

[0007] Abundant and selective expression of cathepsin K in osteoclasts strongly suggests that the enzyme is essential for bone resorption. Thus, selective inhibition of cathepsin K includes, but is not limited to, gingival diseases such as osteoporosis, gingivitis and periodontitis, Paget disease, malignant hypercalcemia and metabolic bone disease It may provide an effective treatment for diseases of excessive bone loss. Cathepsin K levels have also been shown to be elevated in cartilage resorbing cells of the osteoarthritis synovium. Thus, selective inhibition of cathepsin K may also be useful in treating diseases of excessive cartilage or matrix destruction, including but not limited to osteoarthritis and rheumatoid arthritis. Metastatic tumor cells also typically express high levels of proteolytic enzymes that disrupt the surrounding matrix. Thus, selective inhibition of cathepsin K may also be useful in treating certain neoplastic diseases.

[0008] Several cysteine protease inhibitors are known. Palmer (1995) J.
Med. Chem. 38, 3193 describes cathepsins B, L, S, O2 and cruzain.
Certain vinyl sulfones that irreversibly inhibit cysteine proteases such as in) are disclosed. Other classes of compounds such as aldehydes, nitriles, a-ketocarbonyl compounds, halomethylketones, diazomethylketones, (acyloxy) methylketones, ketomethylsulfonium salts and epoxysuccinyl compounds have also been reported to inhibit cysteine proteases. . See Palmer (ibid.) And references therein.

US Pat. No. 4,518,528 discloses peptidyl fluoromethyl ketone as an irreversible inhibitor of cysteine protease. International Patent Application Publication WO9
4/04172 and European patent applications EP0 525 420 A1, EP0 603 873 A1 and EP0 611 756 A2 are cysteine proteases.
Alkoxymethyl and mercaptomethyl ketones that inhibit cathepsins B, H and L are described. International patent application PCT / US94 / 08868 and European patent application EP 0 623 592 A1 describe cysteine proteases, alkoxymethyl and mercaptomethyl ketones, which inhibit IL-1b convertase. Alkoxymethyl and mercaptomethylketone have also been described as inhibitors of serine proteases, kininogenases (International Patent Application PCT / GB
91/01479).

Azapeptides designed to deliver azaamino acids to the active site of serine proteases and which retain good leaving groups have been described by Elmore et al., Biochem. J., 1968, 107, 103.
Garker et al., Biochem. J., 1974, 139, 555; Gray et al., Tetrahedron, 1977, 33,
837, Gupton et al., J. Biol. Chem., 1984, 259, 4279, Powers et al., J. Biol. Chem.
, 1984, 259, 4288 and are known to inhibit serine proteases. See also Magrath et al., J. Med. Chem., 1992, 35, 4279, Bagg.
io et al., Biochemistry, 1996, 35, 3551 and Xing et al., J. Med. Chem. 1998, 41,
1344 discloses certain azapeptide esters as cysteine protease inhibitors.

Diacylcarbohydrazides have recently been described by Thompson et al., Proc. Natl. Acad. Sci.,
USA, 1997, 94, 14249 and International Patent Application WO 97/16433,
It has been disclosed as an inhibitor of cathepsin K.

[0012] Antipain and leupeptin have been described as reversible inhibitors of cysteine proteases in McConnell et al., J. Med. Chem., 33, 86;
ezawa et al., 45 Meth. Enzymol. 678, disclosed as inhibitors of serine proteases. E64 and its synthetic analogs are also well-known cysteine protease inhibitors (Barrett, Biochem. J., 201, 189 and Grinde, Biochem.
Biophys. Acta., 701, 328).

Accordingly, structurally diverse cysteine protease inhibitors have been identified. However, these known inhibitors have various disadvantages and are not considered suitable for use as therapeutics in animals, especially humans. These disadvantages include lack of selectivity, cytotoxicity, poor solubility and excessively rapid plasma clearance. Accordingly, there is a need for a method of treating diseases caused by cysteine proteases including pathological levels of cathepsin, especially cathepsin K, and novel inhibitory compounds useful in such methods. The inventors have now found a new class of diacyl carbohydrazide compounds that are protease inhibitors, most particularly inhibitors of cathepsin K.

SUMMARY OF THE INVENTION It is an object of the present invention to provide inhibitors of diacylcarbohydrazide proteases, especially such inhibitors of cysteine and serine proteases, more particularly such compounds that inhibit cysteine proteases, and even more particularly of the papain superfamily. Compounds that inhibit cysteine proteases, even more particularly such compounds that inhibit the cysteine proteases of the cathepsin family, most particularly those that inhibit cathepsin K, which are therapeutically modified by altering the activity of such proteases It is an object of the present invention to provide a compound which is useful for treating a possible disease.

Thus, in a first aspect, the present invention provides a compound of formula I: In another aspect, the invention provides a pharmaceutical composition comprising a compound of Formula I and a pharmaceutically acceptable carrier, diluent or excipient. In yet another aspect, the invention provides intermediates useful in preparing compounds of formula I.

In yet another aspect, the invention relates to proteases, particularly cysteine and serine proteases, more particularly cysteine proteases, even more particularly cysteine proteases of the papain superfamily, and even more particularly of the cathepsin family. By inhibiting cysteine proteases, most particularly cathepsin K, there is provided a method of treating diseases whose pathology can be therapeutically modified.

In a particular embodiment, the compounds of the invention are used in particular for bone loss such as osteoporosis and gingival diseases such as gingivitis and periodontitis, or excess cartilage or matrix such as osteoarthritis and rheumatoid arthritis. It is useful for treating diseases characterized by destruction.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a compound of formula I:

Embedded image

[Wherein, R¹And R⁶Is independently C_5-6Alkyl; C_2-6Alkenyl; C_3-11Cycloa
Lequil-C_0-6Alkyl; Ar-C_0-6Alkyl; Het-C_0-6Alkyl; Ar
-C_2-6Alkenyl; Het-C_2-6Alkenyl; Het-C_2-6Alkynyl; A
r-C_2-6Alkynyl; CH (R⁷) Ar; CH (R⁷) OAr; NR⁷R⁸and
CH (R⁷) NR⁸R⁹Selected from the group consisting of: R ', R^Two, R^Three, R^Four, R^Five, R⁸, R^Ten, R¹¹And R¹⁴Is independently H; C_{1 -6} Alkyl; C_2-6Alkenyl; Ar-C_0-6Alkyl; Het-C_0-6Alkyl
And C_3-11Cycloalkyl-C_0-6R is selected from the group consisting of alkyl;⁷And R¹³Is independently H; C_1-6Alkyl; C_2-6Alkenyl; C_2-6Al
Quinyl; C_3-11Cycloalkyl-C_0-6Alkyl; Ar-C_0-6Alkyl; Ar-
C_2-6Alkenyl; Ar-C_2-6Alkynyl; Het-C_0-6Alkyl; Het-
C_2-6Alkenyl; Het-C_2-6Alkynyl; OR^Ten, SR^TenAnd NR^TenR¹¹ C optionally substituted by_1-6Alkyl; N (R ') CO_TwoR '; CO_TwoR
’; CONR^TenR¹¹And N (C = NH) NH_TwoSelected from the group consisting of: R⁷And R⁸Together form a pyrrolidine or piperidine ring,
Well; R^TenAnd R¹¹Together form a pyrrolidine, piperidine or morpholine ring
May be formed; R⁹Is H; R¹²R¹²C (O); R¹²C (S); R¹²OC (O); R¹²OC (
O) NR¹¹CH (R¹³) C (O); R¹²SO_TwoR¹²SO_TwoNR¹¹CH (R¹³) C
(O); R¹²R'NC (O); R¹²R'NCS or COCH (R¹³) NR¹⁴R ^Fifteen R¹²Is NR^TenR¹¹, C_2-6Alkenyl or C_2-6Substituted by alkynyl
C that may be_1-6Alkyl; Ar-C_0-6Alkyl; Ar-C_2-6Alkenyl
Ar-C;_2-6Alkynyl; Het-C_0-6Alkyl; Het-C_2-6Alkenyl
Het-C_2-6Alkynyl; C_1-6Alkyl, (CH_Two)_1-6CO_TwoR 'or a
C optionally substituted with damantyl_3-11R is cycloalkyl;^FifteenIs R¹²R¹²C (O); R¹²C (S); R¹²OC (O); R¹²OC (O)
NR¹¹CH (R¹³) C (O); R¹²SO_TwoR¹²SO_TwoNR¹¹CH (R¹³) C (O
); R¹²R'NC (O) or R¹²R'NCS] and pharmaceutically acceptable salts, hydrates and solvates thereof.
I do.

Compounds of formula I wherein R ² , R ³ , R ⁴ or R ⁵ are H are preferred. Also, in the formula: R ¹ is

[0021]

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[0022]

Embedded image

[0023]

Embedded image

[0024]

Embedded image Is;

R ⁶ is

Embedded image Are preferred.

2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-valinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl) )]-2 '-[N'-(N-benzyloxycarbonyl-L-phenylalanyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 '-[N'-(N -Benzyloxycarbonyl-L-isoleucinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 '-[N'-(N-benzyloxycarbonyl-L-methionyl)] carbohydrazide;

2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′-[
N '-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide; (2S) -2- [N- (N-benzyloxycarbonyl-2-aminobutyryl)]-2'-[N '-(N- Benzyloxycarbonyl-L-leucinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-lydinyl-L-leucinyl)] carbo Hydrazide; (2R) -2- [N- (3-benzyloxybenzoyl)]-2 ′-[N′-
[4-methyl-2- (3-phenylphenyl) pentanoyl] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-arginyl- L-leucinyl)] carbohydrazide;

2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl) )]-2 '-[N'-(N-benzyloxycarbonyl-L-cyclohexylglycinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 '-[N'-(N -Benzyloxycarbonyl-L-cyclohexylalanyl)] carbohydrazide; 2,2 '-[N, N'-bis [N- (3-nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L-leucinyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 '-[
N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide;

2- [NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (3,4-dimethoxybenzoyl) ) -L-Leucinyl]]-
2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 '-[N'-
[N- (4-trifluoromethylbenzoyl) -L-leucinyl]] carbohydrazide; 2- [N- [N- (3,4-dichlorobenzoyl) -L-leucinyl]]-2
'-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzofuran-2-ylcarbonyl-L-leucinyl)]-
2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide;

2- [N- [N- (5,6-dimethoxybenzofuran-2-ylcarbonyl)
-L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-methyl-2-phenyloxazol-4-yl) Carbonyl) -L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1 '-N
'-Methyl-2'-[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- [4-methyl-2- (4-trifluoromethylphenyl) thiazole-5] -Ylcarbonyl] -L-leucinyl]]-2 '-[N'-[3-
(2-pyridinyl) phenylacetyl]] carbohydrazide;

2- [N- [N- (3-isoquinolinoyl) -L-leucinyl] -2 ′-[N
'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-chlorobenzofuran-2-ylcarbonyl) -L-leucinyl]]-2'-[N'- [3- (2-pyridinyl) phenylacetyl]]
Carbohydrazide; 2- [N- [N- (3,5-difluorobenzoyl) -L-leucinyl]]-
2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzothiazol-6-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1- (N
-Methyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide;

2- [N- (N-picolinoyl-L-leucinyl)]-2 ′-[N ′-[3-
(2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (2,6-dimethoxynicotinoyl) -L-leucinyl]]
-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (4-methanesulfonylbenzoyl) -L-leucinyl]]
-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 '-[N'-
[N- (3,4,5-trimethoxybenzoyl) -L-leucinyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 ′-[N′-
[N- (8-quinolinoyl) -L-leucinyl]] carbohydrazide;

2- [N- [N- [3,4- (1,3-propylenedioxy) benzoyl]-
L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 '-[ N'-
(N-thieno [2,3-b] thiophen-2-ylcarbonyl-L-leucinyl)] carbohydrazide; 2- [N- [N- (4-methoxybenzoyl) -L-leucinyl]]-2′-
[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- [4-methoxy-3- [2- (4-morpholino) ethoxy]]
Benzoyl] -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl)
Phenylacetyl]] carbohydrazide; 2- [N- [N- [2- (4-morpholino) pyrimidin-4-ylcarbonyl] -L-leucinyl]]-2 ′-[N ′-[3- (2- Pyridinyl) phenylacetyl]] carbohydrazide;

2- [N- [N- (2,3-dihydrobenzofuran-5-ylcarbonyl)-
L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-carboxybenzofuran-2-ylcarbonyl) -L
-Leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (3-benzyloxybenzoyl) -2 '-[N'-[N- (3-
Nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L
-Leucinyl]] carbohydrazide;

2- [N- [N- (3-nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L-leucinyl]]-2- [N- [3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-oxo-2-pyridinyl) phenylacetyl]
A carbohydrazide; and a compound of formula I selected from the group consisting of 2,2 ′-[N, N′-bis [N- (3-iodo-4-methoxy) -L-leucinyl]] carbohydrazide, It is a more preferred embodiment of the present invention.

Definitions The present invention includes all hydrates, solvates, complexes and prodrugs of the compounds of the present invention. Prodrugs are any covalently bonded compounds that release the active parent drug according to Formula I in vivo. When another form of a chiral or isomer center is present in the compounds of the invention, all forms of such isomers or isomers, including enantiomers and diastereomers, may be encompassed herein. Is intended. Invention compounds containing chiral centers can be used as racemic mixtures, enantiomeric enriched mixtures, or racemic mixtures can be separated using well-known techniques, and the individual enantiomers can be used alone. Where a compound has an unsaturated carbon-carbon bond, both the cis (Z) and trans (E) isomers are within the scope of the invention. Where the compounds exist in tautomeric forms, such as keto-enol tautomers, each tautomer is encompassed within the present invention and the equilibrium or one form predominates.

In Formula I or any of its supplements, the meaning of any substituent at any one event is, unless otherwise specified, its meaning at any other event or any other substituent. Independent of the meaning. Abbreviations and symbols are commonly used in peptides, and chemical formulas are used herein to describe compounds of the invention. Generally, abbreviations for amino acids are
Eur.Biochem., 158, 9 (1984).
Follow ical Nomenclature.

As used herein, the term “amino acid” refers to alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, Refers to the D or L isomer of serine, threonine, tryptophan, tyrosine and valine.

As applied herein, “C_1-6The term "alkyl" is substituted or unsubstituted
Methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and
t-butyl, pentyl, n-pentyl, isopentyl, neopentyl and hexyl
It is meant to include syl as well as its simple aliphatic isomers. Any C _1-6 Alkyl groups have 1 to 5 halogens, SR¹⁶, OR¹⁶, N (R¹⁶)_Two, C
(O) N (R¹⁶)_Two, Carbamyl or C_1-4Alkyl (where R¹⁶Is C_1-6A
Which is independently alkyl. C₀The alkyl is
It means that there is no alkyl group in the moiety. Therefore, Ar-C₀Archi
Is equal to Ar.

As applied herein, the term “C _3-11 cycloalkyl” refers to substituted and unsubstituted cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, cyclodecane, cycloundecane Is meant to be included. As applied herein, the term “C _2-6 alkenyl” refers to an alkyl group of 2 to 6 carbons in which a carbon-carbon single bond is replaced by a carbon-carbon double bond. . _C2-6 alkenyl includes ethylene, 1-propene, 2-propene, 1-butene, 2-butene, isobutene and some isomers pentene and hexene. Both cis and trans isomers are included.

The term “C _2-6 alkynyl” refers to a group consisting of 2 to 6 carbons in which a carbon-carbon single bond is replaced by a carbon-carbon triple bond. _C2-6 alkynyl includes acetylene, 1-propyne, 2-propyne, 1-butyne, 2-butyne, 3-butyne and the simple isomers of pentyne and hexyne. The term "halogen" means F, Cl, Br and I.

Ar is phenyl; one or more Ph-C_0-6Alkyl, Het-C_0-6Alkyl, C _1-6 Alkyl, C_1-6Alkoxy, Ph-C_0-6Alkoxy, Het-C_0-6Arco
Xy, OH, (CH_Two)_1-6NR^TenR¹¹, O (CH_Two)_1-6NR^TenR¹¹, (CH_Two)_{0 -6} CO_TwoR^', O (CH_Two)_1-6CO_TwoR^', SR^Ten, SO_TwoR^Ten, CF_Three, Nitro
Or naphthyl optionally substituted by halogen; Ph is one or more
C_1-6Alkyl, C_1-6Alkoxy, OH, (CH_Two)_1-6NR^TenR¹¹, O (CH_Two
)_1-6NR^TenR¹¹, (CH_Two)_0-6CO_TwoR^', O (CH_Two)_1-6CO_TwoR^', SR^Ten
, SO_TwoR^Ten, CF_Three, Cyano or halogen; two C _1-6 Alkyl or C_1-6An alkoxy group binds to a saturated or unsaturated Ar ring
To form a 5- to 7-membered ring.

As used herein, the term “Het” or “heterocyclic” is either saturated or unsaturated and is a carbon atom and a 1 selected from the group consisting of N, O and S. Or three heteroatoms, wherein nitrogen and sulfur heteroatoms may be oxidized and nitrogen heteroatoms may be quaternized.
A 7 to 7 membered monocyclic, stable 7 to 10 membered bicyclic, or stable 11 to 18 membered tricyclic heterocycle, wherein any of the above heterocycles is fused to a benzene ring. And any bicyclic group having the formula: Heterocycles may be attached at any heteroatom or carbon atom and may be substituted with Ar (including on nitrogen) to yield certain stable structures. Examples of such heterocycles are piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl,
-Piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl,
Pyridyl, pyrazinyl, oxazolinyl, oxazolyl, isoxazolyl, morpholinyl, thiazolidinyl, thiazolinyl, thiazolyl, quinuclidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl,
Includes benzoxazolyl, furyl, pyranyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzoxazolyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone and oxadiazolyl.

The term “5-7 membered ring fused on a saturated or unsaturated Ar ring” refers to fused bicyclic ring systems such as indane, 1,2,3,4-tetrahydrodecalin, methylenedioxy It means phenyl, 1,2-ethylenedioxyphenyl and 1,3-propylenedioxyphenyl.

Here and throughout the application, the term “C ₀ ” indicates that the immediately following substituent is absent; for example, in the partial ArC _0-6 alkyl, when C is 0, the substituent is Ar , For example, phenyl. Conversely, if the partial ArC _0-6 alkyl is identified as a particular aromatic group, eg, phenyl, C is understood to be 0.

Certain groups are abbreviated herein. t-Bu represents a tertiary butyl group;
oc represents a t-butyloxycarbonyl group, Fmoc represents a fluorenylmethoxycarbonyl group, Ph represents a phenyl group, and Cbz represents a benzyloxycarbonyl group.

Certain reagents are abbreviated herein. DCC stands for dicyclohexylcarbodiimide, DMAP stands for 2,6-dimethylaminopyridine, and EDC stands for N-
Ethyl-N '(dimethylaminopropyl) -carbodiimide is shown. HOBT indicates 1-hydroxybenzotriazole, DMF indicates dimethylformamide, and BOP indicates benzotriazol-1-yloxy-tris (dimethylamino).
Denotes phosphonium hexafluorophosphate, DMAP is dimethylaminopyridine, NMM is N-methylmorpholine, TFA indicates trifluoroacetic acid, and THF indicates tetrahydrofuran. Jones reagent is a solution of chromium trioxide, water and sulfuric acid well known in the art.

Preparation Methods The compounds of the present invention can be conveniently prepared by the methods shown in Schemes 1-3 below. Compounds of formula I wherein R ¹ ≠ R ⁶ and R ² , R ³ , R ⁴ and R ⁵ are H are prepared by methods analogous to those described in Scheme 1.

[0049]

Embedded image

[0050]1-Scheme 1In a protic solvent (eg, methanol or ethanol)
Treated with hydrazine hydrate in2-Scheme 1And put it in toluene
Treated with phosgene3-Scheme 1I got The substance is converted to a protic solvent (eg,
Treated with hydrazine hydrate in methanol or ethanol)4-Scheme 1
I got4-Scheme 1In an aprotic solvent (eg, DMF)
Bonic acid (for example, N-benzyloxycarbonyl-L-valine, N-benzylo
Xycarbonyl-L-phenylalanine, N-benzyloxycarbonyl-L-
Isoleucine, N-benzyloxycarbonyl-L-methionine, N-benzyl
Oxycarbonyl-L-norvaline, (S) -N-benzyloxycarbonyl-
2-aminobutyric acid, N-tert-butoxycarbonyl-L-leucine, (R)-
4-methyl-2- (3-phenylphenyl) pentanoic acid, N-benzyloxyca
Rubonyl-L-cyclohexylglycine or N-benzyloxycarbonyl-
L-cyclohexylalanine) and a peptide coupling reagent (eg, ED
C · HCl / 1-HOBT)5-Scheme 1I got R is tert
-Butyl, by treatment with trifluoroacetic acid in dichloromethane.
What5-Scheme-1To6-Scheme 1Was converted to Alternatively, R is benzyl
In some cases, by treatment with hydrobromic acid in acetic acid,5-Scheme 1To6-
Scheme 1Was converted to6-Scheme 1With an aprotic solvent (eg, DMF)
In a carboxylic acid (eg, N_α-Benzyloxycarbonyl-N_ε−te
rt-butoxycarbonyl-L-lysine, N_α-Benzyloxycarbonyl-N _{φ Two} -Bis (tert-butoxycarbonyl) -L-arginine, 3,4-dim
Toxic benzoic acid, 4-trifluoromethyl benzoic acid, 3,4-dichlorobenzoic acid
, Benzofuran-2-carboxylic acid, 5,6-dimethoxybenzofuran-2-cal
Bonic acid, 5-methyl-2-phenyloxazole-4-carboxylic acid, benzothio
Phen-2-carboxylic acid, 4-methyl-2- (4-trifluoromethylphenyl
) Thiazole-5-carboxylic acid, isoquinoline-3-carboxylic acid, 5-chlorobe
Benzofuran-2-carboxylic acid benzothiazole-6,5-difluorobenzoate
-Carboxylic acid, picolinic acid, 2,6-dimethoxynicotinic acid, 4-methanesulfo
Nylbenzoic acid, 3,4,5-trimethoxybenzoic acid, quinoline-8-carboxylic acid
, 3,4- (1,3-propylenedioxy) benzoic acid, thieno [2,3-b] thi
Offen-2-carboxylic acid, 4-methoxybenzoic acid, 3- [2- (4-morpholy
No) ethoxy] -4-methoxybenzoic acid, 2- (4-morpholino) pyrimidine-
4-carboxylic acid, 2,3-dihydrobenzofuran-5-carboxylic acid, 5-carbo
Xybenzofuran-2-carboxylic acid) and peptide coupling reagents (eg,
, EDC · HCl / 1-HOBT)7-Scheme-1I got Alternative
Then6-Scheme 1In dichloromethane with an acyl chloride (e.g.
Toro-1,2,7-benzoxadiazol-6-yl-L-prolinyl chloride
) And N-methylmorpholine7-Scheme 1Was prepared. R¹²C
O_TwoH is N_α-Benzyloxycarbonyl-N_ε-Tert-butoxycarboni
Le-L-lysine or N_α-Benzyloxycarbonyl-N_{φ Two}-Screw (ter
(t-butoxycarbonyl) -L-arginine in dichloromethane
The tert-butoxycarbonyl group is removed by treatment with trifluoroacetic acid in
I left.

[0051]

Embedded image

R¹≠ R⁶And R^ThreeOr R^FourCompounds of formula I where ≠ H are described in Scheme 2
It is prepared by the same method as described above.1-Scheme 2In dichloromethane
Treated with benzyl chloroformate,2-Scheme 2I got Alternatively,1-
Scheme 2With di-tert-butyl dicarboxylate in dioxane
hand,2-Scheme 2I got2-Scheme 2With an aprotic solvent (eg, DM
F) In carboxylic acid (for example, N-benzyloxycarbonyl-L-leu)
Syn or 3- (2-pyridinyl) phenylacetic acid) and peptide coupling
Treating with a reagent (eg, EDC.HCl / 1-HOBT)3-Scheme 2To
Obtained. When R is benzyl, by treatment with hydrobromic acid in acetic acid
,3-Scheme 2To4-Scheme 2Was converted to Alternatively, R is tert-butyl
When treated with trifluoroacetic acid in dichloromethane, 3-Scheme 2 To4-Scheme 2Was converted to4-Scheme 2The dichloromethane
In phosgene and N-methylmorpholine in
Acylhydrazine (eg, N-benzyloxycarbonyl-L
-Leucinyl hydrazide or 3- (2-pyridinyl) phenylacetylhydra
Treatment with N-methylmorpholine5-Scheme 2I got

[0053]

Embedded image

Compounds of formula I where R ¹ ＲR ⁶ ＲR ¹² CONHCH (R ⁷ ) CO ₂ H are prepared by methods analogous to those described in Scheme 3. 1-Scheme 3 is tert-butoxycarbonyl protected amino acid (eg, tert-butoxycarbonyl-L-leucine) and peptide coupling reagent (eg, EDC.HCl / 1-HOBT) in an aprotic solvent (eg, DMF) 2 ) Skiing
To give 3 -methine which is treated with trifluoroacetic acid in dichloromethane to give 3-s
You have a chime 3 . 3- Scheme 3 is treated with a carboxylic acid (eg, 3-iodo-4-methoxybenzoic acid) and a peptide coupling reagent (eg, EDC · HCl / 1-HOBT) in an aprotic solvent (eg, DMF). And 4-
Scheme 3 was obtained. Alternatively, 3-Scheme 3 can be prepared using an acyl chloride (e.g., 3-nitro-1,2,7-benzoxadiazol-6-yl-) in dichloromethane.
4-Scheme 3 was prepared by treatment with L-prolinyl chloride) and N-methylmorpholine.

Referring to the methods of preparing compounds of Formula I shown in Schemes 1-3 above, one skilled in the art will appreciate that the present invention encompasses all novel intermediates required for the preparation of compounds of Formula I. You will understand. More specifically, the present invention provides compounds:

2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-te
rt-butoxycarbonyl-L-leucinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(L-leucinyl)] carbohydrazide; 2- [N- (3 - benzyloxy benzoyl)] - 2 '- [N ' - (N α - benzyloxycarbonyl -N _{epsilon-tert-butoxycarbonyl} -L- lysinyl -L- leucinyl)] carbohydrazide; 2- [N- (3- benzyloxy benzoyl)] - 2 '- [N ' - (N α - benzyloxycarbonyl -N _{phi 2} - bis (tert- butoxycarbonyl) -L
-Lidinyl-L-leucinyl)] carbohydrazide; 2,2 '-[N, N'-bis- (N-tert-butoxycarbonyl-L-leucinyl)] carbohydrazide;

2,2 ′-[N, N′-bis- (L-leucinyl)] carbohydrazide; 2-hydroxy-4,5-dimethoxybenzaldehyde; 4,5-dimethoxy-2-ethoxycarbonylmethoxybenzaldehyde; 5,6-dimethoxybenzofuran-2-carboxylic acid ethyl; 5,6-dimethoxybenzofuran-2-carboxylic acid;

1- (N-benzyloxycarbonyl) -1- (N-methyl) -2- [N- [
3- (2-pyridyl) phenylacetyl]] hydrazine; 1- (N-methyl) -2- [N- [3- (2-pyridyl) phenylacetyl]
Hydrazine; 1- (N-tert-butoxycarbonyl) -1 (-N-methyl) -2- (N
-(Benzyloxycarbonyl-L-leucinyl) hydrazine; 2- (N-benzyloxycarbonyl-L-leucinyl) -1- (N-methyl) hydrazine; 3- (2-pyridyl) phenylacetylhydrazine;

Ethyl 2- (4-morpholino) pyrimidine-4-carboxylate; 2- (4-morpholino) pyrimidine-4-carboxylic acid; 4-benzyloxycarbonylmethoxy-3-formylbenzaldehyde; 5-formylbenzofuran-2 -Benzyl carboxylate; benzyl 5-carboxybenzofuran-2-carboxylate;

Benzyl 5-tert-butoxycarbonylbenzofuran-2-carboxylate; 5-tert-butoxycarbonylbenzofuran-2-carboxylic acid; 2- [N- [N- (5-tert-butoxycarbonylbenzofuran-2-yl) Carbonyl) -L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 '-[
N '-[3- (2-oxo-2-pyridinyl) phenylacetyl]] carbohydrazide; and 2- (NL-leucinyl) -2'-[N '-[3- (2-oxo-2- Pyridinyl) phenylacetyl]] carbohydrazide.

The starting materials used herein are commercially available amino acids, or are prepared by conventional methods well known to those of ordinary skill in the art and are available from COMPENDIUM OF
It can be found in standard reference books such as ORGANIC SYNTHETIC METHODS, Vol. I-VI (published by Wiley-Interscience).

The coupling methods for forming amide bonds herein are generally well-known in the art. Bodansky et al., THE PRACTICE OF PEPTIDE SYNTHESIS, Springe
r-Verlag, Berlin, 1984; E. Gross and J. Meienhofer, THE PEPTIDES, Vol.
1, 1-284 (1979); and JM Stewart and JD Young, SOLID PHASE PEPTI
The method of peptide synthesis set forth by DE SYNTHESIS, 2d Ed., Pierce Chemical Co., Rockford, Ill., 1984, is generally exemplary of the art and is hereby incorporated by reference.

Synthetic methods for preparing compounds of the present invention often employ protective groups to mask reactive functionality or to minimize unnecessary side reactions. Such protecting groups are generally described in Green, TW, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, John Wiley & So
ns, New York (1981). The term "amino protecting group" is generally
Refers to Boc, acetyl, benzoyl, Fmoc and Cbz groups and derivatives thereof as known in the art. Methods of protection and deprotection, and replacement of an amino protecting group with another group are well known.

The acid addition salt of a compound of Formula I is prepared in a suitable solvent with the parent compound and an excess of an acid such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, acetic acid, trifluoroacetic acid. , Maleic acid, succinic acid or methanesulfonic acid in a standard manner. Certain compounds form acceptable internal salts or zwitterions. A cation salt is an excess of an alkaline reagent containing the appropriate cation containing the parent compound,
For example, by treatment with a hydroxide, carbonate or alkoxide; or by treatment with a suitable organic amine. Cations such as Li ⁺ , Na ⁺ , K ⁺ , Ca ⁺⁺ , Mg ⁺⁺ and NH ₄ ⁺ are particular examples of cations present in pharmaceutically acceptable salts. The halides, sulfates, phosphates, alkanates (eg, acetate and trifluoroacetate), benzoates, and sulfonates (eg, mesylate) are anions present in pharmaceutically acceptable salts This is an example.

The present invention also provides a pharmaceutical composition comprising a compound of Formula I and a pharmaceutically acceptable carrier, diluent or excipient. Thus, the compounds of formula I can be used for the manufacture of a medicament. Pharmaceutical compositions of a compound of Formula I, prepared as described herein above, may be formulated as solutions for parenteral administration or as lyophilized powders. The powder is
Prior to use, they may be reconstituted by the addition of a suitable diluent or other pharmaceutically acceptable carrier. The liquid formulation may be a buffered isotonic aqueous solution. Examples of suitable diluents are normal isotonic saline solutions, standard 5% dextrose water or buffered sodium or ammonium acetate solutions. Such formulations are particularly suitable for parenteral administration, but may also be used for oral administration, or may be included in an inhaler or nebulizer with a dosimeter for inhalation. It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxycellulose, gum arabic, polyethylene glycol, mannitol, sodium chloride or sodium citrate.

Alternatively, these compounds may be encapsulated, tableted, or prepared into an emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to extend or stabilize the composition, or to facilitate preparation of the composition. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, magnesium or stearic acid, talc, pectin, acacia, agar or gelatin. The liquid carrier is
Includes syrup, peanut oil, olive oil, saline and water. The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate alone or with a wax. The amount of solid carrier varies but, preferably, will be from about 20 mg to about 1 g per dosage unit. Pharmaceutical preparations are prepared according to the usual techniques of pharmacy, including crushing, mixing, granulating and compressing as needed in tablet form; or crushing, mixing and filling in hard gelatin capsule form. If a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or aqueous or non-aqueous suspension. Such liquid formulations may be administered orally directly or may be filled into soft gelatin capsules.

For rectal administration, the compounds of the present invention may be mixed with excipients such as cocoa butter, glycerin, gelatin or polyethylene glycol and formed into suppositories.

Utility of the Invention The compounds of formula I are useful as protease inhibitors, specifically as inhibitors of cysteine and serine proteases, more particularly as inhibitors of cysteine proteases, and even more particularly of the papain superfamily. And more particularly as inhibitors of the cysteine proteases of the cathepsin family, most particularly as inhibitors of cathepsin K. The present invention also provides useful compositions and formulations of the compounds, including pharmaceutical compositions and formulations of the compounds.

The compounds of the present invention may be used in the form of Pneumocystis carini, Trypanosoma cruzi,
Infection with Trypanosoma brucei and Chrysizia fusicata; and cysteine protease-related diseases including schistosomiasis, malaria, tumor metastasis, metachromatic leukodystrophy, muscular dystrophy, muscular atrophy; and, in particular, cathepsin K Diseases, most particularly gingival diseases including osteoporosis, gingivitis and periodontitis, arthritis, more particularly osteoarthritis and rheumatoid arthritis, Paget disease; malignant hypercalcemia, and metabolic Useful for treating disorders of excessive bone or cartilage loss, including bone disorders.

Also, metastatic tumor cells typically express high levels of proteolytic enzymes that destroy the surrounding matrix, and certain tumors and metastatic neoplasia
It can be effectively treated with the compounds of the present invention.

The invention also relates to proteases at the pathological level, in particular cysteine and serine proteases, more particularly cysteine proteases, even more particularly the cysteine proteases of the papain superfamily, even more particularly the cathepsin family. A method of treating a disease caused by a cysteine protease of the present invention, said method comprising administering a compound of the invention to an animal in need of treatment, particularly a mammal, most particularly a human. I do. The present invention relates to a method for treating diseases caused by cathepsin K, particularly at a pathological level,
The method of treatment provides a method comprising administering to an animal in need of treatment, particularly a mammal, most particularly a human, an inhibitor of cathepsin K, including a compound of the present invention. The present invention specifically relates to infections by Pneumocystis carini, Trypsanoma cruz, Trypsanoma brucei, and Criticia fusicata; and schistosomiasis, malaria, tumor metastasis, metachromatic leukodystrophy, muscular dystrophy, muscular atrophy. Diseases involving cysteine proteases, and particularly those involving cathepsin K, most particularly gingival diseases including osteoporosis, gingivitis and periodontitis, arthritis, more particularly osteoarthritis and rheumatoid arthritis, Methods of treating excessive bone or cartilage loss diseases, including Paget's disease, malignant hypercalcemia, and metabolic bone disease.

The present invention is further directed to a method of treating osteoporosis or inhibiting bone loss, comprising administering an effective amount of a compound of Formula I alone or with another inhibitor of bone resorption, such as a bisphosphonate (ie, alendronate). ), Hormone replacement therapy, in vivo administration to a patient in combination with an antiestrogen or calcitonin. Further, the compounds of the present invention and anabolic substances, such as bone morphogenic proteins,
Treatment with iproflavone may be used to prevent bone loss or increase bone mass.

For acute treatment, parenteral administration of the compounds of formula I is preferred. Intravenous infusion of a similar formulation with 5% dextrose water or a compound in a normal saline or similar excipient is most effective, but intramuscular mass injection is also effective. Typically,
Parenteral administration can be carried out in a manner that maintains the drug concentration in plasma at a concentration sufficient to inhibit cathepsin K, from about 0.01 to about 100 mg / kg; preferably 0.1 to 2 mg / kg.
0 mg / kg. Compounds are administered 1 to 4 times daily, for a total daily dose of about 0.4
It is administered at a level to reach about 400 mg / kg / day. The precise amount of a compound of the present invention that is therapeutically effective and the route by which the compound is best administered will be determined by comparing the blood level of the drug to the concentration required to have a therapeutic effect. Determined easily by the vendor.

The compounds of the invention may also have sufficient drug concentration to inhibit bone resorption or to achieve any other therapeutic indication as disclosed herein. In such a method, it may be administered orally to the patient. Typically, a pharmaceutical composition containing the compound will provide from about 0.1 to about 50 m in a manner consistent with the condition of the patient.
It is administered at an oral dose of g / kg. Preferably, the oral dose will be from about 0.5 to about 20 mg / kg.

When a compound of the present invention is administered in accordance with the present invention, no unacceptable toxicological effects are expected.

Biological Assays The compounds of the invention may be tested in one of several biological assays to determine the concentration of the compound required to have a given pharmacological effect.

Measurement of Cathepsin K Proteolytic Catalytic Activity All assays for cathepsin K were performed using human recombinant enzymes. Standard assay conditions for the determination of rate constants are fluorogenic peptide substrates, typically Cbz-Ph
Using e-Arg-AMC, measurements were taken in 100 mM Na acetate at pH 5.5 containing 20 mM cysteine and 5 mM EDTA. The stock substrate solution is either 10 or 20 mM in DMSO with a final substrate concentration of 20 μM in the assay.
It was prepared at a concentration. All assays contained 10% DMSO. Independent experiments revealed that the levels of DMSO had no effect on enzyme activity or rate constants. All assays were performed at ambient temperature. Product fluorescence (360n
M excitation light; 460 nM emission) was monitored on a Perceptive Biosystems Cytofluor II fluorescence plate reader. The product transition curve occurred over 20-30 minutes after formation of the AMC product.

Inhibition Studies Potential inhibitors were evaluated using the transition curve method. Assays were performed in the presence of various concentrations of test compound. The reaction was started by adding the enzyme to a buffer solution of inhibitor and substrate. Data analysis was performed according to one of two procedures that depended on the appearance of the transition curve in the presence of the inhibitor. For compounds whose transition curve is linear, the apparent inhibition constant (K _i , app) was calculated according to Equation 1 (Brandt et al., Biochemistry, 1989, 28, 140).

V = V_mA / [K_a(1 + I / K_i, App) + A] (1) where v is the reaction rate, the maximum rate is Vm, and A is the Michaelis constant K _a And I is the concentration of the inhibitor.

For compounds whose transition curves showed a downward curve characterized by time-dependent inhibition, data from individual sets was analyzed and k _obs determined according to Equation 2. [AMC] = v _ss t + (v 0 −v _ss ) [1-exp (−k _obs t)] / k _obs (2) where [AMC] is the concentration of product formed over time t , V0
Is the initial reaction rate and _vss is the steady state rate. The value of k _obs is then analyzed as a linear function of inhibitor concentration to yield an apparent second order rate constant (k _obs / inhibitor concentration or k _obs / [I]) representing time-dependent inhibition. Was. A complete discussion of this kinetic process is well documented (Morrison et al., Adv. Enzymol.
Relat. Areas Mol. Biol., 1998, 61, 201).

Human Osteoclast Resorption Assay An aliquot of the cell suspension from osteoclastoma was removed from liquid nitrogen storage and
℃ quickly, centrifugation (1000 rpm, 5 minutes, 4 ℃) RPMI-
Washed once in 1640 medium. The medium is aspirated and in RPMI-1640 medium 1:
Replaced with 3 diluted murine anti-HLA-DR antibodies and incubated on ice for 30 minutes. The cell suspension was mixed frequently.

The cells were centrifuged (1000 rpm, 5 minutes, 4 ° C.) with cold RPMI-164
Washed twice at 0 and then transferred to a 15 mL centrifuge tube. Neub with improved mononuclear cell count
Counted in the auer counting chamber. Sufficient magnetic beads (5 / mononuclear cells) coated with goat anti-mouse IgG were removed from the stock bottle and placed in 5 mL of fresh medium (which removes toxic azide preservatives). The medium was removed by fixing the beads on a magnet and replaced with fresh medium.

The beads were mixed with the cells and the suspension was incubated on ice for 30 minutes. The suspension was mixed frequently. The cells coated with the beads were fixed on a magnet, and the remaining cells (the osteoclast-rich fraction) were decanted into a sterile 50 mL centrifuge tube. Fresh media was added to the cells coated with beads to remove any captured osteoclasts. This washing process was repeated 10 times. Cells coated with beads were discarded.

Samples were loaded into the counting chamber using a large caliber disposable plastic Pasteur pipette and osteoclasts were counted in the counting chamber. The cells were pelleted by centrifugation and the density of the osteoclasts was adjusted to 1.5 × in EMEM medium supplemented with 10% fetal calf serum and 1.7 g / L sodium bicarbonate.
Adjusted to 10 ⁴ / mL. Fifteen 3 mL aliquots of cell suspension (per treatment)
Decant in mL centrifuge tube. These cells were pelleted by centrifugation. 3 mL of the appropriate treatment was added to each tube (diluted to 50 μM in EMEM medium).
Appropriate vehicle control, positive control (87 MEM diluted to 100 μg / mL)
1) and an isotype control (IgG2a diluted to 100 μg / mL) are also included. The tubes were incubated at 37 ° C. for 30 minutes.

A 0.5 mL aliquot of cells was plated on sterile dentin slices in a 48 well plate and incubated at 37 ° C. for 2 hours. Each treatment was screened in quadruplicate. The slices were washed with 6 changes of warm PBS (10 mL / well in a 6-well plate), then placed in a fresh treatment or control and incubated at 37 ° C. for 48 hours. The slices were then washed in phosphate-buffered saline and fixed in 2% glutaraldehyde (in 0.2 M sodium cacodylate) for 5 minutes, after which they were washed with water and 5% at 37 ° C. in buffer. Incubated for minutes. The slices are then washed in cold water and placed in cold acetate buffer / Fast Red Garnet at 4 ° C.
For 5 minutes. Excess buffer was aspirated, and after washing with water, the slices were air-dried.

[0086] TRAP-positive osteoclasts were counted by bright-field microscopy and then removed from the dentin surface by sonication. Fossa volume was measured using a Nikon / Lasertec ILM21W confocal microscope.

General Nuclear magnetic resonance spectra were recorded at 250 or 400 MHz, respectively, using either a Bruker AM 250 or Bruker AC 400 spectrometer. CDCl ₃ is deuteriochloroform, DMSO-d ₆ is hexadeuteriodimethylsulfoxide, and CD ₃ OD is tetradeuteriomethanol. Chemical shifts represent the downfield from the internal standard tetramethylsilane in parts per million (d
). Abbreviations of NMR data are as follows. s = single, d = double, t = triple, q = quadruple, m = multiple, dd = double double, dt = triple double, a
pp = apparent, br = wide. J indicates the NMR coupling constant measured in Hertz. Continuous wave infrared (IR) spectra were recorded on a Perkin-Elmer 683 infrared spectrometer and Fourier transform infrared (FTIR) spectra were recorded on a Nicolet Impact 400 D
Recorded on an infrared spectrometer. IR and FTIR spectra are recorded in transmission mode, and band positions are recorded at the reciprocal wave number (cm ^-1 ). Mass spectra were performed on either a VG 70 FE, PE Syx API III or VG ZAB HF instrument using fast atom bombardment (FAB) or electrospray (ES) ionization techniques. Elemental analysis was obtained using a Perkin-Elmer 240C elemental analyzer. Melting point is Tho
Measured in mas-Hoover melting point apparatus, no correction. All temperatures are recorded in degrees Celsius.

Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin plate were used in thin layer chromatography. Both flash and gravity chromatography were performed on E. Merck Kieselgel 60 (230-400 mesh) silica gel. When indicated, certain substances may be found in Aldrich, Milwaukee, Wisconsin.
Chemical Co., Chemical Dyn, South Plainfield, NJ
Purchased from amics Corp. and Advanced Chemtech, Louisville, KY.

EXAMPLES In the following synthesis examples, temperatures are in degrees Celsius (° C.). Unless otherwise noted, all starting materials were obtained from commercial sources. Without further elaboration, it is believed that one skilled in the art using the above description can utilize the present invention to its fullest extent. These examples are provided to illustrate the present invention and are not intended to limit its scope. What is reserved for the inventors is referred to in the claims.

Example 1 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-valinyl)] carbohydrazide a) 3-benzyloxybenzoate Methyl acrylate To a mixture of methyl 3-hydroxybenzoate (10.0 g, 65.7 mmol) and potassium carbonate (13.6 g, 98.6 mmol) in acetone (125 mL) was added benzyl bromide (11.2 g, 65.85 mmol). 7 mmol) was added. 1 at reflux temperature
After stirring for 6 hours, the mixture was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried (MgSO _4), filtered, and concentrated to give the title compound as a white solid (15.57g, 98%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.67 (m, 2H), 7.48-7.35 (m, 6H), 7.19 (m, 1H
), 5.02 (s, 2H), 3.93 (s, 3H).

B) 3-Benzyloxybenzoylhydrazide To a stirred solution of the compound of Example 1 (a) (10.0 g, 41.3 mmol) in 100 mL of methanol was added hydrazide hydrate (21.0 g, 413 mmol). added. The solution was stirred at room temperature for 16 hours, then concentrated to give the title compound as an off-white solid (9.56 g, 96%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.45-7.27 (m, 9H), 7.24 (m, 1H), 5.12 (s, 2H
), 4.10 (sb, 2H)

C) 5- (3-benzyloxyphenyl) -1,3,4-oxadiazole-
2-one Toluene (125 g) of the compound of Example 1 (b) (7.0 g, 28.9 mmol)
Phosgene (45 mL, 1.93 M in toluene) was added to the stirred solution in (mL).
The solution was heated at reflux for 4 hours and then concentrated to give the title compound as a pale yellow foam (
7.45 g, 96%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.41-7.28 (m, 9H), 7.06 (m, 1H), 5.06 (s, 2H
)

D) 2- [N- (3-benzyloxybenzoyl)] carbohydrazide 5- (3-benzyloxyphenyl) -1,3,4-oxadiazole-2-
Example 1 (b), except that on is used in place of methyl 3-benzyloxybenzoate
) To give the title compound as a white foam (7.73 g, 93%).
). MS (ESI): 301.1 (M + H) ⁺

E) 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-
Benzyloxycarbonyl-L-valinyl)] carbohydrazide The compound of Example 1 (d) (0.125 g, 0.417 mmol), N-benzyloxycarbonyl-L-valine (0.096 g, 0.458 mmol) and To a stirred solution of 1-hydroxybenzotriazole (0.011 g, 0.0834 mmol) in DMF (5 mL) was added 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.088 g, 0.458 mmol). ) Was added. After stirring at room temperature for 16 hours, the solution was poured into water (50 mL). The precipitate was collected by filtration and washed several times with water and then hexane. The solid was air dried for 2 hours to give the title compound as a white solid (0.183g, 89%). MS (ESI): 492.3 (M + H) ⁺

Example 2 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-phenylalanyl)] carbohydrazide In Step (e) Example 1 (a) -1 except that benzyloxycarbonyl-L-phenylalanine was used instead of benzyloxycarbonyl-L-valine
Following the procedure of (e), the title compound was obtained as a white solid (0.186 g,
77%). MS (ESI): 604.3 (M + Na) ⁺

Example 3 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-isoleucinyl)] carbohydrazide Benzyloxy in step (e) Example 1 (a) -1 (e) except that carbonyl-L-isoleucine was used instead of benzyloxycarbonyl-L-valine.
The title compound was prepared as a white solid (0.204 g,
89%). MS (ESI): 570.3 (M + Na) ⁺

Example 4 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-methionyl)] carbohydrazide The title compound was prepared as a white solid according to the procedure of 1 (a) -1 (e), except that carbonyl-L-methionine was used instead of benzyloxycarbonyl-L-valine (0.200 g, 85%).
. MS (ESI): 566.4 (M + H) ⁺

Example 5 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′-[
Preparation of N '-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide a) N-benzyloxycarbonyl-L-leucine methyl ester L-leucine methyl ester hydrochloride (2.0 g, 11.0 mmol) 1,4
- the medium-stirring solution in dioxane _{(20mL), Na 2 CO 3} (12.1ml, 2M in water
) And then benzyl chloroformate (1.96 g, 11.5 mmol) was added. The mixture was stirred at room temperature for 4 hours, then partitioned between ethyl acetate and water.
The organic layer was washed with brine, dried (MgSO _4), filtered and concentrated to give the title compound as a colorless oil (3.1g, 100%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.34 (m, 5H), 5.27 (d, 1H), 5.12 (s, 2H), 4.41 (s, 2H), 3.75 (s, 3H), 1.65 (m, 3H), 0.96 (m, 6H).

B) N-benzyloxycarbonyl-L-leucinyl hydrazide The procedure of Example 1 (b) was followed except that N-benzyloxycarbonyl-L-leucine methyl ester was used instead of methyl 3-benzyloxybenzoate. Therefore, the title compound was prepared as an off-white solid (3.1 g, 100%).
MS (ESI): 280.2 (M + H) ⁺

C) 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′
-[N ′-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide In step (b), N-benzyloxycarbonyl-L-leucine methyl ester is used in place of methyl 3-benzyloxybenzoate, The title compound was prepared as a white solid according to the procedure of 1 (c) -1 (e), except that N-benzyloxycarbonyl-L-norvaline was used in place of N-benzyloxycarbonyl-L-valine in e). (0.085 g, 40%). MS (ESI): 571.3 (M + H) ⁺

Example 6 of (2S) -2- [N- (N-benzyloxycarbonyl-2-aminobutyryl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-leucinyl)] carbohydrazide Preparation The procedure of Example 5 (a) -5 (c) was followed except that in step (c), (S) -N-benzyloxycarbonyl-2-aminobutyric acid was used instead of N-benzyloxycarbonyl-L-norvaline. Therefore, the title compound was prepared as a white solid (0.065 g, 32%). MS (ESI): 579.3 (M + Na) ⁺

Example 7 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-ridinyl-L-leucinyl)] carbohydrazide a) 2 -[N- (3-benzyloxybenzoyl)]-2 '-[N'-(N-
tert-butoxycarbonyl-L-leucinyl)] carbohydrazide Follow the procedure of Example 1 (e) except that N-tert-butoxycarbonyl-L-leucine is used in place of N-benzyloxycarbonyl-L-valine. The compound was prepared as a white solid (0.800 g, 94%). MS (ESI): 536.3 (M + Na) ⁺

B) 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(L-
Leucinyl)] carbohydrazide To a stirred solution of the compound of Example 7 (a) (0.800 g, 1.55 mmol) in dichloromethane (15 mL) was added trifluoroacetic acid (4 mL). 2 at room temperature
After stirring for an hour, the solution was concentrated. The residue was dissolved in ethyl acetate and washed with saturated aqueous sodium bicarbonate and brine. The organic layer was dried (MgSO _4), filtered, and concentrated to give the title compound as a white solid (0.640g, 100%). MS (ESI): 414.2 (M + H) ⁺

[0104] c) 2- [N- (3- benzyloxy-benzoyl)] - 2 '- [N ' - (N α - benzyloxycarbonyl -N _{epsilon-tert-butoxycarbonyl} -L- lysinyl -L- leucinyl) ] carbohydrazide N _alpha - benzyloxycarbonyl -N _{epsilon-tert-butoxycarbonyl} -L
The title compound was prepared as a white solid according to the procedure of Example 1 (e) except that -lysine was used in place of N-benzyloxycarbonyl-L-valine.
. 158 g, 67%). MS (ESI): 799.4 (M + Na) ⁺

D) 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-
Benzyloxycarbonyl -L- lysinyl -L- leucinyl)] carbohydrazide 2- [N- (3- benzyloxy-benzoyl)] - 2 '- [N ' - (N α - benzyloxycarbonyl -N _{epsilon-tert-butoxy} Carbonyl-L-lysinyl-L-leucinyl)] carbohydrazide is converted to 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-tert-butoxycarbonyl-L-leucinyl)] carbohydrazide The title compound was prepared as a white solid (0.126 g, 93%) according to the procedure of Example 7 (b) except for substituting. MS (ESI): 676.5 (M + H) ⁺

Example 8 (2R) -2- [N- (3-benzyloxybenzoyl)]-2 ′-[N′-
Preparation of [4-methyl-2- (3-phenylphenyl) pentanoyl] carbohydrazide a) Methyl 3-bromophenylacetate 3-Bromophenylacetic acid (2.15 g, 10 mmol) was dissolved in ether and then yellow Was maintained with a solution of diazomethane. Next, AcO
The reaction was quenched with H, concentrated in vacuo and used for the next reaction without further purification.

B) Methyl 3-phenylphenylacetate The compound of Example 8 (a) (2.29 g, 10 mmol) was dissolved in toluene (30 mL).
). Then phenylboronic acid (1.46 g, 12 mmol), then aqueous sodium carbonate (2 M, 4.24 mL, 40 mmol), then tetrakis (triphenylphosphine) palladium (0) (0.35 g, 0.3 mmol) ) Was added and the mixture was heated at reflux overnight. The reaction was cooled to room temperature, diluted with saturated ammonium chloride, and then extracted with EtOAc (2x10mL). The combined organic layers were dried (MgSO _4), filtered, concentrated and chromatographed (silica gel, 5% EtOAc: hexanes) to give to give the title compound as a white solid (
1.93 g, 84%). MS (ESI): 263 (M + H) ⁺

C) 3-Phenylphenylacetic acid The compound of Example 8 (b) (1.90 g, 8.4 mmol) was treated with MeOH (40 m
L) and water (6 mL), then LiOH.H ₂ O (0.7 g, 1
6.8 mmol) and the reaction was stirred at room temperature for 2 hours. The reaction was diluted with water, acidified with 6N HCl (1 mL), and then extracted with EtOAc (2 × 10 mL). The combined organic layers were dried (MgSO ₄ ), filtered and concentrated to give the title compound as a white solid (1.66 g, 93%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.6-7.25 (m, 9H), 3.7 (s, 2H)

D) 4-Methyl-2- (3-phenylphenyl) penta-4-enoic acid n-BuLi (5.2 mmol, 3.26 mL, 1.6 M in hexane) was treated with TH.
Diisopropylamine (0.54 g, 5.3 mmol, 0.7 mmol) in F (6 mL)
4 mL) was added dropwise to the solution at 0 ° C. The reaction was stirred for 15 minutes and then cooled to -78 ° C. The compound of Example 8 (c) (0.5 g, 2.35 mmol) in THF (
(2 mL) was added dropwise. The reaction was warmed to 0 ° C. and stirred for 40 minutes at −78 ° C.
And cooled. 3-Bromo-2-methylpropene (0.475 g, 3.52 mmol) was added and the reaction was stirred for 1 hour. Water (2 mL) was added and volatiles were removed in vacuo. The reaction was diluted with water, acidified with 6N HCl (1 mL), and
Extracted with tOAc (2 × 10 mL). Dry the combined organic layers (MgSO ₄
), Filtered, concentrated and chromatographed (silica gel, 5% MeOH: methylene chloride) to give the title compound as a white solid (0.582g, 93%).
. ¹ H NMR (400 MHz, CDCl ₃ ) d 7.6-7.3 (m, 9H), 4.75 (d, 2H), 3.87 (t, 1H), 2
.87 (dd, 1H), 2.50 (dd, 1H), 1.70 (s, 3H)

E) 4-Methyl-2- (3-phenylphenyl) pentanoic acid The compound of Example 8 (d) (0.5 g, 1.87 mmol) was treated with EtOAc (25
mL). Then 10% Pd / C (60 mg) was added and the reaction was stirred under a balloon of hydrogen gas for 2.5 hours. The mixture was filtered, concentrated in vacuo, and then redissolved in 1: 5 EtOAc: EtOH (15 mL). Then 10
% Pd / C (80 mg) was added and the reaction was stirred overnight under a balloon of hydrogen gas. The mixture was filtered, concentrated in vacuo, and chromatographed (silica gel, 5% MeO
H: methylene chloride) to give the desired product as a white solid (0.5 g,
100%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.6-7.3 (m, 9H), 3.7 (t, 1H), 2.07-1.95 (m, 1H
), 1.8-1.7 (m, 1H), 1.6-1.45 (m, 1H)

F) (R) -4-Methyl-2- (3-phenylphenyl) pentanoic acid The compound of Example 8 (e) (16.6 g, 62 mmol) was added to EtOH (100 m
L) and EtOAc (200 mL). (S) -p-bromo-α-
Methylbenzylamine (12.31 g, 62 mmol) was added and the solution was heated at 65 ° C. until the solid was completely dissolved. The solution was cooled in a refrigerator and white crystals were collected overnight.
Formed. The crystals were collected and then dried under vacuum. 1: 2 EtOAc / E
Recrystallized four times from tOH to give a crystalline white solid (3.05 g, 21% recovery). Chiral HPLC showed enantiomeric ratios of 99.3% (R) and 0.7% (S). The solid was then dissolved in EtOAc, extracted with 1N aqueous HCl, and the combined organic layers were dried (MgSO ₄ ), filtered, concentrated in vacuo and used in the next reaction without further purification.

G) (2R) -2- [N- (3-benzyloxybenzoyl)]-2 ′-[N
'-[4-Methyl-2- (3-phenylphenyl) pentanoyl] carbohydrazide In step (e), (R) -2- (3-phenylphenyl) -4-methylpentanoic acid was converted to N-benzyloxycarbonyl-L The title compound was prepared as a white solid (0.106 g, 36%) according to the procedure of Example 1 (a) -1 (e) except using -valine. MS (ESI): 573.3 (M + Na) ⁺

Example 9 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-arginyl-L-leucinyl)] carbohydrazide Step (c) )), N _α -benzyloxycarbonyl-N _{φ 2} -bis (ter
The t- butoxycarbonyl) -L- arginine N _alpha - except using in place of benzyloxycarbonyl -N _{epsilon-tert-butoxycarbonyl} -L- lysine according to the procedure of Example 7 (a) -7 (d) , The title compound was prepared as a white solid (0.364 g, 100%). MS (ESI): 704.4 (M + H) ⁺

Example 10 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide Example 1 (a)-
The title compound was prepared as a white solid according to the procedure of 1 (e) (0.19
5g, 88%). MS (ESI): 556.2 (M + Na) ⁺

Example 11 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-cyclohexylglycinyl)] carbohydrazide In Step (e) The title compound was prepared as a white solid according to the procedure of Example 1 (a) -1 (e) except that N-benzyloxycarbonyl-L-cyclohexylglycine was used in place of N-benzyloxycarbonyl-L-valine. (0.217 g, 91%). MS (ESI): 596.2 (M + Na) ⁺

Example 12 Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-cyclohexylalanyl)] carbohydrazide In Step (e) The title compound was prepared as a white solid according to the procedure of Example 1 (a) -1 (e) except that N-benzyloxycarbonyl-L-cyclohexylalanine was used instead of N-benzyloxycarbonyl-L-valine. (0.246 g, 80%). MS (ESI): 588.5 (M + H) ⁺

Example 13 2,2 ′-[N, N′-bis [N- (3-nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L-leucinyl] Preparation of Carbohydrazide a) 2,2 ′-[N, N′-bis- (N-tert-butoxycarbonyl-L
-Leucinyl)] carbohydrazide carbohydrazide (0.200 g, 2.22 mmol), N-tert-butoxycarbonyl-L-leucine (1.2 g, 4.88 mmol) and 1-hydroxybenzotriazole (0.060 g, 0.444 mmol) of DMF (2
(0 mL) to the stirred solution was added 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.935 g, 4.88 mmol). After stirring at room temperature for 16 hours, the solution was poured into water (200 mL). The precipitate was filtered and washed with water and hexane. The solid was air dried for 2 hours to give the title compound as a white solid (0.993 g, 86%). MS (ESI): 539.4 (M + Na) ⁺

B) 2,2 ′-[N, N′-bis- (L-leucinyl)] carbohydrazide
Bis (trifluoroacetate) salt 2,2 ′-[N, N′-bis- (N-tert-butoxycarbonyl-L-leucinyl)] carbohydrazide is converted to 2- [N- (3-benzyloxybenzoyl)
] -2 '-[N'-(N-tert-butoxycarbonyl-L-leucinyl)]
Following the procedure of Example 7 (b), but using instead of carbohydrazide,
The title compound was prepared as a white solid and used without further characterization (1.0
g, 100%).

C) 2,2 ′-[N, N′-bis [N- (3-nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L-leucinyl]] Carbohydrazide The compound of Example 13 (b) (0.021 g, 0.0383 mmol) and N
-Methylmorpholine (0.019 g, 0.192 mmol) in dichloromethane (
2 mL) and a stirred solution in DMF (0.50 mL) were added with N- (3-nitro-chloride).
1,2,7-benzoxadiazol-6-yl) -L-prolinyl (0.025
g, 0.0843 mmol). After stirring at room temperature for 16 hours, the solution was diluted with ethyl acetate and washed successively with saturated aqueous sodium bicarbonate, water (2x) and brine. The organic layer was dried (MgSO ₄ ), filtered and concentrated. The residue was purified by column chromatography (silica gel; methanol / dichloromethane) to give the title compound as an orange solid (0.007 g, 22%). MS (ESI): 837.4 (M + H) ⁺

Example 14 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′-[
Preparation of N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide a) Methyl 3- (trifluoromethylsulfonyloxy) phenylacetate Sodium hydride (2.54 g, 60% dispersion in mineral oil) under argon atmosphere , 63
. 5 mmol) in an oven-dried flask.
mL) was added. The slurry was stirred for 5 minutes, allowed to settle, much of the pentane was removed, and anhydrous THF (40 mL) was added. To the suspension was added a solution of methyl 3-hydroxyphenylacetate (9.99 g, 60.1 mmol) in anhydrous THF (20 mL) and the reaction was stirred at room temperature for 20 minutes. The mixture was then added to N-phenyltrifluoromethanesulfonimide (22.53 g, 63.1 mmol) in anhydrous TH.
A solution in F (40 mL) was added until TLC analysis indicated complete consumption of starting material (
The reaction was stirred at room temperature for 1.5 hours. The reaction was quenched by the addition of H ₂ O (10 mL), concentrated to half its original volume, then diluted with CHCl ₃ (200 mL) and washed with H ₂ O. The aqueous layer was washed with fresh CHCl ₃ (50 mL) and the combined organic layers were washed with 10% Na ₂ CO ₃ , water and saturated brine, then dried (MgSO ₄ ), filtered and concentrated. The residue was subjected to column chromatography (silica gel, 5:95 EtOAc: hexane, then 10:90 EtOAc: hexane) to give 17.47 g of the title compound. ¹ H NMR (400 MHz, CDCl ₃ ) 7.42 (m, 1H), 7.31-7.19 (m, 3H), 3.72 (s, 3H), 3
.68 (s, 2H)

B) Methyl 3- (2-pyridyl) phenylacetate To a solution of the compound of Example 14 (a) (6.86 g, 23.0 mmol) in anhydrous dioxane (100 mL) was added 2-pyridyltributylstannane ( 8.89 g,
24.1 mmol), LiCl (2.94 g, 69.3 mmol), 2,6-di-tert-butyl-4-methylphenol (slight crystals) and Pd (PP
h _{3) 4} (632.1mg, was added 0.55 mmol). The reaction was protected from light with foil and heated to reflux overnight. The reaction was cooled to room temperature and concentrated. The residue was purified by column chromatography (silica gel, 1: 3 EtOAc: hexane, then 1
: 2 EtOAc: Hexanes) to give 3.85 g of the title compound. MS (ESI): 228.1 (M + H) ⁺

C) 3- (2-Pyridyl) phenylacetic acid The compound of Example 14 (b) (3.8 g, 16.7 mmol) in THF (50 m
To the solution in L) was added an aqueous solution (10 mL) of LiOH.H ₂ O (780.2 mg, 18.6 mmol). The reaction was stirred at room temperature until TLC analysis showed complete consumption of starting material (2 hours). The reaction mixture was concentrated to remove THF, then neutralized to pH 7 by the addition of 1N HCl, diluted with brine (50 mL) and washed with CHCl ₃ (100 mL). The aqueous layer was returned to pH 7 by the addition of 1N NaOH and washed with fresh CHCl ₃ (100 mL). After repeating this procedure one or more times, the organic layers were combined, dried (MgSO ₄ ), filtered and concentrated to give 3.79 g of the title compound. MS (ESI): 214.3 (M + H) ⁺

D) 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′
-[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide The procedure was performed except that 3- (2-pyridyl) phenylacetic acid was used in place of N-benzyloxycarbonyl-L-norvaline in step (c). Example 5 (a) -5 (c)
The title compound was prepared as a white solid (1.03 g, 65
%). MS (ESI): 533.3 (M + H) ⁺

Example 15 Preparation of 2- (N-L-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- [N- (N-benzyloxycarbonyl) -L-leucinyl)]-2 '-[
N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide (1.
To a stirred solution of 5 g (2.8 mmol) in acetic acid (15 mL) was added 30% HBr in acetic acid (45 mL). After stirring at room temperature for 1 hour, diethyl ether (50
0 mL) was added and the precipitate was filtered. Dry the precipitate under high vacuum to give the title compound as a pale pink solid (1.4 g, 89%). MS (ESI): 399.4 (M + H) ⁺

Example 16 2- [N- [N- (3,4-dimethoxybenzoyl) -L-leucinyl]]-
Preparation of 2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl Example 1 except that carbohydrazide was used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and 3,4-dimethoxybenzoic acid was used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to the procedure of (e) (0.038 g, 30%).
. MS (ESI): 563.2 (M + H) ⁺

Example 17 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 ′-[N′-
Preparation of [N- (4-trifluoromethylbenzoyl) -L-leucinyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] Example 1 (e) except that carbohydrazide was used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and 4-trifluoromethylbenzoic acid was used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid (0.057 g, 45).
%). MS (ESI): 571.2 (M + H) ⁺

Example 18 2- [N- [N- (3,4-dichlorobenzoyl) -L-leucinyl]]-2
Preparation of '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl] Example 1 (except that carbohydrazide is used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and 3,4-dichlorobenzoic acid is used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to the procedure of e) (0.087 g, 68%).
MS (ESI): 571.2 (M + H) ⁺

Example 19 2- [N- (N-benzofuran-2-ylcarbonyl-L-leucinyl)]-
Preparation of 2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl Example 1 (except that carbohydrazide was used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and benzofuran-2-carboxylic acid was used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to the procedure of e) (0.078 g, 65%
). MS (ESI): 543.2 (M + H) ⁺

Example 20 2- [N- [N- (5,6-dimethoxybenzofuran-2-ylcarbonyl)]
-L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide a) 2-Hydroxy-4,5-dimethoxybenzaldehyde 2-benzyloxy-4,5 -Dimethoxybenzaldehyde (1.0 g, 3.
To a stirred solution of (67 mmol) in ethyl acetate (25 mL) was added 10% palladium on carbon (0.50 g). The mixture was stirred under a hydrogen atmosphere for 4 hours, then
Filtered through Celite. The filtrate was concentrated to give the title compound as a pale yellow solid (0.
632 g, 95%). ¹ H NMR (400 MHz, CDCl ₃ ) d 11.41 (s, 1H), 9.72 (s, 1H), 6.89 (s, 1H), 6.4
8 (s, 1H), 3.91 (s, 3H), 3.88 (s, 3H)

B) 4,5-Dimethoxy-2-ethoxycarbonylmethoxybenzaldehyde Except that 2-hydroxy-4,5-dimethoxybenzaldehyde is used instead of methyl 3-hydroxybenzoate and ethyl bromoacetate is used instead of benzyl bromide Prepared the title compound according to the procedure of Example 1 (a) (0.758 g,
82%). ¹ H NMR (400 MHz, CDCl ₃ ) d 10.39 (s, 1H), 7.30 (s, 1H), 6.41 (s, 1H), 4.7
2 (s, 2H), 4.22 (q, 2H), 3.90 (s, 3H), 3.83 (s, 3H), 1.26 (t, 3H)

C) Ethyl 5,6-dimethoxybenzofuran-2-carboxylate A mixture of the compound of Example 20 (b) (0.758 g, 2.8 mmol) and potassium carbonate (0.975 g, 7.1 mmol) At 80-C in DMF (20
(mL) for 5 hours. The mixture was cooled and partitioned between ethyl acetate and water.
The organic layer was washed with water and brine, then dried (MgSO _4), filtered, and concentrated. The residue was purified by column chromatography (silica gel, ethyl acetate / hexane) to give the title compound as a white solid (0.405 g, 5
8%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.45 (s, 1H), 7.10 (s, 1H), 7.04 (s, 1H), 4.41 (q, 2H), 3.93 (s, 3H), 3.91 (s, 3H), 1.41 (t, 3H)

D) 5,6-Dimethoxybenzofuran-2-carboxylic acid Example 14 (c) except that ethyl 5,6-dimethoxybenzofuran-2-carboxylate was used in place of methyl 3- (2-pyridyl) phenylacetate )). The title compound was prepared as a white solid (0.263 g, 73%). ¹ H NMR (400 MHz, CDCl ₃ ) d 7.40 (s, 1H), 7.03 (s, 1H), 7.01 (s, 1H), 3.90 (s, 3H), 3.88 (s, 3H)

E) 2- [N- [N- (5,6-dimethoxybenzofuran-2-ylcarbonyl) -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl ]] Carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide is converted to 2- [N- (3-benzyloxybenzoyl)] carbo Prepare the title compound as a white solid according to the procedure of Example 1 (e) except that 5,6-dimethoxybenzofuran-2-carboxylic acid is used instead of hydrazide in place of N-benzyloxycarbonyl-L-valine. (0
. 110 g, 68%). MS (ESI): 603.2 (M + H) ⁺

Example 21 2- [N- [N- (5-methyl-2-phenyloxazol-4-ylcarbonyl) -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl) ) Phenylacetyl]] carbohydrazide Preparation of 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2- [N- (3-benzyl Oxybenzoyl)] Instead of carbohydrazide, 5-methyl-2-phenyloxazole-
The title compound was prepared as a white solid according to the procedure of Example 1 (e) except that 4-carboxylic acid was used instead of N-benzyloxycarbonyl-L-valine (0.107 g, 68%). MS (ESI): 584.3 (M + H) ⁺

Example 22 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1′-N
Preparation of '-methyl-2'-[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide a) 1- (N-benzyloxycarbonyl) -1- (N-methyl) hydrazine methylhydrazine ( 10.6 g, 230 mmol) of dichloromethane (100
Benzyl chloroformate (3.9 g, 23 mmol) was added dropwise at 0-C to the stirred solution in mL). After stirring at room temperature for 1 hour, the solution was concentrated, and the residue was purified by column chromatography (silica gel; ethyl acetate / hexane) to give the title compound as a colorless oil (3.9 g, 94%). MS (ESI): 181.0 (M + H) ⁺

B) 1- (N-benzyloxycarbonyl) -1- (N-methyl) -2- [N
-[3- (2-pyridyl) phenylacetyl]] hydrazine 1- (N-benzyloxycarbonyl) -1- (N-methyl) hydrazine was converted to 2
Instead of-[N- (3-benzyloxybenzoyl)] carbohydrazide, 3
The title compound was prepared as a white solid according to the procedure of Example 1 (e) except that-(2-pyridinyl) phenylacetic acid was used instead of N-benzyloxycarbonyl-L-valine (0.373 g, 96%). ). MS (ESI): 376.3 (M + H) ⁺

C) 1- (N-methyl) -2- [N- [3- (2-pyridyl) phenylacetyl]] hydrazine bis (hydrobromide) salt 1- (N-benzyloxycarbonyl)- 1- (N-methyl) -2- [N- [
3- (2-Pyridyl) phenylacetyl]] hydrazine is converted to 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] The title compound was prepared as a light orange solid according to the procedure of Example 15 except that carbohydrazide was used (0.454 g). MS (ESI): 242.1 (M + H) ⁺

D) 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1 ′
-N'-methyl-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]
] Carbohydrazide phosgene (0.327 mL, 1.93 M in toluene) in dichloromethane (2 m
L) The solution in Example 22 (c) (0.200 g, 0.49
A solution of 7 mmol) and N-methylmorpholine (0.161 g, 1.6 mmol) in dichloromethane (2 mL) was added dropwise. After stirring for 15 minutes, Example 5 (b)
(0.139 g, 0.497 mmol) and N-methylmorpholine (
A solution of 0.055 g (0.547 mmol) in dichloromethane (2 mL) was added dropwise. After stirring overnight at room temperature, the solution was diluted with ethyl acetate and washed successively with saturated aqueous sodium bicarbonate, water (2x) and brine. Dry the organic layer (Mg
SO ₄ ), filtered and concentrated. The residue was subjected to column chromatography (silica gel;
Purification by methanol / dichloromethane) afforded the title compound as a white solid (0.028 g, 10%). MS (ESI): 547.3 (M + H) ⁺

Example 23 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)
Preparation of] -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) Phenylacetyl]] carbohydrazide is replaced with 2- [N- (3-benzyloxybenzoyl)] carbohydrazide, and benzothiophen-2-carboxylic acid is replaced with N-
Example 1 (e) except that it is used instead of benzyloxycarbonyl-L-valine
The title compound was prepared as a white solid (0.089 g, 6
0%). MS (ESI): 559.1 (M + H) ⁺

Example 24 2- [N- [N- [4-methyl-2- (4-trifluoromethylphenyl) thiazol-5-ylcarbonyl] -L-leucinyl]]-2 ′-[N′- [3-
Preparation of (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2- [N- (3-benzyloxybenzoyl)] Instead of carbohydrazide, 4-methyl-2- (4-trifluoromethylphenyl) thiazole-5-carboxylic acid was converted to N-benzyloxycarbonyl-L
The title compound was prepared as a white solid (0.074 g, 41%) following the procedure of Example 1 (e) except substituting valine. MS (ESI): 668.1 (M + H) ⁺

Example 25 2- [N- [N- (3-isoquinolinoyl) -L-leucinyl] -2 ′-[N
Preparation of '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2'-[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide The procedure of Example 1 (e) was followed except that isoquinoline-3-carboxylic acid was used instead of N-benzyloxycarbonyl-L-valine instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide. Therefore, the title compound was prepared as a white solid (0.067 g, 45%
). MS (ESI): 554.3 (M + H) ⁺

Example 26 2- [N- [N- (5-chlorobenzofuran-2-ylcarbonyl) -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl] ]
Preparation of Carbohydrazide 2- (N-L-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2- [N- (3-benzyloxybenzoyl)] carbo. Example 1 except that 5-chlorobenzofuran-2-carboxylic acid is used instead of hydrazide in place of N-benzyloxycarbonyl-L-valine.
The title compound was prepared as a white solid according to the procedure of (e) (0.035
g, 23%). MS (ESI): 577.1 (M + H) ⁺

Example 27 2- [N- [N- (3,5-difluorobenzoyl) -L-leucinyl]]-
Preparation of 2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl Example 1 except that carbohydrazide was used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and 3,5-difluorobenzoic acid was used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to the procedure of (e) (0.064 g, 44%).
. MS (ESI): 539.3 (M + H) ⁺

Example 28 2- [N- (N-benzothiazol-6-ylcarbonyl-L-leucinyl)
Preparation of] -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) Phenylacetyl]] carbohydrazide is replaced with 2- [N- (3-benzyloxybenzoyl)] carbohydrazide, and benzothiazole-6-carboxylic acid is replaced with N-
Example 1 (e) except that it is used instead of benzyloxycarbonyl-L-valine
The title compound was prepared as a white solid (0.078 g, 5
2%). MS (ESI): 560.1 (M + H) ⁺

Example 29 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1- (N
Preparation of -methyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide a) 1- (N-tert-butoxycarbonyl) -1- (N-methyl) hydrazine methylhydrazine (0.866 g, 18.8 mmol) of dioxane (1.8
mL) at 0-C to the stirred solution in di-tert-butyl dicarboxylate (2.0 mL).
A solution of 5 g (9.4 mmol) in dioxane (2.8 mL) was added slowly. After stirring at room temperature for 2 hours, the mixture was filtered and the filtrate was concentrated to give the title compound as a colorless oil (0.258 g). MS (ESI): 146.8 (M + H) ⁺

B) 1- (N-tert-butoxycarbonyl) -1 (-N-methyl) -2
-(N-benzyloxycarbonyl-L-leucinyl) hydrazine 1- (N-tert-butoxycarbonyl) -1- (N-methyl) hydrazine is substituted for 2- [N- (3-benzyloxybenzoyl)] carbohydrazide The title compound was prepared as a colorless oil according to the procedure of Example 1 (e) except that N-benzyloxycarbonyl-L-leucine was used instead of N-benzyloxycarbonyl-L-valine (0.595 g). , 85%). MS (ESI): 394.4 (M + H) ⁺

C) 2- (N-benzyloxycarbonyl-L-leucinyl) -1- (N-methyl) hydrazine 1- (N-tert-butoxycarbonyl) -1 (-N-methyl) -2- (N
-Benzyloxycarbonyl-L-leucinyl) hydrazine was converted to 2- [N- (3-
Example 7 except that benzyloxybenzoyl)]-2 ′-[N ′-(N-tert-butoxycarbonyl-L-leucinyl)] carbohydrazide was used instead.
The title compound was prepared as a white solid according to the procedure of (b) (0.310
g, 70%). MS (ESI): 294.3 (M + H) ⁺

D) 3- (2-pyridyl) phenylacetylhydrazide According to the procedure of Example 1 (b), but using methyl 3- (2-pyridyl) phenylacetate instead of methyl 3-benzyloxybenzoate. The title compound was prepared as an off-white solid (0.590 g, 100%). MS (ESI): 228.0 (M + H) ⁺

E) 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1-
(N-methyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]
Carbohydrazide 2- (N-benzyloxycarbonyl-L-leucinyl) -1- (N-methyl) hydrazine is converted to 1- (N-methyl) -2- [N- [3- (2-pyridyl) phenylacetyl] Example 22 (d) except that 3- (2-pyridyl) phenylacetylhydrazide is used instead of N-benzyloxycarbonyl-L-leucinylhydrazide instead of hydrazine bis (hydrobromide) salt. Following the procedure, the title compound was prepared as a white solid (0.034 g, 7%). MS (ESI): 548.3 (M + H) ⁺

Example 30 2- [N- (N-picolinoyl-L-leucinyl)]-2 ′-[N ′-[3-
Preparation of (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2- [N- (3-Benzyloxybenzoyl)] The title compound was prepared as a white solid according to the procedure of Example 1 (e) except that picolinic acid was used instead of N-benzyloxycarbonyl-L-valine instead of carbohydrazide. (0.068 g, 50%). MS (ESI): 504.3 (M + H) ⁺

Example 31 2- [N- [N- (2,6-dimethoxynicotinoyl) -L-leucinyl]]
Preparation of -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenyl [Acetyl]] carbohydrazide was used in place of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide, and 2,6-dimethoxynicotinic acid was used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to the procedure of 1 (e) (0.086 g, 57%
). MS (ESI): 564.2 (M + H) ⁺

Example 32 2- [N- [N- (4-methanesulfonylbenzoyl) -L-leucinyl]]
Preparation of -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- (2-pyridinyl) phenyl Example 1 except that acetyl]] carbohydrazide was used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and 4-methanesulfonylbenzoic acid was used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to the procedure of (e) (0.078 g, 50%
). MS (ESI): 581.2 (M + H) ⁺

Example 33 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 ′-[N′-
Preparation of [N- (3,4,5-trimethoxybenzoyl) -L-leucinyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenyl [Acetyl]] carbohydrazide is replaced with 2- [N- (3-benzyloxybenzoyl)] carbohydrazide instead of 3,4,5-trimethoxybenzoic acid with N-
Example 1 (e) except that it is used instead of benzyloxycarbonyl-L-valine
The title compound was prepared as a white solid (0.072 g, 4
5%). MS (ESI): 593.2 (M + H) ⁺

Example 34 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 ′-[N′-
Preparation of [N- (8-quinolinoyl) -L-leucinyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide The procedure of Example 1 (e) was followed, except that quinoline-8-carboxylic acid was used instead of N-benzyloxycarbonyl-L-valine instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide. Therefore, the title compound was prepared as a white solid (0.075 g, 51%).
MS (ESI): 554.3 (M + H) ⁺

Example 35 2- [N- [N- [3,4- (1,3-propylenedioxy) benzoyl]-
Preparation of L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- ( 2-pyridinyl) phenylacetyl]] carbohydrazide instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide instead of 3,4- (1,3-propylenedioxy)
The title compound was prepared as a white solid according to the procedure of Example 1 (e), except that benzoic acid was used in place of N-benzyloxycarbonyl-L-valine.
. 091 g, 59%). MS (ESI): 575.2 (M + H) ⁺

Example 36 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 ′-[N′-
Preparation of (N-thieno [2,3-b] thiophen-2-ylcarbonyl-L-leucinyl)] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2- Pyridinyl) phenylacetyl]] carbohydrazide is replaced with 2- [N- (3-benzyloxybenzoyl)] carbohydrazide, and thieno [2,3-b] thiophene-2-carboxylic acid is replaced with N-benzyloxycarbonyl-L The title compound was prepared as a white solid according to the procedure of Example 1 (e) except substituting valine for (0
. 072 g, 48%). MS (ESI): 565.0 (M + H) ⁺

Example 37 2- [N- [N- (4-methoxybenzoyl) -L-leucinyl]]-2′-
Preparation of [N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2'-[N '-[3- (2-pyridinyl) phenylacetyl]] carbo The procedure of Example 1 (e) except that hydrazide is used instead of 2- [N- (3-benzyloxybenzoyl)] carbohydrazide and 4-methoxybenzoic acid is used instead of N-benzyloxycarbonyl-L-valine. The title compound was prepared as a white solid according to (0.086 g, 60%). MS (ESI): 533.2 (M + H) ⁺

Example 38 2- [N- [N- [4-methoxy-3- [2- (4-morpholino) ethoxy]]
Benzoyl] -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl)
Phenylacetyl]] Preparation of carbohydrazide a) Methyl 4-methoxy-3- (2-N-morpholinoethoxy) benzoate Methyl 3-hydroxy-4-methoxybenzoate was substituted for methyl 3-hydroxybenzoate in place of methyl 3-hydroxybenzoate. The title compound was prepared as a pale yellow oil (5.71 g, 65%) following the procedure of Example 1 (a) except that (2-chloroethyl) morpholine hydrochloride was used in place of benzyl bromide. MS (ESI): 296.2 (M + H) ⁺

B) 4-Methoxy-3- [2- (4-morpholino) ethoxy] benzoic acid Methyl 4-methoxy-3- (2-N-morpholinoethoxy) benzoate is converted to 3- (
The title compound was prepared as a white solid according to the procedure of Example 14 (c), except substituting for methyl 2-pyridyl) phenylacetate (5.4 g, 100%).
). MS (ESI): 282.2 (M + H) ⁺

C) 2- [N- [N- [4-methoxy-3- [2- (4-morpholino) ethoxy] benzoyl] -L-leucinyl]]-2 ′-[N ′-[3- ( 2-pyridinyl) phenylacetyl]] carbohydrazide 2- (N-L-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide is 2- [N- (3- Example 1 (e) except that 4-methoxy-3- [2- (4-morpholino) ethoxy] benzoic acid was used in place of N-benzyloxycarbonyl-L-valine in place of benzyloxybenzoyl)] carbohydrazide. The title compound was prepared as a white solid (0.068 g, 38%). MS (ESI): 662.4 (M + H) ⁺

Example 39 2- [N- [N- [2- (4-morpholino) pyrimidin-4-ylcarbonyl] -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl) ) Phenylacetyl]] Preparation of carbohydrazide a) Potassium salt of 2-methylthiopyrimidine-4-carboxylic acid 5-Bromo-2-methylthiopyrimidine-4-carboxylic acid (1.
To a suspension of 25 g, 5.0 mmol) in methanol (60 mL) was added potassium hydroxide (0.630 g, 11.2 mmol) followed by 10% palladium on BaSO ₄ (0.630 g, 50% w / w) was added. After shaking under hydrogen at 35 psi on a Parr shaker for 3 hours, the mixture was filtered through celite. The filtrate was concentrated to give the title compound. ¹ H NMR (400 MHz, CD ₃ OD) d 8.59 (d, 1H), 7.48 (d, 1H), 2.60 (s, 3H)

B) Ethyl 2-methylthiopyrimidine-4-carboxylate To a suspension of the compound of Example 39 (a) in ethanol was added concentrated HCl (6 mL).
I got it. After stirring at reflux for 16 hours, the solution was concentrated, dissolved in ethyl acetate and saturated
Washed with aqueous sodium bicarbonate and brine. Dry the organic layer (MgSO 4 _Four ), Filtered and concentrated to give the title compound as an oily yellow solid (0.85).
1 g, 86%).¹ H NMR (400 MHz, CDCl_Three) d 8.72 (d, 1H), 7.58 (d, 1H), 4.44 (q, 2H), 2.62 (s, 3H), 1.45 (t, 3H)

C) Ethyl 2-methanesulfonylpyrimidine-4-carboxylate To a solution of the compound of Example 39 (b) (0.851 g, 4.3 mmol) in dichloromethane (50 mL) was added m-chloroperoxybenzoic acid ( 2.0 g, 11.6
Mmol) was added slowly. After stirring at room temperature for 3 hours, the solution was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate. Dry the organic layer (MgSO ₄ )
, Filtered and concentrated. The residue was purified by column chromatography (silica gel; ethyl acetate / hexane) to give the title compound as a white solid (0.78
8g, 80%). ¹ H NMR (400 MHz, CDCl ₃ ) d 9.16 (d, 1H), 8.17 (d, 1H), 4.50 (q, 2H), 3.43 (s, 3H), 1.45 (t, 3H)

D) Ethyl 2- (4-morpholino) pyrimidine-4-carboxylate The compound of Example 39 (c) in morpholine (2 mL) (0.200 g,
(0.869 mmol) was stirred at 80-C for 16 hours. The solution was diluted with ethyl acetate and washed with water and brine. Dry the organic layer (MgSO ₄ ), filter,
Concentrated. The residue was purified by column chromatography (silica gel; ethyl acetate / hexane) to give the title compound as a white solid (0.118 g, 57
%). ¹ H NMR (400 MHz, CDCl3) d 8.51 (d, 1H), 7.24 (d, 1H), 4.40 (q, 2H), 3.87 (t, 4H), 3.77 (t, 4H), 1.45 (t, 3H )

E) 2- (4-morpholino) pyrimidine-4-carboxylic acid Example 14 except that ethyl 2-N-morpholinopyrimidine-4-carboxylate was used in place of methyl 3- (2-pyridyl) phenylacetate The title compound was prepared as a white solid according to the procedure of (c) (0.125 g, 100%). ¹ H NMR (400 MHz, CDCl ₃ ) d 8.48 (d, 1H), 7.17 (d, 1H), 3.82 (t, 4H), 3.72 (t, 4H)

F) 2- [N- [N- [2- (4-morpholino) pyrimidin-4-ylcarbonyl] -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl) Phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide is converted to 2- [N- (3-benzyloxybenzoyl) ] Instead of carbohydrazide, 2- (4-morpholino) pyrimidine-4-
The title compound was prepared as a white solid according to the procedure of Example 1 (e) except that the carboxylic acid was used in place of N-benzyloxycarbonyl-L-valine (
0.056 g, 44%). MS (ESI): 590.3 (M + H) ⁺

Example 40 2- [N- [N- (2,3-dihydrobenzofuran-5-ylcarbonyl)-
Preparation of L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[3- ( 2-pyridinyl) phenylacetyl]] carbohydrazide is replaced with 2- [N- (3-benzyloxybenzoyl)] carbohydrazide, and 2,3-dihydrobenzofuran-5-carboxylic acid is replaced with N-benzyloxycarbonyl-L- The title compound was prepared as a white solid according to the procedure of Example 1 (e) except substituting for valine (0.
070g, 47%). MS (ESI): 545.4 (M + H) ⁺

Example 41 2- [N- [N- (5-carboxybenzofuran-2-ylcarbonyl) -L
-Leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide a) 4-Benzyloxycarbonylmethoxy-3-formylbenzaldehyde 5-formylsalicylaldehyde (2.2 g) , 14.7 mmol) and potassium bromide (5.0 g, 36.8 mmol) in acetone (50 mL).
Benzyl bromoacetate (4.8 g, 16.1 mmol) was added. After stirring at reflux for 6 hours, the mixture was diluted with ethyl acetate and washed with water and brine. Dry the organic layer (MgSO ₄ ), filter and concentrate to give the title compound (4.13 g)
, 94%). ¹ H NMR (400 MHz, CDCl ₃ ) d 10.56 (s, 1H), 9.95 (s, 1H), 8.38 (s, 1H), 8.0
7 (d, 1H), 7.38 (m, 5H), 6.95 (d, 1H), 5.26 (s, 2H), 4.91 (s, 2H)

B) Benzyl 5-formylbenzofuran-2-carboxylate 4-benzyloxycarbonylmethoxy-3-formylbenzaldehyde
The title compound was prepared as a white solid (1.78 g, 46%) according to the procedure of Example 20 (c) except using instead of, 5-dimethoxy-2-ethoxycarbonylmethoxybenzaldehyde. ¹ H NMR (400 MHz, CDCl ₃ ) d 10.09 (s, 1H), 8.24 (s, 1H), 8.05 (d, 1H), 7.7
1 (d, 1H), 7.68 (s, 1H), 7.42 (m, 5H), 5.43 (s, 2H)

C) Benzyl 5-carboxybenzofuran-2-carboxylate The compound of Example 41 (b) (0.380 g, 1.36 mmol) in THF (5
solution in water (2 mL) and a solution of sodium chlorite (0.245 g, 2.71 mmol) and sulfamic acid (0.277 g, 2.86 mmol) in tert-butanol (1 mL). added. After stirring at room temperature for 3 hours, the solution was partitioned between ethyl acetate and water. The organic layer was washed successively with water, saturated aqueous sodium bicarbonate and brine, then dried (MgSO ₄ )
Filtration and concentration gave the title compound as an off-white solid (0.272 g
, 68%). ¹ H NMR (400 MHz, CDCl ₃ ) d 8.52 (s, 1H), 8.23 (d, 1H), 7.67 (m, 2H), 7.49 (m, 2H), 7.41 (m, 3H), 5.46 (s, 2H)

D) Benzyl 5-tert-butoxycarbonylbenzofuran-2-carboxylate To a solution of the compound of Example 41 (c) (0.272 g, 0.919 mmol) in toluene (3 mL) was added N, N-dimethyl. Formamide di-tert-butyl acetal (0.748 g, 3.68 mmol) was added. After stirring at 80-C for 4 hours, the solution was concentrated, and the residue was dissolved in ethyl acetate. The organic layer was washed with water and brine, then dried (MgSO _4), filtered, and concentrated. The residue was purified by column chromatography (silica gel, ethyl acetate / hexane) to give the title compound as a white solid (0.144 g, 45%). ¹ H NMR (400 MHz, CDCl ₃ ) d 8.38 (s, 1H), 8.12 (d, 1H), 7.62 (m, 2H), 7.50 (m, 2H), 7.41 (m, 3H), 5.46 (s, 2H) 1.65 (s, 9H)

E) 5-tert-Butoxycarbonylbenzofuran-2-carboxylate Example except that benzyl 5-tert-butoxycarbonylbenzofuran-2-carboxylate was used in place of 2-benzyloxy-4,5-dimethoxybenzaldehyde The title compound was prepared as a white solid according to the procedure of 20 (a) (0.098 g, 91%). MS (ESI): 261.2 (MH) ^-

F) 2- [N- [N- (5-tert-butoxycarbonylbenzofuran-2)
-Ylcarbonyl) -L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 '-[N'-[ 3- (2-pyridinyl) phenylacetyl]] carbohydrazide is replaced with 2- [N- (3-benzyloxybenzoyl)] carbohydrazide, and 5-tert-butoxycarbonylbenzofuran-2-carboxylic acid is replaced with N-benzyloxy. The title compound was prepared as a white solid (0.123 g, 56%) following the procedure of Example 1 (e) except for substituting carbonyl-L-valine. MS (ESI): 643.2 (M + H) ⁺

G) 2- [N- [N- (5-carboxybenzofuran-2-ylcarbonyl)
-L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- [N- [N- (5-tert-butoxycarbonylbenzofuran-2-ylcarbonyl)] -L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2- [N- (3-benzyloxybenzoyl)]-2 '-[N'- (N-tert-butoxycarbonyl-L-leucinyl)] The title compound was prepared as a white solid according to the procedure of Example 7 (b), except substituting for carbohydrazide (0.160 g, 100%).
MS (ESI): 587.2 (M + H) ⁺ Example 42 2- [N- (3-benzyloxybenzoyl) -2 '-[N'-[N- (3-
Nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L
-Leucinyl]] carbohydrazide Preparation of 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(L-leucinyl)] carbohydrazide by 2,2 ′-[N, N′-bis -(L-leucinyl)] carbohydrazide The title compound was prepared as an orange solid according to the procedure of Example 13 (c) except that bis (trifluoroacetate) salt was used (0.012 g, 5%). ). MS (ESI): 674.2 (M + H) ⁺

Example 43 2- [N- [N- (3-nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L-leucinyl]]-2- [N- Preparation of [3- (2-pyridinyl) phenylacetyl]] carbohydrazide 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2, The title compound was converted to an orange solid according to the procedure of Example 13 (c) except that 2 '-[N, N'-bis- (L-leucinyl)] carbohydrazide was used instead of bis (trifluoroacetate) salt. (0.038 g, 19%). MS (ESI): 659.2 (M + H) ⁺

Example 44 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-oxo-2-pyridinyl) phenylacetyl]
Preparation of carbohydrazide a) 3- (2-oxo-2-pyridinyl) phenylacetic acid The compound of Example 14 (c) (1.2 g, 5.6 mmol) in diethyl ether (500 mL) and dichloromethane (10 mL) To the solution was added m-chloroperoxybenzoic acid (2.5 g, 8.5 mmol). After standing at room temperature for 36 hours,
The collected white needles were air dried for 2 hours to give the title compound (1.1 g, 86%).
. MS (ESI): 230.1 (M + H) ⁺

B) 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′
-[N '-[3- (2-oxo-2-pyridinyl) phenylacetyl]] carbohydrazide In the step (c), 3- (2-oxo-2-pyridinyl) phenylacetic acid is converted to N-
Example 5 (except that benzyloxycarbonyl-L-norvaline was used instead of
The title compound was prepared as a white solid according to the procedure of a) -5 (c) (0
. 842 g, 33%). MS (ESI): 549.2 (M + H) ⁺

C) 2- (NL-leucinyl) -2 ′-[N ′-[3- (2-oxo-2-
Pyridinyl) phenylacetyl]] carbohydrazide 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 ′-[
N '-[3- (2-oxo-2-pyridinyl) phenylacetyl]] carbohydrazide is converted to 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2.
The title compound was prepared as an off-white solid according to the procedure of Example 15 except for substituting '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide (0.152 g, 100 %). MS (ESI): 415.2 (M + H) ⁺

D) 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)]-2 ′-[N ′-[3- (2-oxo-2-pyridinyl) phenylacetyl]] Carbohydrazide 2- (N-L-leucinyl) -2 '-[N'-[3- (2-oxo-2-pyridinyl) phenylacetyl]] carbohydrazide was converted to 2- [N- (3-benzyloxybenzoyl) The title compound was prepared as a white solid according to the procedure of Example 1 (e) except that benzothiophene-2-carboxylic acid was used instead of N-benzyloxycarbonyl-L-valine instead of carbohydrazide ( 0.
085 g, 47%). MS (ESI): 575.2 (M + H) ⁺

Example 45 Preparation of 2,2 ′-[N, N′-bis [N- (3-iodo-4-methoxy) -L-leucinyl]] carbohydrazide The compound (0) of Example 13 (b) .200 g, 0.368 mmol), 3-iodo-4-methoxybenzoic acid (0.225 g, 0.809 mmol), triethylamine (0.082 g, 0.809 mmol) and 1-hydroxybenzotriazole (0.020 g , 0.147 mmol) in DMF (4 mL)
1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.
155 g, 0.809 mmol). After stirring at room temperature for 16 hours, the solution was diluted with ethyl acetate and washed successively with saturated aqueous sodium bicarbonate, water (2x) and brine. The organic layer was dried (MgSO ₄ ), filtered and concentrated. The residue was purified by column chromatography (silica gel; methanol / dichloromethane) to give the title compound as a white solid (0.147 g, 48%). MS (ESI): 882.2 (M + 2Na) ⁺

The above specification and examples fully disclose the preparation and use of the compounds of the present invention. However, the invention is not limited to the specific embodiments described hereinabove, but encompasses all modifications thereof within the scope of the claims. Various references to publications, patents and other publications cited herein, including the state of the art, are hereby incorporated by reference as if fully set forth by reference.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 31/4406 A61K 31/4406 4C056 31/443 31/443 4C063 31/4436 31/4436 4C071 31/4439 31 / 4439 4C083 31/444 31/444 4C086 31/4709 31/4709 4C206 31/4725 31/4725 4H006 31/5377 31/5377 38/00 A61P 1/02 A61P 1/02 3/12 3/12 19/02 19/02 19/10 19/10 29/00 101 29/00 101 C07D 213/56 C07D 213/56 213/81 213/81 213/82 213/82 215/48 215/48 217/26 217/26 239 / 34 239/34 263/34 263/34 277/56 277/56 277/62 277/62 295/08 Z 295/08 307/84 307/84 321/10 321/10 401/12 401/12 405 / 12 405/12 409/12 409/12 413/12 413/12 417/12 417/12 495/04 101 495/04 101 A61K 37/02 (81) Designated country EP (AT, BE, CH, C , DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW) , ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AU, BA, BB, BG, BR, CA, CN, CZ, EE, GE, HU, ID, IL, IS, JP, KP, KR, LC, LK, LR, LT, LV, MG, MK, MN, MX, NO, NZ, PL, RO, SG, SI, SK, SL, TR, TT, UA, US, UZ, VN, YU (72) Inventor Scott Kevin Thompson, Guildford, Phoenixville, PA 19460 Kul 75 (72) Inventor Daniel Frank Baber United States 19002 Ambler, PA, No. 290 F-Term (reference) 4C031 MA02 4C033 AD08 AD09 AD17 AD20 4C034 AN01 4C037 QA10 4C055 AA01 BA02 BA03 BA02 BA58 BA02 CA02 CA58 CB02 DA01 4C056 AA01 AB01 AC02 AD01 AE03 BA03 BA04 BB14 BC01 4C063 AA01 BB02 CC14 CC29 CC52 CC58 CC62 CC76 DD03 DD12 EE01 4C071 AA01 BB01 CC22 EE13 FF23 HH28 LL01 4C084 AA2 ZA1A14 ZA1A17 ZA1A19 BC30 BC69 BC73 BC82 GA02 GA03 GA06 GA09 GA10 GA12 MA04 NA14 ZA67 ZA96 ZA97 ZB11 ZB15 ZC20 4C206 AA01 AA02 AA03 HA26 MA04 NA14 ZA67 ZA96 ZA97 ZB11 ZB15 ZC20 4H006 AA01 AB20 AB27

Claims (28)

[Claims] 1. Formula I: ## STR1 ##[Where R¹And R⁶Is independently C_5-6Alkyl; C_2-6Alkenyl; C_3-11Cycloa
Lequil-C_0-6Alkyl; Ar-C_0-6Alkyl; Het-C_0-6Alkyl; Ar
-C_2-6Alkenyl; Het-C_2-6Alkenyl; Het-C_2-6Alkynyl; A
r-C_2-6Alkynyl; CH (R⁷) Ar; CH (R⁷) OAr; NR⁷R⁸and
CH (R⁷) NR⁸R⁹Selected from the group consisting of: R ', R^Two, R^Three, R^Four, R^Five, R⁸, R^Ten, R¹¹And R¹⁴Is independently H; C_{1 -6} Alkyl; C_2-6Alkenyl; Ar-C_0-6Alkyl; Het-C_0-6Alkyl
And C_3-11Cycloalkyl-C_0-6R is selected from the group consisting of alkyl;⁷And R¹³Is independently H; C_1-6Alkyl; C_2-6Alkenyl; C_2-6Al
Quinyl; C_3-11Cycloalkyl-C_0-6Alkyl; Ar-C_0-6Alkyl; Ar-
C_2-6Alkenyl; Ar-C_2-6Alkynyl; Het-C_0-6Alkyl; Het-
C_2-6Alkenyl; Het-C_2-6Alkynyl; OR^Ten, SR^TenAnd NR^TenR¹¹ C optionally substituted by_1-6Alkyl; N (R ') CO_TwoR '; CO_TwoR
’; CONR^TenR¹¹And N (C = NH) NH_TwoSelected from the group consisting of: R⁷And R⁸Together form a pyrrolidine or piperidine ring,
Well; R^TenAnd R¹¹Together form a pyrrolidine, piperidine or morpholine ring
May be formed; R⁹Is H; R¹²R¹²C (O); R¹²C (S); R¹²OC (O); R¹²OC (
O) NR¹¹CH (R¹³) C (O); R¹²SO_TwoR¹²SO_TwoNR¹¹CH (R¹³) C
(O); R¹²R'NC (O); R¹²R'NCS or COCH (R¹³) NR¹⁴R ^Fifteen R¹²Is NR^TenR¹¹, C_2-6Alkenyl or C_2-6Substituted by alkynyl
C that may be_1-6Alkyl; Ar-C_0-6Alkyl; Ar-C_2-6Alkenyl
Ar-C;_2-6Alkynyl; Het-C_0-6Alkyl; Het-C_2-6Alkenyl
Het-C_2-6Alkynyl; C_1-6Alkyl, (CH_Two)_1-6CO_TwoR 'or a
C optionally substituted with damantyl_3-11R is cycloalkyl;^FifteenIs R¹²R¹²C (O); R¹²C (S); R¹²OC (O); R¹²OC (O)
NR¹¹CH (R¹³) C (O); R¹²SO_TwoR¹²SO_TwoNR¹¹CH (R¹³) C (O
); R¹²R'NC (O) or R¹²R'NCS] and pharmaceutically acceptable salts, hydrates and solvates thereof. 2. The compound according to claim 1, wherein R ² , R ³ , R ⁴ and R ⁵ are independently H. 3. R ¹ is independently: Embedded image Embedded image Embedded image Embedded image R ⁶ is independently: The compound according to claim 1, which is 4. 2- [N- (3-benzyloxybenzoyl)]-2 ′-[
N '-(N-benzyloxycarbonyl-L-valinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2'-[N '-(N-benzyloxycarbonyl-L-phenylala) 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-isoleucinyl)] carbohydrazide; 2- [N- (3- Benzyloxybenzoyl)]-2 '-[N'-(N-benzyloxycarbonyl-L-methionyl)] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-2 '-[
N '-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide; (2S) -2- [N- (N-benzyloxycarbonyl-2-aminobutyryl)]-2'-[N '-(N- Benzyloxycarbonyl-L-leucinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-lydinyl-L-leucinyl)] carbo Hydrazide; (2R) -2- [N- (3-benzyloxybenzoyl)]-2 ′-[N′-
[4-methyl-2- (3-phenylphenyl) pentanoyl] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 ′-[N ′-(N-benzyloxycarbonyl-L-arginyl- L-leucinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 '-[N'-(N-benzyloxycarbonyl-L-norvalinyl)] carbohydrazide; 2- [N- ( 3-benzyloxybenzoyl)]-2 '-[N'-(N-benzyloxycarbonyl-L-cyclohexylglycinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2 '-[ N ′-(N-benzyloxycarbonyl-L-cyclohexylalanyl)] carbohydrazide; 2,2 ′-[N, N′-bis [N- (3-nitro- , 2,7 benzoxadiazole-6-yl) -L- prolinyl -L- leucinyl]] carbohydrazide; 2- [N- (-L- N- benzyloxycarbonyl leucinyl)] - 2 '- [
N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- (NL-leucinyl) -2'-[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (3,4-dimethoxybenzoyl) -L-leucinyl]]-
2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 '-[N'-
[N- (4-trifluoromethylbenzoyl) -L-leucinyl]] carbohydrazide; 2- [N- [N- (3,4-dichlorobenzoyl) -L-leucinyl]]-2
'-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzofuran-2-ylcarbonyl-L-leucinyl)]-
2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5,6-dimethoxybenzofuran-2-ylcarbonyl)
-L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-methyl-2-phenyloxazol-4-yl) Carbonyl) -L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1 '-N
'-Methyl-2'-[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- [4-methyl-2- (4-trifluoromethylphenyl) thiazole-5] -Ylcarbonyl] -L-leucinyl]]-2 '-[N'-[3-
(2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (3-isoquinolinoyl) -L-leucinyl] -2 ′-[N
'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-chlorobenzofuran-2-ylcarbonyl) -L-leucinyl]]-2'-[N'- [3- (2-pyridinyl) phenylacetyl]]
Carbohydrazide; 2- [N- [N- (3,5-difluorobenzoyl) -L-leucinyl]]-
2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzothiazol-6-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl)]-1- (N
-Methyl) -2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-picolinoyl-L-leucinyl)]-2 '-[N'-[ 3-
(2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (2,6-dimethoxynicotinoyl) -L-leucinyl]]
-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (4-methanesulfonylbenzoyl) -L-leucinyl]]
-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 '-[N'-
[N- (3,4,5-trimethoxybenzoyl) -L-leucinyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 ′-[N′-
[N- (8-quinolinoyl) -L-leucinyl]] carbohydrazide; 2- [N- [N- [3,4- (1,3-propylenedioxy) benzoyl]-
L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [3- (2-pyridinyl) phenylacetyl]]-2 '-[ N'-
(N-thieno [2,3-b] thiophen-2-ylcarbonyl-L-leucinyl)] carbohydrazide; 2- [N- [N- (4-methoxybenzoyl) -L-leucinyl]]-2′-
[N '-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- [4-methoxy-3- [2- (4-morpholino) ethoxy]]
Benzoyl] -L-leucinyl]]-2 ′-[N ′-[3- (2-pyridinyl)
Phenylacetyl]] carbohydrazide; 2- [N- [N- [2- (4-morpholino) pyrimidin-4-ylcarbonyl] -L-leucinyl]]-2 ′-[N ′-[3- (2- Pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (2,3-dihydrobenzofuran-5-ylcarbonyl)-
L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-carboxybenzofuran-2-ylcarbonyl) -L
-Leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (3-benzyloxybenzoyl) -2 '-[N'-[N- (3-
Nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L
-Leucinyl]] carbohydrazide; 2- [N- [N- (3-nitro-1,2,7-benzoxadiazol-6-yl) -L-prolinyl-L-leucinyl]]-2- [N -[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzothiophen-2-ylcarbonyl-L-leucinyl)
] -2 '-[N'-[3- (2-oxo-2-pyridinyl) phenylacetyl]
The compound according to claim 1, which is selected from the group consisting of: carbohydrazide; and 2,2 '-[N, N'-bis [N- (3-iodo-4-methoxy) -L-leucinyl]] carbohydrazide. . 5. 5- [N- (N-benzothiophen-2-ylcarbonyl-)
L-leucinyl)]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- [N- (5-chlorobenzofuran-2-ylcarbonyl) -L- Leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]]
5. The compound of claim 4, wherein the compound is selected from the group consisting of carbohydrazide; and 2,2 '-[N, N'-bis [N- (3-iodo-4-methoxy) -L-leucinyl]] carbohydrazide. 6. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable carrier, diluent or excipient. 7. A pharmaceutical composition comprising the compound of claim 4 and a pharmaceutically acceptable carrier, diluent or excipient. 8. An effective amount for a patient in need of inhibiting a protease selected from the group consisting of cysteine protease and serine protease.
A method of inhibiting a protease selected from the group consisting of a cysteine protease and a serine protease, comprising administering the compound as described above. 9. An effective amount for a patient in need of inhibiting a protease selected from the group consisting of cysteine protease and serine protease.
A method of inhibiting a protease selected from the group consisting of a cysteine protease and a serine protease, comprising administering the compound as described above. 10. The method according to claim 8, wherein the protease is a cysteine protease. 11. The method according to claim 9, wherein the protease is a cysteine protease. 12. The cysteine protease is cathepsin K.
The described method. 13. The cysteine protease is cathepsin K.
The described method. 14. A method for treating a disease characterized by bone loss, comprising inhibiting bone loss by administering an effective amount of a compound of claim 1 to a patient in need of treatment for the disease characterized by bone loss. Treatment. 15. The method according to claim 14, wherein the disease is osteoporosis. 16. The method according to claim 14, wherein the disease is periodontitis. 17. The method according to claim 14, wherein the disease is gingivitis. 18. Inhibiting excessive cartilage or matrix destruction by administering an effective amount of a compound of claim 1 to a patient in need of treatment for a disease characterized by excessive cartilage or matrix destruction. , Treatment of diseases characterized by excessive cartilage or matrix destruction. 19. The method according to claim 18, wherein the disease is osteoarthritis. 20. The method according to claim 18, wherein the disease is rheumatoid arthritis. 21. A method for treating a disease characterized by bone loss, comprising inhibiting bone loss by administering an effective amount of a compound of claim 4 to a patient in need of treatment for the disease characterized by bone loss. Treatment. 22. The method according to claim 21, wherein the disease is osteoporosis. 23. The method according to claim 21, wherein the disease is periodontitis. 24. The method according to claim 21, wherein the disease is gingivitis. 25. Inhibiting excessive cartilage or matrix destruction by administering an effective amount of a compound of claim 4 to a patient in need of treatment for a disease characterized by excessive cartilage or matrix destruction. , Treatment of diseases characterized by excessive cartilage or matrix destruction. 26. The method according to claim 25, wherein the disease is osteoarthritis. 27. The method according to claim 25, wherein the disease is rheumatoid arthritis. 28. 2- [N- (3-benzyloxybenzoyl)]-2′-
[N '-(N-tert-butoxycarbonyl-L-leucinyl)] carbohydrazide; 2- [N- (3-benzyloxybenzoyl)]-2'-[N '-(L-leucinyl)] carbohydrazide; 2- [N- (3- benzyloxy-benzoyl)] - 2 '- [N ' - (N α - benzyloxycarbonyl -N _{epsilon-tert-butoxycarbonyl} -L- lysinyl -L- leucinyl)] carbohydrazide; 2 -[N- (3-benzyloxybenzoyl)]-2 '-[N'-(N [ _alpha ] -benzyloxycarbonyl-N [ _{phi] 2} -bis (tert-butoxycarbonyl) -L
-Lidinyl-L-leucinyl)] carbohydrazide; 2,2 '-[N, N'-bis- (N-tert-butoxycarbonyl-L-leucinyl)] carbohydrazide; 2,2'-[N, N ' -Bis- (L-leucinyl)] carbohydrazide; 2-hydroxy-4,5-dimethoxybenzaldehyde; 4,5-dimethoxy-2-ethoxycarbonylmethoxybenzaldehyde; ethyl 5,6-dimethoxybenzofuran-2-carboxylate; , 6-Dimethoxybenzofuran-2-carboxylic acid; 1- (N-benzyloxycarbonyl) -1- (N-methyl) -2- [N- [
3- (2-pyridyl) phenylacetyl]] hydrazine; 1- (N-methyl) -2- [N- [3- (2-pyridyl) phenylacetyl]
Hydrazine; 1- (N-tert-butoxycarbonyl) -1 (-N-methyl) -2- (N
-(Benzyloxycarbonyl-L-leucinyl) hydrazine; 2- (N-benzyloxycarbonyl-L-leucinyl) -1- (N-methyl) hydrazine; 3- (2-pyridyl) phenylacetylhydrazine; 2- (4- Ethyl morpholino) pyrimidine-4-carboxylate; 2- (4-morpholino) pyrimidine-4-carboxylic acid; 4-benzyloxycarbonylmethoxy-3-formylbenzaldehyde; benzyl 5-formylbenzofuran-2-carboxylate; 5-carboxy Benzyl benzofuran-2-carboxylate; benzyl 5-tert-butoxycarbonylbenzofuran-2-carboxylate; 5-tert-butoxycarbonylbenzofuran-2-carboxylic acid; 2- [N- [N- (5-tert-butoxycarbonyl) Benzofuran- 2- [yl])-L-leucinyl]]-2 '-[N'-[3- (2-pyridinyl) phenylacetyl]] carbohydrazide; 2- [N- (N-benzyloxycarbonyl-L-leucinyl) ] -2 '-[
N '-[3- (2-oxo-2-pyridinyl) phenylacetyl]] carbohydrazide; and 2- (NL-leucinyl) -2'-[N '-[3- (2-oxo-2- Pyridinyl) phenylacetyl]] carbohydrazide.