JPWO2007083689A1 - Plasminogen activator inhibitor-1 inhibitor - Google Patents

Abstract

The PAI-1 inhibitor and pharmaceutical composition of the present invention have the following formula (1): [Chemical formula 1] [wherein R1 is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R2 represents an optionally substituted phenyl group or thienyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms; R3 represents a hydrogen atom. Or R2 and R3 may be bonded to each other to form a 5- to 6-membered ring; A is a linear or branched alkylene, alkenylene or alkynylene having 1 to 7 carbon atoms, or a single bond; R4 Is a hydrogen atom, an optionally substituted phenyl group or furyl group, or the following formula (2); [Chemical formula 2] wherein R1 ′, R2 ′ and R3 ′ are R1, R2 and R3, respectively. Is the same meaning as.). ] Or a salt thereof as an active ingredient.

Description

  The present invention relates to a plasminogen activator inhibitor-1 (hereinafter also referred to as “PAI-1”) inhibitor. The present invention also relates to a pharmaceutical composition having an action of inhibiting PAI-1 activity and effective for the prevention or treatment of various diseases in which PAI-1 activity is involved in the onset. Furthermore, this invention relates to the novel compound which has PAI-1 inhibitory activity.

  Thrombus is the cause of ischemic heart disease such as cerebral embolism, cerebral infarction, ischemic cerebrovascular disorder such as transient cerebral ischemic attack, angina, myocardial infarction, atrial thrombosis in atrial fibrillation, heart failure obtain. Blood circulation requires fluidity for transporting oxygen and nutrients to body tissues and collecting unwanted substances, but also requires coagulation to stop blood and prevent blood loss during trauma. When the contradictory functions of fluidity and coagulation are imbalanced and tilted toward the coagulation side, it is thought that blood clots are formed in the blood vessels, causing ischemic cerebrovascular disorders and heart diseases.

  The fibrinolytic system (hereinafter referred to as “dietary system”) plays an important role in thrombolysis, tissue destruction and repair, cell migration, and the like. The fibrinolytic system is activated when a plasminogen activator (hereinafter referred to as “PA”) converts plasminogen to plasmin. On the other hand, plasminogen activator inhibitor-1 (PAI-1) inhibits PA.

  A tissue plasminogen activator (hereinafter referred to as “t-PA”) converts plasminogen, which is a precursor of plasmin, to plasmin. Plasmin breaks down fibrin into a fibrin degradation product.

  PAI-1 is a serine protease inhibitor that specifically inhibits t-PA and urokinase-type plasminogen activator (hereinafter referred to as “u-PA”), suppresses the production of plasmin, and thus degrades fibrin. Inhibits.

  PAI-1 is classified into an active form that exhibits PA inhibitory activity and a latent form that does not exhibit PA inhibitory activity, depending on the three-dimensional structure. Usually, 20 ng / mL PAI-1 is present in plasma, and it is also known that it is produced by hepatocytes, megakaryocytes, and adipocytes in addition to vascular endothelial cells, which are the main production cells. Yes.

  PAI-1 is an acute phase protein, and its production is enhanced by various cytokines and growth factors. It is considered as one of the causes of ischemic organ damage in sepsis and disseminated intravascular coagulation syndrome (DIC). Yes. In addition, genetic polymorphism due to single base substitution of the PAI-1 gene promoter is known, and it has been clarified that plasma PAI-1 concentration increases due to the genetic polymorphism.

  In addition, with regard to diabetes, it is believed that accelerated arteriosclerosis and microvascular complications cause ischemic heart disease, diabetic retinopathy and kidney damage, which are important complications of diabetes. For example, in diabetic nephropathy, an increase in extracellular matrix in the glomerulus and interstitial fibrosis are characteristically observed, and the expression of PAI-1 in the glomeruli and tubules is enhanced. In proximal tubule culture, increased production of PAI-1 is observed under hyperglycemic conditions. Further, in an experiment using a renal interstitial fibrosis model mouse, a correlation between expression of PAI-1 in the renal tissue and macrophage infiltration has been confirmed (see Non-Patent Document 1).

  Moreover, from the result of concentrating the daily urine collection of patients with nephrotic syndrome and measuring the amount of PAI-1 in urine, it has been reported that the concentration of PAI-1 in urine of patients with nephrotic syndrome is high (see Non-Patent Document 2). ).

  As a result of administering an inactive PAI-1 mutant (Non-patent document 3) or t-PA (Non-patent document 4) as a PAI-1 antagonist to a Thy-1 nephritis model, reduction of inflammation (cell infiltration), TGF- There is a report that a decrease in β and a decrease in mesangial substrate were confirmed, and an improvement in Thy-1 nephritis was observed.

  The decrease in fibrinolytic activity due to increased plasma concentration of PAI-1 has been associated with deep vein thrombosis, ischemic heart disease and diabetic vasculopathy. In addition to reduced fibrinolytic activity, several other thrombogenic abnormalities, including hypercoagulability and platelet hyperaggregability, have also been shown in diabetic patients, which contribute to microthrombosis and are diabetic It plays an important role in the progression of microangiopathy and diabetic macrovascular disorders.

  Thus, PAI-1 is considered to be involved in the formation and development of various pathologies such as various thrombosis, cancer, diabetes, and arteriosclerosis. Therefore, compounds that inhibit PAI-1 activity are useful as prophylactic and therapeutic agents for diseases associated with decreased fibrinolytic activity such as thrombosis, cancer, diabetic complications, and arteriosclerosis. (Non-patent document 5).

In addition, tissue fibrosis occurs in many tissues and organs such as the lung, heart, blood vessels, liver, and kidneys, but there are no drugs that can fundamentally treat them, such as prednisolone and corticosteroids. At present, cytotoxic drugs such as cyclophosphamide (alkylating agent) and azathioprine (antimetabolite, immunosuppressive agent) are used for symptomatic treatment.
Aya N, et al., J. Pathol., The Journal of Pathology, 166, 289-295, 1992 Yoshida Y, et al., Nephron, 88, 24-29, 2001 WA Border, et al., J. Clin. Invest. (The Journal of Clinical Investigation), 112, 379, 2003 WA Border, et al., Kidney Int. (Kidney International), 59, 246, 2001 Egelund R, et al. (Egelund et al.), J. Biol. Chem. (The Journal of Biological Chemistry), 276, 13077-13086, 2001

  Until now, urokinase, which is u-PA, is known as a fibrinolytic promoter, but it is obtained by purification from human urine, and it cannot be said that production efficiency and safety are high. Furthermore, urokinase is a high molecular compound having a molecular weight of about 54,000. Other fibrinolytic promoters include tisokinase, alteplase (genetical recombination), nasarplase (cell culture), nateplase (genetic recombination), monteplase (genetic recombination), pamitepase (genetic recombination) and batroxobin. However, both are high molecular compounds. Therefore, there is a demand for a low molecular weight compound-derived drug that can be synthesized in large quantities and is highly safe as a fibrinolytic promoter. There is also a need for the development of effective drugs that fundamentally treat and improve tissue fibrosis.

  In view of such conventional problems, the present invention provides a highly safe pharmaceutical composition comprising a low-molecular compound capable of mass synthesis as an active ingredient, particularly a pharmaceutical composition useful as a fibrinolytic promoter or antifibrosis agent. The purpose is to do. It is another object of the present invention to provide a novel compound useful as an active ingredient of a pharmaceutical composition such as a fibrinolytic accelerator or an antifibrotic agent.

  As a result of diligent research to solve the above problems, the present inventors have found that the compound represented by the following formula (1) or a salt thereof, or a solvate thereof (hereinafter collectively referred to as “the present invention”). Of compound (1) "or" compound (1) ") has a high inhibitory activity against plasminogen activator inhibitor-1 (PAI-1), which is further a PAI-1 inhibitor It was confirmed that it was useful as an active ingredient of a fibrinolytic accelerator. In addition, the present inventors have found that PAI-1 is a main cause of tissue fibrosis, particularly pulmonary fibrosis, and confirmed that this tissue fibrosis is significantly improved by the compound (1) of the present invention. In addition, the present inventors also include a compound represented by the formula (3) described later (hereinafter, these compounds (1) are collectively referred to as “compound (3) of the present invention” or “compound (3)”). In particular, it was confirmed that the compounds (4) to (16) are novel compounds not described in any literature. The present invention has been completed based on such findings.

That is, the present invention includes the following embodiments:
(I) Plasminogen activator inhibitor-1 (PAI-1) inhibitor Item 1. The following formula (1):

[Wherein, R ₁ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, or carbon A linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ is a hydrogen atom, an optionally substituted phenyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms; Or a halogen atom. R ₂ and R ₃ may be bonded to each other to form a 5- to 6-membered ring; A is a linear or branched alkylene, alkenylene or alkynylene having 1 to 7 carbon atoms, 3 carbon atoms -8 cycloalkylene, or single bond; R ₄ is a hydrogen atom, an optionally substituted phenyl group or furyl group, or the following formula (2);

(Wherein R ₁ ′ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ ′ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, Or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ ′ is a hydrogen atom, an optionally substituted phenyl group, or a linear or branched chain group having 1 to 6 carbon atoms. And R ₂ ′ and R ₃ ′ may be bonded to each other to form a 5- to 6-membered ring. ]
Or a salt thereof, or a solvate thereof, as an active ingredient.

Item 2. In the general formula (1), A is a linear or branched alkylene having 1 to 7 carbon atoms, or a cycloalkylene having 3 to 8 carbon atoms; R ₄ is the following formula (2);

(Wherein R ₁ ′ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ ′ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, Or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ ′ is a hydrogen atom, an optionally substituted phenyl group, or a linear or branched chain group having 1 to 6 carbon atoms. And R ₂ ′ and R ₃ ′ may be bonded to each other to form a 5- to 6-membered ring.)
The PAI-1 inhibitor of claim | item 1 which uses the compound shown by these, its salt, or these solvates as an active ingredient.

Item 3. R ₂ and R ₂ ′ are the same or different and may be a phenyl group, a 2-thienyl group, —CH ₂ CH (CH ₃ ) ₂ which may have a substituent, or the same or different, Item 3. The PAI-1 inhibitor according to Item 1 or 2, wherein ₂ and R ₃ or R ₂ 'and R ₃ ' are bonded to form a condensed 6-membered ring.

Item 4. R ₃ and R ₃ ′ are the same or different and each is a hydrogen atom, a linear or branched alkyl group having 1 to 3 carbon atoms, a phenyl group, or a halogen atom, The PAI-1 inhibitor described.

Item 5. Item 5. The PAI-1 inhibition according to any one of Items 1 to 4, wherein A is a linear alkylene having 3 to 5 carbon atoms, —CH ₂ —C (CH ₃ ) ₂ —CH ₂ —, or cyclohexylene. Agent.

Item 6. Item 6. The PAI-1 inhibitor according to any one of Items 1 to 5, wherein R ₁ is a hydrogen atom, R ₂ and R ₂ ′ are a phenyl group or a 2-thienyl group, and A is butylene.

(II) Pharmaceutical composition Item 7. Item 7. A pharmaceutical composition comprising the PAI-1 inhibitor according to any one of Items 1 to 6, and a pharmaceutically acceptable carrier or additive. In other words, a pharmaceutical composition comprising the compound described in the general formula (1) or a salt thereof, or a solvate thereof, and a pharmaceutically acceptable carrier or additive.

  Item 8. Item 8. The pharmaceutical composition according to Item 7, which is a preventive or therapeutic agent for a disease in which PAI-1 activity is involved in the onset.

Item 9. Item 10. The pharmaceutical composition according to Item 7 or 8, which is a fibrinolytic promoter. Diseases related to PAI-1 activity include angina, myocardial infarction or atrial thrombosis in atrial fibrillation, ischemic heart disease, ischemic cerebrovascular disorder, arteriosclerosis, pulmonary embolism, during surgery Restenosis after deep vein thrombosis (DVT), disseminated intravascular coagulation syndrome (DIC), vascular disorders as diabetic complications, neuropathy, retinopathy or nephropathy, or percutaneous coronary angioplasty (PTCA) Item 10. The pharmaceutical composition according to Item 9, wherein

  Item 11. Item 9. The pharmaceutical composition according to Item 8, wherein the disease associated with the onset of PAI-1 activity is a disease associated with tissue fibrosis.

  Item 12. Item 12. The pharmaceutical composition according to Item 11, wherein the disease involving tissue fibrosis is pulmonary fibrosis.

  Item 13. Item 13. The pharmaceutical composition according to any one of Items 7 to 12, which has an oral dosage form.

  Item 14. The following formula (3):

[Wherein, R ₁ and R ₁ ′ are the same or different, a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ and R ₂ ′ are the same or different; A hydrogen atom, an optionally substituted phenyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ and R ₃ ′ may be the same or different, A phenyl group which may have a group, a linear or branched alkyl group having 1 to 6 carbon atoms, or a halogen atom; A is a linear or branched alkylene having 1 to 7 carbon atoms, Or a C3-C8 cycloalkylene is shown. However, when R ₂ and R ₂ ′ are phenyl groups having no substituent and R ₃ and R ₃ ′ are hydrogen atoms, A represents butylene and —CH ₂ —C (CH ₃ ) ₂ —CH _2. When neither R nor R ₂ and R ₂ ′ are isobutyl groups and R ₃ and R ₃ ′ are hydrogen atoms, A is not butylene. ]
Or a salt thereof.

  Item 15. A compound represented by any of the following formulas (4) to (16) or a salt thereof:

2- [5- (3'-carboxy-4 '-(p-chlorophenyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid

2- [4- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -butyrylamino] -4-isobutylthiophene-3-carboxylic acid

2- [4- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -3,3-dimethylbutyrylamino] -4-isobutylthiophene-3-carboxylic acid

2- [6- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -hexanoylamino] -4-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-methyl-4'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-chlorothiophene-2'-ylcarbamoyl) -pentanoylamino] -5-chlorothiophene-3-carboxylic acid

2- [4- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3-carboxylic acid

2- [5- (3'-carboxy-5'-isopropyl-4'-methylthiophen-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid

2- [3- (3'-Carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -propanoylamino] -4-isobutylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-4'-isopropylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid

2- [5- (3'-Carboxy-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

2- [5- (3'-tert-butoxycarbonyl-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

  Item 16. Item 16. A pharmaceutical composition comprising as an active ingredient at least one selected from the group consisting of the compound according to Item 14 or 15, or a pharmaceutically acceptable salt thereof, or a solvate thereof.

  In addition, the use of the said pharmaceutical composition is not specifically limited, It is not necessarily limited to what is based on PAI-1 inhibitory action, such as the prevention or treatment use of the disease in which PAI-1 activity is related to onset.

  INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a pharmaceutical composition comprising a low-molecular compound that can be synthesized in large quantities and is highly safe as an active ingredient. The pharmaceutical composition comprises a compound having a high inhibitory action on PAI-1 (PAI-1 inhibitor) as an active ingredient, and prevents or treats various diseases caused by PAI-1 activity. It can be used effectively as an agent. In particular, the pharmaceutical composition of the present invention is used as a fibrinolytic promoter as angina, myocardial infarction or atrial thrombosis in atrial fibrillation, ischemic heart disease, ischemic cerebrovascular disorder, arteriosclerosis, pulmonary embolism, After surgical deep vein thrombosis (DVT), disseminated intravascular coagulation syndrome (DIC), vascular disorders as diabetic complications, neuropathy, retinopathy or nephropathy, or percutaneous coronary angioplasty (PTCA) It is useful for the prevention or treatment of restenosis. The pharmaceutical composition of the present invention is useful as an antifibrotic agent for the prevention or treatment of various diseases related to tissue fibrosis, particularly pulmonary fibrosis.

  Moreover, according to this invention, the novel compound which has a high inhibitory effect with respect to PAI-1 can be provided. Such a compound is useful as an active ingredient of a pharmaceutical composition such as a prophylactic or therapeutic agent for various diseases caused by PAI-1 activity.

(I) PAI-1 inhibitor The PAI-1 inhibitor provided by the present invention has the following formula (1):

Or a salt thereof as an active ingredient.

In formula (1), R ₁ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms. Here, as the alkyl group having 1 to 4 carbon atoms, straight chain such as methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, and ter-butyl group Or a branched alkyl group can be mentioned, and R ₁ is preferably a hydrogen atom and a t-butyl group.

In Formula (1), R ₂ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms. Preferred are a phenyl group, a thienyl group, a methyl group, an isobutyl group, an isopropyl group, or a hydrogen atom, which may have a substituent, and more preferred are a thienyl group, a phenyl group, and an isobutyl group.

Preferred examples of the thienyl group include a 2-thienyl group. Examples of the substituent of the phenyl group or thienyl group include a carboxyl group, an amino group, a halogen atom, and a heterocyclic group. Preferably, it is a halogen atom (for example, chlorine atom, fluorine atom, iodine atom, etc.).
Examples of the linear or branched alkyl group having 1 to 6 carbon atoms include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, and ter-butyl group. Pentyl group, isopentyl group, neopentyl group, ter-pentyl group, hexyl group, isohexyl group, neohexyl group, ter-hexyl group. Preferably it is a C1-C4 linear or branched alkyl group, More preferably, they are a methyl group, an isopropyl group, and an isobutyl group.

In formula (1), R ₂ and R ₃ may be bonded to each other together with the carbon atom to which they are bonded to form a 5- to 6-membered ring. Examples of such 5-6 membered rings include cyclohexane, cyclohexene, 1,3-cyclohexadiene, cyclopentane, cyclopentene and benzene. Cyclohexane is preferred.

R ₃ can also be a hydrogen atom, a phenyl group which may have a substituent, a linear or branched alkyl group having 1 to 6 carbon atoms, or a halogen atom. Examples of the substituent of the phenyl group include a carboxyl group, an amino group, a halogen atom, and a heterocyclic group as described above. Moreover, what is mentioned above similarly can be mentioned also as a C1-C6 linear or branched alkyl group. R ₃ is preferably a hydrogen atom, a phenyl group, a methyl group, an isopropyl group, or a halogen atom.

A is a single bond; a linear or branched alkylene, alkenylene, or alkynylene having 1 to 7 carbon atoms; a cycloalkylene having 3 to 8 carbon atoms. Preferred are straight-chain or branched alkylene having 2 to 5 carbon atoms, cycloalkylene having 6 carbon atoms (cyclohexanediyl), and vinylene. Specific examples of the linear or branched alkylene having 2 to 5 carbon atoms include an ethylene group, a propylene group, a butylene group, a pentylene group, and —CH ₂ C (CH ₃ ) ₂ CH ₂ —. . A butylene group, —CH ₂ C (CH ₃ ) ₂ CH ₂ —, and cyclohexanediyl are preferred.

In the formula (1), examples of R ₄ include a hydrogen atom or a phenyl group or a furyl group which may have a substituent.

  Here, the furyl group is preferably a 2-furyl group. When the phenyl group or furyl group has a substituent, the substituent is a chlorine atom, an iodine atom, a bromine atom or a halogen atom of a fluorine atom; a linear or branched alkyl group having 1 to 6 carbon atoms; Can be mentioned. A halogen atom is preferably a fluorine atom or a chlorine atom. Specific examples of the linear or branched alkyl group having 1 to 6 carbon atoms include those described above, preferably a ter-butyl group and an n-butyl group.

R ₄ may be a group represented by the following formula (2):

Here, as R ₁ ′, a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms can be exemplified as in R ₁ described above. When the compound (1) has the above group (2) as R ₄ , R ₁ ′ and R ₁ may be different, but are preferably the same.

As R ₂ ′, as in R ₂ described above, a hydrogen atom, an optionally substituted phenyl group or thienyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms. The group can be mentioned. When the compound (1) has the above group (2) as R ₄ , R ₂ ′ and R ₂ may be different but are preferably the same.

Similarly to R ₃ described above, R ₃ ′ is a hydrogen atom, a phenyl group which may have a substituent, a linear or branched alkyl group having 1 to 6 carbon atoms, or a halogen atom. Alternatively, R ₂ ′ may be combined with the carbon atom to which they are bonded to form a 5- to 6-membered ring. When the compound (1) has the above group (2) as R ₄ , R ₃ and R ₃ ′ may be different from each other, but are preferably the same, and R ₂ and R ₂ ′ are simultaneously the same. And may form a 5- to 6-membered ring.

As a preferred compound, in formula (1), A is a linear or branched alkylene having 1 to 7 carbon atoms, or a cycloalkylene having 3 to 8 carbon atoms, and R ₄ is the group (2 ). Examples of such suitable compounds include compounds represented by the following formula (17):

(Wherein R ₁ , R ₂ , R ₃ , R ₁ ′, R ₂ ′ and R ₃ ′ are as defined above.)
The compound (1) (particularly the compound (17)) targeted by the present invention is preferably a compound in which R ₁ and R ₁ ′ are hydrogen atoms, and R ₁ and R ₁ ′ are the same or different and are ter-butyl groups Can be mentioned. More preferably, R ₂ and R ₂ ′ are a hydrogen atom, a phenyl group having a substituent or an unsubstituted group, a 2-thienyl group, an isopropyl group, an isobutyl group [—CH ₂ CH (CH ₃ ) ₂ ]. Or R ₂ and R ₃ and R ₂ ′ and R ₃ ′ are bonded to each other to form a 6-membered ring. Even more preferably, A is ethylene, propylene, butylene, pentylene, —CH ₂ —C (CH ₃ ) ₂ —CH ₂ —, or cyclohexanediyl. As a more preferred compound (1) of the present invention (particularly compound (17)), R ₁ , R ₁ ′, R ₃ and R ₃ ′ are all hydrogen atoms, and R ₂ and R ₂ ′ are phenyl groups or A compound that is a 2-thienyl group and A is butylene or —CH ₂ —C (CH ₃ ) ₂ —CH ₂ — can be mentioned.

  In addition, although these compounds (1) which this invention makes object may have a free form as mentioned above, they may have a salt form. Here, examples of the salt include pharmaceutically acceptable salts such as salts with inorganic bases or organic bases, salts with basic amino acids, and the like. Examples of the inorganic base include alkali metals such as sodium and potassium; alkaline earth metals such as calcium and magnesium; aluminum and ammonium. Examples of the organic base include primary amines such as ethanolamine; secondary amines such as diethylamine, diethanolamine, dicyclohexylamine, N, N′-dibenzylethylenediamine; trimethylamine, triethylamine, pyridine, picoline, triethanolamine and the like. And tertiary amines. Examples of basic amino acids include arginine, lysine, ornithine and the like.

  Specific examples of the preferred compound (1) targeted by the present invention include the following compounds an to n and o to zz.

Compound a
2- [3- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid

Compound b and its salt
2- [5- (3'-carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid

2- [5- (3'-carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid disodium salt

Compound c
2- [3- (3'-carboxy-4 ', 5', 6 ', 7'-tetrahydro-benzo [b] thiophen-2'-ylcarbamoyl) -pentanoylamino] -4,5,6,7 -Tetrahydro-benzo [b] thiophene-3-carboxylic acid

Compound d and its salt
2- [5- (3'Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid disodium salt

Compound e
2- [3- (3′-carboxy-4 ′, 5 ′, 6 ′, 7′-tetrahydro [b] thiophen-2′-ylcarbamoyl) -propanoylamino] -4,5,6,7-tetrahydro -Benzo [b] thiophene-3-carboxylic acid

Compound f
2-Hexanoylamino-4-p-tolylthiophene-3-carboxylic acid

Compound g
2- (3-Furan-2-yl-acryloylamino) -4- (2-thienyl) -3-carboxylic acid

Compound h and its salt
2- [4- (3'-Carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl)-3,3-dimethylbutyrylamino] -4- (2-thienyl) thiophene-3-carbon acid

2- [4- (3'-Carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl)-3,3-dimethylbutyrylamino] -4- (2-thienyl) thiophene-3-carbon Acid disodium salt

Compound i
4-Isobutyl-2-cinnamoylthiophene-3-carboxylic acid

Compound j
2- [4- (3′-carboxy-4 ′, 5 ′, 6 ′, 7′-tetrahydro-benzo [b] thiophen-2-ylcarbamoyl) -3,3-dimethyl] -butyrylamino) -4,5 , 6,7-Tetrahydro-benzo [b] thiophene-3-carboxylic acid

Compound k
2- [5- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl)-(3,3-dimethyl)) butylamino] -4-phenyl-thiophene-3-carboxylic acid

Compound l
2- (4-tert-Butylbenzoylamino) -4- (2-thienyl) -3-carboxylic acid

Compound m
2- (3-Phenylpropionylamino) -4- (2-thienyl) -3-carboxylic acid

Compound n
2- (4-Fluorobenzoylamino) -4- (2-thienyl) -3-carboxylic acid

Compound o
2- [5- (3'-Carboxy-4 '-(p-chlorophenyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid

Compound p
2- [4- (3'-Carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -butyrylamino] -4-isobutylthiophene-3-carboxylic acid

Compound q
2- [4- (3'-Carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -3,3-dimethylbutyrylamino-dimethylbutyrylamino] -4-isobutylthiophene-3-carboxylic acid

Compound r
2- [6- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -hexanoylamino] -4-phenylthiophene-3-carboxylic acid

Compound s
2- [5- (3'-Carboxy-5'-methyl-4'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3-carboxylic acid

Compound t
2- [5- (3'-Carboxy-5'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-phenylthiophene-3-carboxylic acid

Compound u
2- [5- (3'-Carboxy-5'-chlorothiophene-2'-ylcarbamoyl) -pentanoylamino] -5-chlorothiophene-3-carboxylic acid

Compound v
2- [4- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3-carboxylic acid

Compound w
2- [5- (3'-Carboxy-5'-isopropyl-4'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid

Compound x
2- [3- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -propanoylamino] -4-isobutylthiophene-3-carboxylic acid

Compound y
2- [5- (3'-Carboxy-4'-isopropylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid

Compound z
2- [5- (3'-Carboxy-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

Compound zz
2- [5- (3'-tert-Butoxycarbonyl-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

  These compounds a to n and o to zz are all included in the compound (1) represented by the general formula (1), and among them, the compounds a, b, c, d, e, h, j , K, o to zz are compounds included in the compound represented by the general formula (17). Of the compounds (1), compounds a, b, c, d, e, g, h, i, j, k, l, s, t, v, and r are more preferable, and compounds a, b, d, e, h, s and t, particularly preferably compounds a, b, d and h.

  Further, the compound (1) targeted by the present invention may be a compound formed by solvating a free substance or a salt thereof. Such solvates include hydrates.

Compound (1), particularly compounds a to n, are commercially available, and can be produced by a method known per se. For example, among the compounds (1), a compound having a group represented by the formula (2) as R ₄ can be produced by the production method 1 of the following (A) or a method analogous thereto. In addition, in the compound (1), compounds o to zz are novel compounds not yet described in the literature, and the production method thereof will be described in detail in Production Examples 5 to 17.

(A) Manufacturing method 1
Formula (18):

(In the formula, R ₁ ^″ is a linear or branched alkyl group having 1 to 4 carbon atoms, and R ₂ and R ₃ are as defined above.)
A compound represented by the formula (19)

(Wherein A is as defined above and X is a leaving group.)
Then, the resulting compound is hydrolyzed as necessary to produce a compound (1) (especially compound (17)) in which R ₁ is a hydrogen atom.

A compound having a hydrogen atom as R ₄ or a phenyl group or a furyl group which may have a substituent can be produced by the production method 2 of the following (B) or a method analogous thereto.

(B) Manufacturing method 2
Formula (20):

(In the formula, A is as defined above, X is a leaving group, R ₄ ^′ is a hydrogen atom, a phenyl group or a furyl group which may have a substituent.)
A compound represented by formula (18)

(Wherein R ₁ ^″ , R ₂ and R ₃ are as defined above.)
And then the resulting compound is optionally hydrolyzed, and R ₁ is a hydrogen atom, R ₄ is a hydrogen atom, or a phenyl group or a furyl group optionally having a substituent. Compound (1) is produced.

  Here, in the formulas (19) and (20), examples of the leaving group represented by X include a halogen atom. Specific examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom and an iodine atom. Preferably it is chlorine.

The reaction of the compound (18) with the compound (19) or the compound (18) and the compound (20) is carried out in accordance with a normal acylation reaction known to those skilled in the art, and an appropriate base in an appropriate solvent. Can be done in the presence. As the base, an organic base such as triethylamine or pyridine can be suitably used. The object compound (1) (amide-carboxylic acid form) in which R ₁ is hydrogen can be produced by subjecting the amide-ester form thus obtained to an alkaline hydrolysis reaction as necessary. This alkaline hydrolysis reaction can be carried out by an ordinary method known to those skilled in the art, and as the alkali to be used, potassium hydroxide, sodium hydroxide, barium hydroxide or the like is used.

  The PAI-1 inhibitory activity of compound (1) can be evaluated by an in vitro assay system. As such an in vitro assay system, for example, a method for measuring a change in PAI-1 activity relative to tissue plasminogen activator (t-PA) in the presence of compound (1) can be mentioned. In addition, the activity fluctuation | variation of this PAI-1 can be measured by making into a parameter | index the reaction product produced by the effect | action of t-PA with respect to a substrate. For example, in Experimental Example 1, which will be described later, in vitro measurement of PAI-1 activity variation using the amount of p-nitroanilide (reaction product) generated by the action of t-PA on the chromogenic substrate (S-2288) as an index. 1 illustrates an assay system. It can be judged that the smaller the amount of reaction product produced, the higher the PAI-1 inhibitory activity.

  Further, the PAI-1 inhibitory activity of the compound (1) is the change in the formation of a complex of PAI-1 and t-PA (PAI-1 / t-PA complex) in the presence of the compound (1), for example. It can also be evaluated by measuring by Western blotting or the like (for example, see Experimental Example 2). Here, it can be judged that the smaller the amount of PAI-1 / t-PA complex formed (PAI-1 / t-PA complex formation inhibition), the higher the PAI-1 inhibitory activity.

  Compound (1) has an action of inhibiting the activity of PAI-1. Among them, compounds a to l, preferably compounds a, b, d, e and h, particularly preferably compounds a, b, d and h have an excellent PAI-1 activity inhibitory action as shown in the experimental examples described later. is doing. By this action, it is possible to enhance the degradation of fibrin and fibrinogen by plasmin, promote the fibrinolytic system of the living body, and improve the degradation of the fibrinolytic system of the living body.

  In addition, it has now been found that one of the causes of tissue fibrosis is PAI-1. Therefore, according to compound (1), it is possible to prevent or ameliorate tissue fibrosis and diseases related to tissue fibrosis based on the action of inhibiting the activity of PAI-1.

  The PAI-1 inhibitor of the present invention comprises such a compound (1) as an active ingredient. The PAI-1 inhibitor of the present invention may be composed of 100% of the compound (1), or contains an effective amount of the compound (1) that exhibits a PAI-1 inhibitory action even if it is not. Anything is acceptable. Although not limited, the PAI-1 inhibitor usually contains compound (1) in the range of 0.1 to 99% by weight, preferably 1 to 80% by weight.

(II) Pharmaceutical Composition The present invention provides a pharmaceutical composition containing the aforementioned PAI-1 inhibitor as an active ingredient. In other words, the pharmaceutical composition of the present invention contains the aforementioned compound (1) as an active ingredient. The pharmaceutical composition of the present invention has a PAI-1 inhibitory action by containing an effective amount of compound (1). As a result, the degradation of fibrin and fibrinogen by plasmin is enhanced, and the fibrinolytic system of the living body. Has the effect of promoting or improving the decreased fibrinolytic system of the living body.

  For this reason, the pharmaceutical composition of the present invention can be used as a fibrinolytic promoter. Specifically, the pharmaceutical composition of the present invention is useful as a prophylactic or therapeutic agent for a thrombotic disease or condition in which PAI-1 activity is involved in the onset, or a disease or condition caused by a decrease in fibrinolytic system. . Examples of such diseases or conditions include ischemic brain such as angina pectoris, myocardial infarction, intra-atrial thrombus in atrial fibrillation, ischemic heart disease such as heart failure, cerebral embolism, cerebral infarction, transient ischemic attack, etc. Vascular disorders, arteriosclerosis, pulmonary embolism, deep vein thrombosis (DVT) during surgery, disseminated intravascular coagulation syndrome (DIC), diabetic complications (vascular disorders, neuropathy, retinopathy, nephropathy, etc.) In addition, various diseases or conditions in which thrombus formation such as restenosis after percutaneous coronary angioplasty (PTCA) is involved can be mentioned.

  Moreover, the pharmaceutical composition of this invention has a PAI-1 inhibitory effect by containing an effective amount of compound (1), As a result, it has the effect | action which prevents or improves the fibrosis of a tissue or an organ. Therefore, the pharmaceutical composition of the present invention is useful as a prophylactic or therapeutic agent for diseases and pathological conditions related to tissue or organ fibrosis caused by PAI-1 activity. Examples of such diseases or conditions include pulmonary fibrosis, tissue fibrosis associated with myocardial infarction, tissue fibrosis associated with nephropathy, and the like.

  The pharmaceutical composition of the present invention is usually prepared by blending a pharmaceutically acceptable carrier or additive in addition to an effective amount of compound (1) for promoting (or improving) the fibrinolytic system or antifibrosis. The The compounding amount of the compound (1) in the pharmaceutical composition is appropriately selected according to the type of disease or pathology to be targeted and the dosage form. Usually, in the case of a systemic preparation, the total weight of the pharmaceutical composition (100% by weight) of 0.001 to 50% by weight, particularly 0.01 to 10% by weight.

  Examples of the administration method of the pharmaceutical composition of the present invention include oral administration and parenteral administration such as intravenous administration, intramuscular administration, subcutaneous administration, transmucosal administration, transdermal administration, and rectal administration. Oral administration and intravenous administration are preferable, and oral administration is more preferable. The pharmaceutical composition of the present invention can be prepared into various forms of preparations (dosage forms) depending on the administration method. The preparations (dosage forms) will be described below, but the dosage forms used in the present invention are not limited to these, and various dosage forms that are usually used in the pharmaceutical preparation field can be used.

  Examples of dosage forms for oral administration include powders, granules, capsules, pills, tablets, elixirs, suspensions, emulsions, and syrups, and can be appropriately selected from these. . Moreover, modifications such as sustained release, stabilization, easy disintegration, poor disintegration, enteric solubility, easy absorption and the like can be applied to these preparations.

  In addition, dosage forms for intravenous administration, intramuscular administration, or subcutaneous administration include injections and infusions (including dried products prepared at the time of use), and can be selected as appropriate.

  In addition, dosage forms for transmucosal administration, transdermal administration, or rectal administration include mastication agents, sublingual agents, buccal agents, troches, ointments, patches, liquids, etc. You can select here as appropriate. These formulations can also be modified such as sustained release, stabilization, easy disintegration, difficulty disintegration, and easy absorption.

  The pharmaceutical composition of the present invention can contain pharmaceutically acceptable carriers and additives depending on the dosage form (oral administration or various parenteral administration dosage forms). Pharmaceutically acceptable carriers and additives include solvents, excipients, coating agents, bases, binders, lubricants, disintegrants, solubilizers, suspending agents, thickeners, emulsifiers, Stabilizers, buffers, tonicity agents, soothing agents, preservatives, flavoring agents, fragrances, and coloring agents can be mentioned. Specific examples of pharmaceutically acceptable carriers and additives are listed below, but the present invention is not limited thereto.

  Examples of the solvent include purified water, sterilized purified water, water for injection, physiological saline, peanut oil, ethanol, glycerin and the like. Excipients include starches (eg potato starch, wheat starch, corn starch), lactose, glucose, sucrose, crystalline cellulose, calcium sulfate, calcium carbonate, sodium bicarbonate, sodium chloride, talc, titanium oxide, trehalose, xylitol Etc.

  Examples of binders include starch and derivatives thereof, cellulose and derivatives thereof (for example, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose), gelatin, sodium alginate, tragacanth, gum arabic and other natural polymer compounds, polyvinyl pyrrolidone, polyvinyl alcohol, etc. Synthetic polymer compounds, dextrin, hydroxypropyl starch and the like.

  Lubricants include light anhydrous silicic acid, stearic acid and its salts (eg, magnesium stearate), talc, waxes, wheat denbun, macrogol, hydrogenated vegetable oil, sucrose fatty acid ester, polyethylene glycol, silicone oil, etc. be able to.

  Examples of disintegrants include starch and derivatives thereof, agar, gelatin powder, sodium bicarbonate, calcium carbonate, cellulose and derivatives thereof, hydroxypropyl starch, carboxymethylcellulose and salts thereof, and cross-linked products thereof, and low-substituted hydroxypropylcellulose. be able to.

  Examples of the solubilizer include cyclodextrin, ethanol, propylene glycol, polyethylene glycol and the like. Examples of the suspending agent include sodium carboxymethylcellulose, polypinyl pyrrolidone, gum arabic, tragacanth, sodium alginate, aluminum monostearate, citric acid, various surfactants and the like.

  Examples of the thickener include sodium carboxymethyl cellulose, polypinyl pyrrolidone, methyl cellulose, hydroxypropyl methyl cellulose, polyvinyl alcohol, tragacanth, gum arabic, sodium alginate and the like.

  Examples of the emulsifier include gum arabic, cholesterol, tragacanth, methyl cellulose, lecithin, various surfactants (for example, polyoxyl 40 stearate, sorbitan sesquioleate, polysorbate 80, sodium lauryl sulfate).

  Stabilizers include tocopherols, chelating agents (eg, EDTA, thioglycolic acid), inert gases (eg, nitrogen, carbon dioxide), reducing substances (eg, sodium bisulfite, sodium thiosulfate, ascorbic acid, Rongalite) and the like. be able to.

  Examples of the buffer include sodium hydrogen phosphate, sodium acetate, sodium citrate, boric acid and the like.

  Examples of isotonic agents include sodium chloride and glucose. Examples of soothing agents include local anesthetics (procaine hydrochloride, lidocaine), pendyl alcohol, glucose, sorbitol, amino acids and the like.

  Examples of the corrigent include sucrose, saccharin, licorice extract, sorbitol, xylitol, glycerin and the like. Examples of fragrances include spruce tincture and rose oil. Examples of the colorant include water-soluble food dyes and lake dyes.

  Examples of the preservative include benzoic acid and its salts, paraoxybenzoic acid esters, chlorobutanol, reverse soap, benzyl alcohol, phenol, thimerosal, dehydroacetic acid, boric acid, and the like.

  Coating agents include sucrose, hydroxypropylcellulose (HPC), shellac, gelatin, glycerin, sorbitol, hydroxypropylmethylcellulose (HPMC), ethylcellulose, polyvinylpyrrolidone (PVP), hydroxypropylmethylcellulose phthalate (HPMCP), cellulose acetate phthalate (CAP) ), Methyl methacrylate-methacrylic acid copolymer and the above-described polymers.

  Bases include petrolatum, liquid paraffin, carnauba wax, beef tallow, hardened oil, paraffin, beeswax, vegetable oil, macrogol, macrogol fatty acid ester, stearic acid, sodium carboxymethylcellulose, bentonite, cacao butter, witepsol, gelatin, stearyl Alcohol, hydrolanolin, cetanol, light liquid paraffin, hydrophilic petrolatum, simple ointment, white ointment, hydrophilic ointment, macrogol ointment, hard fat, oil-in-water emulsion base, water-in-oil emulsion glaze etc. Can do.

  In addition, about each said dosage form, the technique of a well-known drug delivery system (DDS) is employable. The DDS preparations referred to in this specification include sustained release preparations, topical preparations (troches, buccal tablets, sublingual tablets, etc.), drug release control preparations, enteric preparations and gastric preparations, etc., administration routes, bioavailability Considering side effects and the like, it is a preparation in an optimal preparation form.

  When the pharmaceutical composition of the present invention is used as a prophylactic or therapeutic agent for a disease state associated with a decrease in fibrinolytic system (thrombosis formation), the oral dose is 0.03 in terms of the amount of compound (1). The range of 300 mg / kg body weight is preferable, and 0.1 to 50 mg / kg body weight is more preferable. In the case of intravenous administration, a dose such that the effective blood concentration of compound (1) is in the range of 0.2 to 50 μg / mL, more preferably 0.5 to 20 μg / mL can be mentioned.

  When the pharmaceutical composition of the present invention is used as a prophylactic or therapeutic agent for a disease state associated with tissue fibrosis, the oral dose is 0.03 to 300 mg / kg body weight in terms of the amount of compound (1). The range is preferably 0.1 to 50 mg / kg body weight. In the case of intravenous administration, a dose such that the effective blood concentration of compound (1) is in the range of 0.2 to 50 μg / mL, more preferably 0.5 to 20 μg / mL can be mentioned. Note that these doses may vary depending on age, sex, body type and the like.

(III) Novel compound and use thereof As described above, among the compounds represented by the general formula (1), the compound represented by the following formula (3) and a salt thereof are novel compounds not described in any literature.

[Wherein, R ₁ and R ₁ ′ are the same or different, a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ and R ₂ ′ are the same or different; A hydrogen atom, an optionally substituted phenyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ and R ₃ ′ may be the same or different, A phenyl group which may have a group, a linear or branched alkyl group having 1 to 6 carbon atoms, or a halogen atom; A is a linear or branched alkylene having 1 to 7 carbon atoms, Or a C3-C8 cycloalkylene is shown. However, when R ₂ and R ₂ ′ are phenyl groups having no substituent and R ₃ and R ₃ ′ are hydrogen atoms, A represents butylene and —CH ₂ —C (CH ₃ ) ₂ —CH _2. When neither R nor R ₂ and R ₂ ′ are isobutyl groups and R ₃ and R ₃ ′ are hydrogen atoms, A is not butylene. ]
Or a salt thereof.

  The present invention provides such novel compounds.

Here, in the above formula (3), specific definitions of R ₁ , R ₂ , R ₃ , R ₁ ′, R ₂ ′, R ₃ ′ and A are as described for the compound (1) described above. It is.

Examples of the novel compound (3) targeted by the present invention include those in which R ₁ and R ₁ ′ are the same or different and are a hydrogen atom or a ter-butyl group. Preferably, R ₁ and R ₁ ′ are hydrogen atoms. In addition, as the novel compound (3), R ₂ and R ₂ ′ are the same or different and each has a hydrogen atom, a halogen atom as a substituent, or a phenyl group, a methyl group, an isopropyl group, an isobutyl group [ may be mentioned those which are -CH _{2 CH} (CH _{3) 2].} Further, examples of the novel compound (3) include those in which R ₃ and R ₃ ′ are the same or different and are a hydrogen atom, a methyl group, an isopropyl group, a phenyl group, or a halogen atom. Examples of A include propylene, butylene, pentylene, —CH ₂ —C (CH ₃ ) ₂ —CH ₂ —, and cyclohexylene.

  Specific examples of the new compound (3) include the following compounds o to zz. The method for producing such a compound will be described in detail in Production Examples 5 to 17.

Compound o
2- [5- (3'-Carboxy-4 '-(p-chlorophenyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid

Compound p
2- [4- (3'-Carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -butyrylamino] -4-isobutylthiophene-3-carboxylic acid

Compound q
2- [4- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -3,3-dimethylbutyrylamino] -4-isobutylthiophene-3-carboxylic acid

Compound r
2- [6- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -hexanoylamino] -4-phenylthiophene-3-carboxylic acid

Compound s
2- [5- (3'-Carboxy-5'-methyl-4'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3-carboxylic acid

Compound t
2- [5- (3'-Carboxy-5'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-phenylthiophene-3-carboxylic acid

Compound u
2- [5- (3'-Carboxy-5'-chlorothiophene-2'-ylcarbamoyl) -pentanoylamino] -5-chlorothiophene-3-carboxylic acid

Compound v
2- [4- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3-carboxylic acid

Compound w
2- [5- (3'-Carboxy-5'-isopropyl-4'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid

Compound x
2- [3- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -propanoylamino] -4-isobutylthiophene-3-carboxylic acid

Compound y
2- [5- (3'-Carboxy-4'-isopropylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid

Compound z
2- [5- (3'-Carboxy-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

Compound zz
2- [5- (3'-tert-Butoxycarbonyl-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

  These compounds have a PAI-1 inhibitory action as described above, and diseases or pathologies related to the onset of PAI-1, such as pathologies or tissue fibers related to a decrease in the fibrinolytic system (thrombus formation) It is useful as an active ingredient of a prophylactic or therapeutic agent (pharmaceutical composition) for pathological conditions related to chemicalization. Among these compounds (compounds o to zz), compounds o to x are preferable, compounds s, t, r, q and w are more preferable, and compounds s, t and r are more preferable.

  Therefore, the present invention provides a pharmaceutical composition comprising the above compound (3), particularly compounds o to zz (preferably compounds o to x) or a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient. To do. The pharmaceutical composition can be prepared in a suitable dosage form according to the dosage form by a conventional method according to the description in (II).

Experimental example

  Hereinafter, the present invention will be described more specifically with reference to production examples and experimental examples, but the present invention is not limited to these examples.

Production Example 1
In accordance with the scheme (Step 1, Step 2, Step 3) shown in FIG.

(In the formula, R is a phenyl group or a 2-thienyl group.)
The compound shown in was synthesized.

In the formula, the compound in which R is a phenyl group is 2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid Hereinafter, it is also referred to as “compound a”. On the other hand, the compound in which R ₂ and R ₂ ^′ are 2-thienyl groups is 2- [3- (3′-carboxy-4′-thienylthiophen-2′-ylcarbamoyl) -pentanoylamino] -4-thienyl. Thiophene-3-carboxylic acid, hereinafter also referred to as “compound b”.

(1) Step 1: Synthesis of amide-ester (3)
A mixture of 10 g (54.6 mmol) adipic acid chloride anhydride (1) 114 mmol aminoester compound (2) (R = phenyl or 2-thienyl group) and 11 ml (114 mmol) pyridine in 70 ml dry dioxane. For 2.5 hours. The reaction mixture was poured into cold water while warm, and then alkalized by adding 15 g of potassium carbonate. The precipitate was filtered off and washed with water. The precipitate was dried and recrystallized using a mixed solution of MeOH / THF (50:50, 400 ml) to give 16-21 g of amide-ester (3) [R = phenyl group: 2- [3- ( 3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid diethyl ester, R = 2-thienyl group: 2- [3- (3 ′ -Carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid diethyl ester] (yield 70%) .

(2) Step 2: hydrolysis of amide-ester (3)
42.3 mmol of amide-ester (3) was dissolved in 870 mL of THF, and 0.91 mmol of aqueous sodium hydroxide solution (4) dissolved in 115 mL of water was added thereto. Heated with stirring for 3.5 hours and then left at room temperature overnight. Non-soluble impurities were filtered off and the mixture was then evaporated until the volume was approximately 250 ml (T _bath = 40 ° C.). Residual salt (5) [R = Phenyl group: 2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid Disodium salt, R = 2-thienyl group: 2- [3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl ) Thiophene-3-carboxylic acid disodium salt] was collected by filtration and washed with methylene chloride and methanol (yield 30%).

(3) Step 3: Synthesis of amide-carboxylic acid (6) (compounds a and b) The salt (5) obtained above was dissolved in a mixed solution of THF / H ₂ O (150 ml / 200 ml), and 20% Acidified with aqueous acetic acid. The precipitate was collected by filtration and washed with water. Next, the obtained acid (6) [R = phenyl group: 2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3 -Carboxylic acid (molecular weight: 552.67) (compound a), R = 2-thienyl group: 2- [3- (3'-carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl) -pentanoyl Amino] -4- (2-thienyl) thiophene-3-carboxylic acid (molecular weight: 560.69) (compound b)] was dried under vacuum at room temperature. The overall yield was about 15%. The NMR data of compounds a and b are shown in FIGS. 2 and 3, respectively.

Production Example 2
2- [5- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid (compound b) Of disodium salt

Compound b synthesized in Production Example 1 was converted to a disodium salt using an aqueous NaOH solution in THF to prepare the title compound.
mp: 258-260 ℃
¹ H-NMR (DMSO-d6) δ: 1.60-1.80 (4H, m), 2.35-2.60 (4H, m), 6.76 (2H, s), 6.95 (1H, dd, J = 3.4, 3.4Hz), 7.30 (1H, dd, J = 1.2, 3.4Hz), 7.35 (1H, dd, J = 1.2, 3.4Hz), 14.1 (2H, s).

Production Example 3
Preparation of disodium salt of 2- [5- (3'-carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid (compound d)

Compound d (Specs, The Netherlands) was converted to the disodium salt using aqueous NaOH in THF to prepare the title compound.
mp: 248-251 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 0.81 (12H, d, J = 6.8Hz), 1.55-1.70 (4H, m), 1.90 (2H, septet, J = 6.8Hz), 2.24-2.43 (4H, m), 2.72 (2H, d, J = 6.8 Hz), 6.26 (2H, s).

Production Example 4
2- [4- (3'-Carboxy-4 '-(2-thienyl) thiophen-2'-ylcarbamoyl)-3,3-dimethylbutyrylamino] -4- (2-thienyl) thiophene-3-carbon Production of disodium salt of acid (compound h)

Compound h (Specs, The Netherlands) was converted to the disodium salt using aqueous NaOH in THF to prepare the title compound.
mp: 245-248 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.14 (6H, s), 2.48 (4H, s), 6.76 (2H, s), 6.95 (1H, dd, J = 3.4, 5.2Hz), 7.29 (1H, dd, J = 1.2, 3.4 Hz), 7.35 (1H, dd, J = 1.2, 5.2 Hz), 14.1 (2H, s).

Production Example 5
2- [5- (3'-carboxy-4 '-(p-chlorophenyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid (compound o) accordance Step 1 to 3 to be described later fabrication, shown in the following formula, to synthesize a compound of the title.

(1) Step 1
7.73 g (50 mmol) p-chloroacetophenone, 4.96 g (50 mmol) methyl cyanoacetate, 0.45 g (7.5 mmol) acetic acid and 0.19 g (2.5 mmol) ammonium acetate were dehydrated and heated to reflux in 30 ml toluene for 3 hours. did. After cooling, the reaction mixture was diluted with ethyl acetate and washed with water. The obtained organic layer was dried over magnesium sulfate, and then the solvent was distilled off under reduced pressure. To the obtained residue, 60 ml of methanol, 1.6 g (50 mmol) of sulfur and 4.4 g (50 mmol) of morpholine were added and heated under reflux for 4 hours. Thereafter, the reaction solution was cooled and then filtered to concentrate the filtrate. The obtained residue was diluted with ethyl acetate, washed with water, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 1.1 g of 2-amino-4- (p-chlorophenyl) thiophene-3-carboxylic acid methyl ester (yield 7%).

(2) Step 2
A mixture of 0.16 ml (1.1 mmol) adipic acid chloride and 0.64 g (2.3 mmol) 2-amino-4- (4′-chlorophenyl) -3-thiophenecarboxylic acid methyl ester was stirred in 5 ml DMF at room temperature overnight. did. The reaction mixture was poured into ice water. The precipitate was filtered off, washed with water and diethyl ether and dried to give 0.66 g of 2- [5- (3′-carboxy-4 ′-(p-chlorophenyl) thiophen-2′-ylcarbamoyl)- Pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid dimethyl ester was obtained (yield 92%).

(3) Step 3
0.61 g (0.9 mmol) of 2- [5- (3′-carboxy-4 ′-(p-chlorophenyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3 -Carboxylic acid dimethyl ester was dissolved in 10 ml of THF solution, then 2.2 ml of 1N aqueous sodium hydroxide solution was added, and the mixture was stirred at 50-60 ° C for 2.5 hours, and further stirred at room temperature for 15 hours. The reaction mixture was concentrated and the residue was collected by filtration and washed with water. After adding THF-ethyl acetate-water to the obtained residue, separating the acidified organic layer by adding 1N hydrochloric acid and concentrating, the resulting residue was washed with water, ethyl acetate, and then dried. 2- [5- (3′-carboxy-4 ′-(p-chlorophenyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid ( Compound o) was obtained (yield 45%).
mp: 271-273 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.60-1.80 (4H, m), 2.50-2.65 (4H, m), 6.89 (2H, s), 7.29-7.41 (8H, m), 11.2 (2H, s ).

Production Example 6
Preparation of 2- [4- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -butyrylamino] -4-isobutylthiophene-3-carboxylic acid (Compound p) According to Steps 1 and 2 described below, The title compound, shown below, was synthesized.

(1) Step 1
A mixture of 0.2 ml (1.5 mmol) glutaric chloride and 0.64 g (3 mol) 2-amino-4-isobutylthiophene-3-carboxylic acid methyl ester was stirred overnight at room temperature in 6 ml DMA. The reaction mixture was extracted with ethyl acetate, dried over magnesium sulfate and concentrated to 0.73 g of 2- [4- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -butyrylamino] -4 -Dibutyl thiophene-3-carboxylic acid dimethyl ester was obtained (yield 93%).

(2) Step 2
Dissolve 730 mg (1.4 mmol) 2- [4- (3′-carboxy-4′-isobutylthiophene-2′-ylcarbamoyl) -butyrylamino] -4-isobutylthiophene-3-carboxylic acid dimethyl ester in THF; Subsequently, 1N sodium hydroxide aqueous solution was added and it stirred at 50-60 degreeC for 16 hours. After cooling, THF was distilled off under reduced pressure. The residue was collected by filtration and washed with water and ethyl acetate. The obtained residue was suspended in water, 1N hydrochloric acid was added, the mixture was extracted with ethyl acetate, and the organic layer was washed with water. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The obtained residue was collected by filtration, washed with diethyl ether and then dried to give the title 2- [4- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -butyrylamino] -4- Isobutylthiophene-3-carboxylic acid (compound p) was obtained (yield 17%).
mp: 217-218 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 0.84 (12H, d, J = 7.4Hz), 1.60-2.10 (4H, m), 2.40-2.74 (4H, m), 2.60 (4H, d, J = 7.4 Hz), 6.60 (2H, s), 11.3 (2H, s).

Production Example 7
Production of 2- [4- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -3,3-dimethylbutyrylamino] -4-isobutylthiophene-3-carboxylic acid (compound q) According to Steps 1 and 2, the title compound represented by the following formula was synthesized.

(1) Step 1
A mixture of 0.46 g (2.34 mmol) 3,3-dimethylpentanedioyl dichloride 1.0 g (4.69 mol) and 2-amino-4-isobutylthiophene-3-carboxylic acid methyl ester was stirred in DMA solution at room temperature overnight. . The reaction mixture was poured into ice water and alkalized by adding sodium bicarbonate. The mixture was extracted with ethyl acetate, dried over magnesium sulfate and concentrated. The obtained crude product was separated and purified by silica gel column chromatography to obtain 1.29 g of 2- [4- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -3,3-dimethylbutyrate. Rylamino] -4-isobutylthiophene-3-carboxylic acid dimethyl ester was obtained (yield 92%).

(2) Step 2
1.19 g (2.17 mmol) 2- [4- (3′-carboxy-4′-isobutylthiophene-2′-ylcarbamoyl) -3,3-dimethylbutyrylamino] -4-isobutylthiophene-3-carboxylic acid The dimethyl ester was dissolved in THF, 1N sodium hydroxide was then added, and the mixture was stirred at 50-60 ° C. for 4 hours. Next, the mixture was stirred overnight at room temperature, and the reaction mixture was evaporated under reduced pressure, and the resulting residue was acidified with 1N hydrochloric acid and stirred. This was filtered and the mother liquor was concentrated. The precipitated crystals were filtered and dried to give the title 2- [4- (3′-carboxy-4′-isobutylthiophene-2′-ylcarbamoyl) -3,3-dimethylbutyrylamino] -4-isobutyl Thiophene-3-carboxylic acid (compound q) was obtained (22% yield).
mp: 198-201 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 0.84 (12H, d, J = 6.6Hz), 1.12 (6H, s), 1.70-1.95 (2H, m), 2.53-2.64 (8H, m), 6.60 ( 1H, s), 11.4 (1H, s).

Production Example 8
Synthesis of 2- [6- (3′-carboxy-4′-phenylthiophen-2′-ylcarbamoyl) -hexanoylamino] -4-phenylthiophene-3-carboxylic acid (compound r) Steps 1 to 3 described later According to the procedure, the title compound shown in the following formula was synthesized.

(1) Step 1
10.6 g (88.2 mmol) benzophenone, 20 g (141.6 mmol) cyanoacetic acid tert-butyl ester, 5.3 g (88.2 mmol) acetic acid and 6.2 g (70.8 mmol) morpholine were dehydrated and heated to reflux for 12.5 hours in 50 ml toluene. did. After cooling, the reaction solution was washed with water. The obtained organic layer was dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. To the obtained residue, 100 ml of DMF, 2.8 g (88.2 mmol) of sulfur and 7.7 g (88.2 mmol) of morpholine were added and stirred overnight. Thereafter, ethyl acetate was added to the reaction solution, followed by washing with water. The obtained organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography and then recrystallized from hexane to obtain 14.7 g of 2-amino-4-phenylthiophene-3-carboxylic acid tert-butyl ester (yield) 60.5%).

(2) Step 2
A mixture of 0.36 g, (1.82 mmol) pimeloyl chloride and 1.0 g (3.63 mmol) 2-amino-4-phenylthiophene-3-carboxylic acid tert-butyl ester was stirred in a DMF solution at room temperature overnight. The reaction solution was poured into ice water and alkalized by adding sodium hydrogen carbonate. The precipitate was filtered off, washed with water, ethyl acetate-IPE mixture, dried and 0.68 g of 2- [6- (3'-carboxy-4'-phenylthiophen-2'-ylcarbamoyl)- Hexanoylamino] -4-phenylthiophene-3-carboxylic acid di-tert-butyl ester was obtained (yield 55%).

(3) Step 3
0.66 g (0.98 mmol) 2- [6- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -hexanoylamino] -4-phenylthiophene-3-carboxylic acid di-tert-butyl The ester and TFA were stirred overnight in methylene chloride solvent at room temperature. After concentrating the reaction solution, the residue was collected by filtration, washed with ethyl acetate-IPE, and dried to give the title 2- [6- (3′-carboxy-4′-phenylthiophen-2′-ylcarbamoyl) -Hexanoylamino] -4-phenylthiophene-3-carboxylic acid (compound r) was obtained (yield 85%).
mp: 204-206 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.24-1.46 (2H, m), 1.68 (4H, q, J = 7.6Hz), 2.75 (4H, t, J = 7.6Hz), 6.84 (1H, s) , 7.23-7.40 (10H, m), 11.3 (2H, s), 12.9 (2H, br).

Production Example 9
2- [5- (3′-Carboxy-5′-methyl-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3-carboxylic acid (compounds The title compound shown in the following formula was synthesized according to Steps 1 to 3 described later.

(1) Step 1
10 g (74.5 mmol) propiophenone, 14.9 g (105.5 mmol) cyanoacetic acid tert-butyl ester, 4.51 g (74.5 mmol) acetic acid and 5.19 g (59.5 mmol) morpholine dehydrated in 100 ml toluene for 3.5 hours Heated to reflux. After cooling, the reaction solution was washed with water. The obtained organic layer was dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. To the obtained residue, 100 ml of DMF, 2.4 g (74.5 mmol) of sulfur and 6.5 g (74.5 mmol) of morpholine were added and stirred overnight, and then ethyl acetate was added to the reaction solution and washed with water. The obtained organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 8.38 g of 2-amino-5-methyl-4-phenylthiophene-3-carboxylic acid tert-butyl ester (yield 39%). .

(2) Step 2
Mixture of 0.32 g (1.7 mmol) adipic acid chloride and 1.0 g (3.6 mmol) 2-amino-5-methyl-4-phenylthiophene-3-carboxylic acid tert-butyl ester in 5 ml DMA at room temperature for 2 hours Stir. The reaction mixture was poured into ice water and then alkalinized by adding sodium bicarbonate. The precipitate was filtered off, washed with water and IPE and dried to give 1.2 g of 2- [5- (3′-carboxy-5′methyl-4′-phenylthiophen-2′-ylcarbamoyl) -pentanoylamino ] -5-Methyl-4-phenylthiophene-3-carboxylic acid di-tert-butyl ester was obtained (yield 87%)
(3) Step 3
0.97 g (1.4 mmol) 2- [5- (3′-carboxy-5′methyl-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3 -Carboxylic acid di-tert-butyl ester and 2 ml TFA were stirred in 3 ml methylene chloride at room temperature overnight. After the reaction mixture was concentrated, the residue was collected by filtration, washed with ethyl acetate-IPE and dried to give the title 2- [5- (3′-carboxy-5′methyl-4′-phenylthiophene-2 '-Ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3-carboxylic acid (compound s) was obtained (yield 98%).
mp: 250-252 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.59-1.80 (4H, m), 2.07 (3H, s), 2.55-2.63 (4H, m), 7.14-7.40 (10H, m), 11.3 (2H, s ), 12.6 (2H, br).

Production Example 10
Preparation of 2- [5- (3′-carboxy-5′-phenylthiophen-2′-ylcarbamoyl) -pentanoylamino] -5-phenylthiophene-3-carboxylic acid (compound t) Steps 1 to 3 described later According to the procedure, the title compound shown in the following formula was synthesized.

(1) Step 1
10.6.g (88.2 mmol) phenylaldehyde, 20 g (141.1 mmol) cyanoacetic acid tert-butyl ester, 5.3 g (88.2 mmol) acetic acid and 6.2 g (70.6 mmol) morpholine in 50 ml toluene for 6.5 hours Heated to reflux. After cooling, the reaction solution was washed with water. The obtained organic layer was dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. To the obtained residue, 100 ml of DMF, 2.83 g (88.2 mmol) of sulfur and 7.68 g (88.2 mmol) of morpholine were added and stirred overnight. Thereafter, ethyl acetate was added to the reaction solution, followed by washing with water. The obtained organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 2.69 g of 2-amino-5-phenylthiophene-3-carboxylic acid tert-butyl ester (yield 11.1%).

(2) Step 2
A mixture of 0.42 g (2.3 mmol) adipic acid chloride and 1.0 g (3.6 mmol) 2-amino-5-phenylthiophene-3-carboxylic acid tert-butyl ester was stirred overnight at room temperature in 10 ml DMA. The reaction mixture was diluted with ethyl acetate, washed with aqueous sodium hydrogen carbonate solution, brine and water, the organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 0.37 g of 2- [5- (3′-carboxy-5′-phenylthiophen-2′-ylcarbamoyl) -pentanoylamino] -5 -Phenylthiophene-3-carboxylic acid di-tert-butyl ester was obtained (yield 31%).

(3) Step 3
0.37 g (0.56 mmol) 2- [5- (3′-carboxy-5′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -5-phenylthiophene-3-carboxylic acid di-tert-butyl The ester and 2 ml TFA were stirred in 3 ml methylene chloride under ice cooling for 2 hours and then at room temperature for 1 hour. After the reaction mixture was concentrated, the residue was collected by filtration, washed with ethyl acetate and dried. The title 2- [5- (3′-carboxy-5′-phenylthiophen-2′-ylcarbamoyl) -pentanoylamino] -5-phenyl was then recrystallized using a mixture of THF / hexane. Thiophene-3-carboxylic acid (compound t) was obtained (52% yield).
mp: 289-291 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.61-1.80 (4H, m), 2.57-2.71 (4H, m), 7.20-7.69 (10H, m), 7.51 (2H, s), 11.1 (2H, s ), 13.3 (2H, br).

Production Example 11
Production of 2- [5- (3′-carboxy-5′-chlorothiophene-2′-ylcarbamoyl) -pentanoylamino] -5-chlorothiophene-3-carboxylic acid (compound u) Steps 1 to 3 described later According to the procedure, the title compound shown in the following formula was synthesized.

(1) Step 1
A mixture of 2.2 ml (15.4 mmol) adipic acid chloride and 5.3 g (33.9 mmol) 2-aminothiophene-3-carboxylic acid methyl ester was stirred in 30 ml DMA at room temperature for 1.5 hours, then at 50-60 ° C. for 1 hour. Stir for hours. The reaction mixture was added to ice water and then made alkaline with an aqueous sodium hydrogen carbonate solution. The precipitated precipitate was collected by filtration, washed with water and ethyl acetate, and dried to give 6.5 g of 2- [5- (3′-carboxythiophene-2′-ylcarbamoyl) -pentanoylamino] -thiophene-3. -Carboxylic acid dimethyl ester was obtained (yield 99%).

(2) Step 2
To an ethanol solution of 0.85 g (2 mmol) 2- [5- (3′-carboxythiophene-2′-ylcarbamoyl) -pentanoylamino] -thiophene-3-carboxylic acid dimethyl ester was added 1N aqueous sodium hydroxide solution. After stirring at room temperature for 0.5 hour, the mixture was stirred at 50-60 ° C for 3 hours. After cooling, ethanol was distilled off under reduced pressure. Water was added to the resulting residue and the mixture was washed with ethyl acetate, and then a 1N hydrochloric acid aqueous solution was added to the aqueous layer to obtain a precipitate. The resulting precipitate was collected by filtration, washed with water and dried to obtain 0.73 g of 2- [5- (3′-carboxythiophene-2′-ylcarbamoyl) -pentanoylamino] -thiophene-3-carboxylic acid. Obtained (yield 91%).

(3) Step 3
0.5 g (1.26 mmol) 2- [5- (3′-carboxythiophen-2′-ylcarbamoyl) -pentanoylamino] -thiophene-3-carboxylic acid and 0.34 g (2.52 mmol) NCS in THF / DMF The mixture was stirred overnight at room temperature. After pouring the reaction mixture into ice water, the precipitate was collected by filtration, washed with methanol and ethyl acetate, dried and recrystallized with a mixture of THF / hexane to give 2- [5- (3'-carboxy-5 '-Chlorothiophen-2'-ylcarbamoyl) -pentanoylamino] -5-chlorothiophene-3-carboxylic acid was obtained (yield 53%).
mp: 282-284 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.46-1.78 (4H, m), 2.50-2.72 (4H, m), 7.12 (2H, s), 11.0 (2H, s), 11.3 (2H, br).

Production Example 12
Preparation of 2- [4- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3-carboxylic acid (compound v) Steps 1 to 2 described below According to the procedure, the title compound shown in the following formula was synthesized.

(1) Step 1
A mixture of 0.52 g (3.0 mmol) 1,4-cyclohexanedicarboxylic acid (cis, trans mixture) and 2.2 ml (30 mmol) thionyl chloride was stirred overnight at room temperature and then concentrated. The obtained residue and 1.49 g (5.4 mmol) of 2-amino-5-phenylthiophene-3-carboxylic acid tert-butyl ester were stirred in DMA at room temperature for 4 hours. The reaction mixture was poured into ice water and alkalized by adding sodium bicarbonate. The precipitate was collected by filtration, washed with water, diethyl ether, dried, and 0.97 g of 2- [4- (3′-carboxy-4′-phenylthiophen-2′-ylcarbamoyl) -cyclohexylcarbonylamino] -4 -Phenylthiophene-3-carboxylic acid di-tert-butyl ester was obtained (yield 52%).

(2) Step 2
0.41 g (0.6 mmol) 2- [4- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3-carboxylic acid di-tert-butyl The ester and 1 ml TFA were stirred in 3 ml methylene chloride at room temperature for 5 hours. The reaction mixture was concentrated and the resulting residue was collected by filtration, washed with IPE and dried. Then recrystallized using a mixture of THF / hexane to give 2- [4- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3- Carboxylic acid was obtained (yield 73%).
mp:> 290 ℃
¹ H-NMR (DMSO-d6) δ: 1.34-2.10 (8H, m), 2.51-2.68 (2H, m), 6.69 (2H, s), 7.22-7.40 (10H, m), 13.3 (2H, br ).

Production Example 13
Preparation of 2- [5- (3′-carboxy-5′-isopropyl-4′-methylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid (compound w) According to Steps 1 to 3, the title compound shown in the following formula was synthesized.

(1) Step 1
54 ml (430 mmol) of 4-methyl-2-pentanone, 25 ml (285 mol) of cyanoacetic acid methyl ester, 3.5 ml (61 mmol) of acetic acid and 1.5 g (19 mmol) of ammonium acetate in 90 ml of toluene for 2.5 hours did. After cooling, the reaction mixture was diluted with ethyl acetate and washed with aqueous sodium hydrogen carbonate solution and aqueous NaCl solution. The obtained organic layer was dried over magnesium sulfate, and then the solvent was distilled off under reduced pressure. To the obtained residue, 80 ml of methanol, 8.8 g (276 mmol) of sulfur and 7.4 ml (85 mmol) of morpholine were added and heated under reflux for 6 hours. The reaction solution was cooled and then filtered, and the filtrate was concentrated. The obtained residue was diluted with ethyl acetate, washed with water and aqueous NaCl solution, dried over magnesium sulfate, evaporated under reduced pressure, separated and purified by silica gel column chromatography, 2-amino-5-isopropyl-4- A mixture of methylthiophene-3-carboxylic acid methyl ester and 2-amino-4-isobutylthiophene-3-carboxylic acid methyl ester was obtained.

(2) Step 2
4.27 g of the above mixture and 0.7 ml (5 mmol) of adipic acid chloride were stirred in 20 ml of DMA at room temperature for 15 hours. The reaction mixture was poured into ice water and then neutralized by the addition of sodium bicarbonate. The resulting mixture was extracted with ethyl acetate, dried over magnesium sulfate and concentrated. The obtained crude product was separated and purified by silica gel column chromatography, crystals were precipitated with diethyl ether / hexane and separated by filtration. The obtained filtrate was concentrated and crystallized with diethyl ether to give 0.81 g of 2- [5- (3′-carboxy-5′-isopropyl-4′-methylthiophen-2′-ylcarbamoyl) -pentanoyl Amino] -4-isobutylthiophene-3-carboxylic acid dimethyl ester was obtained.

(3) Step 3
0.3 g (0.56 mmol) of 2- [5- (3′-carboxy-5′-isopropyl-4′-methylthiophen-2′-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid The dimethyl ester was dissolved in ethanol, then 2 ml of 1N sodium hydroxide was added and heated at 60-65 ° C. for 3 hours. The reaction mixture was concentrated, ice water was added to the residue, and the mixture was washed with ethyl acetate. Next, 1N hydrochloric acid was added to the aqueous layer under ice-cooling, and the mixture was concentrated under reduced pressure. Et ₂ O was added to the residue, and the precipitated crystals were collected by filtration and washed with Et ₂ O and water. 5- (3′-carboxy-5′-isopropyl-4′-methylthiophen-2′-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid (compound w) was obtained (yield) 47%).
mp: 190-192 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 0.84 (6H, d, J = 6.6Hz), 1.20 (6H, d, J = 6.8Hz), 1.15-1.70 (1H, m), 1.70-1.90 (1H, m), 2.22 (3H, s), 2.40-2.60 (6H, m), 3.25 (1H, septet, J = 6.6Hz), 6.59 (1H, s), 11.2 (1H, s), 11.3 (1H, s ).

Production Example 14
Preparation of 2- [3- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -propanoylamino] -4-isobutylthiophene-3-carboxylic acid (compound x) Steps 1 to 2 described later The title compound x shown in the following formula was synthesized.

(1) Step 1
A mixture of 0.17 ml (1.5 mmol) succinic chloride and 640 mg (3 mol) 2-amino-4-isobutylthiophene-3-carboxylic acid methyl ester was stirred in 6 ml DMA at room temperature for 18 hours. The reaction mixture was poured into ice water and alkalized by adding sodium bicarbonate. The precipitate was filtered off, washed with water, IPE and then dried to yield 646 mg of 2- [3- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -propanoylamino] -4 -Dibutyl ester of isobutylthiophene-3-carboxylic acid was obtained (yield 84%).

(2) Step 2
610 mg (1.2 mmol) 2- [3- (3'-carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -propanoylamino] -4-isobutylthiophene-3-carboxylic acid dimethyl ester dissolved in THF Then, 1N aqueous sodium hydroxide solution was added, and the mixture was stirred at 50 to 60 ° C. for 6 hours. After cooling, THF was distilled off under reduced pressure. The residue was collected by filtration and washed with water and ethyl acetate. The obtained residue was suspended in water, 1N hydrochloric acid was added, and the mixture was extracted with ethyl acetate-THF. The organic layer was washed with an aqueous NaCl solution, and then the organic layer was dried over anhydrous magnesium sulfate and concentrated. The obtained residue was collected by filtration and recrystallized with ethanol to give the title 2- [3- (3′-carboxy-4′-isobutylthiophen-2′-ylcarbamoyl) -propanoylamino] -4-isobutyl. Thiophene-3-carboxylic acid was obtained (yield 26%).
mp: 278-280 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 0.84 (12H, d, J = 6.6Hz), 1.81 (2H, septet, J = 6.8Hz), 2.61 (4H, d, J = 6.8Hz), 2.86 (4H , s), 6.60 (2H, s), 11.3 (2H, br).

Production Example 15
Preparation of 2- [5- (3′-carboxy-4′-isopropylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid (Compound y) Steps 1 to 3 described later According to the procedure, the title compound shown in the following formula was synthesized.

(1) Step 1
A mixture of 15.2 g (176 mmol) 3-methyl-2-butanone, 24.9 g (176 mmol) cyanoacetic acid tert-butyl ester, 5.7 g (176 mmol) sulfur and 15.4 g (176 mmol) morpholine is added to 75 ml DMF. The mixture was stirred at room temperature for 15 hours under an argon atmosphere. The reaction mixture was then stirred at 70-80 ° C for 8 hours. The reaction mixture was poured into aqueous NaCl solution and extracted with ethyl acetate. The organic layer was then washed with water, and the solvent was distilled off under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 18.3 g of 2-amino-4-isopropylthiophene-3-carboxylic acid tert-butyl ester (43% yield).

(2) Step 2
A mixture of 1.1 ml (7.5 mmol) adipic acid chloride and 3.63 g (15 mmol) 2-amino-4-isopropylthiophene-3-carboxylic acid tert-butyl ester was stirred in 20 ml DMA at room temperature for 15 hours. The reaction mixture was poured into ice water. The resulting precipitate was collected by filtration, washed with water and hexane, and 4.19 g of 2- [5- (3′-carboxy-4′-isopropylthiophen-2′-ylcarbamoyl) -pentanoylamino] -4 -Isopropyloffene-3-carboxylic acid di-tert-butyl ester was obtained (yield 93%).

(3) Step 3
1.2 g (2 mmol) 2- [5- (3′-carboxy-4′-isopropylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid di-tert-butyl ester Was dissolved in chloroform, then 2 ml of TFA was added, and the mixture was stirred at room temperature for 16 hours. After concentrating the reaction mixture, water was added to the residue and the precipitate was collected by filtration and washed with water and Et ₂ O to give the title 2- [5- (3′-carboxy-4′-isopropylthiophene- 2'-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid (compound x) was obtained (yield 75%).
mp: 247-249 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.17 (12H, d, J = 6.8Hz), 1.66 (4H, m), 2.50-2.60 (4H, m), 3.50 (2H, septet, J = 6.8Hz) , 6.68 (2H, s), 11.4 (2H, s).

Production Examples 16 and 17
2- [5- (3′-carboxy-5′-methylthiophen-2′-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid (compound z), and 2- [5- ( Preparation of 3′-tert-butoxycarbonyl-5′-methylthiophen-2′-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid (compound zz) According to Steps 1 to 3 described below, The title compound z shown in the formula was synthesized, and then the title compound zz was synthesized in Step 4.

(1) Step 1
A mixed solution of 5.8 g (99.9 mmol) propionaldehyde, 14.1 g (176 mmol) tert-butyl cyanoacetate, 59 mg (0.68 mmol) morpholine, 41 mg (0.68 mmol) acetic acid and molecular sieve 4A in toluene for 3 days Stir. Thereafter, the reaction solution was filtered, washed with water, dried over magnesium sulfate, and the solvent was concentrated under reduced pressure. A mixture of the obtained residue, 3.2 g (99.9 mmol) of sulfur and 8.7 g (99.9 mmol) of morpholine was added to 100 ml of DMF and stirred overnight. The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was then dried over magnesium sulfate and the solvent was removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 4.93 g of 2-amino-4-methylthiophene-3-carboxylic acid tert-butyl ester (yield 23%).

(2) Step 2
A mixture of 0.47 g (2.58 mmol) adipic acid chloride and 1.0 g (4.69 mmol) 2-amino-4-methylthiophene-3-carboxylic acid di-tert-butyl ester was stirred in 10 ml DMA at room temperature for 3 hours. . The reaction mixture was poured into ice water and then alkalinized by adding sodium bicarbonate. The resulting precipitate was collected by filtration and recrystallized from ethyl acetate / THF / hexane to give 0.96 g of 2- [5- (3′-carboxy-5′-methylthiophen-2′-ylcarbamoyl) -pentanoyl Amino] -5-methylthiophene-3-carboxylic acid di-tert-butyl ester was obtained (77% yield).

(3) Step 3
0.96 g (1.8 mmol) 2- [5- (3′-carboxy-5′-methylthiophene-2′-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid di-tert-butyl It was dissolved in ester methylene chloride, then 2 ml of TFA was added and stirred at room temperature for 3 hours. The reaction mixture was concentrated, hexane was added to the residue, and the mixture was filtered and washed with IPE and ethyl acetate. It was then recrystallized from THF / hexanes and the title 2- [5- (3′-carboxy-5′-methylthiophene-2′-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid ( Compound z) was obtained (yield 75%).
mp: 260-262 ° C (dec.)
¹ H-NMR (DMSO-d6) δ: 1.59-1.75 (4H, m), 2.32 (6H, d, J = 1.2Hz), 2.50-2.61 (4H, m), 6.82 (2H, d, J = 1.2 Hz), 10.9 (2H, s), 13.0 (2H, br).

(4) Step 4
Next, all of the filtrate and washing solution were combined and concentrated under reduced pressure, and the resulting crude product was separated and purified by silica gel column chromatography to give the title 2- [5- (3′-tert-butoxycarbonyl-5 ′]. -Methylthiophen-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid (compound zz) was obtained (yield 16%).
mp: 175-177 ℃
¹ H-NMR (DMSO-d6) δ: 1.54 (9H, s), 1.60-1.71 (4H, m), 2.30-2.33 (6H, m), 2.49-2.63 (4H, m), 6.78 (1H, d , J = 1.3Hz), 6.82 (1H, d, J = 1.3Hz), 10.7 (1H, s), 11.4 (1H, br).

Experimental Example 1 Measurement of PAI-1 Inhibitory Activity of Compounds a and b The inhibitory action of compounds a and b (test compounds) on human PAI-1 (Molecular Innovations Inc. (USA), the same applies hereinafter) was evaluated. Specifically, human-derived PAI-1 was placed in a 100 mM Tris-HCl (pH 8) solution containing 0.1% Tween 80 containing each of the above-mentioned test compounds at various concentrations (20, 35, 50 and 100 μM). Incubated for 15 minutes. Next, human-derived tissue plasminogen activator (t-PA) (American Diagnostica, Inc. (USA), the same applies hereinafter) adjusted to 0.53 pmol / μL was added thereto, followed by incubation at 37 ° C. for 15 minutes. Subsequently, a chromogenic substrate, 1.25 mM S-2288 synthetic substrate (Chromogenix (Italy), hereinafter the same) was added thereto. The final mixture was 100 mM Tris-HCl (pH 8), 30 mM NaCl, 1% DMSO, 0.1% Tween 80, 67 nM PAI-1, 9.8 nM t-PA, 1 mM S-2288 and each test compound a or b (20, 35 , 50 or 100 μM).

  P-nitroanilide cleaved and released from the chromogenic substrate (S-2288) by the action of t-PA was measured at an absorbance of 405 nm every 5 minutes for 30 minutes using a spectrophotometer. A system not containing the test compound was also tested in the same manner, and the PAI-1 activity when each test compound was added was evaluated with the PAI-1 activity 30 minutes after this system (control system) as 100%.

  As a comparative test, the same test was conducted on the following compound (tiplaxtinin) (concentrations of 20, 35, and 50 μM) introduced in clinical trials in the United States as an antithrombotic agent instead of the test compound.

  The results are shown in FIGS. FIGS. 4 (A), (B) and (C) show the compounds a [2- [3- (3′-carboxy-4′-phenylthiophen-2′-ylcarbamoyl) -pentanoylamino] -4- Phenylthiophene-3-carboxylic acid] (concentration 20, 35, 50, 100 μM), compound b [2- [3- (3′-carboxy-4′-thienylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-thienylthiophene-3-carboxylic acid] (concentration 20, 35, 50, 100 μM) and tiplaxtinin (comparative compound) (concentration 20, 35, 50 μM) show PAI-1 activity (%) . As a result, it was found that at concentrations of 35 μM and 50 μM, compounds a and b were stronger than tiplaxtinin (comparative compound) and suppressed PAI-1 activity (PAI-1 inhibitory activity).

Experimental Example 2 Complex formation inhibitory action of PAI-1 and t-PA For each test compound (compounds a and b), a complex of PAI-1 and t-PA (PAI-1 / t-PA complex) The formation inhibitory activity was tested by Western blotting.

  Specifically, the PAI-1 solution adjusted to 6.7 pmol / μL and each test compound were mixed in 20 mM Tris-HCl (pH 7.8) containing 0.1% Tween 80 so that the molar ratio was 1500 times. And incubated at 37 ° C. for 15 minutes. The test compound was removed from the reaction solution using a 10,000 cut filter (Millipore), t-PA adjusted to 1.4 pmol / μL was added, and the mixture was reacted at 37 ° C. for 15 minutes.

  This was subjected to SDS-polyacrylamide electrophoresis and Western blotting was performed. Specifically, the reaction solution was prepared with PBS so that the protein amount was 0.3 μg / lane, mixed with loading buffer (Daiichi Chemical), and heated at 100 ° C. for 5 minutes to make a sample solution. . The sample solution was electrophoresed on a 4-20% polyacrylamide gel (Daiichi Chemical) using an electrophoresis apparatus (Daiichi Chemical) and Tris-Glycine buffer (Daiichi Chemical). . After completion of electrophoresis, Western blotting was performed. Specifically, the electrophoresed protein is transferred to a polyvinylidene difluoride membrane (PVDF membrane, manufactured by Biorad) (100 V for 1 hour), and then the PVDF membrane is shaken with Block Ace (Snow Brand Milk Products) for 2 hours at room temperature. And blocked. Then, it was made to react overnight at 4 degreeC with the t-PA antibody (Cedarlane Laboratories Ltd. (Canada)) diluted 1000 times. Thereafter, an alkaline phosphatase-labeled sheep IgG antibody was added, reacted for 1 hour at room temperature, and then developed with an NBT-BCIP solution.

  The results are shown in FIG. In FIG. 5, lane 1 is an electrophoretic image of t-PA, lanes 2 and 3 are electrophoretic images of sample solutions when compounds a and b are used as test compounds, respectively, and lane 4 is a test compound (compounds a and b). The electrophoretic images of the sample solution in which PAI-1 and t-PA were reacted without using () are shown.

  As can be seen from the results, compound a [2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid] and Compound b [2- [3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid] Both inhibited the formation of complexes between PAI-1 and t-PA.

Experimental Example 3 Evaluation of fibrinolytic activity The fibrinolytic activity of each test compound (compounds a and b) was evaluated according to Matsuo et al. (Matsuo, O. et al., Haemostasis 16, 43-50 (1986)). did.

Specifically, an aqueous solution containing fibrinogen (Organon Teknica) at a rate of 1.5 mg / ml on a 9 cm plate of thrombin (10NIH U / ml: Mochida Pharmaceutical) dissolved in 0.2 ml of physiological saline. 25 mM sodium barbital, 50 mM NaCl, and 25 mM CaCl ₂ ) was added, and the mixture was allowed to stand at room temperature for 2 hours. This was used to perform a fibrinolysis assay.

  That is, a mixture of PAI-1, t-PA and each test compound was dropped on the plate, incubated at room temperature for 18 hours, and fibrinolysis due to plasminogen activation was measured by the dissolution area on the plate. did. In addition, after making it react like Example 2 as a mixture of PAI-1, t-PA and each test compound, what was adjusted so that tPA might be set to 50 NIH / mL was used. The results are shown in FIG.

  As a result, compound a [2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid] and compound b [2 -[3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid] is It was found to inhibit the suppression of fibrinolysis by PAI-1.

Experimental Example 4 Examination of inhibitory action of test compound on antiplasmin (AP) activity The inhibitory action of each test compound on antiplasmin (AP) activity was evaluated.

  Specifically, each test compound was added to AP (manufactured by Sigma-Aldrich Co.) adjusted to 0.2 pmol / μL in an excess amount corresponding to a molar ratio of 6000 to 60000 times, and 20 mM Tris-containing 0.1% Tween80. Incubation was carried out in HCl (pH 7.8) at 37 ° C. for 15 minutes. Then, it was made to react with plasmin adjusted to 0.2 pmol / microliter for 15 minutes at 37 degreeC. Subsequently, 1.25 mM plasmin synthetic substrate (Peptide Institute, Japan) was added, and AMC (7-amino-4-methylcoumarin) released from the substrate by the cleavage action of plasmin was detected at a fluorescence wavelength of 380 nm and an excitation wavelength of 460 nm. Measured every minute for 30 minutes. As a control test, the same test was conducted on a system not containing the test compound, and the AP activity of each test compound was evaluated with the AP activity 30 minutes after the reaction as 100%.

  FIG. 7 (A) shows the results regarding the compound a [2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid]. And compound b [2- [3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid The results regarding [acid] are shown in FIG. As a result, neither compound a nor b inhibited AP activity. From this result, it was confirmed that compounds a and b did not act on serine prostheses different from PAI-1 activity but acted specifically on PAI-1 activity.

Experimental Example 5 Examination of inhibition of complex formation of plasmin and antiplasmin The inhibitory action of compound b on antiplasmin (AP) activity was evaluated.

  Specifically, an AP solution adjusted to 7.5 pmol / μL and 1 mM [2- [3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] ] -4- (2-thienyl) thiophene-3-carboxylic acid (compound b) was added at a molar ratio of 3,000 times or 15,000 times, and 20 mM Tris-HCl (pH 7.8) containing 0.1% Tween 80 Incubated for 15 minutes at 37 ° C. Similarly, the PAI-1 solution adjusted to 6.7 pmol / μL and 1 mM compound b were mixed so that the molar ratio was 3,000 times or 15,000 times. Compound b was removed from the reaction solution using a 10,000 cut filter (Millipore), plasmin adjusted to 12.6 pmol / μL was added, and the mixture was reacted at 37 ° C. for 15 minutes.

  The reaction solution was mixed with loading buffer (Daiichi Kagaku Kagaku) and heated at 100 ° C. for 5 minutes to obtain a sample solution. The sample solution was electrophoresed on a 4-20% polyacrylamide gel (Daiichi Kagaku) using an electrophoresis apparatus (Daiichi Kagaku) and Tris-Glycine buffer (Daiichi Kagaku). After the electrophoresis, the band was detected by a usual staining method (Coomassie Brilliant Blue). The results are shown in FIG. In FIG. 8, lanes 1 to 3 are electrophoresis images of plasmin, antiplasmin (AP), and PAI-1, respectively, and lane 4 is a sample solution obtained by reacting plasmin and antiplasmin without using compound b. Electrophoresis images, lanes 5 and 6 are electrophoretic images of sample solutions obtained by reacting plasmin and antiplasmin using compound b in molar ratios of 3,000 times and 15,000 times, respectively, and lane 7 is plasmin without using compound b. Images of sample solution reacted with PAI-1 and lanes 8 and 9 are electrophoresis of sample solution reacted with plasmin and PAI-1 using compound b in molar ratios of 3,000 and 15,000, respectively. Each image is shown.

  As shown in FIG. 8, 2- [3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3 -Carboxylic acid (compound b) inhibited the complex formation of plasmin and PAI-1 at a molar ratio of 3,000 times. On the other hand, the complex formation between plasmin and AP was not inhibited at molar ratios of 3,000 times and 15,000 times.

Experimental Example 6
(1) Compound a [2- [3- (3′-carboxy-4′-phenylthiophene-2′-ylcarbamoyl) -pentanoylamino] -4-phenylthiophene-3-carboxylic acid] and compound b [ 2- [3- (3′-carboxy-4 ′-(2-thienyl) thiophen-2′-ylcarbamoyl) -pentanoylamino] -4- (2-thienyl) thiophene-3-carboxylic acid], and table According to the method described in Experimental Example 1, the inhibitory activity against human PAI-1 was investigated for the compounds c to 1 shown in 1. When the test compound is not mixed, the same test is performed, and each test compound (compound cl, concentration 20, 35, and 50 μM) is added with the PAI-1 activity 30 minutes after this system as 100%. PAI-1 activity was evaluated.

  These compounds a to l are available from Ambinter (46 quai Louis Bleriot Paris, F-75016 France: httP:https://www.ambinter.com.) And Interchim (213 Avenue Kennedy BP 1140 Montlucon, Cedex, 03103 France : https://WWW.interchim.com.), Enamine (I.Kudri str.41 Apt. 29 Kiev 042, 01042 Ukraine: https://www.enamie.relc.com), SPEC and BioSPECS (Fleminglaan l6 CP Rijswijk, 2289 Netherlands: https://www.specs.net).

  The results are shown in Table 1.

From this result, it can be seen that all of the compounds a to l have an inhibitory action on PAI-1 activity. Of these, compounds a, b, d and e, particularly compound d, have excellent inhibitory activity.
(2) The compounds ox prepared in Preparation Examples 5 to 14 were also examined for inhibitory activity against human PAI-1 according to the method described in Experimental Example 1. Similarly to the above, the PAI-1 activity when each compound ox (concentrations 40, 50 and 100 μM) was added was evaluated with the PAI-1 activity 30 minutes after the system not containing the test compound as 100%. .

  The results are shown in Table 2.

  From this result, it can be seen that all of the compounds ox have an inhibitory action on PAI-1 activity. Among them, compounds s, t, v, r, q and w, especially compounds s, t, v and r have high inhibitory activity.

Experimental Example 7 Evaluation of the effect on bleomycin-induced pulmonary fibrosis In order to evaluate the in vivo anti-fibrotic effect of the compound (1) of the present invention, a model animal (mouse) in which pulmonary fibrosis was artificially induced with bleomycin was used. The following experiment was conducted using this.

  First, pentobarbital was intraperitoneally administered to C57BL / 6 mice (male, body weight 19-21 g) and anesthetized, and the cervical organ was incised. Ten mice were used as controls. Control mice (n = 10) were intratracheally administered with bleomycin (manufactured by Nippon Kayaku) (1.5 U / kg) dissolved in physiological saline twice a day for 14 days. On the other hand, in addition to the above-mentioned intratracheal administration, the test subject mice were treated with compound b (2- [3- (3) suspended in 0.5% aqueous carboxymethylcellulose twice a day for 14 days. '-Carboxy-4'-thienylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-thienylthiophene-3-carboxylic acid) (200 mg / kg) was orally administered by gavage. Subsequently, the lung tissues of these control mice and test mice were subjected to histological analysis, and the amount of hydroxyproline was measured. The amount of hydroxyproline in lung tissue was measured as the amount in the hydrolyzate of lung tissue according to the method of Kivirikko et al. (Anal. Biochem. 19, 249-255 (1967)). The level (severity) of pulmonary fibrosis was scored from 0 to 8 using the method of Ashcort et al. (J. Clin. Pathol. 41,467-470, (1988)). In addition, plasma PAI-1 activity (ng / ml) was measured for these control mice and test subject mice. The results of histological analysis of lung tissue are shown in FIG. 9 (a is a fibrosis score, b is a tissue staining image), and the amount of hydroxyproline and plasma PAI-1 activity in lung tissue are shown in Table 3.

  From this result, it can be seen that the amount of hydroxyproline in lung tissue significantly increased by administration of bleomycin is significantly decreased by administration of compound b. It was also found that plasma PAI-1 activity was significantly increased by bleomycin administration, and that this increase in plasma PAI-1 activity was also significantly decreased by administration of compound b.

  As can be seen from FIG. 9, pulmonary fibrosis induced by bleomycin administration (fibrosis score: 4.7 ± 0.17, control group: 0.5 ± 0.17, P <0.001) was significantly improved by administration of compound b (fibrosis). Syndrome score: 2.9 ± 0.42, P <0.01), which was consistent with the above PAI-1 activity results.

  These results show that the compound (1) of the present invention having a PAI-1 inhibitory action including the compound b has an action of preventing the pulmonary fibrosis process in addition to the fibrinolytic system promoting action. Yes. It has already been reported by Eitzman et al. That there is a strong relationship between PAI-1 expression and collagen accumulation in the lung tissue of mice that overexpress or lack the PAI-1 gene (J .Chin.Invest. 97,232-237 (1996)). The above results showing that compound b with strong PAI-1 inhibitory activity ameliorates pulmonary fibrosis suggests that PAI-1 is not a mere indicator of pulmonary fibrosis but its main factor Yes. This finding is extremely important because fibrosis occurs not only in the lungs, but also in many tissues and organs including the heart, blood vessels, liver, and kidneys. That is, by inhibiting PAI-1 activity, it is considered possible to prevent or treat various diseases related to fibrosis (heart disease, cirrhosis, kidney disease or radiation injury).

Experimental Example 8
Antithrombotic action of 2- [3- (3'-carboxy-4'-thienylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-thienylthiophene-3-carboxylic acid (compound a) on thrombus model animals In order to confirm the effectiveness of the compound of the present invention against antithrombosis, a rat AV shunt model, which is a thrombus model animal, was used to produce 2- [3- (3′-carboxy-4′-thienylthiophene-2′-ylcarbamoyl]. ) -Pentanoylamino] -4-thienylthiophene-3-carboxylic acid (compound a) was investigated for its antithrombotic activity. In addition to this, a blood coagulation test was also performed in parallel.

  Here, the AV shunt model test is performed by publicly known literature (RFPeters, et al., Thromb Haemost 1991; 65, 268-274: Yoshiyuki Morishima, et al., Thromb Haemost 1997; 78, 1366-1371: Atsuhiro Sugidachi, et al., British Journal of Phermacology 2000; 129, 1349-1446).

(1) Preparation of test compound and administration solution In addition to the compound a of the present invention, as a test compound, as a comparative compound (positive control compound), warfarin (manufactured by Wako Pure Chemical Industries, Ltd.) and ticlopidine hydrochloride (Sigma- Aldrich Co.) was used.

  These test compounds consist of a solvent (0.5% CMC-Na solution) prepared by dissolving 0.5 g of carboxymethylcellulose (manufactured by Nacalai Tesque) (hereinafter referred to as “CMC-Na”) in 100 mL of water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.). ) To prepare an administration solution. Specifically, Compound a is suspended in a 0.5% CMC-Na solution to a concentration of 75 mg / mL, and ticlopidine hydrochloride is suspended in a 0.5% CMC-Na solution to a concentration of 142.3 mg / mL. In addition, warfarin was suspended in a 0.5% CMC-Na solution to a concentration of 0.3 mg / mL to prepare an administration solution.

(2) Test animal As an animal used for the AV shunt model test, an 8-week-old CD (SD) rat having a body weight gain within the normal range of the mean value ± 3SD in acclimated breeding and having a weight of 270 to 300 g (Japan) Charles River Co., Ltd.) was used. These rats (total 28 animals) were divided into the solvent administration group (1 group: n = 7), the compound a administration group (2 groups: n = 7), the ticlopidine hydrochloride administration group (3 groups: n = 7), And warfarin administration group (4 groups: n = 7).

(3) Test method
(3-1) AV Shunt Model Test Each test compound was administered to the test animal so that the maximum blood concentration was reached at the time of performing the AV shunt operation in consideration of the pharmacokinetics of each compound in the test animal. Specifically, test compound a was administered at a rate of 300 mg / kg 17.5 hours before anesthesia / shunt surgery, warfarin at a rate of 1.2 mg / kg 23.5 hours before, and ticlopidine hydrochloride at a rate of 500 mg / kg 1.5 hours before. . The AV shunt model was performed when the maximum blood concentration was reached in consideration of the pharmacokinetics of each test compound.

  The operation for creating an AV shunt model was performed according to the following method.

  First, pass a 6.5cm silk thread (Matsuda Medical 1-0) through an 8cm No. 7 polyethylene tube (Hibiki), and pass a No. 3 tube (12.5cm) through a No. 5 tube (1.5cm) at both ends. And a shunt catheter was created. A parafilm was wrapped around the connection through the silk thread to prevent blood from leaking.

  Rats were anesthetized with pentobarbital (Dainippon Sumitomo Pharma Co., Ltd.) administered intraperitoneally at a rate of 50 mg / kg, filled with physiological saline in the catheter, and both ends of the catheter were connected to the right carotid artery and It was inserted into the left jugular vein and perfused with blood. 30 minutes after the start of perfusion, the blood flow was stopped with a forceps between the catheter and the tube part through which the silk thread was passed was cut out. Subsequently, the weight of the thrombus formed by blood perfusion was measured. Specifically, the silk thread was carefully taken out from the cut out tube, the liquid phase was removed with a filter paper, the remaining wet weight was measured, and the weight of the 6.5 cm silk thread was further subtracted from this to obtain the thrombus weight.

(3-2) Blood coagulation test At the same time that the tube was cut out when measuring the above thrombus weight, blood was collected from the abdominal aorta for coagulation test. 1.8 mL of whole blood was added to a vacuum blood collection tube containing 3.13% citrate and centrifuged to obtain plasma. In addition, 0.5 mL of whole blood was added to a micro blood collection tube containing a serum separating agent and a coagulation promoter to obtain serum. The activated partial thromboplastin time (APTT) and prothrombin time (PT) were measured about plasma using the fully automatic blood coagulation measuring apparatus.

(4) Results Table 4 shows the thrombus weight obtained for the vehicle administration group and each test compound administration group (Groups 1 to 4).

  As can be seen from the results, in the rat AV shunt model, the thrombus weight of the compound a administration group of the present invention is significantly higher than that of the vehicle administration group as in the positive control compound (warfarin, ticlopidine hydrochloride) administration group. Decreased. In addition, the prothrombin time (PT) and the activated partial thromboplastin time (APTT) were not significantly changed in the compound a administration group of the present invention as compared with the vehicle administration group.

  On the other hand, the warfarin administration group decreased the thrombus weight and prolonged PT and APTT. The ticlopidine hydrochloride administration group reduced the thrombus weight, but had no effect on PT and APTT as in the case of Compound a of the present invention. Here, both PT and APTT are indicators for determining the coagulation system. That is, in this test, the result that the compound a of the present invention did not affect either PT or APTT means that the compound a is a compound having a property different from that of the anticoagulant.

  From the test results using the rat AV shunt model described above, 2- [3- (3'-carboxy-4'-thienylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-thienylthiophene-3-carvone It has been clarified that the acid (compound a) exhibits an antithrombotic action by an action mechanism different from that of the anticoagulant.

Experimental Example 9 Cytotoxicity Test In general, it is known that lactate dehydrogenase (LDH), which is a cytoplasmic localized enzyme, is released from cells lysed by injury. Since the LDH is relatively stable, the cytotoxicity of the drug can be indirectly examined by quantifying the LDH.

  Compounds a and b of the present invention are added to HeLa cells (cervical cancer-derived epithelial cell line) at a ratio of 250 μM, 125 μM or 50 μM, and LDH eluted from the cells is mixed with INT (tetrazolium salt) and 30 The cytotoxicity of compounds a and b was tested by performing an enzymatic reaction for 1 min and quantifying red formazan converted from INT at 490 nm. As a control test, the same test was performed using 0.5% DMSO instead of compounds a and b. In both compounds a and b of the present invention, no cytotoxicity was observed up to 250 μM.

Experimental Example 10 Acute Toxicity Test ICR mice (obtained from CLEA Japan, male, 19-21 g) (n = 5) were each administered a single oral dose of 1 g / kg, followed by 2-week follow-up Went. There were no deaths in either compound.

Experimental Example 11 Absorbency Test
Wister rats (obtained from CLEA Japan, male, 160-180 g) (n = 5) were orally administered compounds a and b at 50 mg / kg, respectively 1, 2, 6, 18, 24 hours later and Seven days later, blood was collected from the vein, the amount of the compound in the blood was measured, and the maximum blood concentration (Cmax) and half-life (T1 / 2) were examined. As a comparative control test, the same test was performed using tiplaxtinin instead of compounds a and b. Table 5 shows the half-life (T1 / 2) of compounds a and b and tiplaxtinin.

  As can be seen from the results, compounds a and b showed significantly longer blood persistence than tiplaxtinin.

It is a figure which shows the synthetic scheme of the compound (1) (especially compound a and b) of this invention. 1 shows NMR data of Compound a of the present invention (N, N′-bis [3,3′-carboxy-4,4′-phenyl-2,2′-thienyl] hexanedicarboxamide). NMR data of the compound b (N, N′-bis [3,3′-carboxy-4,4 ′-(2,2′-thienyl) -2,2′-thienyl] hexanedicarboxamide) of the present invention Show. (A) N, N′-bis [3,3′-carboxy-4,4′-phenyl-2,2′-thienyl] hexanedicarboxamide (compound a), (B) N, N′-bis [ The figure which shows the PAI-1 inhibitory activity of 3,3'-carboxy-4,4 '-(2,2'-thienyl) -2,2'-thienyl] hexane dicarboxamide (compound b) and (C) tiplaxtinin It is. The vertical axis shows PAI-1 activity (%) (Experimental Example 1). It is an electrophoresis image which shows the complex formation inhibitory effect of the structure | tissue plasminogen activator (t-PA) and PAI-1 of each compound by a western blotting method (Experimental example 2). It is a figure which shows the PAI-1 inhibitory effect of each compound by the fibrin plate method (natural substrate method) (Experimental example 3). N, N'-bis [3,3'-carboxy-4,4'-phenyl-2,2'-thienyl] hexanedicarboxamide (compound a) (Figure (A)), and N, N'-bis Antiplasmin (AP) activity of [3,3′-carboxy-4,4 ′-(2,2′-thienyl) -2,2′-thienyl] hexanedicarboxamide (compound b) (FIG. (B)) It is a figure which shows the effect with respect to (Experimental example 4). Plasmin of N, N′-bis [3,3′-carboxy-4,4 ′-(2,2′-thienyl) -2,2′-thienyl] hexanedicarboxamide (compound b) by Western blotting It is an electrophoretic image which shows the complex formation inhibitory effect of an antiplasmin. Of N, N′-bis [3,3′-carboxy-4,4 ′-(2,2′-thienyl) -2,2′-thienyl] hexanedicarboxamide (compound b) against bleomycin-induced pulmonary fibrosis An anti-fibrous effect is shown, a is a fibrosis score, and b is a tissue stained image (Experimental Example 7).

Claims (16)

The following formula (1):

[Wherein, R ₁ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, or carbon A linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ is a hydrogen atom, an optionally substituted phenyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms; Or a halogen atom. R ₂ and R ₃ may be bonded to each other to form a 5- to 6-membered ring; A is a linear or branched alkylene, alkenylene or alkynylene having 1 to 7 carbon atoms, 3 carbon atoms -8 cycloalkylene, or single bond; R ₄ is a hydrogen atom, an optionally substituted phenyl group or furyl group, or the following formula (2);

(Wherein R ₁ ′ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ ′ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, Or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ ′ is a hydrogen atom, an optionally substituted phenyl group, or a linear or branched chain group having 1 to 6 carbon atoms. And R ₂ ′ and R ₃ ′ may be bonded to each other to form a 5- to 6-membered ring. ]
Or a salt thereof, or a solvate thereof, as an active ingredient, a plasminogen activator inhibitor-1 inhibitor. In the general formula (1), A is a linear or branched alkylene having 1 to 7 carbon atoms, or a cycloalkylene having 3 to 8 carbon atoms; R ₄ is the following formula (2);

(Wherein R ₁ ′ is a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ ′ is a hydrogen atom, an optionally substituted phenyl group or thienyl group, Or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ ′ is a hydrogen atom, an optionally substituted phenyl group, or a linear or branched chain group having 1 to 6 carbon atoms. And R ₂ ′ and R ₃ ′ may be bonded to each other to form a 5- to 6-membered ring.)
The plasminogen activator inhibitor-1 inhibitor according to claim 1, wherein the compound is a compound represented by the formula (1) or a salt thereof, or a solvate thereof. R ₂ and R ₂ ′ are the same or different and may be a phenyl group, a 2-thienyl group, —CH ₂ CH (CH ₃ ) ₂ which may have a substituent, or the same or different, _The plasminogen activator inhibitor-1 inhibitor according to claim 1, wherein ₂ and R ₃ or R ₂ 'and R ₃ ' are bonded to each other to form a condensed 6-membered ring. The plasminoid according to claim 1, wherein R ₃ and R ₃ 'are the same or different and each is a hydrogen atom, a linear or branched alkyl group having 1 to 3 carbon atoms, a phenyl group, or a halogen atom. Genactivator inhibitor-1 inhibitor. A is straight-chain alkylene of 3-5 carbon _{atoms, -CH 2 -C (CH 3)} 2 -CH 2 -, or cyclohexylene, plasminogen activator inhibitor-1 according to claim 1 Inhibitor. The plasminogen activator inhibitor-1 inhibitor according to claim 1, wherein R ₁ is a hydrogen atom, R ₂ and R ₂ 'are a phenyl group or a 2-thienyl group, and A is butylene.   A pharmaceutical composition comprising the plasminogen activator inhibitor-1 inhibitor according to any one of claims 1 to 6, and a pharmaceutically acceptable carrier or additive.   The pharmaceutical composition according to claim 7, which is a prophylactic or therapeutic agent for a disease associated with plasminogen activator inhibitor-1 activity.   The pharmaceutical composition according to claim 8, which is a fibrinolytic accelerator.   Diseases associated with the onset of plasminogen activator inhibitor-1 activity are angina, myocardial infarction or atrial thrombosis in atrial fibrillation, ischemic heart disease, ischemic cerebrovascular disorder, arteriosclerosis, pulmonary embolism , Deep vein thrombosis (DVT) during surgery, disseminated intravascular coagulation syndrome (DIC), vascular disorders as diabetic complications, neuropathy, retinopathy or nephropathy, or percutaneous coronary angioplasty (PTCA) 9) The pharmaceutical composition according to claim 8, which is subsequent restenosis.   The pharmaceutical composition according to claim 8, wherein the disease associated with the onset of plasminogen activator inhibitor-1 activity is a disease associated with tissue fibrosis.   The pharmaceutical composition according to claim 11, wherein the disease involving tissue fibrosis is pulmonary fibrosis. 8. A pharmaceutical composition according to claim 7 having an oral dosage form. The following formula (3):

[Wherein, R ₁ and R ₁ ′ are the same or different, a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R ₂ and R ₂ ′ are the same or different; A hydrogen atom, an optionally substituted phenyl group, or a linear or branched alkyl group having 1 to 6 carbon atoms; R ₃ and R ₃ ′ may be the same or different, A phenyl group which may have a group, a linear or branched alkyl group having 1 to 6 carbon atoms, or a halogen atom; A is a linear or branched alkylene having 1 to 7 carbon atoms, Or a C3-C8 cycloalkylene is shown. However, when R ₂ and R ₂ ′ are phenyl groups having no substituent and R ₃ and R ₃ ′ are hydrogen atoms, A represents butylene and —CH ₂ —C (CH ₃ ) ₂ —CH _2. When neither R nor R ₂ and R ₂ ′ are isobutyl groups and R ₃ and R ₃ ′ are hydrogen atoms, A is not butylene. ]
Or a salt thereof. Compounds represented by the following formulas (4) to (15) or salts thereof:
2- [5- (3'-Carboxy-4 '-(p-chlorophenyl) thiophen-2'-ylcarbamoyl) -pentanoylamino] -4- (p-chlorophenyl) thiophene-3-carboxylic acid

2- [4- (3'-Carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -butyrylamino] -4-isobutylthiophene-3-carboxylic acid

2- [4- (3'-Carboxy-4'-isobutylthiophen-2'-ylcarbamoyl) -3,3-dimethylbutyrylamino] -4-isobutylthiophene-3-carboxylic acid

2- [6- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -hexanoylamino] -4-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-methyl-4'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methyl-4-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-phenylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-chlorothiophene-2'-ylcarbamoyl) -pentanoylamino] -5-chlorothiophene-3-carboxylic acid

2- [4- (3'-Carboxy-4'-phenylthiophene-2'-ylcarbamoyl) -cyclohexylcarbonylamino] -4-phenylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-5'-isopropyl-4'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isobutylthiophene-3-carboxylic acid

2- [3- (3'-Carboxy-4'-isobutylthiophene-2'-ylcarbamoyl) -propanoylamino] -4-isobutylthiophene-3-carboxylic acid

2- [5- (3'-Carboxy-4'-isopropylthiophene-2'-ylcarbamoyl) -pentanoylamino] -4-isopropylofen-3-carboxylic acid

2- [5- (3'-Carboxy-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

2- [5- (3'-tert-Butoxycarbonyl-5'-methylthiophene-2'-ylcarbamoyl) -pentanoylamino] -5-methylthiophene-3-carboxylic acid

  A pharmaceutical composition comprising as an active ingredient at least one selected from the group consisting of the compound according to claim 14 or 15 or a pharmaceutically acceptable salt thereof, or a solvate thereof.

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