KR19990036271A - C-proteinase inhibitors used to treat diseases associated with collagen overproduction - Google Patents

Abstract

The present invention relates to new uses of organic molecules capable of inhibiting C-proteinase activity to modulate, modify and inhibit abnormal collagen formation.

Description

C-proteinase inhibitors used to treat diseases associated with collagen overproduction

Collagen Structure: To date, 19 kinds of collagen have been identified. Such collagen, including fibrillar collagen I, II, III, is synthesized from procollagen promolecules comprising amino and carboxylate peptide extensions. Such peptide extensions, called "pro-parts", are termed N- and C-propeptides, respectively.

In general, when secreting procollagen triple helix precursors from a cell, the pro-part is cleaved to produce a finished triple helix collagen molecule. Upon cleavage, "mature" collagen molecules have the ability to associate with highly structured collagen fibers (Fessler and Fessler, 1978, Annu. Rev. Biochem. 47: 129-162; Bornstein and Traub, 1979, in: The Proteins (eds. Neurath, H. and Hill, RH), Academic Press, New York, pp. 412-632; Kivirikko et al., 1984, in: Extracellular Matrix Biochemistry (eds. Piez, KA and Reddi, AH), Elsevier Science Publishing Co., Inc., New York, pp. 83-118; Prockop and Kivirikko, 1984, N. Engl. J. Med. 311: 376-383; Kuhn, 1987, in: Structure and Function of Collagen Types (eds. Mayne, R. and Burgeson, RE), Academic Press, Inc., Orlando, Florida, pp. 1-42).

Diseases associated with abnormal production of collagen: Diseases in which abnormal production or production of collagen are not controlled include pathological fibrosis or scars, including endocardiosclerosis, spontaneous interstitial fibrosis, interstitial pulmonary fibrosis, myofascial fibrosis, cymus fibrosis, central Peripheral fibrosis, hepatitis, fibrofibrosis, biliary cirrhosis, alcoholic cirrhosis, acute pulmonary fibrosis, spontaneous pulmonary fibrosis, acute respiratory fatigue syndrome, renal fibrosis / glomerulonephritis, renal fibrosis / diabetic nephropathy, scleroderma / systemic, scleroderma / Local, pedigree, hypertrophic scar, severe joint adhesion / arthritis, bone fibrosis, corneal scar, bladder fibrosis, duchenne's, cardiac fibrosis, muscle fibrosis / retinal detachment, esophageal stricture, payronles disease and the like. Fibrotic diseases also include scar correction / plastic surgery, glaucoma, cataract fibrosis, corneal scar, joint adhesion, host disease for transplantation, tendon surgery, nerve fixation, dupuytren buildup, OB / GYN adhesion / fibrosis, pelvic adhesion, duodenal fibrosis, re It may be induced or initiated by surgery, including stenosis and the like. One way to treat such a disease is to prevent the pathological overproduction of collagen. Thus, a major concern of the medical community is in the identification and isolation of molecules that regulate, inhibit or modulate the production of collagen.

Relationship between collagen formation and C-proteinases: According to recent studies, C-proteinase is a fundamental essential enzyme that catalyzes the cleavage of C-propeptides of fibrillar collagen, including collagen types I, II and III. See U.S. Application 60 / 002,038 (August 8, 1995).

C-proteinase was first identified in culture media of human and mouse fibroblasts (Goldberg et al., 1975, Cell 4: 45-50; Kessler and Goldberg, 1978, Anal. Biochem. 86: 463-469) , and chick tendon fibroblasts (Duskin et al., 1978, Arch. Biochem. Biophys. 185: 326-332; Leung et al., 1979, J. Biol. Chem. 254: 224-232). Acidic proteases have also been identified that remove C-end propeptide from type I procollagen (Davidson et al., 1979, Eur. J. Biochem. 100: 551).

In 1982, some purified proteins with C-proteinase activity were obtained from chick cranial tubes (Njieha et al., 1982, Biochemistry 23: 757-764.).

In 1985, chick C-proteinase was isolated and purified from the conditioned medium of chick embryos and characterized (Hojima et al., 1985, J. Biol. Chem. 260: 15996-16003). Murine C-proteinase was purified from the culture of cultured mouse fibroblasts (Kessler et al., 1986, Collagen Relat. Res. 6: 249-266; Kessler and Adar, 1989, Eur. J. Biochem. 186: 115-121). Finally, cDNA encoding human C-proteinase was identified as set forth in the US related application presented above.

Experiments in tablet form of chick and mouse C-proteinases have shown that enzymes are essential for the formation of functional collagen fibers (Fertala et al., 1994, J. Biol. Chem. 269: 11584).

C-proteinase inhibitors: Following the conclusion that enzymes play an important role in collagen production, scientists have identified a number of C-proteinase inhibitors (Hojima et al., Supra). For example, several metal chelates have been described as C-proteinase inhibitors. Similarly, chymostatin and pepstatin A have been found to be relatively potent C-proteinase inhibitors. In addition, α ₂ -macroglobulin, ovostatin and fetal bovine serum have been shown to partially inhibit C-proteinase activity.

Dithiotretol, SDS, Concanavalin A, Zn ²⁺ , Cu ²⁺ and Cd ²⁺ are reported to have inhibitory activity even at low concentrations. Similarly, some reducing agents, some amino acids, phosphates and ammonium sulfate were shown to have inhibitory activity at concentrations of 1-10 mM. In addition, enzymes have been shown to be inhibited by basic amino acids lysine and arginine (Leung et al., Supra; Ryhanen et al., 1982, Arch. Biochem. Biophys. 215: 230-236). Finally, high concentrations of NaCl or Tris-HCl buffer have been found to interfere with C-proteinase activity. For example, the activity of C-proteinase was reduced by 66, 38 and 25% by 0.2, 0.3, 0.5M NaCl, respectively. Activity is rapidly inhibited by Tris-HCl buffer at a concentration of 0.2-0.5 M (Hojima et al., Supra). In contrast, microbial inhibitors such as lupeptin, phosphoramidone, antipine, bestatin, elastinal and amazatin had little or no effect on the activity of C-proteinase.

Various assays were used to determine C-proteinase activity and its inhibition (kessler and Goldberg, 1978, Anal. Biochem. 86: 463; Njieha et al., 1982, Biochemistry 21: 757-764). Despite the usefulness of these tests, large-scale testing and testing of potential C-proteinase inhibitors could not be performed because of limitations in the use of human C-proteinases. As described in many literatures, enzymes are difficult to isolate by conventional biochemical means, and cDNA sequences encoding such enzymes have not yet been reported.

Development of compounds that inhibit C-proteinase activity: In view of its fundamental role in the formation and maturation of collagen, C-proteinase is an ideal target for treating diseases associated with improper or unregulated production of collagen. However, none of the inhibitors identified to date have demonstrated a therapeutic effect in the treatment of diseases associated with collagen or in inhibiting C-proteinase activity.

It is an object of the present invention to identify effective compounds that specifically inhibit C-proteinases in order to modulate or modify the production of inappropriate collagen.

3. Summary of the Invention

The present invention relates to organic molecules that can affect C-proteinase activity to regulate, modify and inhibit collagen production. In particular, the compounds of the present invention are as follows.

a. Inhibitor A

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroallyl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxy Alkyl, acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulf Phonyl) -alkyl;

R ₂ is selected from H, lower alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl;

R ₄ is selected from aryl, heteroaryl, alkyl, aralkyl, heteroaralkyl, alkylamino, arylalkylamino,

X is selected from SO ₂ , C═O;

Y is selected from OH, HOHN (hydroxyamine), H ₂ N, alkylamino;

Z is directly bonded at methylene, oxygen, sulfur, amino;

n is 0 or 1; or

b. Inhibitor B

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl , Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl ) -Alkyl;

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or

c. Inhibitor C

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become;

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or

d. Inhibitor D

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become;

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or

e. Inhibitor E

Wherein R ₁ is selected from OH, alkoxyl, lower alkyl, alkylamino, peptide,

X is selected from N, C,

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl.

The present invention relates to a pharmaceutical composition consisting of a pharmaceutically effective amount of the compound and a pharmaceutically acceptable carrier or excipient. Such compositions inhibit C-proteinase activity to regulate production and maturation of collagen.

The present invention relates to the use of such compounds and compositions for modifying, inhibiting, and modulating C-proteinase activity to treat diseases associated with abnormal production of collagen.

More specifically, the compounds of the present invention include methods for treating diseases including but not limited to rheumatoid arthritis, scleroderma, pathological fibrosis or scars.

4. Definition

"C-proteinase" refers to an enzyme capable of treating collagen molecules, derivatives or fragments thereof, or precursors thereof by cleavage through -Ala ↓ Asp-Asp- and / or -Gly ↓ Asp-Glu-. The term includes human C-proteinases and derivatives, analogs, fragments and variants thereof having C-proteinase activity.

"Pharmaceutically acceptable salts" have the properties and biological effects of free acids, with sodium hydroxide, magnesium hydroxide, ammonia, trialkylamines, dialkylamines, monoalkylamines, dibasic amino acids, acetate sodium, benzoate potassium, triethanolamine It refers to a salt that can be obtained by reaction with an inorganic or organic base such as.

"Alkyl" refers to saturated aliphatic hydrocarbons comprising straight, branched and cyclic alkyl groups. Suitably the alkyl group has 1 to 12 carbons. More preferably, it is a low cost alkyl having 1 to 7 carbons, suitably 1 to 4 carbons. Typical alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, quaternary butyl, pentyl, hexyl and the like. Alkyl groups may be substituted or unsubstituted. Substituted groups are suitably carboxyl, hydroxyl, mercapto, cycloalkyl, heterocycloalkyl, halo, alkoxyl, alkylamino.

"Aryl" refers to an aromatic group having at least one ring having a conjugated pi electron system, which includes carbocyclic aryl, heterocyclic aryl, and biaryl groups, all of which may be optionally substituted. Suitably aryl is substituted or unsubstituted phenyl or pyridyl. Suitable aryl substituents are halogen, trihalomethyl, hydroxyl, SH, NO ₂ , amine, iether, cyano, alkoxy and the like for phenyl or pyridyl.

This application is a continuous application of US Application 08 / 601,203 filed February 14, 1996 entitled "C-Protease Inhibitors Used in the Treatment of Diseases Associated with Collagen Overproduction". And this is a serial application of US application 60 / 002,038, filed August 8, 1995.

1. Field of Invention

Collagen enters the titration of connective tissue. Thus, many connective tissue diseases and disorders result from excessive or underproduction of collagen or abnormal production of collagen (including poorly treated collagen). C-proteinases are key enzymes essential for the proper maturation of collagen and thus are ideal targets for inhibition, regulation or modulation of collagen formation.

The present invention relates to organic molecules that can interfere with C-proteinase activity to modulate, modulate or inhibit abnormal collagen formation. More specifically, the present invention relates to the use of compounds and pharmaceutical compositions to treat various diseases associated with the production of inappropriate or uncontrolled collagen.

The present invention relates to compounds capable of modulating the formation of collagen by interfering with C-proteinase activity.

More specifically, the present invention relates to a compound which inhibits C-proteinase activity in a method of treating or modulating various connective tissue diseases including fibrotic disease, arthritis disease or surgically induced or disclosed diseases. Related.

5.1. compound

The present invention generally relates to compounds having the formula:

a. Inhibitor A

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxy Alkyl, acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulf Phonyl) -alkyl;

R ₂ is selected from H, lower alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl;

R ₄ is selected from aryl, heteroaryl, alkyl, aralkyl, heteroaralkyl, alkylamino, arylalkylamino,

X is selected from SO ₂ , C═O;

Y is selected from OH, HOHN (hydroxyamine), H ₂ N, alkylamino;

Z is directly bonded at methylene, oxygen, sulfur, amino;

n is 0 or 1;

b. Inhibitor B

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl , Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl ) -Alkyl;

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or

c. Inhibitor C

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become;

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or

d. Inhibitor D

Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become;

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or

e. Inhibitor E

Wherein R ₁ is selected from OH, alkoxyl, lower alkyl, alkylamino, peptide,

X is selected from N, C,

R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl;

R ₄ is selected from H, lower alkyl;

R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl;

In an embodiment of the invention, the compound of the present invention has the formula:

And pharmaceutically acceptable salts thereof.

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

One

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof;

or

And pharmaceutically acceptable salts thereof.

The chemical formula referred to here represents the phenomenon of totomary. Since the formulas in the specification represent only one of the possible totomeric forms, it is to be understood that the present invention also encompasses other totomeric forms that have the ability to inhibit C-proteinase activity to regulate collagen production and maturation. In addition, it should be understood that the present invention also encompasses all isomers of the respective compounds detailed herein.

In addition to the compounds and their pharmaceutically acceptable salts, the present invention also provides a dissolved form of a compound having the ability to inhibit C-proteinase activity to modulate, inhibit or modify the production and maturation of collagen, or It relates to the undissolved form (eg hydrated form).

The aforementioned compounds may be prepared by any process known to be used for the preparation of chemically related compounds. The titration process is described in the following examples. Essential starting materials can be obtained by standard processes of organic chemistry.

As the substances affecting C-proteinase activity, the related activities and effects of each compound are determined using the available techniques. Suitably, the compound undergoes a series of screening processes to determine the compound's ability to modulate, modify and inhibit the production and maturation of collagen. Such screening includes biochemistry, cell culture, and animal models.

The compounds of the present invention can be synthesized according to known techniques. Provided are methods for synthesizing and testing the compounds of the claimed invention.

6.1. Example 1: Compound Synthesis

6.1.1. Synthesis of N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-chlorobenyl)] amino] -acetamide.

N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N' (4-chlorobenzyl)] amino] -acetamide (5) designated FG047 (Inhibitor A) Certain compounds in the specification are referred to as "C #" for naming, where "#" is an Arabic numeral).

Synthesis of ethyl-2 [(4-chlorobenzyl) amino] -acetate (3).

Anhydrous ZnCl ₂ solid (75 mg) in a cooled solution of glycine ethyl ester hydrogen chloride (1) (1.00 g, 7.16 mmol) and 4-chlorobenzylaldehyde (2) (1.00 g, 7.16 mmol) in anhydrous MeOH (10 mL). , 0.55 mmol) and NaBCNH ₃ solid (0.45 g, 7.16 mmol) are added. After stirring for 18 hours at room temperature, the reaction mixture is cooled with 1N HCl (20 mL) aqueous solution and stirred for an additional 30 minutes. The mixture is concentrated in a rotary evaporator to remove most of the MeOH solvent and extract with ether (20 mL). The aqueous layer is basified to pH 10 with 45% (w / w) KOH aqueous solution in an ice bath and extracted with EtOAc (2 × 50 mL). The combined EtOAc organic layer is washed with salt, dried over Na ₂ SO ₄ and concentrated. The product is purified by silica gel chromatography (4/1 hexanes / EtOAc) to give ethyl-2 [(2-chlorobenzyl) amino] -acetate (3) in oil form.

Synthesis of ethyl-2-[[N- (4-methoxybenzene-sulfonyl) -N- (4-chlorobenzyl)] amino] acetate (4).

Ethyl-2 [(4-chlorobenzyl) amino] -acetate (3) (600 mg, 2.64 mmol) and p-methoxybenzenesulfonyl chloride (545 mg, 2.64 m) in anhydrous CH ₂ Cl ₂ (7 mL). Mol) triethylamine (294 mg, 2.90 mmol) was added dropwise to the solution. The mixture is stirred at rt for 15 h and treated with 1N HCl (20 mL) solution. The two phases produced are separated and the aqueous layer is extracted with CH ₂ Cl ₂ . The combined organic layers are washed with brine, dried over MgSO ₄ and concentrated. The product was purified by silica gel chromatography (3/1 to 2/1 hexanes / EtOAc) to give ethyl-2-[[N- (4-methoxybenzene-sulfonyl) -N- (4-chlorobenzyl) in oil form. )]-Amino] acetate (4) is obtained.

Synthesis of N-hydroxy-2 [[N '-(4-methoxy-benzenesulfonyl) -N' (4-chlorobenzyl)] amino] -acetaamide (5).

A separate solution of hydrogen chloride hydroxyamine (171 mg, 2.46 mmol) in MeOH (1.3 mL) and KOH (207 mg, 3.69 mmol) in MeOH (1.3 mL) was prepared at boiling point, cooled to 40 ° C. and the latter Add the solution to the electronic solution. After cooling the reaction mixture in an ice bath for 30 minutes, the calcium chloride solid is removed by filtration. To the filtrate is added ethyl ester (4) (487 mg, 1.23 mmol). After stirring for 6 hours at room temperature, the reaction mixture is treated with 1N HCl (20 mL) solution and extracted with CH ₂ Cl ₂ . The organic layer is washed with brine, dried over MgSO ₄ and concentrated to give the product. The residue was triturated with ether and a white solid was obtained which gave N-hydroxy-2 [[N '-(4-methoxy-benzenesulfonyl) -N' (4-chloro designated as hydroxyxamate or FG-047). Benzyl)]-amino] -acetamide (5).

mp: 124-125 ° C .; MS (ES) (M + H) ⁺ : 385; 1 H NMR (360 MHz, DMSO-d 6) δ 10.47 (s, 1H, OH), 8.81 (s, 1H, NH), 7.81-7.08 (m, 8H, Ph), 4.34 (s, 2H, CH ₂ CO) , 3.85 (s, 3H, OMe), 3.63 (s, 2H, CH ₂ Ph).

6.1.2. Synthesis of N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-fluorobenzyl)] amino] -acetamide.

N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-fluorobenzyl)] amino] -acetamide (6) designated as FG053 (Inhibitor A) A suitable method of synthesis is N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N' (4-chloro using 4-chlorobenzaldehyde dash in 4-chlorobenzaldehyde dash as starting material. Benzyl)] amino] -acetamide is essentially identical to that described above (paragraph 6.1.1).

MS (ES) (M-1): 367; ¹ H NMR (360 MHz, DMSO-d6) δ 10.45 (s, 1H, OH), 8.83 (s, 1H, NH), 7.81-7.08 (m, 8H, Ph), 4.33 (s, 2H, CH ₂ CO ), 3.85 (s, 3H, OMe), 3.62 (s, 2H, CH ₂ Ph).

6.1.3. Synthesis of Hydroxamate.

Suitable methods for synthesizing the hydroxamate 11 (inhibitor B) are as follows.

Tetraester (8) is provided by alkylation of tertiary butyl bromoacetate with malon ester (7). In the saponification and decarboxylation reactions, the resulting adjuvant (9) is combined with Glu (OBn) NHMe using a peptide binding reaction to obtain compound (10). After successive hydrolysis, compound 10 is converted to the desired hydroxamate 11 by anhydride formation and addition of NH ₂ OH.

6.1.4. Synthesis of N-carboxymethyl digapeptide.

A suitable method of synthesizing the N-carboxymethyl divalent peptide 16 designated as FG057 (inhibitor D) is as follows.

Block compounds (12) and (13), which can be used according to the procedures in the literature, are combined by the conventional DCC / HOBt method to obtain the divalent peptide (14) with a yield of 78%. Deprotection of compound (14) with TFA / CH ₂ Cl ₂ and alkylation with benzyl bromoacetate in the presence of NMM result in the formation of N-carboxymethyl divalent peptide (15), which is catalyzed by hydrogenation with a catalyst. The free acid 16 is converted.

Boc-Glu (OBn) OH and Boc-Asp (OBn) OH were synthesized using methods known in the art.

Synthesis of Boc-Glu (OBn) NHMe (13).

Triethylamine (2.0 g, 20 mmol) is added to a solution of Boc-Glu (OBn) OH (6.7 g, 20 mmol) in anhydrous THF (100 mL). The solution is cooled to −78 ° C. under argon, and dropwise with ethyl chloroformate (2.2 g, 20 mmol). The reaction mixture is raised to −30 ° C. for at least 2 hours, 40% aqueous methyl amine solution (22 mmol) is added, and the reaction mixture is brought to room temperature. The reaction was stirred for an additional 4 hours before adding diethyl ether (50 mL) and water (70 mL). The organic layer is separated, washed with 1M NaHCO ₃ , 10% citric acid, saturated NaCl solution and dried over MgSO ₄ . The solvent is evaporated in vacuo to give a white solid (5.0 g, 72% yield).

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.38 (s, 9H, CH ₃ ); 1.85-2.59 (m, 4H, CH ₂ ); 2.75 (d, 3H, NCH ₃ ); 4.17 (m, 1 H, CH); 5.08 (δ, 2H, OCH ₂ ); 5.33 (bδ, 1 H, NH); 6.30 (bs, 1 H, NH); 7.31 (s, 5 H, Ph-H).

Synthesis of Boc-Asp (OBn) -Glu (OBn) NHMe (14).

1.5 ml TFA is added to a tert-butoxycarbonyl aminoester (13) (700 mg, 2 mmol) solution in 10 ml CH ₂ Cl ₂ , and the reaction mixture is stirred at room temperature under argon for 1 hour. The excess acid is evaporated in vacuo and the residue is treated several times with diethyl ether and concentrated under reduced pressure to give a colorless oil which is not further purified. TFA-salt, Boc-Asp (OBn) OH (446 mg, 2 mmol), HOBt (170 mg, 2 mmol) and NMM (202 mg, ₂ mmol) / CH ₂ Cl ₂ (10 mL) were added and the reaction Is stirred under argon gas for 12 hours at room temperature. The dicyclourea precipitate is removed by filtration and the filtrate is concentrated under reduced pressure. The residue is diluted with EtOAc (30 mL), filtered again and washed with 1M NaHCO ₂ , 10% citric acid, saturated NaCl solution. The organic layer is dried over MgSO ₄ , concentrated in vacuo and a white solid (78% yield 867 mg) is obtained.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.41 (s, 9H, CH ₃ ); 1.85-3.05 (m, 9H, 3CH ₂ , NCH ₃ ); 4.40 (m, 2 H, CH); 5.01 (m, 4H, OCH ₂ ); 5.53 (d, 1 H, NH); 6.48 (bs, 1 H, NH); 7.15-7.38 (m, 10H, Ph-H).

Synthesis of N-carboxymethyl-dipeptide (15).

1 ml TFA is added to a solution of Boc-Asp (OBn) -Glu (OBn) NHMe (14) (555 mg, 1 mmol) in 10 ml CH ₂ Cl ₂ and the reaction mixture is stirred at room temperature under argon for 1 hour. The excess acid is evaporated in vacuo and the residue is treated several times with diethyl ether and concentrated under reduced pressure to give a colorless oil which is dissolved in anhydrous THF (20 mL). NMM (101 mg, 1 mmol) and benzyl bromoacetate (230 mg, 1 mmol) are added to this solution, and the reaction is stirred under argon gas for 12 hours at room temperature. The reaction is concentrated under reduced pressure. The residue is diluted with EtOAc (20 mL), filtered again and washed with 1M NaHCO ₂ , 10% citric acid, saturated NaCl solution. The organic layer was dried over MgSO ₄ , concentrated, and chromatographed on silica gel (ethyl acetate / MeOH 10: 1) to yield 88% of a white solid compound (15) (410 mg).

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.83-2.95 (m, 9H, 3CH ₂ , NCH ₃ ); 3.30-3.60 (m, 4H, CH & N CH ₂ ); 4.39 (m, 2 H, CH); 5.05 (m, 6H, OCH ₂ ); 6.64 (bs, 1 H, NH); 7.29- (m, 15H, Ph-H). 8.02 (bs.-1 H, NH); ¹³ C-NMR (60 MHz, CDCl ₃ ); δ 26.17, 27.24, 30.59, 36.57 (3CH ₂ , NCH ₃ ); 49.59 (NCH ₂ ); 52.54, 59.07 (2CH); 66.46, 66.71, 66.86 (3OCH); 128.17, 128.26, 128.34, 128.55, 128.80, 135.27, 135.44, 135.79 (Ph-C); 171.25, 171.34, 172.01, 172,61, 173.05 (5C = O).

Synthesis of N-carboxymethyl-Asp-Glu-NMe (16).

To a portion of the benzyl-protected N-carboxymethyl dipeptide (15) (68 mg, 0.113 mmol) in methanol (5 mL) was added Pd / C powder (50 mg) in portions. The mixture is stirred for 20 hours at room temperature under H ₂ atmosphere (gut pressure). The catalyst is filtered through a pad of celite and rinsed with methanol. The filtrate was concentrated and the solid was re-blooded from EtOAc / MeOH at −20 ° C. to give the product N-carboxymethyl-Asp-Glu-NMe (16) (23 mg, 0.065 mmol) (white solid, 58% yield). Get

mp: 147-149 ° C .; NMR (360 MHz, DMSO-d6) δ8.19 (d, J = 8.5 Hz, 1H), 7.72 (m, 1H), 4.20 (m, 1H), 3.45-3.20 (m, 4H), 2.63-2.44 ( m, 5H), 2.20 (m, 2H), 1.93 (m, 1H), 1.72 (m, 1H).

6.1.5. Synthesis of Mercapto Compounds.

The titration method for synthesizing the mercapto compound (22) designated as FG-074 (inhibitor C) is as follows.

Tert-butyl bromoacetate and diethyl phosphate (17) are alkylated under NaH to give 90% yield of phosphate (18). Compound (18) is reacted with formaldehyde and Horner-Emmons using K ₂ CO ₃ as a base to form unsaturated ester (19). The saponification reaction between LIOH and compound (19) is carried out and Michael is added to obtain acid (20). The acid (20) is combined with Glu (OBn) NHMe using the DCC / HOBt method to give an isomeric mixture (ipeptide 21) by flash chromatography. Monovalent acid 22 is obtained by deprotection of divalent peptide 21 with TFA in CH ₂ Cl ₂ .

Synthesis of 1-ethyl-2-diethylphosphono-succinic acid-4-tert-butyl ester (18).

To anhydrous THF (80 mL) was added dropwise a triethyl phosphonoacetate (4.48 g, 20 mmol) solution to THF (20 mL) in a suspension of NaH (0.48 g, 20 mmol), and THF (20 mL) after 3 hours at room temperature. Tert-butyl bromoacetate is added dropwise. The resulting suspension is stirred for 3 hours at room temperature, water (50 mL) is added and the reaction mixture is acidified to pH 3 with 1M HCl. After addition of diethyl ether (70 mL), the organic layer is separated, washed with saturated NaCl solution, dried over MgSO ₄ and the solvent is evaporated in vacuo to give a colorless oil (90% yield).

Synthesis of 1-ethyl-2-methylene-succinic acid-4-tert-butyl ester (19).

The mixture (18) (5.4 g, 16 mmol), K ₂ CO ₃ (6.9 g, 50 mmol) and formaldehyde (3.2 g, 10 mmol) 30% aqueous solution mixture is refluxed for 3 hours. After cooling, the mixture is extracted with hexane and the organic layer is washed with water and brine and dried over MgSO ₄ . After evaporation under filtration and vacuum, an oil residue is obtained (yield 80%, 2.7 g).

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.29 (t, J = 7 Hz, 3H, CH ₂ ); 1.43 (s, 9 H, CH ₃ ); 3.24 (s, 2 H, CH ₂ ); 4.20 (q, J = Hz, 2H, OCH ₂ ); 5.63, 6.28 (s, 2H, = CH ₂ ).

Synthesis of 2-acetylsulfanylmethyl-succinic acid-4-tert-butyl ester (20).

Unsaturated ester 19 (2.3 g, 11 mmol) is dissolved in THF (50 mL) and 0.2 M aqueous solution of LIOH (264 mg, 11 mmol) is added at 0 ° C. The mixture is brought to room temperature and stirred for an additional 3 hours. The organic solvent is evaporated and the aqueous layer is extracted with diethyl ether (30 mL). The aqueous layer is acidified to pH 3 with 1M HCl and extracted with EtOAc. The organic layer is washed with water and brine and dried over MgSO ₄ . After filtration and evaporation of the solvent, a white solid acid is obtained (yield 60%).

¹ H-NMR (200 MHz, CDCl ₃ ); δ 1.44 (s, 9H, CH ₃ ); 3.26 (s, 2 H, CH ₂ ); 5.78, 6.42 (s, 2H, = CH ₂ ); 10.01 (bs, 1 H, COOH).

Acid (1.0 g, 5.4 mmol) is dissolved in CHCl ₃ and thioacetic acid (1.45 g, 19 mmol) is added. The mixture is stirred at 60 ° C. for 49 h. The solvent is evaporated in vacuo and compound (20) is obtained as a colorless oil.

¹ H-NMR (200 MHz, CDCl ₃ ); δ 1.44 (s, 9H, CH ₃ ); 2.34 (s, 3 H, CH ₃ ); 2.45-2.75 (m, 2H, CH ₂ ); 2.96-3.33 (m, 3H, CH ₂ , CH); 9.74 (bs, 1 H, COOH).

Synthesis of 3-acetylthiomethyl-4-oxo-5-aza-6- (R) -methylcarbamoyl-8-benzyloxycarbonyl-octanoic acid, tert-butyl ester (21).

To a solution of tert-butoxycarbonyl amino ester (13) (700 mg, 2 mmol) in 10 mL CH ₂ Cl ₂ was added 1 mL TFA, and the reaction mixture was stirred at room temperature under argon for 1 hour. The excess acid is evaporated under vacuum, the residue is treated several times with diethyl ether (15 mL) and concentrated under reduced pressure to give a colorless oil which can be used without further purification. TFA-salt, acid (20) (525 mg, 2 mmol), HOBt (170 mg, 2 mmol) and NMM (202 mg, 2 mmol) are dissolved in CH ₂ Cl ₂ (40 mL). To the CH ₂ CH ₂ (10 mL) was added a DCC (412 mg, 2 mmol) solution and the reaction was stirred for 12 h at room temperature under argon. The dicyclohexylurea precipitate is removed by filtration and the filtrate is concentrated under reduced pressure. The residue is diluted with EtOAc (30 mL), filtered again and washed with 1M NaHCO ₃ , 10% citric acid and saturated NaCl solution. The organic layer is dried over MgSO ₄ and concentrated in vacuo to give a colorless oil (21) which is purified by silica gel chromatography using EtOAc as eluent. Yield as an isomer mixture is 72% (710 mg). ¹ H-NMR (200 MHz, CDCl ₃ ); δ 1.36, 1.39 (2s, 9H, CH ₃ ); 1.87-3.2 (m, 14H, 4CH ₃ , CH ₃ , NCH ₃ ); 4.40 (m, 2 H, CH); 5.08, 5.11 (2s, 2H, OCH ₂ ); 6.43, 6.76, 7.07 (bs, -2H, NH); 6.76 (2d, 1 H, NH); 7.31 (m, 15 H, Ph-H).

Synthesis of 3-thiomethyl-4-oxo-5-aza-6- (R) -methylcarbamoyl-8-carboxy-octanoic acid (22) (FG 074).

TFA (1.5 mL) was added to a tert-butoxycarbonyl amino ester (21) (495 mg, 1 mmol) solution in 15 mL CH ₂ Cl ₂ , and the reaction mixture was stirred for 1 h at room temperature under argon gas. . The solvent and excess acid are evaporated under vacuum, the residue is treated several times with diethyl ether (10 ml) and concentrated under reduced pressure to give a colorless oil which is instantaneous in silica gel (ethyl acetate / MeOH 10: 1; with 1% acetic acid) Purification by chromatography yields the intermediate product, a white solid (84% yield 370 mg).

To a solution of intermediate (30 mg, 0.07 mmol) in 1 mL (1.5 / 1) methanol / H ₂ O at room temperature is added LiOH.H ₂ O (12 mg, 0.27 mmol). After stirring for 3 hours at room temperature the reaction mixture is cooled with 0.5 mL 1N NaOH aqueous solution and extracted with EtOAc (2 × 10 mL). The combined organic layer was washed with brine, dried over MgSO ₄ and concentrated to give thio compound (22) as a gum solid (19 mg, 0.06 mmol) (yield 86%).

MS (ES) (M + H) ⁺ : 307

6.1.6. Synthesis of 2-[[N- (4-methoxybenzenesulfonyl) -N- (4-chlorobenzyl)] amino] acetic acid.

A suitable method for synthesizing 2-[[N- (4-methoxybenzenesulfonyl) -N- (4-chlorobenzyl)]-amino] acetic acid (23) designated FG046 (inhibitor A) is as follows.

Synthesis of 2-[[N- (4-methoxybenzenesulfonyl) -N- (4-chlorobenzyl)] amino] acetic acid (23).

LiOH / H ₂ O is added to ethyl ester suspension mixture 4 (300 mg, 0.75 mmol) in 1.5: 1 MeOH / H ₂ O (4 mL). After stirring at room temperature for 5 hours, the mixture is cooled with 1N HCl solution (20 mL) and extracted with CH ₂ Cl ₂ (2 × 20 mL). The combined organic layer is washed with brine, dried over MgSO ₄ and concentrated. The solid is recrystallized from hot ether to give a white solid (23).

mp: 139.5-140 ° C .; ¹ H NMR (360 MHz, DMSO-d6) δ 7.79-7.08 (m, 4H, Ph), 4.37 (s, 2H, CH ₂ ), 3.85 (s, 3H, OCH ₃ ), 3.83 (s, 2H, CH ₂ ).

6.1.7. Synthesis of N-hydroxy-2 [[N '-(4-methoxybenzenesulfonyl) -N' (carboxymethyl)] amino] acetaamide.

A suitable method for synthesizing N-hydroxy-2 [[N '-(4-methoxybenzenesulfonyl) -N' (carboxymethyl)] amino] acetaamide (30), called FG055 (inhibitor A) As follows.

Synthesis of ethyl 2-[[N- (4-methoxybenzene-sulfonyl)] amino] acetate (26).

Triethylamine in a mixture of glycine ethyl ester hydrogen chloride (24) (3.0 g, 21.5 mmol) and 4-methoxybenzenesulfonyl chloride (25) (4.4 g, 21.3 mmol) in anhydrous CH ₂ Cl ₂ (60 mL). (4.79 g, 47.3 mmol) is added. After stirring at room temperature for 15 hours, the reaction mixture is cooled with 1N HCl (120 mL) and extracted with CH ₂ Cl ₂ (2 × 100 mL). The combined organic layer is washed with brine, dried over MgSO ₄ and concentrated. The solid is recrystallized in EtOAc / hexanes to give a white solid sulfonamide (26).

Synthesis of ethyl-2-[[N- (4-methoxybenzene-sulfonyl) -N-tert-butyloxycarbonylmethyl)] amino] acetate (27).

To a slurry of hydride sodium (60% dispersed in mineral oil) (162 mg, 4.03 mmol) in anhydrous THF (10 mL) was added sulfonamide (26) (1.0 g, 3.66 mmol) in an ice bath, Parent tert-butyl acetate (785 mg, 4.03 mmol) is added. The mixture is vigorously stirred at 0 ° C. for 30 minutes and then for 15 hours. After cooling the reaction mixture in an ice bath, it was quenched with water (25 mL) and extracted with ether (2 × 50 mL). The combined organic layer is washed with brine, dried over MgSO ₄ and concentrated. The residue is purified using silica gel chromatography (2: 1 hexanes: EtOAc) to give a colorless syrup (27).

Synthesis of N-hydroxy-2 [[N'-4-methoxybenzenesulfonyl) -N '-(tert-butyloxycarbonyl-methyl)] amino] acetaamide (28).

Prepare a separate solution of hydrogen chloride hydroxylamine (377 mg, 5.43 mmol) in MeOH (2.7 mL) and KOH (456 mg, 8.13 mmol) in MeOH (2.7 mL) at room temperature, cool to 40 ° C, and use the latter solution. Is added to the electronic solution. After cooling in an ice bath for 30 minutes, the potassium chloride solids are removed by filtration. To the filtrate is added ethyl ester 27 (1.05 g, 2.71 mmol). After stirring for 6 hours at room temperature the reaction mixture is neutralized to pH 4 with 1N HCl solution and then partitioned between CH ₂ Cl ₂ (60 mL) and water (20 mL). The resulting two phases are separated and the aqueous layer is extracted with CH ₂ Cl ₂ (60 mL). The combined organic layers are washed with brine, dried over MgSO ₄ and concentrated. The residue is purified by silica gel chromatography (11: 1 CH ₂ Cl ₂ : MeOH) to give ester hydroxamate (28) which is a white solid, FG-058.

Synthesis of N-hydroxy-2 [[N '-(4-methoxybenzenesulfonyl) -N' (carboxymethyl)]-amino] acetamide (29).

A tertiary butyl ester hydroxamate (28) solution (520 mg, mmol) in 35% CF ₃ COOH / CH ₂ Cl ₂ (9 mL) was stirred at 0 ° C. for 10 minutes and at room temperature for 1.5 hours. The mixture is concentrated and dried in vacuo. The residue is humidified with EtOAc and the solid is recrystallized from EtOAc / MeOH / hexanes to give the acid hydroxamate (29) as a white solid.

mp: 160-161 ° C .; MS (ES) (M + H) ⁺ : 319; ¹ H NMR (360 MHz, DMSO-d 6) δ 12.08 (brs. 1 H, CO ₂ H), 10.69 (s, 1H, OH), 8.96 (s, 1H, NH), 7.76 (d, J = 8.7 Hz , 2H, Pb), 7.09 (d, J = 8.7 Hz, 2H, Ph), 4.01 (s, 2H, CH ₂ ), 3.84 (s, 3H, OMe), 3.83 (s, 2H, CH ₂ ).

6.1.8. Synthesis of N- (4-methoxybenzene-sulfonyl) -L-proline hydroxamate.

A suitable method for synthesizing N- (4-methoxybenzenesulfonyl) -L-proline hydroxamate designated FG054 (inhibitor A) is as follows.

Synthesis of N- (4-methoxybenzenesulfonyl) -L-proline methyl ester.

L-proline methyl ester hydrochloride (30) (1.00 g, 6.03 mmol) in anhydrous CH ₂ Cl ₂ (17 mL) and 4-methoxybenzenesulfonyl chloride (31) (1.19 g, 5.75 mmol) Ethylamine (1.22 g, 12.06 mmol) is added. After stirring at room temperature for 15 hours, the reaction mixture is cooled with 1N HCl (30 mL) in aqueous solution and extracted with EtOAc (2 × 100 mL). The combined EtOAc organic layer is washed with brine, dried over Mg ₂ SO ₄ and concentrated to give (32). This product is used directly in the next reaction without further purification.

Synthesis of N- (4-methoxybenzenesulfonyl) -L-proline hydroxamate (33).

Prepare a separate solution of hydrogen chloride hydroxylamine (465 mg, 6.68 mmol) in MeOH (3.4 mL) and KOH (561 mg, 10.0 mmol) in MeOH (3.4 mL) at boiling point and then cool to 40 ° C. The latter solution is added to the former. After cooling in an ice bath for 30 minutes, the potassium chloride solids are removed by filtration. To the filtrate is added ethyl ester 32 (1.0 g, 3.34 mmol). After stirring at room temperature for 15 hours, the reaction mixture is treated with 1N HCl (40 mL) solution and extracted with CH ₂ Cl ₂ / MeOH (10: 1). The organic layer is washed with brine, dried over MgSO ₄ and concentrated to yield the product. The solid is recrystallized in hot MeOH / EtOAc to give the hydroxamate (33) as a solid.

6.1.9. Synthesis of N- (hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-trifluoromethylbenzyl)] amino] -acetamide.

N- (hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-trifluoromethylbenzyl)] amino] -aceta designated FG-066 (inhibitor A) A suitable method for synthesizing the mead is to use N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl)-by using tritrimethylmethylbenzaaldehyde instead of 4-chlorobenzenealdehyde. It is basically the same as synthesizing N '-(4-chlorobenzyl)] amino] -acetamide (see 6.1.1.).

¹ H NMR (360 MHz, DMSO-d 6) δ 10.50 (s, 1H, OH), 8.83 (s, 1H, NH), 7.81-7.08 (m, 8H, Ph), 4.45 (s, 2H, CH ₂ CO), 3.86 (s, 3H, OMe), 3.67 (s, 2H, CH ₂ Ph).

6.1.10. Synthesis of N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-methoxybenzyl) 'amino] -acetamide.

Synthesis of N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-methoxybenzyl)] amino] -acetamide designated FG-067 (inhibitor A) An appropriate method is N-hydroxy-2-[[N '-(4-methoxybenzsulfonyl) -N- (4-chlorobenzyl) using 4-methoxybenzaldehyde instead of 4-chlorobenzaldehyde as a starting material. Basically the same as synthesizing] amino] -acetamide (see 6.1.1.).

¹ H NMR (360 MHz, DMSO-d 6) δ 10.42 (s, 1H, OH), 8.79 (s, 1H, NH), 7.81-6.86 (m, 8H, Ph), 4.29 (s, 2H, CH ₂ CO), 3.85 (s, 3H, OMe), 3.73 (s, 3H, OMe), 3.28 (s, 2H, CH ₂ Ph).

6.1.11. Synthesis of N-hydroxy-2-[[N '-(4-benzenesulfonyl) -N'-(4-chlorobenzyl)] amino] -acetamide.

Suitable method for synthesizing N-hydroxy-2-[[N '-(4-benzenesulfonyl) -N'-(4-chlorobenzyl)] amino] -acetamide designated FG-080 (Inhibitor A) N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(4-chloro using silver benzenesulfonyl chloride instead of 4-methoxybenzenesulfonyl chloride as a starting material It is basically the same as synthesizing benzyl)] amino] -acetaamide.

MS (ES) (M + H) ⁺ : 355.

6.1.2. Synthesis of N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(benzyl)] amino] -acetamide.

Suitable method for synthesizing N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(benzyl)] amino] -acetamide known as FG-061 (inhibitor A) N-hydroxy-2-[[N '-(4-methoxybenzenesulfonyl) -N'-(carboxymethyl)] amino] using benzyl bromide instead of bromo tert-butyl acetate as a silver starting material It is basically the same as synthesizing acetamide (see 6.1.7).

MS (ES) (M-H) ⁺ : 349.

6.1.3. Synthesis of N- (4-methoxybenzenesulfonyl) -β-benzyl- (L) -aspartic acid.

A suitable method for synthesizing N- (4-methoxybenzenesulfonyl) -β-benzyl- (L) -aspartic acid designated FG084 (inhibitor A) is as follows.

N- (4-methoxybenzenesulfonyl) -β-benzyl- (L) -aspartic acid (40).

Of b-benzyl- (L) -aspartic acid HCl salt (38) (2.00 g, 8.96 mmol) and p-methoxybenzenesulfonyl chloride (39) (1.76 g, 8.53 mmol) in anhydrous CH ₂ Cl ₂ To the suspension mixture is added triethylamine (1.81 g, 17.91 mmol) at room temperature. After stirring for 15 hours, the reaction mixture is cooled with 1N HCl (60 mL) and extracted with CH ₂ Cl ₂ (3 × 50 mL). The combined organic layers were washed with brine, dried over MgSO ₄ and concentrated to give N- (4-methoxybenzenesulfonyl) -b-benzyl- (L) -aspartic acid (3.14 g, 7.99 mmol, 94% yield). Get

N-benzyloxy-N '-(4-methoxybenzenesulfonyl) -β-benzyl- (L) -aspartamide (41)

(7/3) N- (4-methoxybenzenesulfonyl) -b-benzyl- (L) -aspartic acid (300 mg, 0.76 mmol) and O-benzylhydride in THF / DMF (10 mL) anhydrous solution N-hydroxybenzotriazole (HOBT) (103 mg, 0.76 mmol), N-ethylmorpholine (204 mg, 1.68 mmol) and diisopropylcarbodiimide (106 mg, 0.84) in a loxylamine / HCl mixture mmol) is added at room temperature. After stirring on holiday (2,5 days), the reaction mixture was diluted with (1/1) hexanes / EtOAc (40 mL), 1N HCl (2 × 20 mL), saturated aqueous NaHCO ₃ solution (2 × 20 mL) and brine. Rinse continuously with The organic layer is dried over MgSO ₄ and concentrated. The residue was purified by silica gel flash chromatography ((1/1) EtOAc / hexane) to give N-benzyloxy-NO- (4-methoxybenzenesulfonyl) -b-benzyl- (L) -aspartamide (114). Mg, 0.22 mmol, 30% yield, white solid).

mp: 128-129. C; MS (ES) (M + H) ⁺ : 499

N-hydroxy-N '-(4-methoxybenzenesulfonyl)-(L) -aspart amide (42).

N-benzyloxy-NO- (4-methoxybenzenesulfonyl) -b-benzyl- (L) -aspartamide (102 mg, 0.20 mmol) and 10% Pd / C (43 mg) in methanol (7 mL) The mixture is vigorously stirred under hydrogen gas for 20 hours. The catalyst is filtered through a pad of celite and the filtrate is concentrated. The residue is lyophilized from water to give N-hydroxy-NO- (4-methoxybenzenesulfonyl)-(L) -aspartamide (50 mg, 0.16 mmol, 77% yield) (hygroscopic powder). .

¹ H NMR (360 MHz, DMSO-d6) δ10.60 (s, 1H), 8.78 (s, 1H), 7.87 (d, J = 8.2 Hz, 1H), 3.93 (m, 1H), 3.82 (s, 3H), 2.50 (m, 1H), 2.21 (dd, J = 16.0, 6.5 Hz, 1H).

6.2. Example 2: C-proteinase test.

6.2.1. Determination of C-proteinase Activity and Inhibitor IC ₅₀ in Vitro

The following tests are used to determine the activity levels and effects of the various compounds of the invention on C-proteinase activity.

Add approximately 125 μg radiolabeled (14C) procollagen to 10 Units / ml chick C-proteinase in 0.1 M Tris-HCl, 0.1 M NaCl, 0.02% Brij-35 and 5 mM CaCl ₂ solution in a total volume of 10 μL. do. The reaction proceeds for 15 minutes at 35 ° C. and stops the reaction using 1/3 volume 3 × stop / drop buffer (30 mM EDTA, 30% glycerol, 6% SDS, 0.006% bromophenol-blue). The sample was subsequently heated to 100 ° C. for 4 minutes and dissolved in SDS-PAGE (Novex) using 6% polyacrylamide gel. Protein bands can be detected by autoradiography. The amount of enzymatic activity is based on the disappearance of the band corresponding to the uncleaved procollagen.

The IC ₅₀ of the inhibitor is determined by plotting the inhibitor concentration against% active and measuring the inhibitor concentration with 50% activity.

IC ₅₀ values of the tested inhibitors are shown in Table 1.

IC ₅₀ of various identified C-proteinase inhibitors Inhibitor General classification IC ₅₀ FG-047 A 50 μM FG-061 A 100 μM FG-053 A 100 μM FG-052 ¹ B 125 μM FG-066 A 150 μM FG-067 A 150 μM FG-086 (cbz-Pro-Leu-Gly-hydroxyxamate ² ) A 200 μM FG-087 (Ac-PYYG-hydroxyxamate) A 335 μM FG-088 (Actinine ³ ) B 350 μM FG-054 A 400 μM FG-057 D 1500 μM FG-058 ⁴ A 2100 μM FG-055 A 2600 μM

FG-051 ⁵ C >> 100 μM FG-046 A >> 1000μM 1 Purchase from Peptides International (IHN-3850-PI) 2 Purchase from Sigma (C-8537) 3 Purchase from Sigma (A-6671) 4 Intermediate product (28) (6.1.7) 5 Peptides International (ISN-3835-PI Buy at)

6.2.2. ELISA assay to determine C-proteinase activity and IC ₅₀ of inhibitor in vitro.

IC ₅₀ values of inhibitors can also be determined by filtration ELISA assay. About 25 mg of unlabeled human procollagen I in this test is incubated with C-proteinase for 1 hour (see 6.2.1.). The reaction is stopped by addition of 40 μl precipitation buffer (0.5 × reaction buffer, 0.1 mgml chick collagen II, 10 μg / ml BSA, 7.5 mM EDTA). Add 25 μl 75% ethanol and mix the reaction and leave on ice for 1 hour to precipitate procollagen. Soluble C-propeptide is isolated from precipitated collagen by filtration through a Millipore multiscreen vacuum manifold HV-0.45 μm hydrophilic plate using Millipore multiscreen. 20 μl filtrate is removed and the amount of cleaved C-propeptide is determined using Takara Biomedicals's procollagen type I C-peptide (PIP).

Recombinant hBMP-1 cell medium (Kessler et al. (1996) was concentrated with about 20ng of radiolabeled ( ¹²⁵ I) human procollagen I in a total volume of 10 μl in reaction buffer to hBMP-1 inhibition. ) Science 271: 360). The reaction proceeds at 35 ° C. for 1 hour, the reaction is stopped with 1/3 volume of 3 × stop / drop buffer and analyzed on SDS-PAGE.

IC ₅₀ of inhibitors is determined by plotting inhibitor concentrations relative to% active and measuring inhibitor concentrations with 50% activity. Table 2 shows the IC ₅₀ values.

IC ₅₀ of various identified C-proteinase inhibitors determined by ELISA Inhibitor General group IC ₅₀ ELISA FG-061 A 12 μM FG-047 A 13 μM FG-053 A 22 μM FG-067 A 37 μM FG-066 A 48 μM

6.2.3. Tissue culture assay to determine C-proteinase activity and IC ₅₀ of inhibitor

The production of procollagen and mature collagen in conditioned media before and after treatment with the compound determines the IC ₅₀ of C-proteinase activity and inhibitor in vivo. The ratio of collagen and procollagen is directly related to cellular transition from precursors to mature collagen material, which indicates C-proteinase activity.

Or determine the media content of C-propeptide / cell and compare the untreated and inhibitor treated cells.

6.3. Animal model for determination of C-proteinase activity and determination of inhibitor effect.

Several animal models with clinical diseases associated with abnormalities in collagen production are known and can be used to determine the effects in vivo of the compounds of the present invention. Animal models such as ventricular wounds in mice (Schilling et al., 1959, Surgery 46: 702-710);

Estradiol stimulated uterine dilatation model (Mandell et al., 1982, The Journal of Biological Chemistry 257: 5268-5273), inducible angiogenesis model (Matrigel) (Passaniti et al., 1992, Laboratory Investigation 67: 519-528 ) Are included. Animal models also include liver fibrosis models (Tsukamoto et al., 1990, Seminar in Liver Disease 10: 56-65; Kock-Weser, 1952, Laboratory Investigation 1: 324-331; Marrione, 1949, American Journal of Pathology 25: 273 -285; Tams, 1957, American Journal of Pathology 33: 13-27; Wahl et al., 1986, Journal of Experimental Medicine 163: 884-902; Pulmonary fibrosis model (Kelly et al., 1980, Journal of Laboratory Clinical Medicine 96: 954-964); Arterial restenosis model (Jackson, 1994, Trends of Cardiovascular Medicine 4: 122-130; Clowes et al., 1983, Laboratory Investigation 49: 327-333); Renal fibrosis model (Yamamoto et al., 1987, Kidney International 32: 514-525); Gun regeneration model (Franklin et al., 1986, The Journal of Laboratory and Clinical Medicine 108: 103-108); Tumor growth model (Kiohs, et al., 1985, JNCL 75: 353-359); Clinical disease models such as models (Lahery et al., 1989, Journal of Ocular Pharmacology 5: 155-179) and abdominal adhesion models (Williams et al., 1992, Journal of Surgical Research 52: 65-70). .

6.4. Example 4: Determination of Cytotoxicity.

Inhibitors were tested for cytotoxicity to determine if they were effective in survival or proliferation of cells. Such tests multiply rapidly or use resting cells. Known numbers of cells are seeded and exposed to increasing inhibitor concentrations with time. Cell number is counted by cell number counting or staining (crystal violet).

Cytotoxicity is assessed as a function of cell survival and cell proliferation. Cell survival is determined by the number of cells using resting cells. Reduction of cell number indicates loss of cells and thus affects cell survival. Cell proliferation utilizes rapidly proliferating cells, which are also determined by cell number. Reduction in cell number relative to untreated criteria indicates an effect on cell proliferation.

5.2. Applications

The compounds and compositions of the invention can be used to treat diseases associated with inappropriate production or maturation of collagen, such as arthritis diseases, fibrosis diseases and other connective tissue diseases.

Such diseases or disorders include spontaneous interstitial fibrosis, interstitial pulmonary fibrosis, myofiber fibrosis, cymus fibrosis, peripheral fibrosis, hepatitis, cutaneous fibrosis, biliary cirrhosis, alcoholic cirrhosis, acute pulmonary fibrosis, spontaneous pulmonary fibrosis, Acute respiratory fatigue syndrome, renal fibrosis / glomerulonephritis, renal fibrosis / diabetic nephropathy, scleroderma / systemic, scleroderma / topical, pedigree, hypertrophic scar, severe joint adhesion / arthritis, bone fibrosis, corneal scar, bladder fibrosis, muscle Duchenne's, cardiac fibrosis, myofiber / retinal detachment, esophageal stricture, payronles disease, and the like. Fibrotic diseases also include scar correction / plastic surgery, glaucoma, cataract fibrosis, corneal scars, joint adhesion, host disease by transplantation, tendon surgery, nerve fixation, dupuytren construction, OB / GYN adhesion / fibrosis, pelvic adhesion, duodenal fibrosis, re Stenosis, and the like. It may also be induced or initiated by surgery, for example pulmonary fibrosis.

5.3. Pharmaceutical Compositions and Routes of Administration thereof

The compounds identified above may be administered to a patient in need thereof, or as a pharmaceutical composition in admixture with an excipient or a suitable carrier in an amount effective for treating a variety of diseases. A therapeutically effective amount refers to a sufficient amount of compound that can alleviate the symptoms. Techniques for administration and preparation of the compounds of the present application are referenced in "Remingron's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA, latest edition.

5.3.1. Route of administration

Suitable routes of administration include oral, rectal, transmucosal, or visceral administration; Intramuscular, subcutaneous, bone marrow, spinal cord and the like, and non-oral transport such as intravesical, direct intraventricular, intravenous, intraperitoneal, intranasal or intraocular administration.

Alternatively, the compound may be administered locally instead of systemically, for example, the compound may be administered directly to joint or fibrotic tissue, which may be depot or sustained release. These compounds may be administered as eye drops to prevent the scar process often caused by complications of glaucoma surgery.

The drug can also be administered by a target drug transport system such as liposomes coated with specific antibodies targeting arthritis or fibrosis tissue. Liposomes can be selectively taken in diseased tissue.

5.3.2. Compound / Composition

The pharmaceutical composition of the present invention may be made by means of conventional mixing, dissolving, granulating, sugar-making, water blasting, emulsifying, capturing, collecting or lyophilizing processes.

Thus, pharmaceutical compositions which can be used according to the invention can be prepared in a conventional manner using one or more physiologically acceptable carriers which can treat the active compound in a pharmaceutically usable preparation consisting of excipients and auxiliaries. Can be. Properly prepared according to the selected route of administration.

At the time of injection, the substances of the invention can be prepared in aqueous solutions, such as physiologically available buffers such as Hans' solution, Ringer's solution or physiological salt buffer. In transmucosal administration, a suitable penetrant may be used in the composition that can penetrate the barrier. Such penetrants are known in the art.

In the case of oral administration, it may be prepared by combining the active compound with a pharmaceutically acceptable carrier known in the art. Such compounds make the compounds of the present invention into tablets, pills, sugars, capsules, liquids, gels, syrups, slurries, suspensions, and the like, which can be administered orally to patients in need thereof. Pharmaceutical compositions which can be used orally are obtained as solid excipients which can optionally be ground the resulting mixture and, if necessary, added to the appropriate adjuvant and then treated with the granular mixture to obtain tablets or sugars. Suitable excipients include fillers such as sugar, including sugar, lactose, sucrose, mannitol or sorbitol; Corn starch, wheat starch, rice starch, potato starch, gelatin, gum tracatan, mail cellulose, hydroxypropylmethyl-cellulose, carboxymethylcellulose sodium and / or cellulose preparations such as polyvinylpyrrolidone (PVP) have. If necessary, crosslinked polyvinyl pyrrolidone, agar or salts thereof, such as alginic acid or sodium alginate, can be added as a disintegrant.

The sugar core can be made by appropriate coating. For this purpose, concentrated sugar solutions can be used, optionally with gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures, etc. It may include. Pigments or dyes may be added to identify the active compound and to characterize different combinations.

Pharmaceutical compositions that can be used orally include push-fit capsules made of gelatin and soft capsules made of plasticizers such as gelatin and glycerol or sorbitol. Push-fit capsules may include the active ingredient and fillers such as lactose, binders such as starch, lubricants such as talc or magnesium stearate or, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Stabilizers may also be added. All compositions for oral administration may be in dosage forms suitable for such administration.

When administered to the cheek, it may be in the form of a tablet or a sugar prepared in a conventional manner.

When administered by inhalation, the compounds to be used for use according to the invention are pressurized packs using suitable propellants such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. Or from a nebulizer, usually in an aerosol spray form. In the case of a pressurized aerosol, the weak unit is determined by providing a valve to transport the measured amount. Gelatin capsules and cartridges for use in an inhaler or nebulizer are formulated to contain a mixture of a suitable powder and compound, such as lactose or starch.

The compound may be formulated for parenteral administration by buccal administration or continuous infusion. Injectable compositions may be presented in unit dosage form, such as in ampoules or in multidose form. The composition may be in the form of an emulsion in suspension, solution or oil or in an aqueous carrier, and may include composition agents such as suspending agents, stabilizers, dispersants.

Pharmaceutical compositions for parenteral administration include solutions of the active compounds in aqueous form. Alternatively, suspensions of the active compounds may be made of suitable oil injection suspensions. Suitable lipophilic solvents or carriers include fatty oils such as sesame oil or synthetic fatty acid esters such as ethyl oleate or triglycerides or liposomes. Aqueous injection suspensions may contain suitable stabilizers or substances that increase the solubility of the compounds that allow for the preparation of significantly concentrated solutions.

Alternatively, the active ingredient may be in powder form, which may be formulated before use in a suitable vehicle, such as sterile pyrogen-free water.

The compounds may be formulated in rectal compositions such as suppositories or enemas, including conventional suppository bases such as cocoa butter or other glycerides.

In addition to the compositions described above, it may also be made as a depot preparation. Such long acting compositions can be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be prepared from suitable polymers or hydrophobic materials (eg, suspensions in acceptable oils) or ion exchange resins or sprayable solubilizers or salts.

Pharmaceutical carriers that can be used in the hydrophobic compounds of the present invention are cosolvent systems consisting of benzyl alcohol, nonpolar surfactants, organic polymers that are incompatible with water and the aqueous phase. The cosolvent system may be a VPD cosolvent system. VPD is a solution filled with 3% w / v benzyl alcohol, 8% w / v nonpolar surfactant polysorbate 80 and 65% w / v polyethylene glycol 300 and the remaining volume is pure ethanol. The VPD cosolvent system (VPD: 5W) consists of VPV diluted 1: 1 with 5% dextrose in aqueous solution. Such cosolvent systems can change significantly without changing solubility and toxicity characteristics. In addition, the state of the cosolvent component can vary significantly: for example, low toxicity nonpolar surfactants can be used in place of Polysorbate 80; The fractional size of polyethylene glycol can also vary; Other biocompatible polymers may be replaced with polyethylene glycols such as polyvinyl pyrrolidone; Other sugars or polysaccharides may also be used instead of dextrose.

Or other transport systems of hydrophobic pharmaceutical compounds may be used. Liposomes and emulsions are known as transporting carriers or carriers for hydrophobic drugs. Certain organic solvents, such as dimethylsulfoxide, may be used and are generally more toxic. In addition, the compounds may be utilized using sustained release systems such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained release materials are known and are known to those of skill in the art. Depending on the chemical nature, the sustained release capsule releases the compound from several weeks up to 100 days.

Additional strategies for protein stabilization may be used depending on the chemical properties and biological stability of the therapy.

Pharmaceutical compositions consist of a suitable solid or gel carrier or excipient. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, polymers such as gelatin and polyethylene glycol.

Most of the C-proteinase inhibitory compounds of the invention can be provided as pharmaceutically applicable counterions and salts. Such pharmaceutically acceptable salt addition salts include organic or inorganic bases such as sodium hydroxide, magnesium hydroxide, ammonia, trialkylamines, dialkylamines, monoalkylamines, dibasic amino acids, acetate sodium, benzoate calcium, triethanol amines and the like. Salts possessing the properties and biological effects of free acids obtainable by reaction.

5.3.3. Effective dosage

Pharmaceutical compositions that can be used in the present invention have a composition comprising the active ingredient effective to achieve the desired purpose. In particular, a therapeutically effective amount means an amount effective to alleviate or prevent the development of existing symptoms of the individual to be treated. Determination of the effective amount is made according to the ability of those skilled in the art and is included here as provided.

Therapeutic effective amounts in the compounds used in the methods of the invention are first evaluated in cell culture assays. For example, it can be made in an animal model to obtain a calculated range of concentrations including IC ₅₀ as determined in cell culture (e.g., of test compounds to obtain ½ of the maximal inhibition of C-proteinase activity). density). Such information can be used to determine more accurate doses available to humans.

A therapeutically effective dose refers to the amount of a compound that prolongs survival or reduces symptoms in a patient. Toxicity and therapeutic effects of these compounds are standard pharmaceuticals in cell culture or laboratory animals to determine LD ₅₀ (a dose that can kill 50% of the population) and ED ₅₀ (a dose that is effective for 50% of the population). Can be determined by a pharmacological process. The dose ratio between toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD ₅₀ and ED ₅₀ . Compounds with high therapeutic indices are effective. Data obtained from such cell culture assays and animal studies can be prepared in the dosage range that can be used in humans. It is appropriate that a dose of such a compound is within a range of circulating concentrations that include non-toxic or weak ED ₅₀ . The dosage will vary within this range depending on the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage are selected by the attending physician according to the patient's condition (Fingl et al., 1975, in "The Pharmacological Basis of Therapeutics", Ch. 1 p.1).

Dosage and dosing intervals can be adjusted individually to maintain a C-proteinase inhibitory effect or to provide sufficient plasma activity levels to have a minimum effect concentration (MEC). MEC varies for each compound but can be evaluated from in vitro data. For example, it is the essential concentration for inhibiting 50-90% C-proteinase using the tests detailed herein. Dosages necessary to obtain a MEC will depend on the individual's characteristics and route of administration. However, plasma concentrations can be determined using HPLC or biopsy.

Dosage intervals are determined using MEC values. The compound is administered using a method that maintains plasma levels above MEC for 10-90%, suitably 30-90%, most suitably 50-90% of the time.

In the case of topical administration or screening acquisition, the effective local concentration of the drug is independent of the plasma concentration.

The amount of the administered composition can vary depending on the individual being treated, the weight of the individual, the severity of the condition, the method of administration and the judgment of the physician.

5.3.4. Packing

If desired, the composition may be provided in a package or device containing one or more unit dosage forms containing the active ingredient. For example, the pack consists of a plastic foil such as a metal or blister pack. The pack or dispenser is made by instructions for administration. Compositions composed of the compounds of the present invention formulated in a pharmaceutically acceptable carrier are suitably prepared, placed in a suitable container, and indicated for the treatment of the indicated conditions. Appropriate indications on the label include treatment of arthritis or other fibrotic diseases.

Claims (24)

A compound having inhibitory activity against C-proteinase, wherein the compound has the formula or a pharmaceutically acceptable salt thereof. Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxy Alkyl, acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulf Phonyl) -alkyl; R ₂ is selected from H, lower alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl; R ₄ is selected from aryl, heteroaryl, alkyl, aralkyl, heteroaralkyl, alkylamino, arylalkylamino, X is selected from SO ₂ , C═O; Y is selected from OH, HOHN (hydroxyamine), H ₂ N, alkylamino; Z is directly bonded at methylene, oxygen, sulfur, amino; n is 0 or 1; A compound having inhibitory activity against C-proteinase, wherein the compound has the formula or a pharmaceutically acceptable salt thereof. Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl , Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl ) -Alkyl; R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl. A compound having inhibitory activity against C-proteinase, wherein the compound has the formula or a pharmaceutically acceptable salt thereof. Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become; R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl. A compound having inhibitory activity against C-proteinase, wherein the compound has the formula or a pharmaceutically acceptable salt thereof. Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become; R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl. A compound having inhibitory activity against C-proteinase, wherein the compound has the formula or a pharmaceutically acceptable salt thereof. Wherein R ₁ is selected from OH, alkoxyl, lower alkyl, alkylamino, peptide, X is selected from N, C, R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound has the formula or consists of a pharmaceutically acceptable salt thereof. Pharmaceutical composition, characterized in that consisting of a compound selected from the formula: a) Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxy Alkyl, acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulf Phonyl) -alkyl; R ₂ is selected from H, lower alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl; R ₄ is selected from aryl, heteroaryl, alkyl, aralkyl, heteroaralkyl, alkylamino, arylalkylamino, X is selected from SO ₂ , C═O; Y is selected from OH, HOHN (hydroxyamine), H ₂ N, alkylamino; Z is directly bonded at methylene, oxygen, sulfur, amino; n is 0 or 1; or b) Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl , Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl ) -Alkyl; R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, halo-substituted aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, Acyloxyalkyl, mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -Selected from alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or c) Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become; R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or d) Wherein R ₁ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, Selected from mercaptoalkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl Become; R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl; or e) Wherein R ₁ is selected from OH, alkoxyl, lower alkyl, alkylamino, peptide, X is selected from N, C, R ₂ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₃ is H, lower alkyl, mono or poly-haloalkyl, carboxyalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, biaryl, biarylalkyl, hydroxyalkyl, alkoxyalkyl, acyloxyalkyl, mercapto Alkyl, (amino, mono or dialkylamino) alkyl, acylaminoalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, alkyl- (thio, sulfinyl or sulfonyl) -alkyl; R ₄ is selected from H, lower alkyl; R ₅ is selected from H, lower alkyl, carboxyalkyl, (mono or dialkylamino) alkyl, alkyl- (thio, sulfinyl or sulfonyl) alkyl, alkoxyalkylacylalkyl. A pharmaceutical composition comprising a compound selected from compounds of the formula: and a pharmaceutically acceptable excipient or carrier. a) or b) or c) or d) or e) or f) or g) or h) or i) or j) or k) or l) or m) A method of treating a disease associated with improper production of collagen, comprising administering an effective amount of the compound according to claim 20. A method for treating a disease associated with improper production of collagen, comprising administering an effective amount of the compound according to claim 21. 23. The method of claim 22, wherein the fibrosis disease is selected from liver cirrhosis and joint name. 24. The method of claim 23, wherein the fibrosis disease is selected from liver cirrhosis and joint name.