JP2011190257A - Preventive or therapeutic agent for tissue fibrotic disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a therapeutic or preventive agent for fibrosis of tissues. <P>SOLUTION: As an active ingredient of the preventive or therapeutic agent for tissue fibrotic diseases, a metal chelate compound represented by formula (1) (wherein R is an OH group, an O-lower alkyl group, an amino group, an N-substituted amino group, an amino acid residue or a peptide residue; and M is a pharmacologically acceptable metal) or a pharmacologically acceptable salt thereof is used. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a prophylactic or therapeutic agent (anti-antigen) for tissue fibrosis diseases caused by pathological fibrosis of tissues such as pulmonary fibrosis, renal fibrosis, nephrosclerosis, liver fibrosis (including NASH) or cirrhosis. Fibrotic agent).

  A tissue constituting a living body often causes a phenomenon of fibrosis with a disease. Fibrosis refers to tissue stiffening accompanied by excessive deposition of fibrous proteins such as collagen and fibroblast infiltration and proliferation as a result of inflammation that has occurred in the tissue.

  Tissue fibrosis can occur in any tissue, and the resulting phenomenon is the same, even though the site of occurrence and the triggering stimulus are different. When parenchymal cells fall off or tissue function declines, the living body may migrate and replenish fibroblasts unrelated to the function of the original tissue, but fibroblasts restore the function of the tissue. It is only responsible for maintaining the physical form, and the accumulation of fibroblasts and the accumulation of intercellular matrices such as collagen produced by the fibroblasts causes tissue stiffening. Such stiffening is fibrosis, and the fibrotic tissue becomes difficult to function normally, often resulting in a condition called fibrosis. Once fibrotic tissue does not return to its original state, the condition progresses further and may cause severe disability.

  Such tissue fibrosis occurs in many tissues such as, for example, lung, bladder, kidney, heart or liver.

  For example, as one of the fibrotic diseases of the lung, pulmonary fibrosis that occurs as a side effect of administration of bleomycin, an anticancer agent, is known. This is thought to be because radicals generated by bleomycin injured lung tissue, and fibroblast infiltration and proliferation occurred to compensate for the injury, collagen protein was deposited, and lung function was impaired. Injuries to the lung tissue that cause pulmonary fibrosis can also be caused by blood pressure overload in pulmonary hypertension and chronic inflammatory reactions in asthma. As a result of such pulmonary fibrosis, the ability of the lungs to ventilate decreases, leading to further deterioration of the pathological condition. Many treatments for suppressing pulmonary fibrosis have been studied so far, but no satisfactory treatment has yet been found.

  The bladder is also a tissue that contacts the outside world through the urethra, and is exposed to various stimuli including infection, and it is known that the fibrosis of the bladder is induced by an inflammatory reaction caused by such a stimulus. Bladder fibrosis makes urine collection and urination difficult, leading to cystitis with chief complaints of frequent urination and difficulty urinating. In other words, bladder fibrosis occurs in cystitis such as urinary tract tuberculosis, interstitial cystitis, and radiation cystitis (Non-patent Document 1). In particular, interstitial cystitis has been shown to be a disease in which fibrosis accompanied by increased chymase is a problem (Non-patent Document 2). However, treatment for interstitial cystitis due to fibrosis of the bladder tissue has not been established, and symptomatic treatment is repeated, and there are many cases that eventually lead to cystectomy surgery. Fibrosis also develops in the kidney due to various factors. Kidney disease is called by various disease names depending on its cause and onset location, such as IgA nephropathy caused by immune complex deposition, diabetic nephropathy resulting from diabetes, pyelonephritis occurring in the renal pelvis. In this process, functional decline accompanying tissue fibrosis is observed. It is also known that as a result of chronic rejection after kidney transplantation, fibrosis occurs and renal dysfunction may occur again. There is currently no effective treatment to prevent such kidney dysfunction.

  When the heart is subjected to anticancer drug treatment such as adriamycin, or stimulation such as infection or pressure load, it may cause loss of myocardium or decrease in cardiac function, migrating fibroblasts in a compensatory manner, and developing fibrosis . And when it progresses excessively, the heart stiffens and its function as a pump decreases and progresses to chronic heart failure caused by fibrosis. As this treatment, ACE inhibitors and β-blockers are used, but they are not satisfactory treatments for suppression and improvement of fibrosis.

  Regarding the liver, it is known that fibrosis occurs due to stimulation with viruses, tumors, and alcohol, and when it progresses, it undergoes hard changes (cirrhosis). Recently, nonalcoholic steatohepatitis (NASH), in which liver tissue findings similar to alcoholic liver damage, such as fattening and fibrosis, has attracted attention as a serious case of lifestyle-related diseases Yes. Fibrosis rapidly degrades the liver. If the cause is a virus or tumor, treatment with an antiviral agent or anticancer agent is performed, and if it is caused by food intake such as alcohol, diet therapy is taken, but active treatment methods for liver fibrosis are known Not.

  As described above, tissue fibrosis occurs in every organ in the living body, and it is closely related to the deterioration of tissue function.

  Fibrosis of tissue inside the living body is confirmed by examination such as echo or by collecting a part of the tissue and detecting a substance involved in fibrosis. An example is a method for quantifying hydroxyproline, which is technically cumbersome and requires a sample amount. In addition, a method of preparing a tissue specimen and analyzing it pathologically has also been performed. In recent years, it has been reported that tissue fibrosis can be measured by using the expression level of a specific messenger RNA (mRNA) of a protein involved in fibrosis (eg, collagen 3) as an index due to the development of gene manipulation technology (non-native). Patent Document 3).

  By the way, 6,8-dimercaptooctanoic acid is a reduced form of α-lipoic acid, a coenzyme present in mitochondria, and has an action of regenerating oxidized glutathione and vitamin C into a reduced form. However, it is known that 6,8-dimercaptooctanoic acid is very unstable in air and oxidized to return to α-lipoic acid.

  Examples of such derivatives of α-lipoic acid include α-lipoyl amino acid in which glycine, methionine, glutamic acid, valine and the like are bonded to α-lipoic acid (Patent Document 1), and imidazole salt of α-lipoylaminoethylsulfonic acid (Patent Document 2). , Lipoyl esters (Non-Patent Document 4), and metal derivatives of dihydrolipoic acid and dihydrolipoamide (Non-Patent Documents 5 and 6) are known.

  These α lipoic acids or derivatives thereof can be stabilized by chelating the reduced form with a metal. Such metal chelate compounds (6,8-dimercaptooctanoic acid metal chelate compounds or derivatives thereof) It is known that there are tyrosinase inhibitory action, elastase inhibitory action, melanin production inhibitory action, and scalp hair loss healing action (see Patent Documents 3 to 5). However, it is not known that such metal chelate compounds have a preventive or therapeutic effect on tissue fibrosis diseases.

Japanese Patent Publication No. 42-1286 (corresponding US Patent No. 3,238,224) JP 2000-169371 International Publication WO 02/076935 A1 International Publication WO 04/024139 A1 JP 2008-174453 A

Urology (Takita Kurita, p.177 (1995), Nanzan-do) T. Yamada et al., Int. J. Urol., 7, 292 (2000) S.N.Iyer et al., J. Pharmacol. Exp. Therap., 289, 211 (1991) Biochem. J., (1990) 271, 45-49 Inorganica Chimica Acta, 192 (1992) 237-242 J. Org. Chem., 1985, 50, 2522-2524

  As described above, an effective preventive or therapeutic agent for a tissue fibrosis disease has not been developed yet, and only a symptomatic treatment is performed on a tissue whose function is reduced by fibrosis. Therefore, a prophylactic or therapeutic agent for tissue fibrosis is desired.

  Therefore, an object of the present invention is to provide a new preventive or therapeutic agent for tissue fibrosis diseases.

  In order to solve the above-mentioned problems, the present inventors have conducted extensive studies using a pulmonary fibrosis model animal in which pulmonary fibrosis is induced with bleomycin. As a result, a metal chelate compound (1) represented by the following general formula: :

(In the formula, R represents an OH group, an O-lower alkyl group, an amino group, an N-substituted amino group, an amino acid residue, or a peptide residue, and M represents a pharmaceutically acceptable metal).
In particular, it has been found that N- (6,8-dimercaptooctanoyl) histidine sodium / zinc chelate compound has an effect of significantly suppressing tissue fibrosis, and the metal chelate compound (1) is effective in treating tissue fibrosis. It was convinced that it was useful as an active ingredient of a preventive or therapeutic agent.

This invention is completed based on the knowledge which concerns, Comprising: The following embodiment is included.
Preventive or therapeutic agent for tissue fibrosis (I) Preventive or therapeutic agent for tissue fibrosis containing a metal chelate compound represented by the following general formula (1) or a pharmaceutically acceptable salt thereof:

(In the formula, R represents an OH group, a 0-lower alkyl group, an amino group, an N-substituted amino group, an amino acid residue, or a peptide residue, and M represents a pharmaceutically acceptable metal).

  (II) The metal chelate compound (1) is a metal chelate compound of 6,8-dimercaptooctanoic acid lower alkyl ester, a metal chelate compound of N- (6,8-dimercaptooctanoyl) amine, N- (6, (8-dimercaptooctanoyl) amino acid metal chelate compound and at least one metal chelate compound selected from the group consisting of N- (6,8-dimercaptooctanoyl) dipeptide metal chelate compounds (I The preventive or therapeutic agent described in).

  (III) A metal chelate compound of N- (6,8-dimercaptooctanoyl) amine is a 6,8-dimercaptooctanoic acid amide metal chelate, N- (6,8-dimercaptooctanoyl) -2-amino Ethanol metal chelates, N- (6,8-dimercaptooctanoyl) isopropylamine metal chelates, N- (6,8-dimercaptooctanoyl) 5-O-methyl serotonin metal chelates and N- (5,8-di The preventive or therapeutic agent according to (II), which is at least one selected from the group consisting of mercaptooctanoyl) -2-aminopyridine metal chelate compounds.

  (IV) The metal chelate compound represented by the general formula (1) has, as R, an amino acid residue (an amino acid residue bonded with a nitrogen atom) or a peptide residue (a peptide acid residue bonded with a nitrogen atom). The preventive or therapeutic agent according to (I).

  (V) N- (6,8-dimercaptooctanoyl) amino acid metal chelate compound is N- (6,8-dimercaptooctanoyl) -α-amino acid metal chelate, N- (6,8-dimercapto The preventive or therapeutic agent according to (II), which is at least one selected from the group consisting of octanoyl) -ω-amino acid metal chelate and N- (6,8-dimercaptooctanoyl) special amino acid metal chelate compound .

  (VI) N- (6,8-dimercaptooctanoyl) -α-amino acid metal chelate is N- (6,8-dimercaptooctanoyl) glycine metal chelate, N- (6,8-dimercaptoocta Noyl) alanine metal chelate, N- (6,8-dimercaptooctanoyl) threonine metal chelate, N- (6,8-dimercaptooctanoyl) serine metal chelate, N- (6,8-dimercaptooctanoyl) Aspartic acid metal chelate, N- (6,8-dimercaptooctanoyl) glutamic acid metal chelate, N- (6,8-dimercaptooctanoyl) phenylalanine metal chelate, N- (6,8-dimercaptooctanoyl) methionine Metal chelate, N- (6,8-dimercaptooctanoyl) norleucine metal chelate, N- (6 , 8-dimercaptooctanoyl) cysteine metal chelate, N- (6,8-dimercaptooctanoyl) hydroxyproline metal chelate, N- (6,8-dimercaptooctanoyl) histidine metal chelate, N- (6 8-Dimercaptooctanoyl) -5-hydroxytryptophan metal chelate, N- (6,8-dimercaptooctanoyl) penicillamine metal chelate and N- (6,8-dimercaptooctanoyl) lysine metal chelate compound The preventive or therapeutic agent according to (V), which is at least one selected from the group consisting of:

    (VII) N- (6,8-dimercaptooctanoyl) -ω-amino acid metal chelate and N- (6,8-dimercaptooctanoyl) special amino acid metal chelate compound are represented by N- (6,8-dimercapto Octanoyl) -3-aminopropionic acid metal chelate, N- (6,8-dimercaptooctanoyl) -4-aminobutyric acid metal chelate, N- (6,8-dimercaptooctanoyl) -6-aminohexanoic acid Metal chelate, N- (6,8-dimercaptooctanoyl) -4-transaminomethyl-1-cyclohexanecarboxylic acid metal chelate, N- (6,8-dimercaptooctanoyl) -2-aminoethanesulfonic acid metal Chelates, N- (6,8-dimercaptooctanoyl) sulfanilic acid metal chelates and N- (6,8-dimercapto Kutanoiru) are those selected from the group consisting of anthranilic acid metal chelate, a prophylactic or therapeutic agent according to (V).

  (VIII) N- (6,8-dimercaptooctanoyl) dipeptide metal chelate compounds are N- (6,8-dimercaptooctanoyl) aspartiylglycine metal chelate and N- (6,8-dimercapto The preventive or therapeutic agent according to (II), which is selected from the group consisting of (octanoyl) threonylglycine metal chelate compounds.

  (IX) The preventive or therapeutic agent according to any one of (I) to (VIII), wherein the metal is zinc.

  (X) The preventive or therapeutic agent according to any one of (I) to (IX), wherein the target organ for preventing or treating tissue fibrosis is lung, bladder, kidney, heart, or liver.

  (XI) Tissue fibrosis is pulmonary fibrosis, renal fibrosis, endocardial fibrosis, liver fibrosis, nonalcoholic steatohepatitis (NASH), cirrhosis, sclerosing peritonitis, prostatic hypertrophy, chronic nephritis, strong The preventive or therapeutic agent according to any one of (I) to (X), which is any one selected from the group consisting of dermatosis, uterine leiomyoma, retroperitoneal fibrosis, bladder fibrosis, myelofibrosis, and keloid.

  According to the present invention, a therapeutic or prophylactic agent for tissue fibrosis comprising a metal chelate compound represented by the general formula (1) or a pharmacologically acceptable salt thereof as an active ingredient, such as lung, bladder, kidney, heart, liver The therapeutic or prophylactic agent for fibrosis of various tissues such as can be provided.

It is an optical microscope image which showed the influence of DHLHZn with respect to the pulmonary fibrosis tissue artificially induced | guided | derived by the bleomycin (Experimental example 1). Specifically, A.I. Normal lung tissue (HE staining) (control group), B. Bleomycin-treated lung tissue (HE staining) (DHLHZn non-administered group), and C.I. The image of the lung tissue (HE dyeing | staining) (DHLHZn administration group) which administered DHLHZn after bleomycin treatment is shown. It is an optical microscope image which showed the influence of DHLHZn with respect to the pulmonary fibrosis tissue forcedly induced by bleomycin (Experimental example 1). Specifically, A.I. Normal lung tissue (Azan staining) (control group); Pulmonary tissue treated with bleomycin (Azan staining) (DHLHZn non-administered group), and C.I. The image of the lung tissue (Azan staining) (DHLHZn administration group) which administered DHLHZn after bleomycin treatment is shown. It is an optical microscope image which showed the influence of DHLHZn with respect to the fibrotic liver tissue of a non-alcoholic steatohepatitis (NASH) model mouse (Experimental example 2). Specifically, upper left figure: control group (high-fat diet intake group), lower left figure: DHLHZn oral administration group, and upper right figure: DHLHZn subcutaneous administration group, hepatic tissues are HE-stained images. In addition, the NASH activity score is also shown. It is an optical microscope image which showed the influence of DHLHZn with respect to the fibrotic tissue of the liver of a non-alcoholic steatohepatitis (NASH) model mouse (Experimental example 2). Specifically, upper left: control group (high fat diet intake group) (× 100 times), lower left: control group (high fat diet intake group) (× 400 times), upper center: DHLHZn subcutaneous administration group ( (× 100 magnification), upper right diagram: DHLHZn oral administration group (× 100 magnification), and lower right diagram: normal group (ordinary food intake group) (× 100 magnification) liver images of Azan stained images. Results of comparing the amount of SREBP-1c and TNF-α mRNA in liver tissues of non-alcoholic steatohepatitis (NASH) model mice (from left, control group, DHLHZn subcutaneous administration group, DHLHZn oral administration group, normal group) This is shown (Experimental Example 2). The results of comparing the amount of 8-OHdG in the liver tissue of non-alcoholic steatohepatitis (NASH) model mice (from left, control group, DHLHZn subcutaneous administration group, DHLHZn oral administration group, normal group) are shown (Experimental Example 2). . The result which contrasted the expression level of UCP-2 in the liver tissue of a non-alcoholic steatohepatitis (NASH) model mouse (from the left, a control group, a DHLHZn subcutaneous administration group, a normal group) is shown (Experimental example 2).

  The agent for preventing or treating tissue fibrosis of the present invention (hereinafter collectively referred to as “the anti-fibrotic agent of the present invention”) is a metal chelate compound represented by the following general formula (1) or a pharmaceutically acceptable salt thereof. Characterized in that it contains:

(In the formula, R represents an OH group, an O-lower alkyl group, an amino group, an N-substituted amino group, an amino acid residue, or a peptide residue, and M represents a pharmaceutically acceptable metal).

  Here, in the above formula, the “pharmaceutically acceptable metal” represented by M may be any metal having no biotoxicity and capable of coordinating or chelating with two thiol groups, such as zinc, cobalt or selenium. And pharmaceutically acceptable metals such as bivalent metals such as iron, divalent or trivalent metals such as iron, and tetravalent metals such as germanium and selenium. Preferably it is a bivalent metal, More preferably, it is zinc.

  In the above formula, the metal chelate compound (1) when R is an O-lower alkyl group is generically referred to as “a metal chelate compound of a 6,8-dimercaptooctanoic acid lower alkyl ester” in the present invention. Examples of the “lower alkyl group” in this case include linear or branched alkyl groups having 1 to 6 carbon atoms. Specifically, methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group, tert-butyl group, pentyl group, isopentyl group, neopentyl group, tert-pentyl group, hexyl group, Examples include 2-methylpentyl group, 3-methylpentyl group, 4-methylpentyl group, 2,2-dimethylbutyl group, 3,3-dimethylbutyl group, 2-ethylbutyl group and the like. A methyl group and an ethyl group are preferable.

  In the above formula, the metal chelate compound (1) when R is an amino group or an N-substituted amino group is generically referred to as “a metal chelate compound of N- (6,8-dimercaptoocnoyl) amine”. In this case, the “N-substituted amino group” is an aliphatic hydrocarbon group in which 1 or 2 hydrogen atoms of the primary amino group or 1 hydrogen atom of the secondary amino group is 1 to 5 carbon atoms; An aliphatic alcohol residue having 1 to 5 carbon atoms; or a group substituted with a heterocycle containing a nitrogen atom such as a pyridine ring, a pyrimidine ring or an indole ring. Here, the aliphatic hydrocarbon group, the aliphatic alcohol residue, and the heterocycle containing a nitrogen atom may each have a substituent, such as a hydroxyl group; a halogen atom (chlorine atom, A fluorine atom, an iodine element, etc.); an O-lower alkyl group having 1 to 6 carbon atoms; an alkyl group having 1 to 6 carbon atoms; and a nitrogen atom such as an optionally modified pyridine ring, pyrimidine ring or indole ring Heterocyclic groups and the like can be mentioned without particular limitation.

  Examples of the aliphatic hydrocarbon group having 1 to 5 carbon atoms include a linear or branched lower alkyl group having 1 to 5 carbon atoms, a linear or branched lower alkenyl group having 2 to 5 carbon atoms, and carbon. Examples thereof include linear or branched lower alkynyl groups of 2 to 5. Preferably it is a lower alkyl group. Examples of such lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert, as described above. -A pentyl group etc. are mentioned. A methyl group and an ethyl group are preferable.

  Examples of the lower alkenyl group having 2 to 5 carbon atoms include vinyl, propenyl, butenyl and pentenyl groups, and examples of the lower alkynyl group having 2 to 5 carbon atoms include ethynyl, propynyl, butynyl and pentynyl groups. Can be mentioned.

  Specific examples of the aliphatic alcohol residue having 1 to 5 carbon atoms include lower alcohols such as methanol, ethanol, propanol, isopropanol, butanol, and isobutanol.

As the metal chelate compound (1) of the present invention in which R is an N-substituted amino group, specifically, N- (6,8-dimercaptooctanoyl) -2-aminoethanol metal chelate (R = —NH ₂ In which one hydrogen atom is substituted with an ethanol residue), N- (6,8-dimercaptooctanoyl) isopropylamine metal chelate (R = -NH ₂ with one hydrogen atom substituted with an isopropyl group) Group), N- (5,8-dimercaptooctanoyl) -2-aminopyridine metal chelate compound (group in which one hydrogen atom of R = -NH ₂ is substituted with 2-pyridyl group), N- (6 , 8-dimercaptooctanoyl) 5-O-methyl serotonin metal chelate (R = 5-O-methyl serotonin, a group in which one of the hydrogen atoms of the NH ₂ group in the side chain is substituted). it can.

  In the above formula, the metal chelate compound (1) in the case where R is an amino acid residue is generically referred to as “a metal chelate compound of 6,8-dimercaptooctanoic acid amino acid” in the present invention. The “amino acid” in this case is an α-amino acid having a carboxyl group and an amino group in the same molecule; β-amino acid, γ-amino acid, δ-amino acid, and ε-amino acid (β-˜ε-amino acids are generic terms) Aminomethylcyclohexanecarboxylic acid, anthranilic acid, ethyl anthranilate, and aminoethanesulfonic acid (taurine) or p-aminobenzenesulfonic acid having a sulfonic acid group and an amino group in the same molecule. (Sulfanylic acid) [Amino acids from aminomethylcyclohexanecarboxylic acid to sulfanilic acid that are neither α-amino acids nor ω-amino acids are collectively referred to as “special amino acids”].

  The α-amino acid refers to a compound in which an amino group is also bonded to a carbon (α-carbon) to which a carboxyl group (or sulfonic acid group) is bonded. For example, glycine, alanine, valine, leucine, isoleucine, norleucine, serine, threonine, Examples include tyrosine, cysteine, methionine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine, histidine, phenylalanine, tryptophan, and penicillamine. Preferred are glycine, alanine, threonine, serine, aspartic acid, glutamic acid, phenylalanine, methionine, cysteine, lysine, histidine, and penicillamine, and more preferred is histidine. In addition, these α-amino acids may have a hydroxyl group as a substituent. Although it does not restrict | limit as an alpha-amino acid which has a hydroxyl group, For example, 4-hydroxyproline, hydroxytryptophan, 5-hydroxytryptophan penicillamine etc. can be mentioned.

  The ω-amino acid is a compound in which an amino group is bonded to the carbon at the end of the carbon chain opposite to the carbon to which the carboxyl group (or sulfonic acid group) is bonded, and includes β-amino acids, γ-amino acids and δ-amino acids. It is. Examples of amino acids belonging to such ω-amino acids include 4-aminobutyric acid (γ-amino-n-butyric acid (GABA)), carnitine, 5-aminolevulinic acid, 5-aminovaleric acid, 6-aminohexanoic acid, 2- Examples include aminoethanesulfonic acid (taurine), 3-aminopropionic acid, 5-aminolevulinic acid, 5-aminovaleric acid, trans-4- (aminomethyl) cyclohexane-1-carboxylic acid, and the like. For amino acids having a plurality of amino groups in the same molecule, such as lysine, when the amino group is bonded to the α-carbon in the formula (I), the amino group bonded to the α-amino acid and the ε-carbon Is bound to an ω-amino acid.

  Special amino acids refer to amino acids that are not classified as α-amino acids or ω-amino acids due to their structure, and examples include anthranilic acid and 4-aminobenzene-1-sulfonsan.

  Although not limited, among these amino acids, α-amino acids such as histidine, glutamic acid, methionine, penicillamine, lysine, and phenylalanine; ω such as γ-amino-n-butyric acid, aminoethanesulfonic acid, and 6-aminohexanoic acid -Amino acids; and special amino acids such as anthranilic acid, more preferably histidine.

  In the above formula, the metal chelate compound (1) when R is a peptide residue is generically referred to as “metal chelate compound of 6,8-dimercaptooctanoic acid peptide” in the present invention. The “peptide” in this case is preferably a dipeptide in which two amino acids of the same or different types (as defined above) are bonded to each other with an acid amide bond between one carboxyl group and the amino group of the other amino acid. . Specific examples of such dipeptides include asparatiyl glycine and threonyl glycine.

  The metal chelate compound (1) in the present invention includes a mixture of stereoisomers or each stereoisomer in a pure or substantially pure state. For example, the metal chelate compound (1) having an asymmetric center at any of the carbon atoms can exist as an enantiomer or diastereomer or a mixture thereof.

  Pharmacologically acceptable salts of the metal chelate compound (1) of the present invention include inorganic salts such as alkali metal salts such as sodium salt and potassium salt, and alkaline earth metal salts such as calcium salt and magnesium salt; Mention may be made of organic salts such as salts with amines such as monoethanolamine. In addition, even if it is a salt other than these, as long as it is a salt accept | permitted pharmacologically, it can use suitably for the objective of this invention. In addition, the metal chelate compound (1) targeted by the present invention includes hydrates thereof.

  A method for synthesizing the metal chelate compound (1) used as an active ingredient of the anti-fibrotic agent of the present invention will be described below by taking as an example the case where the metal represented by M is zinc.

  Specifically, by reducing α-lipoic acid or α-lipoic acid amide with zinc and hydrochloric acid (or acetic acid), in the above formula, R is a hydroxyl group, a 6,8-dimercaptooctanoic acid zinc chelate compound or The amide compound can be synthesized. The 6,8-dimercaptooctanoic acid lower alkyl ester zinc chelate can be obtained by reducing the lower alkyl ester of α-lipoic acid in the same manner as described above.

  Furthermore, N- (6,8-dimercaptooctanoyl) amine, N- (6,8-dimercaptooctanoyl) amino acid or N- (6,8-dimercaptooctanoyl) peptide may be, for example, α-lipoic acid Are dissolved in chloroform or acetonitrile, and amines, amino acids or peptides are respectively coupled by mixed acid anhydride method using ethyl chlorocarbonate in the presence of triethylamine, and N-α-lipoylamine, N-α-lipoylamino acid or N Obtain α-lipoyl peptide. By reducing these with zinc and acetic acid (or hydrochloric acid), the desired compounds can be obtained respectively.

  Further, when the metal chelate compound (1) having an amino acid residue or a peptide residue as R is converted into an alkali salt (alkali metal salt, alkali metal earth salt), for example, the free acid is dissolved or suspended in water. This is neutralized with an alkali hydroxide (for example, sodium hydroxide, magnesium hydroxide, etc.), dissolved, concentrated, added with alcohol, and the precipitated crystals are collected by filtration. Can obtain a salt of the metal chelate compound (1).

  Reducts of α-lipoic acid or α-lipoic acid derivatives, ie, 6,8-dimercaptooctanoic acid or its derivatives are very unstable in air, but are chelated with metals such as zinc. When converted, it becomes a six-membered ring and becomes a stable compound with good crystallinity.

The acute toxicity LD _{50 in} rats of the metal chelate compound (1) of the present invention, particularly N- (6,8-dimercaptooctanoyl) histidine zinc chelate compound, is 2000 mg / kg or more by oral administration and is not toxic. It is a highly safe compound belonging to the classification.

  The tissue fibrosis that is the subject of the present invention is caused by a process that compensates for the loss of tissue parenchymal cells or tissue function due to stress such as chemical stimulation of drugs, excessive pressure load, inflammatory reaction, etc. In addition, it means stiffening accompanied by tissue dysfunction due to excessive fibroblast migration and subsequent synthetic deposition of intercellular matrix proteins, and there is no particular limitation on the induction stimulus or the onset site.

  Specific examples of such tissue fibrosis include fibrosis of visceral tissues such as lung, bladder, kidney, heart, and liver. The tissue fibrosis diseases targeted by the present invention include tissue fibrosis caused by administration of drugs such as antitumor agents, antibiotics, antibacterial agents, antiarrhythmic agents, anti-inflammatory agents, antirheumatic agents, interferon or Shosaikoto. Also included are diseases and tissue fibrosis associated with diseases such as chronic nephritis, interstitial myocarditis and interstitial cystitis. Specifically, pulmonary fibrosis caused by side effects of bleomycin administration, and pulmonary fibrosis occurring during or after interstitial pneumonia; bladder fibrosis and bladder cervical sclerosis occurring during interstitial cystitis; Renal fibrosis caused by genetic abnormalities, etc., and renal failure (renal sclerosis); endocardial fibrosis caused by remodeling after myocardial infarction; liver fibrosis caused by hepatocyte damage, nonalcoholic steatohepatitis (NASH) , Associated portal hypertension and cirrhosis; keloids caused by excessive tissue repair, sclerosing peritonitis, prostatic hypertrophy, scleroderma, uterine leiomyoma, retroperitoneal fibrosis, myelofibrosis, etc. be able to.

  In addition to treating such tissue fibrosis diseases, the anti-fibrotic agent of the present invention can be used for prevention, particularly coronary restenosis after percutaneous transluminal coronary vasodilation, interstitial myocarditis, interstitial cystitis, etc. It can also be used as an agent for preventing or preventing fibrosis associated with a disease.

  The metal chelate compound (1) of the present invention (hereinafter referred to as “the present compound”) is a human or other mammal as a preventive or therapeutic agent for the above-mentioned tissue fibrosis disease (hereinafter referred to as “the present pharmaceutical composition”). (For example, mice, rats, hamsters, rabbits, cats, dogs, cows, sheep, monkeys, etc.) orally or parenterally (intravenous, subcutaneous, transdermal, transpulmonary, transmucosal ( Nasal nose), rectal administration, etc.]. The present pharmaceutical composition comprises a pharmaceutically acceptable carrier (excipient, binder, disintegrant, disintegrating aid, lubricant, wetting agent, etc.) ordinarily used for oral or parenteral administration. Mixed with additives, etc., granules, powders, tablets (including sugar-coated tablets), pills, buccals, capsules, syrups, solutions, emulsions, suspensions, creams, ointments, eye drops, injections It can be prepared by formulating it into a desired form such as a drip, nasal drop, patch, suppository and the like.

  Pharmaceutically acceptable carriers commonly used for formulation, such as binders (eg syrup, gum arabic, gelatin, sorbit, tragacanth, polyvinylpyrrolidone), excipients (eg lactose, sugar, corn starch, phosphorus (Potassium acid, sorbit, glycine), lubricant (eg, magnesium stearate, talc, polyethylene glycol, silica), disintegrant (eg, potato starch), wetting agent (eg, sodium lauryl sulfate), thickener, dispersant In addition, as other additives, a reabsorption accelerator, a pH adjuster, a surfactant, a solubilizer, a preservative, an emulsifier, an isotonic agent, a stabilizer, and the like may be used as appropriate.

  The content of the present compound in the antifibrotic agent varies depending on the form of the preparation, but is usually 0.1 to 100% by weight, preferably 1 to 98% by weight. For example, in the case of an injection, it is preferable that the injection solution contains 0.1 to 30% by weight, preferably 1 to 10% by weight of the active ingredient.

  The dosage when this compound is used as a prophylactic or therapeutic agent for tissue fibrosis varies depending on the type of the compound used, the weight and age of the patient, the type and condition of the target disease, and the administration method. For example, in the case of an injection, the amount of the present compound is about 1 mg to about 500 mg once a day for an adult, and in the case of an oral administration agent such as a tablet, the dose is about 1 mg for an adult several times a day. About 1000 mg, and in the case of external preparations such as ointments, creams and patches, it is preferable to administer a dose of about 1 mg to about 1000 mg.

  An anti-fibrotic agent (a prophylactic or therapeutic agent for tissue fibrosis disease) containing the present compound may appropriately contain other steroids or other types of medicinal ingredients as long as the object of the present invention is not violated.

  Next, although a manufacture example, an experiment example, and an Example are given and this invention is demonstrated, it is not limited to the range of this invention.

[Production Example 1] 6,8-dimercaptooctanoic acid sodium / zinc chelate compound, and
6,8-dimercaptooctanoic acid / zinc chelate monoethanolamine (aminoethanol) salt 6.2 g of DL-α-riboic acid was dissolved in 70 mL of methanol, and 3.0 g of zinc not added and 40 mL of 1N hydrochloric acid were added at 50 ° C. Stir for 1 hour. Next, the zinc in the end reaction is filtered off, the filtrate is concentrated under reduced pressure, and water is added to this, and the precipitated white crystals are collected by filtration. This was suspended in 150 mL of water, dissolved with 2N sodium hydroxide to about pH 9 and dissolved, and the insoluble material was filtered off. The filtrate was concentrated, ethanol was added thereto, and the precipitated white crystals were collected by filtration. This was recrystallized from water / ethanol to obtain 6.0 g of the title sodium 6,8-dimercaptooctanoate / zinc chelate compound. mp. 300 ° C. or higher, TLC, Rf = 0.88 (n-butanol: acetic acid: water = 4: 1: 2).

  In addition, in the above, monoethanolamine was used in place of sodium hydroxide and the others were treated in the same manner to obtain 8.5 g of the title 6,8-dimercaptooctanoic acid / zinc chelate monoethanolamine (aminoethanol) salt. Obtained. mp. 137-139 ° C.

[Production Example 2] 3.5 g of ethyl 6,8-dimercaptooctanoate / zinc chelate compound DL-α-ribonate was dissolved in 60 mL of tetrahydrofuran, and 2.0 g of zinc powder and 40 mL of 70% aqueous acetic acid solution were added thereto. In addition, the mixture was stirred at 50 ° C. for 2 hours. Next, zinc in the end reaction was filtered off, the filtrate was concentrated, water was added to this, and the precipitated white crystals were collected by filtration and recrystallized from acetic acid / water to obtain 3.6 g of the title metal chelate compound. . mp. Decompose gradually from around 290 ° C. TLC, Rf = 0.88 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 3] 4.2 g of 6,8-dimercaptooctanoic acid amide / zinc chelate compound DL-α-lipoic acid amide was dissolved in 70 mL of tetrahydrofuran, and 2.5 g of zinc dust and 30 mL of 50% acetic acid aqueous solution were added thereto. In addition, the mixture was stirred at 50 ° C. for 2 hours. Next, after the solvent was distilled off, the precipitated crystals mixed with zinc were collected by filtration, washed with water and ethanol, and recrystallized from acetic acid / water to obtain 4.5 g of the title metal chelate compound as white crystals. mp. 257-259 ° C., TLC, Rf = 0.80 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 4] Using 4.2 g of N- (6,8-dimercaptooctanoyl) glycine sodium / zinc chelate compound DL-a-lipoic acid and 1.9 g of glycine, N-α-lipoylglycine sodium ( mp.218-220 ° C.), and this was treated in the same manner as in Production Example 3 to obtain 3.9 g of the title metal chelate compound as white crystals. mp. Decomposes from around 297 ° C. TLC, Rf = 0.64 (chloroform: methanol: water = 5: 4: 1).

[Production Example 5] Using 4.2 g of N- (6,8-dimercaptooctanoyl) aspartic acid monosodium / zinc chelate compound DL-α-lipoic acid and 2.9 g of L-aspartic acid, N-α- This was treated with sodium lipoylaspartate (mp. 300 ° C. or higher) in the same manner as in Production Example 3 to obtain 4.2 g of the title metal chelate compound as white crystals. mp. Decomposes from around 295 ° C. TLC, Rf = 0.53 (chloroform: methanol: water = 5: 4: 1).

[Production Example 6] Using 4.2 g of N- (6,8-dimercaptooctanoyl) methionine sodium / zinc chelate compound DL-α-lipoic acid and 3.5 g of L-methionine, N-α-lipoylmethionine (Mp.108-109 ° C.) was treated in the same manner as in Production Example 3 to obtain 2.8 g of the title compound as white crystals. mp. Decomposition from around 260 ° C., TLC, Rf = 0.82 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 7] N- (6,8-dimerkabutoctanoyl) cysteine / chelate compound DL-α-lipoic acid 4.2 g and L-cysteine 2.6 g were used to produce N-α-lipoylcysteine. This was treated with sodium (mp. Decomposed from around 150 ° C.) and treated in the same manner as in Production Example 3 to obtain 4.1 g of the title compound as white crystals. mp. Decomposition from around 280 ° C., TLC, Rf = 0.71 (chloroform: methanol: water = 5: 4: 1).

[Production Example 8] Using 4.2 g of N- (6,8-dimercaptooctanoyl) phenylalanine sodium / zinc chelate compound DL-α-lipoic acid and 3.5 g of L-phenylalanine, N-α-lipoylphenylalanine (Mp.154-156 ° C.), and this was treated in the same manner as in Production Example 3 to obtain 3.9 g of the title compound as white crystals. mp. Decomposition from around 270 ° C., TLC, Rf = 0.82 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 9] Using 4.2 g of sodium N- (6,8-dimerkabutoctanoyl) -4-aminobutyrate / zinc chelate DL-α-lipoic acid and 2.3 g of 4-aminobutyric acid, N -Α-Lipoyl-4-aminobutyric acid (mp. Decomposed from around 235 ° C) was treated in the same manner as in Production Example 3 to obtain 5.2 g of the title compound as white crystals. mp. Decomposes from around 297 ° C. TLC, Rf = 0.70 (chloroform: methanol: water = 5: 4: 1).

[Production Example 10] Using 4.2 g of sodium N- (6,8-dimercaptooctanoyl) -6-aminohexanoate / zinc chelate DL-α-riboic acid and 3.0 g of 6-aminohexanoic acid, This was treated with sodium N-α-lipoyl-6-aminohexanoate (mp. 200-202 ° C.) in the same manner as in Production Example 3 to obtain 2.0 g of the title compound as white crystals. mp. Decomposes from around 295 ° C. TLC, Rf = 0.84 (chloroform: methanol: water = 5: 4: 1).

[Production Example 11] Using N- (6,8-dimercaptooctanoyl) anthranilate sodium / zinc chelate compound DL-α-riboic acid 4.2 g and anthranilic acid 2.9 g, N-α-lipoylanthranyl This was treated with sodium acid (mp. 300 ° C. or higher) in the same manner as in Production Example 3 to obtain 2.1 g of the title compound as white crystals. mp. Decomposes from around 290 ° C. TLC, Rf = 0.88 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 12] Using 6.2 g of sodium N- (6,8-dimercaptooctanoyl) -2-aminoethanesulfonate / zinc chelate DL-α-lipoic acid and 4.5 g of 2-aminoethanesulfonic acid After passing through sodium N-α-lipoylaminoethanesulfonate (mp. 235-237 ° C.), this was treated in the same manner as in Production Example 3 to obtain 4.5 g of the title compound as white crystals. mp. Decomposes from around 293 ° C. TLC, Rf = 0.51 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 13] Similar to Production Example 12 from 4.2 g of N- (6,8-dimercaptooctanoyl) hydroxyproline sodium / zinc chelate compound DL-α-lipoic acid and 2.8 g of L-4-hydroxyproline As a result, 4.9 g of the title compound as white crystals was obtained. mp. 300 ° C or higher. TLC, Rf = 0.66 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 14] From 4.2 g of N- (6,8-dimercaptooctanoyl) histidine sodium / zinc chelate compound DL-α-lipoic acid and 3.4 g of L-histidine, white color was produced in the same manner as in Production Example 12. 5.8 g of the title compound was obtained. mp. 300 ° C or higher. TLC, Rf = 0.39 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 15] N- (6,8-dimercaptooctanoyl) sodium glutamate / zinc chelate compound DL-α-lipoic acid 4.2 g and L-glutamic acid 3.5 g were used in the same manner as in Production Example 12 to produce white There were obtained 5.7 g of the title compound as crystals. mp. 300 ° C or higher. TLC, Rf = 0.74 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 16] From 4.2 g of N- (6,8-dimercaptooctanoyl) threonine sodium zinc chelate compound DL-α-lipoic acid and 2.6 g of L-threonine, white color was produced in the same manner as in Production Example 12. Obtained 5.5 g of the title compound as crystals. mp. 300 ° C or higher. TLC, Rf = 0.73 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 17] From 4.2 g of N- (6,8-dimercaptooctanoyl) alanine sodium zinc chelate compound DL-α-lipoic acid and 2.1 g of L-alanine and in the same manner as in Production Example 12, white 5.4 g of the title compound was obtained. mp. Decomposes from around 290 ° C. TLC, Rf = 0.78 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 18] From 4.2 g of N- (6,8-dimercaptooctanoyl) serine sodium / zinc chelate compound DL-α-lipoic acid and 2.4 g of L-serine in the same manner as in Production Example 12, Obtained 5.0 g of the title compound as crystals. mp. Decompose gradually from around 285 ° C. TLC, Rf = 0.64 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 19] From 4.2 g of N- (6,8-dimercaptooctanoyl) norleucine sodium / zinc chelate compound DL-α-lipoic acid and 3.0 g of L-norleucine, in the same manner as in Production Example 12, N-α-lipoylnorleucine (mp. 120-121 ° C.) was obtained to give 5.1 g of the title compound as white crystals. mp. Decompose gradually from around 295 ° C. TLC, Rf = 0.90 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 20] Production example from 4.2 g of N- (6,8-dimercaptooctanoyl) -5-hydroxytryptophan sodium / zinc chelate compound DL-α-lipoic acid and 5.0 g of L-5-hydroxytryptophan In the same manner as in Example 12, 6.5 g of the title compound as grayish white crystals was obtained. mp. Decomposes from around 290 ° C. TLC, Rf = 0.81 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 21] From 4.2 g of N- (6,8-dimercaptooctanoyl) penicillamine sodium / zinc chelate compound DL-α-lipoic acid and 3.5 g of D-penicillamine in the same manner as in Production Example 12, white 6.0 g of the title compound was obtained. mp. Decompose gradually from around 280 ° C. TLC, Rf = 0.80 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 22] From 4.2 g of sodium N- (6,8-dimercaptooctanoyl) -3-aminopropionate / zinc chelate DL-α-lipoic acid and 2.0 g of β-alanine, Production Example 12 and In the same manner, 5.8 g of the title compound as white crystals was obtained. mp. Decompose gradually from around 295 ° C. TLC, Rf = 0.83 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 23] N- (6,8-dimercaptooctanoyl) -trans-4- (aminomethyl) cyclohexane-1-carboxylate sodium / zinc chelate compound (also known as N- (6,8-dimercaptoocta Noyl) -sodium tranexamic acid / zinc chelate compound)
In the same manner as in Production Example 12, 5.8 g of the title compound as white crystals was obtained from 4.2 g of DL-α-lipoic acid and 3.5 g of 4-transaminomethylcyclohexanecarboxylic acid. mp. Decomposes gradually from around 297 ° C. TLC, Rf = 0.81 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 24] N- (6,8-dimercaptooctanoyl) sodium sulfanilate / zinc chelate compound DL-α-lipoic acid 4.2 g and sulfanilic acid (4-aminobenzene-1-sulfonic acid) 3.8 g Thus, in the same manner as in Production Example 12, 5.4 g of the title compound as white crystals was obtained. mp. 300 ° C. or higher, TLC, Rf = 0.57 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 25] 4.2 g of N- (6,8-dimercaptooctanoyl) isopropylamine / zinc chelate DL-α-lipoic acid and 2.4 g of triethylamine were dissolved in 50 mL of acetonitrile and the mixture was stirred at -5 ° C. After cooling, 2.4 g of ethyl chlorocarbonate was gradually added dropwise. 20 minutes after the completion of the dropwise addition, a solution of 1.5 g of isopropylamine dissolved in 30 mL of acetonitrile was quickly added and stirred for 30 minutes. After returning to room temperature, the mixture was further stirred for 1 hour. The solvent was distilled off under reduced pressure, water was added to the residue and the mixture was cooled, and the precipitated pale yellow crystals were collected by filtration. This was dissolved in 60 mL of tetrahydrofuran (THF), 20 mL of 50% aqueous acetic acid solution and 2.0 g of zinc dust were added, and the mixture was stirred at 50 ° C. for 2 hours. Unreacted zinc was filtered off, and the filtrate was concentrated. Water was added to the residue, and the precipitated white crystals were collected by filtration and recrystallized from THF / acetic acid / water to obtain 5.0 g of the title compound. mp. 271 to 273 ° C., TLC, Rf = 0.89 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 26] Title of white crystals using 4.2 g of N- (6,8-dimerkabutoctanoyl) -2-aminoethanol / zinc chelate compound DL-α-lipoic acid and 1.5 g of monoethanolamine 4.2 g of compound was obtained. mp. Decomposes gradually from around 298 ° C. TLC, Rf = 0.77 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 27] Using 4.2 g of N- (5,8-dimercaptooctanoyl) 5-O-methyl serotonin / zinc chelate DL-α-lipoic acid and 4.0 g of O-methyl serotonin, white crystals Of the title compound was obtained. mp. 210-212 ° C., TLC, Rf = 0.84 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 28] Title of white crystal using 4.2 g of N- (6,8-dimercaptooctanoyl) -2-aminopyridine / zinc chelate compound DL-α-riboic acid and 2.2 g of 2-aminopyridine 5.3 g of compound was obtained. mp. 243-245 ° C, TLC, Rf = 0.87 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 29] N- (6,8-dimercaptooctanoyl) ethyl anthranilate / zinc chelate compound In place of 1.5 g of isopropylamine, 3.6 g of ethyl anthranilate was used in the same manner as in Production Example 25. 4.6 g of the title compound as white crystals (recrystallized from THF-acetic acid-water) was obtained. mp. Decompose gradually from around 290 ° C. TLC, Rf = 0.88 (n-butanol: acetic acid: water = 4: 1: 2).

Production Example 30] N ^epsilon chromatography (6,8-dimercapto-octanoyl) lysine zinc chelate compound DL-alpha-lipoic acid 4.2 g, triethylamine 2.4g and ethyl chloroformate 2.4g acetonitrile 50 mL, cooled A mixed acid anhydride was used under the reaction, and 3.1 g of L-lysine, 5.5 g of copper sulfate (pentahydrate) and 2.0 g of sodium hydroxide dissolved in 60 mL of water were added and reacted. The precipitated copper salt of N ^ε- (α-lipoyl) lysine was collected by filtration, washed with water and methanol, then suspended in a 70% aqueous acetic acid solution, and filtered with copper sulfide as copper sulfide with hydrogen sulfide. did. The filtrate was concentrated, and 3.5 g of pale yellow crystals deposited by adding methanol to the residue were collected by filtration. mp. 254-255 ° C.

  Next, this was dissolved in a 60% aqueous acetic acid solution, 2.0 g of zinc powder was added, and the mixture was stirred at 50 ° C. for 3 hours. After the zinc was filtered off, the filtrate was concentrated, and methanol was added thereto to precipitate it. 3.4 g of the title compound as white crystals was obtained. mp. Decompose gradually from around 295 ° C. TLC, Rf = 0.47 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 31] N- (6,8-dimercaptooctanoyl) aspartiylglycine disodium / zinc chelate compound 2.1 g of DL-α-lipoic acid and 2.1 g of L-aspartiylglycine were used in the same manner. As a result, 3.1 g of white crystals of the title compound were obtained from N- (α-lipoyl) aspartiylglycine sodium. mp. Decompose gradually from around 270 ° C. TLC, Rf = 0.54 (n-butanol: acetic acid: water = 4: 1: 2).

[Production Example 32] Similar to Production Example 25 using 2.1 g of sodium N- (dimercaptooctanoyl) threonylglycine / zinc chelate compound DL-α-lipoic acid and 2.1 g of L-threonylglycine. As a result, 2.6 g of pale yellowish white crystals of the title compound were obtained. mp. Decompose gradually from around 260 ° C. TLC, Rf = 0.60 (n-butanol: acetic acid: water = 4: 1: 2).

  In Experimental Examples 1 and 2 below, the effect of the metal chelate compound (1) of the present invention on fibrotic tissues (lung and liver) was examined. In addition, as the metal chelate compound (1) of the present invention, N- (6,8-dimercaptooctanoyl) -L-histidine sodium / zinc chelate compound (hereinafter simply referred to as “DHLHZn”) prepared in Production Example 14 (Molecular weight 540.17) was used. The study was approved by the Oita University School of Medicine Animal Research Ethics Committee, and all protocols were conducted in accordance with National Institutes of Health (NIH) guidelines.

Experimental example 1
Using a bleomycin-induced pulmonary fibrosis model animal, the effect of the metal chelate compound (1) of the invention on pulmonary fibrosis was evaluated.

(1) Creation of bleomycin-induced pulmonary fibrosis model animal
Among 30 Wistar rats (male, 250 to 300 g), 20 mg of bleomycin was administered into the respiratory tract for 20 animals to induce pulmonary fibrosis artificially. It was created. The remaining 10 animals served as a control group (normal animals). All rats were allowed unlimited access to food and water before and after the experiment.

(2) Administration of test compound (DHLHZn) Rats (20 rats) administered with bleomycin were divided into 10 rats each for 2 minutes to obtain a DHLHZn administration group and a DHLHZn non-administration group. In the DHLHZn administration group, 10 mg / kg DHLHZn (Production Example 14) suspended in physiological saline was subcutaneously administered twice daily for 14 days from the bleomycin administration day. The control group and the DHLHZn non-administered group were subcutaneously administered with the same amount of physiological saline as the DHLHZn-administered group.

(3) Pathological analysis On the 14th day after administration of bleomycin, lungs were removed from each rat (control group, DHLHZn administration group, DHLHZn non-administration group) and fixed with 10% formalin neutral phosphate buffer. The tissue was photographed with a digital camera (Olympus DP11) after preparing a paraffin section, or hematoxylin / eosin staining (HE staining) or Azan staining (Azan staining).

(4) Results The results of HE staining are shown in FIG. 1, and the results of Azan staining are shown in FIG. In each figure, A is an image of lung tissue of the control group, B is an image of lung tissue of the DHLHZn non-administered group, and C is an image of lung tissue of the DHLHZn-administered group.

  As can be seen from this figure, a clear fibrosis image was observed in the lung 14 days after bleomycin administration, but this fibrosis portion was significantly improved by administration of the metal chelate compound (1) (DHLHZn) of the present invention. It was observed that This indicates that the metal chelate compound (1) of the present invention suppresses tissue fibrosis, that is, has an action of preventing or ameliorating tissue fibrosis disease.

Experimental example 2
Using a non-alcoholic steatohepatitis (NASH) model animal, the effect of the metal chelate compound (1) of the invention on fibrosis of the liver was evaluated.

(1) Test compound (DHLHZn) administration NASH model mice (20 mice), 5 each, DHLHZn subcutaneous administration group, DHLHZn oral administration group, control group (high fat diet intake group), and normal group (normal diet intake group) ). In the DHLHZn subcutaneous administration group, 10 mg of DHLHZn suspended in physiological saline was subcutaneously administered daily for 4 weeks while freely taking a high fat diet (HFD-60). The DHLHZn oral administration group was allowed to freely receive a high fat diet (HFD-60) containing 0.5% DHLHZn for 4 weeks. The control group (high fat diet intake group) and the normal group (normal diet intake group) were allowed to freely receive a high fat diet (HFD-60) and a normal diet for 4 weeks, respectively.

(2) Pathological analysis After the above administration was continued for 4 weeks, the liver was removed from each NASH model mouse (subcutaneous administration group, oral administration group, control group, and normal group) and fixed with 10% formalin neutral phosphate buffer. did. The tissue was prepared with paraffin sections, hematoxylin / eosin stained (HE stained) or Azan stained (Azan stained), and then photographed with a digital camera (DP11 manufactured by Olympus). The results of HE staining are shown in FIG. 3, and the results of Azan staining are shown in FIG. The result of HE staining indicates the severity of NASH as the NASH activity score, and the result of Azan staining indicates the degree of liver fibrosis. The NASH activity score (fatting, inflammation, hypertrophy / expansion) evaluated based on the results of HE staining is also shown in FIG.

  From these results, all NASH mice that were administered DHLHZn subcutaneously or orally had improved fatiness, hypertrophy and swelling in the NASH activity score (see Fig. 3), and also suppressed liver fibrosis. It was recognized (see FIG. 4). From this, it is considered that the symptoms of NASH are reduced and liver fibrosis is suppressed by administering the metal chelate compound (1) of the present invention subcutaneously or orally.

(3) mRNA analysis of SREBP-1c and TNF-α in liver tissue Liver tissue extracted from each NASH model mouse (subcutaneous administration group, oral administration group, control group, and normal group) after the above administration was continued for 4 weeks. Was used to measure the amounts of SREBP-1c and TNF-α mRNA. The results are shown in FIGS. 5A and 5B, respectively. As shown in FIGS. 5A and 5B, the amounts of SREBP-1c and TNF-α mRNA in the liver tissue of NASH model mice administered DHLHZn subcutaneously or orally were significantly lower than those in the control group, and almost the same as in the normal group. It was about the same. SREBP-1c is an important transcription factor involved in fat synthesis, and TNF-α is an important factor as an inflammatory mediator. In this experiment, it was observed that these increased in the NASH model mice and that the increase was suppressed by subcutaneous and oral administration of DHLHZn. In addition, an increase in SREBP-1c is a characteristic phenomenon as a NASH model mouse, indicating that it is in a fatty liver state due to a lipid load. In addition, activation of TNF-α reveals that inflammatory stimuli are added, but it was confirmed that DHLHZn controls both of them.

(4) Antioxidant effect In the liver tissue by ELISA method using liver tissue excised from each NASH model mouse (subcutaneous administration group, oral administration group, control group, and normal group) after the above administration was continued for 4 weeks. The amount of 8-OHdG was measured. The results are shown in FIG. As shown in FIG. 6, the amount of 8-OHdG in the liver tissue of NASH model mice administered DHLHZn subcutaneously or orally was significantly lower than that in the control group. Oxidative stress is involved in various organ diseases, particularly playing an important role in fibrosis. In this experiment, it was confirmed that in NASH model mice, 8-OHDG, which is an index of oxidative stress, was increased, and that the increase was significantly suppressed by subcutaneous and oral administration of DHLHZn.

(5) UCP-2 expression in liver tissue Western blotting using liver tissue extracted from each NASH model mouse (subcutaneous administration group, oral administration group, control group, and normal group) after the above administration was continued for 4 weeks. The UCP-2 expression level in liver tissue was measured by the method. The results are shown in FIG. As shown in FIG. 7, it was confirmed that the expression of UCP-2 was enhanced in the liver tissue of NASH model mice administered subcutaneously with DHLHZn. UCP-2 is an important protein of mitocodria, and maintaining this protein can maintain mitochondrial function and prevent cell death. This experiment confirmed that DHLHZn has a function of protecting mitochondria via UCP-2.

  From the above results, it was observed that administration of the metal chelate compound (1) (DHLHZn) of the present invention significantly improved lung tissue fibrosis. This indicates that the metal chelate compound (1) of the present invention suppresses tissue fibrosis, that is, has an action of preventing or ameliorating tissue fibrosis disease.

Example [Formulation Example 1] Oral Tablets Compound of Production Example 14 30 mg
Lactose 80mg
Potato starch 17mg
Polyethylene glycol 6000 3mg
The above ingredients are molded by a conventional method as a material for one tablet.

[Formulation Example 2] Injection Compound of Production Example 14 1.09
Manitol 4.09
Distilled water for injection balance 100mL
The above is mixed and dissolved by a conventional method to prepare an injection.

Claims (5)

A preventive or therapeutic agent for tissue fibrosis, comprising a metal chelate compound represented by the following general formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:

(In the formula, R represents an OH group, an O-lower alkyl group, an amino group, an N-substituted amino group, an amino acid residue, or a peptide residue, and M represents a pharmaceutically acceptable metal).   The agent for preventing or treating tissue fibrosis according to claim 1, wherein R in the formula (I) is a residue of an amino acid selected from the group consisting of α-amino acids, ω-amino acids and special amino acids.   In formula (I), the amino acid residue represented by R is glycine, alanine, threonine, serine, aspartic acid, glutamic acid, phenylalanine, methionine, norleucine, cysteine, hydroxyproline, histidine, 5-hydroxytryptophan, penicillamine, lysine, At least one selected from the group consisting of 3-aminopropionic acid, 4-aminobutyric acid, 6-aminohexanoic acid, trans-4-aminomethylcyclohexane-1-carboxylic acid, 2-aminoethanesulfonic acid, sulfanilic acid and anthranilic acid The preventive or therapeutic agent for tissue fibrosis according to claim 1 or 2, which is a seed.   The preventive or therapeutic agent for tissue fibrosis according to any one of claims 1 to 3, wherein the metal is zinc.   The agent for preventing or treating tissue fibrosis according to any one of claims 1 to 4, wherein the target organ for preventing or treating tissue fibrosis is lung, bladder, kidney, heart or liver.

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