WO2012137885A1 - Agent for inhibiting or preventing renal interstitial fibrosis - Google Patents

Agent for inhibiting or preventing renal interstitial fibrosis Download PDF

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WO2012137885A1
WO2012137885A1 PCT/JP2012/059411 JP2012059411W WO2012137885A1 WO 2012137885 A1 WO2012137885 A1 WO 2012137885A1 JP 2012059411 W JP2012059411 W JP 2012059411W WO 2012137885 A1 WO2012137885 A1 WO 2012137885A1
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international publication
crth2
protein
interstitial fibrosis
suppressing
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PCT/JP2012/059411
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French (fr)
Japanese (ja)
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元昭 佐野
伊藤　秀之
恵一 福田
中村　正孝
博之 平井
欽也 永田
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学校法人 慶應義塾
国立大学法人東京医科歯科大学
株式会社ビー・エム・エル
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Priority to JP2013508930A priority Critical patent/JPWO2012137885A1/en
Publication of WO2012137885A1 publication Critical patent/WO2012137885A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention mainly relates to an inhibitor or preventive agent for renal interstitial fibrosis.
  • the present invention also relates to a method for suppressing or preventing renal interstitial fibrosis.
  • the kidney is an indispensable organ for maintaining the homeostasis of the human body, which has a function of filtering waste products contained in blood and discharging it as urine.
  • Chronic kidney disease chronic kidney disease (CKD)
  • CKD chronic kidney disease
  • CKD includes a wide range of conditions such as diabetic nephropathy, hypertensive nephropathy or nephrosclerosis, and chronic glomerulonephritis.
  • CKD includes a wide range of conditions such as diabetic nephropathy, hypertensive nephropathy or nephrosclerosis, and chronic glomerulonephritis.
  • CKD chronic kidney disease
  • ⁇ Fibrosis of the renal interstitium occurs as a common symptom when CKD progresses or worsens.
  • the kidney can no longer perform its function, and treatment that replaces the kidney function, such as artificial dialysis, is required.
  • the number of dialysis patients is about 300,000, and out of 30 trillion yen in medical expenses, it costs 1 trillion yen for dialysis.
  • CKD itself is an independent risk factor for cardiovascular disease, so CKD has been called a “new national disease”, and prevention of CKD onset and progression is a global issue It has become.
  • CRTH2 Certhelial growth factor receptor 2
  • PGD2 prostaglandin D2 receptor
  • Patent No. 3144805 International Publication WO01 / 014882 JP 2005-87164 A Japanese Patent Laid-Open No. 2002-241282
  • the main object of the present invention is to provide an inhibitor or preventive agent for renal interstitial fibrosis.
  • Another object of the present invention is to provide a method for suppressing or preventing renal interstitial fibrosis.
  • the present inventor made extensive studies to solve the above problems, and surprisingly found that renal interstitial fibrosis can be suppressed by suppressing the functional expression of CRTH2 protein.
  • the present invention has been completed by making further improvements based on such findings.
  • the present invention includes the following aspects of the invention.
  • Item 1 An inhibitor or preventive agent for renal interstitial fibrosis, comprising a compound capable of suppressing the functional expression of CRTH2 protein.
  • Item 2 The therapeutic agent according to Item 1, for administration to a patient with kidney disease.
  • Item 3 The inhibitor or preventive agent according to Item 1 or 2, which suppresses or prevents the progression of renal interstitial fibrosis in kidney disease.
  • Item 4 The inhibitor according to Item 2 or 3, wherein the kidney disease is selected from the group consisting of diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis, chronic interstitial nephritis, and polycystic kidney disease. Or prophylactic agent.
  • Item 5 The compound according to any one of Items 1 to 4, wherein the compound capable of suppressing the functional expression of CRTH2 protein is at least one compound selected from the group consisting of an antagonist of CRTH2 protein and a nucleic acid capable of suppressing the expression of CRTH2 gene.
  • Item 6 A method for suppressing or preventing renal interstitial fibrosis, comprising a step of administering a compound capable of suppressing the functional expression of CRTH2 protein to a patient with renal disease.
  • Item 7 A compound capable of suppressing the functional expression of CRTH2 protein for use in suppressing or preventing renal interstitial fibrosis.
  • Item 8 Use of a compound capable of suppressing the functional expression of CRTH2 protein for producing an inhibitor or preventive agent for renal interstitial fibrosis.
  • the present invention provides an effective inhibitor or preventive agent for renal interstitial fibrosis, and a method for suppressing or preventing renal interstitial fibrosis.
  • a method for suppressing or preventing renal interstitial fibrosis By suppressing or preventing renal interstitial fibrosis according to the present invention, it is highly expected that the patient's quality of life (QOL) will be improved and the required medical expenses will be reduced.
  • QOL quality of life
  • the inhibitor or preventive agent of the invention can be used as a tool for studying the onset mechanism and treatment method of renal interstitial fibrosis, and is expected to contribute to further improvement of medical technology.
  • the schematic diagram of a unilateral ureteral ligation operation is shown.
  • A Azan staining image (100-fold magnification) of the renal cortical tissue of the unilateral ureter-ligated mouse administered CAY10471 on the 10th day after the operation.
  • B The measurement result of the ratio (Fibrotic Area) (%) of the fibrosis region area that becomes positive by Azan staining to the observed stroma region area is shown.
  • A Azan staining image (100 times magnification) of the renal cortical tissue on the 10th day after the operation.
  • B The measurement result of the ratio (Fibrotic Area) (%) of the fibrosis region area that becomes positive by Azan staining to the observed stroma region area is shown.
  • the expression level in brain tissue on day 0 after the treatment is also shown. In kidney tissue, expression of the DP1 gene was undetected.
  • the following describes in detail the inhibitor or preventive agent for renal interstitial fibrosis, which contains a compound capable of suppressing the functional expression of the CRTH2 protein of the present invention.
  • CRTH2 (Chemoattractant-receptor-homologous-molecule-expressed-on-TH2) protein (also known as DP2, GPR44, CD294, DL1R) is a receptor protein for prostaglandin D2 and is a known protein.
  • CRTH2 protein is thought to be a G ⁇ i type receptor protein.
  • CRTH2 protein is known to be expressed in mammals including humans. In humans, CRTH2 protein is known to be expressed in Th2 cells, granulocytes (eosinophils), and basophils (basophils).
  • the CRTH2 protein sequence and the base sequence of the gene encoding the CRTH2 protein sequence are known. For example, they are registered as accession numbers NP_004769 and NM_004778, respectively, in a database published by the National Center for Biotechnology Information (NCBI). Yes.
  • the receptor protein for prostaglandin D2 is considered to be mainly composed of the above-mentioned CRTH2 protein and DP1 protein (G ⁇ i type receptor protein).
  • any compound capable of suppressing the functional expression of CRTH2 protein can be used as long as the functional expression of CRTH2 protein can be suppressed.
  • the compound capable of suppressing the functional expression of CRTH2 protein include a specific inhibitor for CRTH2 protein and a nucleic acid capable of suppressing the expression of CRTH2 gene, but are not limited thereto.
  • suppressing the functional expression of CRTH2 protein refers to all aspects of suppressing the functional expression of CRTH2 protein, for example, inhibiting the function of CRTH2 protein, suppressing the expression of CRTH2 protein (CRTH2 protein Examples include, but are not limited to, suppression of transcription of the encoded gene, suppression of CRTH2 protein translation, and the like. In addition, it is preferable that the suppression of the functional expression is specific to CRTH2 protein.
  • “specific” refers to the ability to suppress the functional expression of CRTH2 protein without exhibiting an action on other proteins.
  • Examples of the mode of inhibiting the function of CRTH2 protein include, but are not limited to, a mode of inhibiting the binding between CRTH2 protein as a receptor and a ligand.
  • a mode of inhibiting the binding between CRTH2 protein as a receptor and a ligand When the binding between the CRTH2 protein and the ligand is inhibited, signal transduction via the CRTH2 protein in the signal transduction pathway is reduced (preferably blocked).
  • “can suppress the expression of CRTH2 gene” means the destruction of mRNA encoding CRTH2 protein transcribed from CRTH2 gene (the gene encoding CRTH2 protein) (RNA interference action), suppression of translation into CRTH2 protein, etc.
  • RNA interference action RNA interference action
  • suppression of translation into CRTH2 protein etc.
  • any known and later-developed antagonists of CRTH2 protein can be used.
  • the antagonist include ramatroban (3-((3R) -3- ⁇ [(4-fluorophenyl) sulfonyl] amino] -1,2,3,4-tetrahydro-9H-carbazol-9-yl) propanoic acid; product name Baynas; Bayer AG), CAY10471 (also known as TM30089, (+)-3-[[(4-fluorophenyl) sulfonyl] methylamino] -1,2,3,4-tetrahydro-9H-carbazole -9-acetic acid; Cayman Chemical), MK-7246 ( ⁇ (7R) -7-[[(4-fluorophenyl) sulfonyl] (methyl) amino] -6,7,8,9-tetrahydropyrido [1,2- a] indo
  • One embodiment of the CRTH2 protein antagonist includes a compound represented by the following general formula (1).
  • Formula (1) [Wherein, X 1 represents a methylene group or an ethylene group. X 2 represents a hydrogen atom or a methyl group. ]
  • Examples of other antagonists of the CRTH2 protein include CRTH2 protein antagonists disclosed in the following documents: Japanese Patent Application Laid-Open No. 08-245587, Japanese Patent Application Laid-Open No. 08-175991, Japanese Patent Application Laid-Open No.
  • another embodiment of the specific inhibitor for the CRTH2 protein includes a specific antibody for the CRTH2 protein.
  • the antibody preferably binds specifically to the extracellular domain of CRTH2 protein, more preferably specifically binds to the extracellular domain of CRTH2 protein, and can inhibit the binding of CRTH2 protein and prostaglandin D2 Although it is an antibody, it is not limited to this.
  • the antibody may be a polyclonal antibody or a monoclonal antibody.
  • the antibody When the antibody is a monoclonal antibody, it may be a chimeric antibody, a humanized antibody, a human antibody, or the like.
  • the above antibody can be prepared by a known technique. Or a commercially available thing can also be used.
  • the nucleic acid capable of suppressing the expression of the CRTH2 gene is preferably an RNA molecule having RNA interference (RNAi) action targeting mRNA encoding CRTH2 protein such as siRNA, shRNA, dsRNA, or mRNA encoding CRTH2 protein.
  • RNAi RNA interference
  • Examples include miRNA that can suppress translation.
  • it may be an RNA molecule having the above RNA interference (RNAi) action or a DNA molecule such as a vector capable of expressing the above miRNA.
  • the nucleic acid capable of suppressing the expression of the CRTH2 gene can be prepared by a known method based on the base sequence of the gene encoding the CRTH2 protein. Or a commercially available thing can also be used.
  • Compounds that can suppress the functional expression of CRTH2 protein include animals that have a defect in the gene encoding CRTH2 protein (CRTH2 gene) in animals (for example, knockout animals), and that the functional expression of CRTH2 protein is suppressed. It is common in. Therefore, as shown in Examples described later, the effect of suppressing or preventing renal interstitial fibrosis is observed even in animals having a CRTH2 gene deficiency.
  • CRTH2 protein functions as a receptor for prostaglandin D2 together with DP1 protein.
  • the inhibitor or preventive agent of the present invention is considered to exert its function in the kidney, CRTH2 protein expression is observed in the kidney, but DP1 protein expression is below the detection limit.
  • DP1 protein expression is below the detection limit.
  • CRTH2 expression is increased, but DP1 protein is not induced. That is, it is considered that the signal from prostaglandin D2 in the kidney is transmitted mainly via the CRTH2 protein. From the above, suppression or prevention of renal interstitial fibrosis is considered to be achieved by suppressing the functional expression of CRTH2 protein.
  • the compounds capable of suppressing the functional expression of the CRTH2 protein can be used singly or in combination of two or more.
  • the inhibitor or preventive agent of the present invention is an inhibitor or preventive agent for renal interstitial fibrosis.
  • the inhibitor or preventive agent is preferably one that suppresses or prevents the progression of renal interstitial fibrosis in kidney disease, and particularly preferably suppresses or prevents the progression of renal interstitial fibrosis in chronic kidney disease. It is not a thing.
  • One preferred embodiment of the inhibitor or preventive agent of the present invention is one that directly suppresses or prevents the progression of renal interstitial fibrosis.
  • directly refers to an embodiment in which the progression of renal interstitial fibrosis is suppressed or prevented regardless of the presence or absence of treatment of the primary disease (for example, chronic kidney disease) in the kidney.
  • the above chronic kidney disease refers to a pathological condition that suggests the presence of renal diseases such as proteinuria, and that moderate or more impaired renal function persists for 3 months or more.
  • Examples of the chronic kidney disease include, but are not limited to, diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis, chronic interstitial nephritis, and polycystic kidney disease. Findings suggesting the presence of kidney disease are known to those skilled in the art and include, for example, detection of urine protein, but are not limited thereto.
  • the decrease in renal function can be confirmed by a known means for estimating renal function, such as creatinine clearance, but is not limited thereto.
  • kidney interstitium In patients with chronic kidney disease, fibrosis of the renal interstitium is induced and progresses in many cases. In renal interstitial fibrosis, extracellular matrix proteins such as collagen and fibrin are excessively deposited in renal cortical tissue. The progression of renal interstitial fibrosis eventually leads to nephrosclerosis. The kidney that has led to nephrosclerosis is no longer functional (end-stage renal failure), and in order to maintain homeostasis and prevent uremia symptoms, treatments that replace the kidney function such as artificial dialysis are required. Become.
  • “suppression of renal interstitial fibrosis” refers to stopping or suppressing further progression of renal interstitial fibrosis in a patient in which renal interstitial fibrosis has already been induced (usually suppression). (But not limited to this). “Preventing renal interstitial fibrosis” refers to preventing or delaying the onset of renal interstitial fibrosis in patients who have not yet been induced. By suppressing or preventing renal interstitial fibrosis with the therapeutic agent of the present invention, it is possible to eliminate the need to replace the renal function such as artificial dialysis, or to delay the start of the treatment. It becomes. The necessity of introducing a treatment that substitutes for the kidney function can be confirmed by means known to those skilled in the art, for example, means for estimating renal function such as creatinine clearance.
  • the form (administration form) of the inhibitor or preventive agent of the present invention is not particularly limited. Preferably it is provided as a pharmaceutical composition.
  • the inhibitor or preventive agent of the present invention can be formulated by a known technique. Specific examples of the preparation include, but are not limited to, solid preparations such as tablets, capsules, pills, powders, granules, and liquid preparations such as liquids, suspensions, emulsions, injections, and the like.
  • pharmaceutically acceptable carriers and additives can be added as appropriate. Specific examples of the carriers and additives include, but are not limited to, excipients, fillers, binders, wetting agents, fragrances, and coloring agents.
  • a known pharmaceutically acceptable solvent such as physiological saline, a solution having a buffering action, or the like can be used.
  • the administration method of the inhibitor or prophylactic agent of the present invention can be appropriately set by those skilled in the art as long as the effect as the inhibitor or prophylactic agent is obtained, and is not particularly limited.
  • Preferred embodiments of the administration method include injection administration (intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, injection into the affected area, etc.), oral administration, suppository administration, transdermal administration (application, etc.) and the like.
  • Oral administration is particularly preferred as an administration method from the viewpoint of ease of administration and less burden on administration subjects, but is not limited thereto.
  • Subjects for administration of the inhibitor or prophylactic agent of the present invention are humans, non-human mammals (dogs, cats, mice, rats, hamsters, rabbits, cows, apes, etc.), birds, etc., whose renal interstitial fibrosis can progress can do.
  • the administration subject is a human
  • the human subject to be administered can be appropriately selected by those skilled in the art.
  • Suitable humans include patients with kidney disease, particularly those with chronic kidney disease, preferably those with chronic kidney disease, in whom renal interstitial fibrosis has not been induced or is at an early stage of progression. is there.
  • a patient in whom renal interstitial fibrosis has not been induced or in an early stage of progression may be a patient who is predicted to need treatment in the future to replace the function of the kidney.
  • Patients who are predicted to need treatment in the future to replace the function of the kidney are preferably patients who do not need treatment to replace the function of the kidney when the inhibitor or prophylactic agent of the present invention is administered.
  • the present invention is not limited to this.
  • Patients whose renal interstitial fibrosis is not induced or in the early stage of progression can be determined by techniques known to those skilled in the art (for example, pathological examination).
  • a patient who is predicted to need treatment in the future to replace the function of the kidney can be determined using the findings suggesting the presence of kidney disease or the results of means for estimating renal function as an index. it can.
  • the administration subject is a human
  • the weight, age, sex, etc. are not particularly limited.
  • the daily dose of the inhibitor or preventive agent of the present invention is not particularly limited.
  • the dose can be appropriately set according to the symptom, body weight, age, sex, etc. of the administration subject (patient), but is usually about 0.001 mg to 10 g per day for an adult, preferably Can be selected from a range of about 0.01 mg to 5 g.
  • the inhibitor or prophylactic agent is not limited to once daily administration, and can be administered in several divided doses.
  • the suppression method or prevention method of the present invention includes a step of administering a compound capable of suppressing the functional expression of the above-mentioned CRTH2 protein to a patient with kidney disease.
  • Specific administration forms, administration methods, administration subjects, dosages and the like in the step of administering the above compound can be preferably those described above.
  • the present invention also provides a compound capable of suppressing the functional expression of CRTH2 protein for use in suppressing or preventing renal interstitial fibrosis.
  • a compound capable of suppressing the functional expression of CRTH2 protein those mentioned above are used.
  • the present invention also provides use of a compound capable of suppressing the functional expression of CRTH2 protein for producing an inhibitor or preventive agent for renal interstitial fibrosis.
  • a compound capable of suppressing the functional expression of CRTH2 protein those mentioned above are used.
  • the medicament may be a pharmaceutical composition, preferably in the aforementioned dosage form.
  • the manufactured medicine is used in the above-mentioned administration forms, administration methods, administration subjects, dosages, etc., to administration subjects such as humans, non-human mammals, birds, etc., in which the fibrosis of the renal stroma can proceed By doing so, renal interstitial fibrosis can be suppressed or prevented.
  • Example 1 In a mouse unilateral ureteral ligation model, the effect of administering a specific inhibitor against CRTH2 protein was examined.
  • CRTH2 protein antagonist CAY10471 (Cayman Chemical, antagonist of CRTH2 protein) was administered to 9-11 week old male C57B6 / J mice. Specifically, CAY10471 added so as to be 10 mg / kg with respect to the body weight of the administered mouse was added to methyl cellulose (MC) as a vehicle, and this was orally administered twice a day. Mice administered with vehicle alone served as the control group.
  • MC methyl cellulose
  • mice 4 days after administration were anesthetized with 2% isoflurane. In the anesthetized mouse, the left ureter was exposed through a midline incision, and this was ligated using silk thread at two locations at the lower renal level.
  • the control group was the sham group that had been treated (sham operation) in the same manner as above except that ligation was not performed.
  • FIG. 1 Schematic diagram of unilateral ureteral ligation surgery is shown in FIG.
  • the mouse unilateral ureter ligation model is a model system established as an animal model for studying the onset and progression mechanism of renal interstitial fibrosis.
  • the left kidney sample of the mouse on the 10th day after the operation was excised, fixed with 4% paraformaldehyde, and embedded in paraffin.
  • a 4 micrometer-thick section was prepared from the paraffin-embedded sample, and this was stained with Azan by a conventional method.
  • the area ratio (Fibrotic® Area) where collagen deposition visualized by Azan staining was observed was calculated using software attached to the microscope system, using a 100-fold magnified stained image. The measurement result of Fibrotic® Area is shown in FIG.
  • Example 2 Observation of renal interstitial fibrosis Similarly to Example 1, the state of renal interstitial fibrosis was observed using collagen deposition as an index. An example of a 100-fold magnified stained image is shown in FIG. 3 (A), and the measurement result of the fibrotic area is shown in FIG. 3 (B).
  • the gene expression level of collagen was measured by a quantitative RT-PCR method. Specifically, first, a left kidney sample of a mouse 10 days after the operation was removed, and cDNA was prepared using SuperScript® One-Cycle® cDNA® kit (Invitrogen) according to the attached instructions. Next, using the prepared cDNA, the expression level of type I collagen gene was measured by quantitative RT-PCR. The expression level of the gene was determined as a relative amount with respect to the expression level of 18S rRNA. The quantitative RT-PCR method was carried out using a commercially available probe labeled with FAM® (6-carboxy-fluorescein) by a 7300 Real-Time PCR System (Applied Biosystems) according to the attached instructions. The measurement result of the expression level of the type I collagen gene is shown in FIG.
  • the protein content of type I collagen was measured using Sircol collagen assay (Biocolor Ltd.) in the left kidney sample of the mouse 10 days after the operation according to the attached instructions.
  • the measurement result of the protein content of type I collagen is shown in FIG.
  • the left kidney cortex sample of the mouse on the 10th day after the operation was removed, first treated with 40 ⁇ g / ml Liberase (Roche Diagnostics) in RPMI1640 culture medium (at 37 ° C for 25 minutes), and the cells were separated. The separated cells were then resuspended in FACS buffer (1% BSA / 2 mM EDTA / PBS). Thereafter, a certain amount of the resuspended cells were labeled with FITC-labeled anti-CD45 antibody, APC-labeled anti-CD3 antibody, and PE-labeled anti-CD4 antibody (all at eBioscience) (30 minutes at 4 ° C.).
  • FIG. 5 (A) shows the FACS results
  • FIG. 5 (B) shows the measurement results of the ratio of CD3 positive CD4 positive cells to total cells.
  • the expression level of DP1 gene was also measured by the same method as the expression level in brain tissue on the 0th day after the treatment.
  • the measurement results of the expression levels of CRTH2 gene and DP1 gene are shown in FIG.
  • CRTH2 protein and DP1 protein are major receptor proteins for prostaglandin D2. Of these two types, CRTH2 protein is thought to function mainly in the kidney. When combined with the results of Example 1 and Reference Example 1, it strongly suggests that renal interstitial fibrosis is further progressed, that is, exacerbated via CRTH2 protein.

Abstract

The purpose of the present invention is to provide: an agent for inhibiting or preventing renal interstitial fibrosis; and a method for inhibiting or preventing renal interstitial fibrosis. As a means for achieving the purpose, provided are: an agent for inhibiting or preventing renal interstitial fibrosis, which comprises a compound capable of inhibiting the development of the function of CRTH2 protein; and a method for inhibiting or preventing renal interstitial fibrosis, which comprises administering a compound capable of inhibiting the development of the function of CRTH2 protein.

Description

[規則37.2に基づきISAが決定した発明の名称]　腎間質繊維化の抑制剤または予防剤[Name of invention determined by ISA based on Rule 37.2] Inhibitor or preventive agent for renal interstitial fibrosis
　本発明は、主に腎間質線維化の抑制剤または予防剤に関する。また、本発明は腎間質線維化の抑制方法または予防方法にも関する。 The present invention mainly relates to an inhibitor or preventive agent for renal interstitial fibrosis. The present invention also relates to a method for suppressing or preventing renal interstitial fibrosis.
　腎臓は、血中に含まれる老廃物などを濾過し、これを尿として排出する機能を担う、人体の恒常性維持のために必須の臓器である。慢性腎臓病（chronic kidney disease（CKD））は、尿たんぱくが出ているなどの腎疾患の存在を示唆する所見があり中等度以上の腎機能低下が3カ月以上持続する病態を指す。CKDには、糖尿病性腎症、高血圧性腎症あるいは腎硬化症、慢性糸球体腎炎などの幅広い病態が含まれる。現在、世界にはCKD患者が5億人いると想定される。日本人のCKD患者は1330万人、予備軍も含めると2000万人になるとされている。 The kidney is an indispensable organ for maintaining the homeostasis of the human body, which has a function of filtering waste products contained in blood and discharging it as urine. Chronic kidney disease (chronic kidney disease (CKD)) is a pathological condition that suggests the presence of kidney disease such as urinary protein, and that moderately impaired renal function persists for more than 3 months. CKD includes a wide range of conditions such as diabetic nephropathy, hypertensive nephropathy or nephrosclerosis, and chronic glomerulonephritis. Currently, there are estimated to be 500 million CKD patients in the world. It is estimated that there are 13.3 million Japanese CKD patients and 20 million including the reserve army.
　CKDが進行・増悪した際の共通な病像として、腎間質の線維化が生じる。間質の線維化が進行し腎不全へと至った患者においては、腎臓はもはやその機能を果たすことはできず、人工透析などの腎臓の機能を代替する処置が必要となる。日本において、人工透析患者数は約30万人に上り、医療費30兆円のうち人工透析医療に1兆円かかっている。たとえ人工透析に至らなくても、CKD自体が心血管疾患の独立した危険因子であることからCKDは「新たな国民病」といわれるようになっており、CKD発症および進展の予防が世界的課題となっている。 腎 Fibrosis of the renal interstitium occurs as a common symptom when CKD progresses or worsens. In patients whose interstitial fibrosis has progressed to renal failure, the kidney can no longer perform its function, and treatment that replaces the kidney function, such as artificial dialysis, is required. In Japan, the number of dialysis patients is about 300,000, and out of 30 trillion yen in medical expenses, it costs 1 trillion yen for dialysis. Even if it does not lead to artificial dialysis, CKD itself is an independent risk factor for cardiovascular disease, so CKD has been called a “new national disease”, and prevention of CKD onset and progression is a global issue It has become.
　人工透析が必要となった患者は、定期的に人工透析を行なう必要が生じる。その結果、患者の生活における自由度が制限され、QOLが著しく低下してしまう。また、人工透析は定期的に行なう必要があり、莫大な医療費が発生する。すなわち、人工透析を必要とする患者の増大は社会保障費の増大につながるため、国家としても重大な問題でもある。 Patients who need artificial dialysis need to perform artificial dialysis regularly. As a result, the degree of freedom in the patient's life is limited, and the QOL is significantly reduced. In addition, artificial dialysis needs to be performed regularly, and enormous medical costs are generated. In other words, the increase in the number of patients requiring artificial dialysis leads to an increase in social security costs, which is a serious problem for the nation.
　CKD患者における間質の線維化は、比較的病期の後期に観察される。従って、CKDの原疾患治療が困難である場合においても線維化の進行を抑制することができれば、当該患者における人工透析の導入を遅らせることができる。その結果、患者が高いQOLを維持する期間を延長することが可能となる。しかしながら、原疾患治療以外のオプションとして腎間質線維化の進展を直接的かつ効果的に抑制しうる薬剤は、臨床応用に至っていない。 Stromal fibrosis in CKD patients is observed relatively late in the disease stage. Accordingly, even if it is difficult to treat the primary disease of CKD, if the progression of fibrosis can be suppressed, the introduction of artificial dialysis in the patient can be delayed. As a result, it is possible to extend the period during which the patient maintains a high quality of life. However, a drug that can directly and effectively suppress the development of renal interstitial fibrosis as an option other than the treatment of the primary disease has not reached clinical application.
　CRTH2（Chemoattractant Receptor homologous molecule expressed on TH2）タンパク質は、プロスタグランジンD2（PGD2）受容体として知られている。PGD2は、CRTH2タンパク質を介して、Th2細胞（T helper type 2 cells）の遊走および活性化に関与することが知られている。 CRTH2 (Chemoattractant-receptor-homologous-molecule-expressed-on-TH2) protein is known as a prostaglandin D2 (PGD2) receptor. PGD2 is known to be involved in the migration and activation of Th2 cells (T helper type 2 cells) via CRTH2 protein.
　CRTH2タンパク質がTh2細胞に発現していることに関連して、PGD2やCRTH2タンパク質について、アレルギー疾患の治療などに関する研究が行なわれている。一方で、腎臓におけるこれらの生物学的意義は、PGD2の合成酵素であるL－PGDS（Lipocalin-type PGD2 Synthase）は腎障害の早期バイオマーカーとして注目されている程度に過ぎず、ほとんど分かっていない。このような状況下で、CRTH2タンパク質の腎臓における具体的な機能、特に腎間質線維化への関与は、一切知られていないのが現状である。 In connection with the expression of CRTH2 protein in Th2 cells, research is being conducted on the treatment of allergic diseases for PGD2 and CRTH2 proteins. On the other hand, their biological significance in the kidney is largely unknown as L-PGDS (Lipocalin-type PGD2 Synthase), a synthesizing enzyme of PGD2, is only attracting attention as an early biomarker of kidney damage. . Under such circumstances, the specific function of CRTH2 protein in the kidney, particularly the involvement of renal interstitial fibrosis, is not known at all.
特許第3144805号Patent No. 3144805 国際公開WO01/014882号International Publication WO01 / 014882 特開2005-87164号公報JP 2005-87164 A 特開2002-241282号公報Japanese Patent Laid-Open No. 2002-241282
　本発明は、腎間質線維化の抑制剤または予防剤を提供することを主な目的とする。また、本発明は腎間質線維化の抑制方法または予防方法を提供することをも目的とする。 The main object of the present invention is to provide an inhibitor or preventive agent for renal interstitial fibrosis. Another object of the present invention is to provide a method for suppressing or preventing renal interstitial fibrosis.
　本発明者は、上記課題を解決すべく鋭意検討したところ、驚くべきことに、CRTH2タンパク質の機能発現を抑制することで、腎間質線維化を抑制できることを見出した。本発明は、かかる知見に基づいてさらに改良を重ねることによって完成したものである。 The present inventor made extensive studies to solve the above problems, and surprisingly found that renal interstitial fibrosis can be suppressed by suppressing the functional expression of CRTH2 protein. The present invention has been completed by making further improvements based on such findings.
　すなわち、本発明は下記態様の発明を包含する。 That is, the present invention includes the following aspects of the invention.
　項１、CRTH2タンパク質の機能発現を抑制できる化合物を含む、腎間質線維化の抑制剤または予防剤。 Item 1. An inhibitor or preventive agent for renal interstitial fibrosis, comprising a compound capable of suppressing the functional expression of CRTH2 protein.
　項２、腎臓病の患者に投与するための、項１に記載の治療薬。 Item 2. The therapeutic agent according to Item 1, for administration to a patient with kidney disease.
　項３、腎臓病における腎間質線維化の進行を抑制又は予防する、項１または２に記載の抑制剤または予防剤。 Item 3. The inhibitor or preventive agent according to Item 1 or 2, which suppresses or prevents the progression of renal interstitial fibrosis in kidney disease.
　項４、前記腎臓病が、糖尿病性腎症、高血圧性腎症、慢性糸球体腎炎、慢性間質性腎炎および多発性嚢胞腎からなる群から選択される、項２または３に記載の抑制剤または予防剤。 Item 4. The inhibitor according to Item 2 or 3, wherein the kidney disease is selected from the group consisting of diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis, chronic interstitial nephritis, and polycystic kidney disease. Or prophylactic agent.
　項５、CRTH2タンパク質の機能発現を抑制できる化合物が、CRTH2タンパク質のアンタゴニストおよびCRTH2遺伝子の発現を抑制できる核酸からなる群から選択される少なくとも１種の化合物である、項１～４のいずれか１項に記載の抑制剤または予防剤。 Item 5. The compound according to any one of Items 1 to 4, wherein the compound capable of suppressing the functional expression of CRTH2 protein is at least one compound selected from the group consisting of an antagonist of CRTH2 protein and a nucleic acid capable of suppressing the expression of CRTH2 gene. The inhibitor or preventive agent according to item.
　項６、CRTH2タンパク質の機能発現を抑制できる化合物を、腎臓病の患者に投与する工程を含む、腎間質線維化の抑制方法または予防方法。 Item 6. A method for suppressing or preventing renal interstitial fibrosis, comprising a step of administering a compound capable of suppressing the functional expression of CRTH2 protein to a patient with renal disease.
　項７、腎間質線維化の抑制または予防に使用するための、CRTH2タンパク質の機能発現を抑制できる化合物。 Item 7. A compound capable of suppressing the functional expression of CRTH2 protein for use in suppressing or preventing renal interstitial fibrosis.
　項８、CRTH2タンパク質の機能発現を抑制できる化合物の、腎間質線維化の抑制剤または予防剤を製造するための使用。 Item 8. Use of a compound capable of suppressing the functional expression of CRTH2 protein for producing an inhibitor or preventive agent for renal interstitial fibrosis.
　本発明により、効果的な腎間質線維化の抑制剤または予防剤、ならびに、腎間質線維化の抑制方法または予防方法が提供される。本発明による腎間質線維化の抑制または予防により、患者のQOL（Quality of Life）の向上および必要となる医療費の低減が大いに期待される。 The present invention provides an effective inhibitor or preventive agent for renal interstitial fibrosis, and a method for suppressing or preventing renal interstitial fibrosis. By suppressing or preventing renal interstitial fibrosis according to the present invention, it is highly expected that the patient's quality of life (QOL) will be improved and the required medical expenses will be reduced.
　また、発明の抑制剤または予防剤は、腎間質線維化の発症メカニズムや治療方法の研究のためのツールとして用いることができ、さらなる医療技術の向上に貢献することも期待される。 In addition, the inhibitor or preventive agent of the invention can be used as a tool for studying the onset mechanism and treatment method of renal interstitial fibrosis, and is expected to contribute to further improvement of medical technology.
片側尿管結紮手術の模式図を示す。The schematic diagram of a unilateral ureteral ligation operation is shown. （A）CAY10471を投与した片側尿管結紮マウスの、施術後10日目の腎臓皮質組織のアザン染色像（100倍拡大）を示す。（B）Azan染色で陽性となる線維化領域面積の、観察間質領域面積に対する割合（Fibrotic Area）（％）の測定結果を示す。(A) Azan staining image (100-fold magnification) of the renal cortical tissue of the unilateral ureter-ligated mouse administered CAY10471 on the 10th day after the operation. (B) The measurement result of the ratio (Fibrotic Area) (%) of the fibrosis region area that becomes positive by Azan staining to the observed stroma region area is shown. （A）施術後10日目の腎臓皮質組織のアザン染色像（100倍拡大）を示す。（B）Azan染色で陽性となる線維化領域面積の、観察間質領域面積に対する割合（Fibrotic Area）（％）の測定結果を示す。(A) Azan staining image (100 times magnification) of the renal cortical tissue on the 10th day after the operation. (B) The measurement result of the ratio (Fibrotic Area) (%) of the fibrosis region area that becomes positive by Azan staining to the observed stroma region area is shown. （A）I型コラーゲン遺伝子の発現量の測定結果を、18S rRNAの発現量1.0に対する相対量（mRNA Collagen I / 18S）により示す。（B）I型コラーゲンのタンパク質含有量（Tissue collagen content）の測定結果を、腎臓皮質組織1mgに対する、I型コラーゲンの含有量（マイクログラム）（μg Collagen / mg Cortex）により示す。(A) The measurement result of the expression level of type I collagen gene is shown by the relative amount (mRNA Collagen I / 18S) with respect to the expression level of 1.0 of 18S rRNA. (B) The measurement result of the protein content (TissueIcollagen content) of type I collagen is shown by the content (microgram) of type I collagen (μg Collagen / mg Cortex) with respect to 1 mg of kidney cortex tissue. （A）FACS解析の結果を示す。（B）CD3陽性CD4陽性細胞（CD3+CD4+ cells）の、腎臓の総細胞に対する割合（%）（% of total kidney cells）を示す。(A) The result of FACS analysis is shown. (B) The ratio (%) of CD3 positive CD4 positive cells (CD3 + CD4 + cells) to total kidney cells (% of total kidney cells). （A）施術後0日目、5日目および10日目（days）の、腎臓組織でのCRTH2遺伝子の発現量の測定結果を、18S rRNAの発現量1.0に対する相対量により示す。（B）施術後0日目、5日目および10日目（days）の、腎臓（Kidney）組織でのDP1遺伝子の発現量の測定結果を、18S rRNAの発現量1.0に対する相対量により示す。対照として、施術後0日目の脳（Brain）組織での発現量も示す。腎臓組織では、DP1遺伝子の発現は検出されなかった（undetected）。(A) The measurement result of the expression level of CRTH2 gene in the kidney tissue on the 0th, 5th and 10th days after the operation is shown by the relative amount with respect to the expression level of 18S rRNA 1.0. (B) The measurement results of the expression level of the DP1 gene in the kidney tissue on the 0th day, the 5th day, and the 10th day (days) after the operation are shown by relative amounts with respect to the expression level of 18S rRNA of 1.0. As a control, the expression level in brain tissue on day 0 after the treatment is also shown. In kidney tissue, expression of the DP1 gene was undetected.
　なお、図中「*」は、t検定によるP<0.05の有意差があることを示す。「N.S.」は有意差がないことを示す。 In the figure, “*” indicates that there is a significant difference of P <0.05 by t test. “N.S.” indicates no significant difference.
　また、図中の英語表記または省略標記は以下の通りである：
WT：野生型（wild type）マウス
CKO：CRTH2ノックアウト（CRTH2 knockout）マウス
UUO：片側尿管結紮（unilateral urethral obstruction）UUO10d：片側尿管結紮施術後10日目（10 days）
sham：sham手術 Also, the English notation or abbreviation in the figure is as follows:
WT: wild type mouse
CKO: CRTH2 knockout mouse
UUO: unilateral urethral obstruction UUO10d: 10 days after unilateral ureteral ligation
sham: sham surgery
　本発明のCRTH2タンパク質の機能発現を抑制できる化合物を含む、腎間質線維化の抑制剤または予防剤について、以下詳細に説明する。 The following describes in detail the inhibitor or preventive agent for renal interstitial fibrosis, which contains a compound capable of suppressing the functional expression of the CRTH2 protein of the present invention.
　CRTH2（Chemoattractant Receptor homologous molecule expressed on TH2）タンパク質（別名：DP2、GPR44、CD294、DL1R）は、プロスタグランジンD2に対する受容体タンパク質であり、公知のタンパク質である。CRTH2タンパク質は、Gαi型の受容体タンパク質であると考えられている。CRTH2タンパク質は、ヒトを含む哺乳類などにおいて発現していることが知られている。ヒトにおいては、CRTH2タンパク質は、Th2細胞、顆粒球（eosinophils）、好塩基球（basophils）において発現していることが知られている。CRTH2タンパク質配列およびこれをコードする遺伝子の塩基配列は公知であり、例えば米国国立生物工学情報センター（National Center for Biotechnology Information；NCBI）が公開するデータベースにおいて、それぞれアクセッション番号NP_004769およびNM_004778として登録されている。 CRTH2 (Chemoattractant-receptor-homologous-molecule-expressed-on-TH2) protein (also known as DP2, GPR44, CD294, DL1R) is a receptor protein for prostaglandin D2 and is a known protein. CRTH2 protein is thought to be a Gαi type receptor protein. CRTH2 protein is known to be expressed in mammals including humans. In humans, CRTH2 protein is known to be expressed in Th2 cells, granulocytes (eosinophils), and basophils (basophils). The CRTH2 protein sequence and the base sequence of the gene encoding the CRTH2 protein sequence are known. For example, they are registered as accession numbers NP_004769 and NM_004778, respectively, in a database published by the National Center for Biotechnology Information (NCBI). Yes.
　なお、プロスタグランジンD2に対する受容体タンパク質は、上記CRTH2タンパク質およびDP1タンパク質（Gαi型の受容体タンパク質）が主要なものであると考えられている。 The receptor protein for prostaglandin D2 is considered to be mainly composed of the above-mentioned CRTH2 protein and DP1 protein (Gαi type receptor protein).
　CRTH2タンパク質の機能発現を抑制できる化合物は、CRTH2タンパク質の機能発現を抑制できる範囲内において、あらゆる化合物を用いることができる。CRTH2タンパク質の機能発現を抑制できる化合物として、CRTH2タンパク質に対する特異的な阻害剤およびCRTH2遺伝子の発現を抑制できる核酸などが例示されるが、これに限定されるものではない。 As the compound capable of suppressing the functional expression of CRTH2 protein, any compound can be used as long as the functional expression of CRTH2 protein can be suppressed. Examples of the compound capable of suppressing the functional expression of CRTH2 protein include a specific inhibitor for CRTH2 protein and a nucleic acid capable of suppressing the expression of CRTH2 gene, but are not limited thereto.
　ここで、「CRTH2タンパク質の機能発現を抑制」とは、CRTH2タンパク質の機能発現を抑制するあらゆる態様を指し、例えばCRTH2タンパク質の機能を阻害すること、CRTH2タンパク質の発現を抑制すること（CRTH2タンパク質をコードする遺伝子の転写の抑制、CRTH2タンパク質の翻訳の抑制など）などが例示されるが、これに限定されるものではない。なお、上記機能発現を抑制は、CRTH2タンパク質に対して特異的な物であることが好ましい。ここで、「特異的」とは、他のタンパク質に対して作用を示すことなく、CRTH2タンパク質の機能発現を抑制することが可能な能力を指す。CRTH2タンパク質の機能を阻害する態様としては、受容体であるCRTH2タンパク質とリガンドとの結合を阻害する態様などが挙げられるが、これに限定されるものではない。CRTH2タンパク質とリガンドとの結合が阻害されると、シグナル伝達経路におけるCRTH2タンパク質を介した情報伝達が低減（好ましくは、遮断）される。 Here, “suppressing the functional expression of CRTH2 protein” refers to all aspects of suppressing the functional expression of CRTH2 protein, for example, inhibiting the function of CRTH2 protein, suppressing the expression of CRTH2 protein (CRTH2 protein Examples include, but are not limited to, suppression of transcription of the encoded gene, suppression of CRTH2 protein translation, and the like. In addition, it is preferable that the suppression of the functional expression is specific to CRTH2 protein. Here, “specific” refers to the ability to suppress the functional expression of CRTH2 protein without exhibiting an action on other proteins. Examples of the mode of inhibiting the function of CRTH2 protein include, but are not limited to, a mode of inhibiting the binding between CRTH2 protein as a receptor and a ligand. When the binding between the CRTH2 protein and the ligand is inhibited, signal transduction via the CRTH2 protein in the signal transduction pathway is reduced (preferably blocked).
　また、「CRTH2遺伝子の発現を抑制できる」とは、CRTH2遺伝子（CRTH2タンパク質をコードする遺伝子）から転写されるCRTH2タンパク質をコードするmRNAの破壊（RNA干渉作用）、CRTH2タンパク質ヘの翻訳の抑制などのCRTH2タンパク質の発現量が低減される態様が例示されるが、これに限定されるものではない。 In addition, “can suppress the expression of CRTH2 gene” means the destruction of mRNA encoding CRTH2 protein transcribed from CRTH2 gene (the gene encoding CRTH2 protein) (RNA interference action), suppression of translation into CRTH2 protein, etc. Although the aspect in which the expression level of CRTH2 protein is reduced is exemplified, the present invention is not limited thereto.
　上記CRTH2タンパク質に対する特異的な阻害剤としては、例えば、公知のおよび今後開発されるあらゆるCRTH2タンパク質のアンタゴニストを用いることができる。該アンタゴニストの具体例として、ラマトロバン（Ramatroban、3-((3R)-3-{[(4-fluorophenyl)sulfonyl]amino]-1,2,3,4-tetrahydro-9H-carbazol-9-yl)propanoic acid；製品名バイナス（Baynas）；Bayer AG）、CAY10471（別名TM30089、(+)-3-[[(4-fluorophenyl)sulfonyl]methylamino]-1,2,3,4-tetrahydro-9H-carbazole-9-acetic acid；Cayman Chemical）、MK-7246（{(7R)-7-[[(4-fluorophenyl)sulfonyl](methyl)amino]-6,7,8,9-tetrahydropyrido[1,2-a]indol-10yl}acetic acid；Merck Frosst ）、TM30642（3-{3-[(4-fluoro-benzenesulfonyl)-methyl-amino]-1,2,3,4-tetrahydro-carbazol-9-yl}-propionic acid；7TM Pharma）、TM30643（[3-(4-fluoro-benzenesulfonylamino)-1,2,3,4-tetrahydro-carbazol-9-yl]-acetic acid；7TM Pharma）などの化合物、および該化合物の薬学的に許容される塩が挙げられるが、これに限定されるものではない。なお、上記に例示したCRTH2タンパク質のアンタゴニストの作用機序は公知であるが、例えば、CRTH2タンパク質とリガンドとの結合を阻害する等の作用機序が含まれる。 As the specific inhibitor for the CRTH2 protein, for example, any known and later-developed antagonists of CRTH2 protein can be used. Specific examples of the antagonist include ramatroban (3-((3R) -3-{[(4-fluorophenyl) sulfonyl] amino] -1,2,3,4-tetrahydro-9H-carbazol-9-yl) propanoic acid; product name Baynas; Bayer AG), CAY10471 (also known as TM30089, (+)-3-[[(4-fluorophenyl) sulfonyl] methylamino] -1,2,3,4-tetrahydro-9H-carbazole -9-acetic acid; Cayman Chemical), MK-7246 ({(7R) -7-[[(4-fluorophenyl) sulfonyl] (methyl) amino] -6,7,8,9-tetrahydropyrido [1,2- a] indol-10yl} acetic acid; Merck Frosst), TM30642 (3- {3-[(4-fluoro-benzenesulfonyl) -methyl-amino] -1,2,3,4-tetrahydro-carbazol-9-yl} -propionic acid; 7TM Pharma), TM30643 ([3- (4-fluoro-benzenesulfonylamino) -1,2,3,4-tetrahydro-carbazol-9-yl] -acetic acid; 7TM Pharma), and the like Examples include, but are not limited to, pharmaceutically acceptable salts of the compound. In addition, although the action mechanism of the antagonist of CRTH2 protein illustrated above is well-known, action mechanisms, such as inhibiting the coupling | bonding of CRTH2 protein and a ligand, are included, for example.
　CRTH2タンパク質のアンタゴニストの一態様として、下記一般式（１）で表される化合物が挙げられる。
式（１）
Figure JPOXMLDOC01-appb-C000001
 ［式中、Ｘ１は、メチレン基又はエチレン基を示す。Ｘ２は、水素原子又はメチル基を示す。］ One embodiment of the CRTH2 protein antagonist includes a compound represented by the following general formula (1).
Formula (1)
Figure JPOXMLDOC01-appb-C000001
 [Wherein, X 1 represents a methylene group or an ethylene group. X 2 represents a hydrogen atom or a methyl group. ]
　なお、上記式（１）でＸ１がエチレン基およびＸ２が水素原子である場合、化合物はラマトロバン（Ramatroban）である。Ｘ１がメチレン基およびＸ２がメチル基である場合、化合物はCAY10471（別名、TM30089）である。Ｘ１がエチレン基およびＸ２がメチル基である場合、化合物はTM30642である。Ｘ１がメチレン基およびＸ２が水素原子である場合、化合物はTM30643である。 In the above formula (1), when X 1 is an ethylene group and X 2 is a hydrogen atom, the compound is Ramatroban. When X 1 is a methylene group and X 2 is a methyl group, the compound is CAY10471 (also known as TM30089). When X 1 is an ethylene group and X 2 is a methyl group, the compound is TM30642. When X 1 is a methylene group and X 2 is a hydrogen atom, the compound is TM30643.
　その他の上記CRTH2タンパク質のアンタゴニストとして、下記の文献に開示されるCRTH2タンパク質のアンタゴニストを挙げるができる：特開平08-245587号公報、特開平08-175991号公報、特開平11-322600号公報、国際公開WO2006/037982号、国際公開WO2009/061676号、国際公開WO2010/057118号、国際公開WO2010/085820号、国際公開WO2010/042652号、国際公開WO2009/140642号、国際公開WO2010/008864号、国際公開WO2010/003127号、国際公開WO2010/054114号、国際公開WO2010/037059号、国際公開WO2010/039977号、国際公開WO2009/108720号、国際公開WO2009/145989号、国際公開WO2009/102893号、国際公開WO2009/099901号、国際公開WO2010/037054号、国際公開WO2010/003120号、国際公開WO2009/099902号、国際公開WO2010/054113号、国際公開WO2009/063202号、国際公開WO2007/107772号、国際公開WO2009/090414号、国際公開WO2009/093026号、国際公開WO2009/093029号、国際公開WO2005/040114号、国際公開WO2008/012511号、国際公開WO2005/044260号、国際公開WO2009/063215号、国際公開WO2005/121141号、国際公開WO2006/092579号、国際公開WO2005/040112号、国際公開WO2006/095183号、国際公開WO2006/063763号、国際公開WO2005/105727号、国際公開WO2005/123731号、国際公開WO2007/065924号、国際公開WO2007/065683号、国際公開WO2006/125596号、国際公開WO2007/065684号、国際公開WO2007/144127号、国際公開WO2007/068418号、国際公開WO2006/125593号、国際公開WO2005/102338号、国際公開WO2010/031184号、国際公開WO2010/031183号、国際公開WO2010/031182号、国際公開WO2007/019675号、国際公開WO2004/035543号、国際公開WO2008/074966号、国際公開WO2006/136859号、国際公開WO2007/045867号、国際公開WO2007/031747号、国際公開WO2007/036743号、国際公開WO2009/044134号、国際公開WO2008/078069号、国際公開WO2009/044147号、国際公開WO2009/060209号、国際公開WO2007/144625号、国際公開WO2007/062678号、国際公開WO2005/116001号、国際公開WO2007/062773号、国際公開WO2005/115374号、国際公開WO2005/115382号、国際公開WO2007/062677号、国際公開WO2007/062797号、国際公開WO2009/004379号、国際公開WO2005/018529号、国際公開WO2005/018529号、国際公開WO2004/106302号、国際公開WO2003/066047号、国際公開WO2003/066046号、国際公開WO2003/101961号、国際公開WO2004/007451号、国際公開WO2003/101981号、国際公開WO2011/017201号、国際公開WO2011/085033号、国際公開WO2009/049021号、国際公開WO2011/055270号、国際公開WO2009/042138号、国際公開WO2009/042139号、国際公開WO2008/156780号、国際公開WO2008/156781号、国際公開WO2010/027448号、国際公開WO2005/073234号、国際公開WO2004/096777号、国際公開WO2008/072784号、国際公開WO2010/055006号、国際公開WO2010/018112号、国際公開WO2010/018109号、国際公開WO2010/018113号、国際公開WO2010/006944号、国際公開WO2006/021418号、国際公開WO2006/070325号、国際公開WO2012/013566号、国際公開WO2008/113965号、国際公開WO2008/119917号、国際公開WO2012/009137号、国際公開WO2012/009134号、国際公開WO2011/079007号、国際公開WO2010/039982号、国際公開WO2006/034419号、国際公開WO2006/034418号、国際公開WO2009/102462号、国際公開WO2010/142934号、国際公開WO2010/099039号、国際公開WO2007/146838号、国際公開WO2007/0143745号、国際公開WO2010/075200号、国際公開WO2009/158426号（これらの開示全体が、参照により本明細書中に援用される。）。なお、これらの文献には、CRTH2タンパク質のアンタゴニストが、腎間質線維化の抑制または予防の効果を奏することについては、一切記載されていない。 Examples of other antagonists of the CRTH2 protein include CRTH2 protein antagonists disclosed in the following documents: Japanese Patent Application Laid-Open No. 08-245587, Japanese Patent Application Laid-Open No. 08-175991, Japanese Patent Application Laid-Open No. 11-322600, International Publication WO2006 / 037982, International Publication WO2009 / 061676, International Publication WO2010 / 057118, International Publication WO2010 / 085820, International Publication WO2010 / 042652, International Publication WO2009 / 140642, International Publication WO2010 / 008864, International Publication WO2010 / 003127, International Publication WO2010 / 054114, International Publication WO2010 / 037059, International Publication WO2010 / 039977, International Publication WO2009 / 108720, International Publication WO2009 / 145989, International Publication WO2009 / 102893, International Publication WO2009 / 099901, International Publication WO2010 / 037054, International Publication WO2010 / 003120, International Publication WO2009 / 099902, International Publication WO2010 / 054113, International Publication WO2009 / 063202, International Publication WO2007 / 107772, International Publication WO2009 / 090414, International Publication WO2009 / 093026, International Publication WO20 09/093029, International Publication WO2005 / 040114, International Publication WO2008 / 012511, International Publication WO2005 / 044260, International Publication WO2009 / 063215, International Publication WO2005 / 121141, International Publication WO2006 / 092579, International Publication WO2005 / 040112, International Publication WO2006 / 095183, International Publication WO2006 / 063763, International Publication WO2005 / 105727, International Publication WO2005 / 123731, International Publication WO2007 / 065924, International Publication WO2007 / 065683, International Publication WO2006 / 125596, International Publication WO2007 / 065684, International Publication WO2007 / 144127, International Publication WO2007 / 068418, International Publication WO2006 / 125593, International Publication WO2005 / 102338, International Publication WO2010 / 031184, International Publication WO2010 / 031183 International Publication WO2010 / 031182, International Publication WO2007 / 019675, International Publication WO2004 / 035543, International Publication WO2008 / 074966, International Publication WO2006 / 136859, International Publication WO2007 / 045867, International Publication WO2007 / 031747 , International Publication WO2007 / 036743, International Publication WO2009 / 044134, International Publication WO2008 / 078069, International Publication WO2009 / 04414 7, International Publication WO2009 / 060209, International Publication WO2007 / 144625, International Publication WO2007 / 062678, International Publication WO2005 / 116001, International Publication WO2007 / 062773, International Publication WO2005 / 115374, International Publication WO2005 / 115382 International publication WO2007 / 062677 International publication WO2007 / 062797 International publication WO2009 / 004379 International publication WO2005 / 018529 International publication WO2005 / 018529 International publication WO2004 / 106302 International publication WO2003 / 066047 International Publication WO2003 / 066046, International Publication WO2003 / 101961, International Publication WO2004 / 007451, International Publication WO2003 / 101981, International Publication WO2011 / 017201, International Publication WO2011 / 085033, International Publication WO2009 / 049021, International Publication WO2011 / 055270, International Publication WO2009 / 042138, International Publication WO2009 / 042139, International Publication WO2008 / 156780, International Publication WO2008 / 156781, International Publication WO2010 / 027448, International Publication WO2005 / 073234, International Publication Publication WO2004 / 096777, International publication WO2008 / 072784, International publication WO2010 / 055006, International publication WO2010 / 018112, International Publication WO2010 / 018109, International Publication WO2010 / 018113, International Publication WO2010 / 006944, International Publication WO2006 / 021418, International Publication WO2006 / 070325, International Publication WO2012 / 013566, International Publication WO2008 / 113965, International Publication Publication WO2008 / 119917, International Publication WO2012 / 009137, International Publication WO2012 / 009134, International Publication WO2011 / 079007, International Publication WO2010 / 039982, International Publication WO2006 / 034419, International Publication WO2006 / 034418, International Publication WO2009 / 102462, International Publication WO2010 / 142934, International Publication WO2010 / 099039, International Publication WO2007 / 146838, International Publication WO2007 / 0143745, International Publication WO2010 / 075200, International Publication WO2009 / 158426 The entirety is incorporated herein by reference. ). These documents do not describe that CRTH2 protein antagonists have the effect of suppressing or preventing renal interstitial fibrosis.
　また、上記CRTH2タンパク質に対する特異的な阻害剤の別の態様として、CRTH2タンパク質に対する特異的な抗体が挙げられる。該抗体は、好ましくはCRTH2タンパク質の細胞外ドメインに特異的に結合するもの、さらに好ましくはCRTH2タンパク質の細胞外ドメインに特異的に結合して、CRTH2タンパク質とプロストグランジンD2との結合を阻害できる抗体であるが、これに限定されるものではない。 In addition, another embodiment of the specific inhibitor for the CRTH2 protein includes a specific antibody for the CRTH2 protein. The antibody preferably binds specifically to the extracellular domain of CRTH2 protein, more preferably specifically binds to the extracellular domain of CRTH2 protein, and can inhibit the binding of CRTH2 protein and prostaglandin D2 Although it is an antibody, it is not limited to this.
　上記抗体は、ポリクローナル抗体またはモノクローナル抗体のいずれであってもよい。上記抗体がモノクローナル抗体である場合、キメラ抗体、ヒト化抗体、ヒト抗体などであってもよい。上記抗体は、公知の手法により作成することができる。あるいは、市販のものを用いることもできる。 The antibody may be a polyclonal antibody or a monoclonal antibody. When the antibody is a monoclonal antibody, it may be a chimeric antibody, a humanized antibody, a human antibody, or the like. The above antibody can be prepared by a known technique. Or a commercially available thing can also be used.
　上記CRTH2遺伝子の発現を抑制できる核酸は、好適にはsiRNA、shRNA、dsRNAなどのCRTH2タンパク質をコードするmRNAを標的としたRNA干渉（RNAi）作用を有するRNA分子や、CRTH2タンパク質をコードするmRNAの翻訳を抑制することができるmiRNAが挙げられる。あるいは、上記のRNA干渉（RNAi）作用を有するRNA分子または上記のmiRNAを発現できるベクター等のDNA分子であってもよい。上記CRTH2遺伝子の発現を抑制できる核酸は、CRTH2タンパク質をコードする遺伝子の塩基配列に基づいて、公知の手法より作成することができる。あるいは、市販のものを用いることもできる。 The nucleic acid capable of suppressing the expression of the CRTH2 gene is preferably an RNA molecule having RNA interference (RNAi) action targeting mRNA encoding CRTH2 protein such as siRNA, shRNA, dsRNA, or mRNA encoding CRTH2 protein. Examples include miRNA that can suppress translation. Alternatively, it may be an RNA molecule having the above RNA interference (RNAi) action or a DNA molecule such as a vector capable of expressing the above miRNA. The nucleic acid capable of suppressing the expression of the CRTH2 gene can be prepared by a known method based on the base sequence of the gene encoding the CRTH2 protein. Or a commercially available thing can also be used.
　なお、CRTH2タンパク質の機能発現を抑制できる化合物は、動物におけるCRTH2タンパク質をコードする遺伝子（CRTH2遺伝子）の欠損を有する動物（例えば、ノックアウト動物）と、CRTH2タンパク質の機能発現が抑制されるとの点で共通する。従って、後述の実施例で示すように、CRTH2遺伝子の欠損を有する動物でも、腎間質線維化の抑制または予防の効果が観察される。 Compounds that can suppress the functional expression of CRTH2 protein include animals that have a defect in the gene encoding CRTH2 protein (CRTH2 gene) in animals (for example, knockout animals), and that the functional expression of CRTH2 protein is suppressed. It is common in. Therefore, as shown in Examples described later, the effect of suppressing or preventing renal interstitial fibrosis is observed even in animals having a CRTH2 gene deficiency.
　また、CRTH2タンパク質は、DP1タンパク質とともに、プロスタグランジンD2に対する受容体として機能する。本発明の抑制剤または予防剤は、腎臓でその機能を発揮すると考えられるが、腎臓ではCRTH2タンパク質の発現は認められるが、DP1タンパク質の発現は検出限界以下である。さらに、尿管結紮後の腎間質の線維化が誘導される過程において、CRTH2の発現は増加するが、DP1タンパク質は発現誘導も認められない。つまり、腎臓におけるプロスタグランジンD2からのシグナルは、主にCRTH2タンパク質を介して伝達されるものと考えられる。以上のことから、腎間質線維化の抑制または予防は、CRTH2タンパク質の機能発現を抑制することで達成されるものと考えられる。 Moreover, CRTH2 protein functions as a receptor for prostaglandin D2 together with DP1 protein. Although the inhibitor or preventive agent of the present invention is considered to exert its function in the kidney, CRTH2 protein expression is observed in the kidney, but DP1 protein expression is below the detection limit. Furthermore, in the process of inducing fibrosis of the renal stroma after ureteral ligation, CRTH2 expression is increased, but DP1 protein is not induced. That is, it is considered that the signal from prostaglandin D2 in the kidney is transmitted mainly via the CRTH2 protein. From the above, suppression or prevention of renal interstitial fibrosis is considered to be achieved by suppressing the functional expression of CRTH2 protein.
　上記CRTH2タンパク質の機能発現を抑制できる化合物は、１種単独でまたは２種以上を組み合わせて用いることができる。 The compounds capable of suppressing the functional expression of the CRTH2 protein can be used singly or in combination of two or more.
　本発明の抑制剤または予防剤は、腎間質線維化の抑制剤または予防剤である。腎間質線維化の治療剤と換言することもできる。上記抑制剤または予防剤は、好ましくは腎臓病における腎間質線維化の進行、特に好ましくは慢性腎臓病における腎間質線維化の進行を抑制または予防するものであるが、これに限定されるものではない。本発明の抑制剤または予防剤の好ましい態様の一つとして、直接的に腎間質線維化の進行を抑制または予防するものが挙げられる。ここで、「直接的に」とは、腎臓における原疾患（例えば、慢性腎臓病）の治療の有無に関わらず、腎間質線維化の進行を抑制または予防する態様を指す。 The inhibitor or preventive agent of the present invention is an inhibitor or preventive agent for renal interstitial fibrosis. In other words, it can be referred to as a therapeutic agent for renal interstitial fibrosis. The inhibitor or preventive agent is preferably one that suppresses or prevents the progression of renal interstitial fibrosis in kidney disease, and particularly preferably suppresses or prevents the progression of renal interstitial fibrosis in chronic kidney disease. It is not a thing. One preferred embodiment of the inhibitor or preventive agent of the present invention is one that directly suppresses or prevents the progression of renal interstitial fibrosis. Here, “directly” refers to an embodiment in which the progression of renal interstitial fibrosis is suppressed or prevented regardless of the presence or absence of treatment of the primary disease (for example, chronic kidney disease) in the kidney.
　上記慢性腎臓病は、タンパク尿など腎疾患の存在を示唆する所見があり、かつ、中等度以上の腎機能低下が3か月以上持続する病態を指す。上記慢性腎臓病としては、糖尿病性腎症、高血圧性腎症、慢性糸球体腎炎、慢性間質性腎炎、多発性嚢胞腎などが挙げられるが、これに限定されるものではない。腎疾患の存在を示唆する所見は、当業者に公知のものであり、例えば尿タンパクが検出されるなどが例示されるが、これに限定されるものではない。腎機能の低下は、例えばクレアチニンクリアランスなどの、公知の腎機能を推定する手段により確認することができるが、これに限定されるものではない。 The above chronic kidney disease refers to a pathological condition that suggests the presence of renal diseases such as proteinuria, and that moderate or more impaired renal function persists for 3 months or more. Examples of the chronic kidney disease include, but are not limited to, diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis, chronic interstitial nephritis, and polycystic kidney disease. Findings suggesting the presence of kidney disease are known to those skilled in the art and include, for example, detection of urine protein, but are not limited thereto. The decrease in renal function can be confirmed by a known means for estimating renal function, such as creatinine clearance, but is not limited thereto.
　慢性腎臓病の患者は、多くの場合において腎間質の線維化が誘導・進行する。腎間質線維化においては、コラーゲン、フィブリンなどの細胞外マトリックスタンパク質が、腎皮質組織に過剰量沈着する。腎間質の線維化が進行すると、最終的には腎硬化症に至る。腎硬化症に至った腎臓は、もはや機能不全（末期腎不全）となり、生体の恒常性を維持し尿毒症症状を予防するためには、人工透析などの腎臓の機能を代替する処置が必要となる。 In patients with chronic kidney disease, fibrosis of the renal interstitium is induced and progresses in many cases. In renal interstitial fibrosis, extracellular matrix proteins such as collagen and fibrin are excessively deposited in renal cortical tissue. The progression of renal interstitial fibrosis eventually leads to nephrosclerosis. The kidney that has led to nephrosclerosis is no longer functional (end-stage renal failure), and in order to maintain homeostasis and prevent uremia symptoms, treatments that replace the kidney function such as artificial dialysis are required. Become.
　ここで、本発明において「腎間質線維化の抑制」とは、腎間質線維化が既に誘導された患者において、腎間質線維化がさらに進行することを停止または抑制（通常は、抑制であるが、これに限定されない。）することを指す。また、「腎間質線維化の予防」とは、腎間質線維化がまだ誘導されていない患者において、腎間質線維化の発症を防ぐまたは発症時期を遅延させることを指す。本発明の治療薬により腎間質線維化の抑制または予防が達成されることで、人工透析などの腎臓の機能を代替する処置をしなくて済む、または、処置の開始時期を遅らせることが可能となる。腎臓の機能を代替する処置の導入の要否は、当業者に公知の手段、例えばクレアチニンクリアランスなどの腎機能を推定する手段により確認することができる。 Here, in the present invention, “suppression of renal interstitial fibrosis” refers to stopping or suppressing further progression of renal interstitial fibrosis in a patient in which renal interstitial fibrosis has already been induced (usually suppression). (But not limited to this). “Preventing renal interstitial fibrosis” refers to preventing or delaying the onset of renal interstitial fibrosis in patients who have not yet been induced. By suppressing or preventing renal interstitial fibrosis with the therapeutic agent of the present invention, it is possible to eliminate the need to replace the renal function such as artificial dialysis, or to delay the start of the treatment. It becomes. The necessity of introducing a treatment that substitutes for the kidney function can be confirmed by means known to those skilled in the art, for example, means for estimating renal function such as creatinine clearance.
　本発明の抑制剤または予防剤は、その形態（投与形態）は特に限定されるものではない。好適には医薬組成物として提供される。さらに本発明の抑制剤または予防剤は、公知の手法により製剤化することができる。上記製剤の具体例として、錠剤、カプセル剤、丸剤、散剤、顆粒剤などの固形製剤、および液剤、懸濁剤、乳剤、注射剤などの液体製剤が挙げられるが、これらに限定されない。製剤の形態に応じて、適宜薬学的に許容される担体および添加剤を加えることができる。上記担体および添加剤の具体例として、賦形剤、充填剤、結合剤、付湿剤、香料、着色剤などが挙げられるが、これらに限定されない。製剤が液体製剤である場合は、公知の薬学的に許容される溶媒、例えば生理食塩水、緩衝作用を有する溶液などを用いることができる。 The form (administration form) of the inhibitor or preventive agent of the present invention is not particularly limited. Preferably it is provided as a pharmaceutical composition. Furthermore, the inhibitor or preventive agent of the present invention can be formulated by a known technique. Specific examples of the preparation include, but are not limited to, solid preparations such as tablets, capsules, pills, powders, granules, and liquid preparations such as liquids, suspensions, emulsions, injections, and the like. Depending on the form of the preparation, pharmaceutically acceptable carriers and additives can be added as appropriate. Specific examples of the carriers and additives include, but are not limited to, excipients, fillers, binders, wetting agents, fragrances, and coloring agents. When the preparation is a liquid preparation, a known pharmaceutically acceptable solvent such as physiological saline, a solution having a buffering action, or the like can be used.
　本発明の抑制剤または予防剤の投与方法は、抑制剤または予防剤としての効果が得られることを限度として、当業者が適宜設定することができ、特に制限されるものではない。投与方法の好ましい態様として、注射投与（静脈注射、皮下注射、筋肉注射、腹腔注射、患部への注射など）、経口投与、座薬投与、経皮投与（塗布など）などが例示される。投与の簡便さや投与対象への負担が少ないとの観点から、投与方法として経口投与が特に好ましいが、これに限定されるものではない。 The administration method of the inhibitor or prophylactic agent of the present invention can be appropriately set by those skilled in the art as long as the effect as the inhibitor or prophylactic agent is obtained, and is not particularly limited. Preferred embodiments of the administration method include injection administration (intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, injection into the affected area, etc.), oral administration, suppository administration, transdermal administration (application, etc.) and the like. Oral administration is particularly preferred as an administration method from the viewpoint of ease of administration and less burden on administration subjects, but is not limited thereto.
　本発明の抑制剤または予防剤の投与対象は、腎間質の線維化が進行し得るヒト、非ヒト哺乳類（イヌ、ネコ、ネズミ、ラット、ハムスター、ウサギ、ウシ、類人猿など）、鳥類などとすることができる。投与対象がヒトである場合は、投与対象となるヒトは当業者が適宜選択することができる。好適なヒトとしては、腎臓病の患者、特に慢性腎臓病の患者が挙げられ、好ましくは慢性腎臓病の患者であって、腎間質線維化が誘導されていないもしくは進行が初期段階の患者である。腎間質線維化が誘導されていないもしくは進行が初期段階の患者は、腎臓の機能を代替する処置の導入が将来必要であることが予見される患者であってもよい。腎臓の機能を代替する処置の導入が将来必要であることが予見される患者は、本発明の抑制剤または予防剤を投与時には腎臓の機能を代替する処置を必要としていない患者であることが好ましいが、これに限定されるものではない。腎間質線維化が誘導されていないもしくは進行が初期段階の患者は、当業者に公知の手法（例えば、病理検査）により判定することができる。腎臓の機能を代替する処置の導入が将来必要であることが予見される患者であることは、腎疾患の存在を示唆する所見や、腎機能を推定する手段の結果を指標として判定することができる。また、投与対象がヒトである場合、体重、年齢、性別等は特に限定されるものではない。 Subjects for administration of the inhibitor or prophylactic agent of the present invention are humans, non-human mammals (dogs, cats, mice, rats, hamsters, rabbits, cows, apes, etc.), birds, etc., whose renal interstitial fibrosis can progress can do. When the administration subject is a human, the human subject to be administered can be appropriately selected by those skilled in the art. Suitable humans include patients with kidney disease, particularly those with chronic kidney disease, preferably those with chronic kidney disease, in whom renal interstitial fibrosis has not been induced or is at an early stage of progression. is there. A patient in whom renal interstitial fibrosis has not been induced or in an early stage of progression may be a patient who is predicted to need treatment in the future to replace the function of the kidney. Patients who are predicted to need treatment in the future to replace the function of the kidney are preferably patients who do not need treatment to replace the function of the kidney when the inhibitor or prophylactic agent of the present invention is administered. However, the present invention is not limited to this. Patients whose renal interstitial fibrosis is not induced or in the early stage of progression can be determined by techniques known to those skilled in the art (for example, pathological examination). A patient who is predicted to need treatment in the future to replace the function of the kidney can be determined using the findings suggesting the presence of kidney disease or the results of means for estimating renal function as an index. it can. In addition, when the administration subject is a human, the weight, age, sex, etc. are not particularly limited.
　本発明の抑制剤または予防剤の1日当りの投与量は、特に限定されるものではない。投与対象がヒトである場合、投与量は当該被投与対象（患者）の症状、体重、年齢、性別等に応じて適宜設定することができるが、通常成人1日当り約0.001mg～10g程度、好ましくは約0.01mg～5g程度の範囲から選ぶことができる。当該抑制剤または予防剤は1日1回投与に限らず、数回に分けて投与することができる。 The daily dose of the inhibitor or preventive agent of the present invention is not particularly limited. When the administration subject is a human, the dose can be appropriately set according to the symptom, body weight, age, sex, etc. of the administration subject (patient), but is usually about 0.001 mg to 10 g per day for an adult, preferably Can be selected from a range of about 0.01 mg to 5 g. The inhibitor or prophylactic agent is not limited to once daily administration, and can be administered in several divided doses.
　本発明の抑制方法または予防方法は、前述のCRTH2タンパク質の機能発現を抑制できる化合物を、腎臓病の患者へ投与する工程を含む。上記化合物を投与する工程の具体的な投与形態、投与方法、投与対象、投与量等は、好適に前述のものとすることができる。 The suppression method or prevention method of the present invention includes a step of administering a compound capable of suppressing the functional expression of the above-mentioned CRTH2 protein to a patient with kidney disease. Specific administration forms, administration methods, administration subjects, dosages and the like in the step of administering the above compound can be preferably those described above.
　本発明は、腎間質線維化の抑制または予防に使用するための、CRTH2タンパク質の機能発現を抑制できる化合物をも提供する。CRTH2タンパク質の機能発現を抑制できる化合物としては、前述のものを用いる。前述の腎間質の線維化が進行し得るヒトを含む哺乳類、鳥類などの投与対象へ、例えば、前述の投与形態、投与方法、投与対象、投与量等にて使用することで、腎間質線維化を抑制または予防することができる。 The present invention also provides a compound capable of suppressing the functional expression of CRTH2 protein for use in suppressing or preventing renal interstitial fibrosis. As the compound capable of suppressing the functional expression of CRTH2 protein, those mentioned above are used. By using the above-mentioned administration form, administration method, administration subject, dosage, etc., to the administration subject such as mammals including humans, birds, etc., where the fibrosis of the renal interstitium can progress, Fibrosis can be suppressed or prevented.
　本発明は、CRTH2タンパク質の機能発現を抑制できる化合物の、腎間質線維化の抑制剤または予防剤を製造するための使用をも提供する。CRTH2タンパク質の機能発現を抑制できる化合物としては、前述のものを用いる。医薬は医薬組成物であってもよく、好適には前述の投与形態のものである。製造される医薬は、前述の腎間質の線維化が進行し得るヒト、非ヒト哺乳類、鳥類などの投与対象へ、例えば、前述の投与形態、投与方法、投与対象、投与量等にて使用することで、腎間質線維化を抑制または予防することができる。 The present invention also provides use of a compound capable of suppressing the functional expression of CRTH2 protein for producing an inhibitor or preventive agent for renal interstitial fibrosis. As the compound capable of suppressing the functional expression of CRTH2 protein, those mentioned above are used. The medicament may be a pharmaceutical composition, preferably in the aforementioned dosage form. The manufactured medicine is used in the above-mentioned administration forms, administration methods, administration subjects, dosages, etc., to administration subjects such as humans, non-human mammals, birds, etc., in which the fibrosis of the renal stroma can proceed By doing so, renal interstitial fibrosis can be suppressed or prevented.
　以下、本発明を更に詳しく説明するため、参考例および実施例を挙げるが本発明はこれに限定されるものではない。 Hereinafter, in order to describe the present invention in more detail, reference examples and examples are given, but the present invention is not limited thereto.
　［実施例１］
マウス片側尿管結紮モデルにおける、CRTH2タンパク質に対する特異的な阻害剤を投与することの効果を検証した。 [Example 1]
In a mouse unilateral ureteral ligation model, the effect of administering a specific inhibitor against CRTH2 protein was examined.
　＜実験方法および結果＞
１．CRTH2タンパク質のアンタゴニストの投与
9～11週齢の雄C57B6/Jマウスに、CAY10471（Cayman Chemical社、CRTH2タンパク質のアンタゴニスト）を投与した。具体的には、被投与マウスの体重に対して10mg/kgとなるように添加したCAY10471を、ビヒクルであるメチルセルロース（methyl cellulose、MC）に添加し、これを1日2回経口投与した。ビヒクルのみを投与したマウスを対照群とした。 <Experimental method and results>
1. Administration of CRTH2 protein antagonist
CAY10471 (Cayman Chemical, antagonist of CRTH2 protein) was administered to 9-11 week old male C57B6 / J mice. Specifically, CAY10471 added so as to be 10 mg / kg with respect to the body weight of the administered mouse was added to methyl cellulose (MC) as a vehicle, and this was orally administered twice a day. Mice administered with vehicle alone served as the control group.
　２．片側尿管結紮
投与後4日目のマウスを、2%イソフルランを用いて麻酔した。麻酔をしたマウスにおいて、正中切開により左尿管を露出させ、これを腎下極レベルにて2か所、絹糸を用いて結紮した。結紮を行なわない以外は上記術式と同様に施術（sham手術）したsham群を対照群とした。 2. Unilateral ureteral ligation Mice 4 days after administration were anesthetized with 2% isoflurane. In the anesthetized mouse, the left ureter was exposed through a midline incision, and this was ligated using silk thread at two locations at the lower renal level. The control group was the sham group that had been treated (sham operation) in the same manner as above except that ligation was not performed.
　片側尿管結紮手術の模式図を、図1に示す。なお、マウス片側尿管結紮モデルは、腎間質線維化の発症・進行機序を研究する動物モデルとして確立されているモデル系である。 Schematic diagram of unilateral ureteral ligation surgery is shown in FIG. The mouse unilateral ureter ligation model is a model system established as an animal model for studying the onset and progression mechanism of renal interstitial fibrosis.
　３．腎間質線維化の観察
　片側尿管結紮マウスの腎臓組織における腎間質線維化の様子を、コラーゲン沈着を指標として観察した。 3. Observation of renal interstitial fibrosis The state of renal interstitial fibrosis in the kidney tissue of unilateral ureteral ligated mice was observed using collagen deposition as an index.
　施術後10日目のマウスの左側腎臓試料を摘出し、4%パラホルムアルデヒド固定の後にパラフィンに包埋した。パラフィン包埋試料から4マイクロメートル厚の切片を調製し、これを常法によりアザン染色した。 The left kidney sample of the mouse on the 10th day after the operation was excised, fixed with 4% paraformaldehyde, and embedded in paraffin. A 4 micrometer-thick section was prepared from the paraffin-embedded sample, and this was stained with Azan by a conventional method.
　無作為に選択された腎臓皮質の組織切片の染色像を、BZ-9000顕微鏡（キーエンス社）により取得した。100倍拡大の染色像の例を、図２（A）に示す。 Stained images of randomly selected tissue sections of kidney cortex were obtained with a BZ-9000 microscope (Keyence). An example of a 100-fold magnified stained image is shown in FIG.
　100倍拡大の染色像を用いて、アザン染色により可視化されたコラーゲン沈着が観察される面積割合（Fibrotic Area）を、顕微鏡システムに付属のソフトウェアにより算出した。Fibrotic Areaの測定結果を、図２（B）に示す。 The area ratio (Fibrotic® Area) where collagen deposition visualized by Azan staining was observed was calculated using software attached to the microscope system, using a 100-fold magnified stained image. The measurement result of Fibrotic® Area is shown in FIG.
　＜考察＞
図２に示されるように、CAY10471を投与したマウスにおいて、コラーゲン沈着が観察される領域が低減した。このことは、CRTH2タンパク質に対する特異的な阻害剤を投与することで、腎間質線維化の誘発および／または進行が抑制できることを示している。 <Discussion>
As shown in FIG. 2, the area where collagen deposition was observed was reduced in mice administered with CAY10471. This indicates that administration and specific progression of CRTH2 protein can suppress the induction and / or progression of renal interstitial fibrosis.
　［参考例１］
マウス片側尿管結紮（unilateral ureteral obstruction（UUO））モデルにおける、CRTH2遺伝子欠損の効果を検証した。 [Reference Example 1]
We examined the effect of CRTH2 gene deletion in a mouse model of unilateral ureteral obstruction (UUO).
　＜実験方法および結果＞
１．片側尿管結紮
野生型マウスとして、実施例１と同様にC57B6/Jマウスを用いた。CRTH2ノックアウトマウス（CKOマウス）は、特開2005-87164号公報に記載のものを用いた。実施例１と同様に、9～11週齢の雄C57B6/JマウスおよびCKOマウスに、片側尿管結紮またはsham手術を施術した。 <Experimental method and results>
1. Unilateral ureteral ligation C57B6 / J mice were used as wild-type mice in the same manner as in Example 1. CRTH2 knockout mice (CKO mice) described in JP-A-2005-87164 were used. In the same manner as in Example 1, male C57B6 / J mice and CKO mice aged 9 to 11 weeks were subjected to unilateral ureteral ligation or sham surgery.
　２．腎間質線維化の観察
実施例１と同様に、腎間質線維化の様子を、コラーゲン沈着を指標として観察した。100倍拡大の染色像の例を、図３（A）、Fibrotic Areaの測定結果を図３（B）にそれぞれ示す。 2. Observation of renal interstitial fibrosis Similarly to Example 1, the state of renal interstitial fibrosis was observed using collagen deposition as an index. An example of a 100-fold magnified stained image is shown in FIG. 3 (A), and the measurement result of the fibrotic area is shown in FIG. 3 (B).
　3．コラーゲンの遺伝子発現量およびタンパク質含有量の測定
片側尿管結紮マウスの腎臓試料におけるコラーゲンの遺伝子発現量およびタンパク質含有量を測定した。 3． Measurement of collagen gene expression and protein content Collagen gene expression and protein content in a kidney sample of a unilateral ureteral ligated mouse were measured.
　コラーゲンの遺伝子発現量は、定量RT-PCR法により測定した。具体的には、まず施術後10日目のマウスの左側腎臓試料を摘出し、SuperScript One-Cycle cDNA kit（インビトロジェン社)を用いて、添付の説明書に従いcDNAを調製した。次いで、調製したcDNAをもちいて、I型コラーゲン遺伝子の発現量を定量RT-PCR法により測定した。遺伝子の発現量は、18S rRNAの発現量に対する相対量として求めた。定量RT-PCR法は、市販のFAM （6-carboxy-fluorescein）で標識されたプローブをもちいて、7300 Real-Time PCR System（アプライドバイオシステムズ社）により、添付の説明書に従い行なった。I型コラーゲン遺伝子の発現量の測定結果を、図４（A）に示す。 The gene expression level of collagen was measured by a quantitative RT-PCR method. Specifically, first, a left kidney sample of a mouse 10 days after the operation was removed, and cDNA was prepared using SuperScript® One-Cycle® cDNA® kit (Invitrogen) according to the attached instructions. Next, using the prepared cDNA, the expression level of type I collagen gene was measured by quantitative RT-PCR. The expression level of the gene was determined as a relative amount with respect to the expression level of 18S rRNA. The quantitative RT-PCR method was carried out using a commercially available probe labeled with FAM® (6-carboxy-fluorescein) by a 7300 Real-Time PCR System (Applied Biosystems) according to the attached instructions. The measurement result of the expression level of the type I collagen gene is shown in FIG.
　I型コラーゲンのタンパク質含有量は、施術後10日目のマウスの左側腎臓試料において、Sircol collagen assay (Biocolor Ltd.)を用いて、添付の説明書に従い測定した。I型コラーゲンのタンパク質含有量の測定結果を、図４（B）に示す。 The protein content of type I collagen was measured using Sircol collagen assay (Biocolor Ltd.) in the left kidney sample of the mouse 10 days after the operation according to the attached instructions. The measurement result of the protein content of type I collagen is shown in FIG.
　４．腎臓皮質へと浸潤したCD4陽性リンパ球の測定
腎臓皮質へと浸潤したCD4陽性リンパ球の割合を、フローサイトメトリー法（FACS解析）により測定した。 4). Measurement of CD4 positive lymphocyte infiltrating into renal cortex The ratio of CD4 positive lymphocyte infiltrating into renal cortex was measured by flow cytometry (FACS analysis).
　施術後10日目のマウスの左側腎臓皮質試料を摘出し、まずRPMI1640培養液中で40 μg/mlのLiberase（ロシュ・ダイアグノスティックス社）で処理（37℃で25分間）し、細胞を分離した。次いで、分離された細胞をFACS buffer （1% BSA / 2mM EDTA / PBS）で再懸濁した。その後、一定量の再懸濁された細胞を、FITC標識抗CD45抗体、APC標識抗CD3抗体およびPE標識抗CD4抗体（いずれもeBioscience社）を用いて標識（4℃で30分間）した。 The left kidney cortex sample of the mouse on the 10th day after the operation was removed, first treated with 40 µμg / ml Liberase (Roche Diagnostics) in RPMI1640 culture medium (at 37 ° C for 25 minutes), and the cells were separated. The separated cells were then resuspended in FACS buffer (1% BSA / 2 mM EDTA / PBS). Thereafter, a certain amount of the resuspended cells were labeled with FITC-labeled anti-CD45 antibody, APC-labeled anti-CD3 antibody, and PE-labeled anti-CD4 antibody (all at eBioscience) (30 minutes at 4 ° C.).
　標識された細胞懸濁液から、FACS Aria II（BD Bioscience社）を用いてCD45陽性細胞、次いでCD3陽性細胞、最後にCD4陽性細胞を分取し、FACS Divaソフトウェア（BD Bioscience社）を用いて解析した。FACSの結果を図５（A）に、およびCD3陽性CD4陽性細胞の、総細胞に対する割合の測定結果を図５（B）に示す。 From the labeled cell suspension, CD45-positive cells, then CD3-positive cells, and finally CD4-positive cells are separated using FACS Aria II (BD Bioscience), and using FACS Diva software (BD Bioscience) Analyzed. FIG. 5 (A) shows the FACS results, and FIG. 5 (B) shows the measurement results of the ratio of CD3 positive CD4 positive cells to total cells.
　＜考察＞
図３および図４に示されるように、片側尿管結紮施術後10日目の野生型マウスにおいては約15％程度の領域でコラーゲン沈着が観察された。これに対して、片側尿管結紮施術後10日目のCRTH2ノックアウトマウス（CKOマウス）では、コラーゲン沈着が観察された領域は約10％程度であった。さらに、図４に示されるように、CRTH2ノックアウトマウスでは、野生型マウスと比べてCKOマウスではコラーゲンの遺伝子発現量およびタンパク質含有量が有意に少なかった。これらの結果は、CRTH2遺伝子の欠損により、腎間質線維化の誘発および／または進行が抑制されたことを示している。 <Discussion>
As shown in FIG. 3 and FIG. 4, collagen deposition was observed in a region of about 15% in wild-type mice 10 days after unilateral ureteral ligation. In contrast, in CRTH2 knockout mice (CKO mice) 10 days after unilateral ureteral ligation, the area where collagen deposition was observed was about 10%. Furthermore, as shown in FIG. 4, in the CRTH2 knockout mouse, the collagen gene expression level and protein content were significantly lower in the CKO mouse than in the wild type mouse. These results show that the induction and / or progression of renal interstitial fibrosis was suppressed by the CRTH2 gene deficiency.
　また、図５に示されるように、片側尿管結紮施術後10日目の野生型マウスとCKOマウスとの間では、腎臓皮質へと浸潤したCD4陽性リンパ球の数に有意差は観察されなかった（それぞれ、腎臓の総細胞に対して0.7％および0.6％）。CRTH2遺伝子の腎臓における生物学的意義は、必ずしも明らかではなく、またこれに拘束されるものではないが、上記の結果は、野生型マウスとCKOマウスとの間で、炎症の程度に差がないことを示唆していると考えられる。また、CRTH2がTh2細胞特異的に発現する分子として同定されている事実などから、単球やマクロファージが腎間質線維化へ関与する可能性は低いと考えられる。 In addition, as shown in FIG. 5, no significant difference was observed in the number of CD4 positive lymphocytes that infiltrated into the renal cortex between wild-type mice and CKO mice 10 days after unilateral ureteral ligation. (0.7% and 0.6% of total kidney cells, respectively). Although the biological significance of the CRTH2 gene in the kidney is not always clear or constrained, the above results do not differ in the degree of inflammation between wild-type and CKO mice This is thought to suggest. In addition, due to the fact that CRTH2 has been identified as a molecule that specifically expresses Th2 cells, monocytes and macrophages are unlikely to be involved in renal interstitial fibrosis.
　［参考例２］
マウス片側尿管結紮（unilateral ureteral obstruction（UUO））モデルにおける、CRTH2遺伝子およびDP1遺伝子の発現量を観察した。 [Reference Example 2]
The expression levels of CRTH2 gene and DP1 gene in a mouse unilateral ureteral obstruction (UUO) model were observed.
　＜実験方法および結果＞
１．片側尿管結紮
実施例１と同様に、9～11週齢の雄C57B6/JマウスおよびCKOマウスに、片側尿管結紮またはsham手術を施術した。 <Experimental method and results>
1. Unilateral ureteral ligation As in Example 1, 9-11 week old male C57B6 / J mice and CKO mice were subjected to unilateral ureteral ligation or sham surgery.
２．CRTH2遺伝子およびDP1遺伝子の発現量の測定
参考例１と同様の手法により、cDNAを調製した。次いで、調製したcDNAをもちいて、CRTH2遺伝子およびDP1遺伝子の発現量を定量RT-PCR法により測定した。遺伝子の発現量は、18S rRNAの発現量に対する相対量として求めた。定量RT-PCR法は、市販のFAM （6-carboxy-fluorescein）で標識されたプローブをもちいて、7300 Real-Time PCR System （アプライドバイオシステムズ社）により、添付の説明書に従い行なった。 2. Measurement of expression levels of CRTH2 gene and DP1 gene cDNA was prepared in the same manner as in Reference Example 1. Subsequently, the expression levels of CRTH2 gene and DP1 gene were measured by quantitative RT-PCR using the prepared cDNA. The expression level of the gene was determined as a relative amount with respect to the expression level of 18S rRNA. Quantitative RT-PCR was carried out using a 7300 Real-Time PCR System (Applied Biosystems) according to the attached instructions using a commercially available probe labeled with FAM (6-carboxy-fluorescein).
　DP1遺伝子の発現量については、対照として、施術後0日目の脳組織での発現量も同様の手法により測定した。
　CRTH2遺伝子およびDP1遺伝子の発現量の測定結果を、図６に示す。 As a control, the expression level of DP1 gene was also measured by the same method as the expression level in brain tissue on the 0th day after the treatment.
The measurement results of the expression levels of CRTH2 gene and DP1 gene are shown in FIG.
＜考察＞
図６（A）に示されるように、片側尿管結紮施術前から腎臓ではCRTH2遺伝子が発現しており、施術後その発現量の増加が観察された。一方、図６（B）に示されるように、片側尿管結紮施術前および施術後のいずれにおいても、DP1遺伝子の発現は発現されなかった。 <Discussion>
As shown in FIG. 6 (A), the CRTH2 gene was expressed in the kidney before the unilateral ureteral ligation operation, and an increase in the expression level was observed after the operation. On the other hand, as shown in FIG. 6 (B), the expression of the DP1 gene was not expressed either before or after the unilateral ureteral ligation.
　CRTH2タンパク質とDP1タンパク質の2つのタンパク質は、プロスタグランジンD2に対する主要な受容体タンパク質である。腎臓では、これら2種のうち、CRTH2タンパク質が主に機能していると考えられる。そして、実施例1及び参考例１の結果と総合すると、CRTH2タンパク質を介して、腎間質線維化がより一層進行、すなわち、増悪されることを強く示唆している。 Two proteins, CRTH2 protein and DP1 protein, are major receptor proteins for prostaglandin D2. Of these two types, CRTH2 protein is thought to function mainly in the kidney. When combined with the results of Example 1 and Reference Example 1, it strongly suggests that renal interstitial fibrosis is further progressed, that is, exacerbated via CRTH2 protein.

Claims (8)

  1. CRTH2タンパク質の機能発現を抑制できる化合物を含む、腎間質線維化の抑制剤または予防剤。 An agent for suppressing or preventing renal interstitial fibrosis, comprising a compound capable of suppressing the functional expression of CRTH2 protein.
  2. 腎臓病の患者に投与するための、請求項１に記載の治療薬。 The therapeutic agent according to claim 1 for administration to a patient with kidney disease.
  3. 腎臓病における腎間質線維化の進行を抑制または予防する、請求項１に記載の抑制剤または予防剤。 The inhibitor or preventive agent according to claim 1, which suppresses or prevents the progression of renal interstitial fibrosis in kidney disease.
  4. 前記腎臓病が、糖尿病性腎症、高血圧性腎症、慢性糸球体腎炎、慢性間質性腎炎および多発性嚢胞腎からなる群から選択される、請求項１に記載の抑制剤または予防剤。 The inhibitor or preventive agent according to claim 1, wherein the kidney disease is selected from the group consisting of diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis, chronic interstitial nephritis, and polycystic kidney disease.
  5. CRTH2タンパク質の機能発現を抑制できる化合物が、CRTH2タンパク質に対するアンタゴニストおよびCRTH2遺伝子の発現を抑制できる核酸からなる群から選択される少なくとも１種の化合物である、請求項１に記載の抑制剤または予防剤。 The inhibitor or preventive agent according to claim 1, wherein the compound capable of suppressing the functional expression of CRTH2 protein is at least one compound selected from the group consisting of an antagonist to CRTH2 protein and a nucleic acid capable of suppressing the expression of CRTH2 gene. .
  6. CRTH2タンパク質の機能発現を抑制できる化合物を、腎臓病の患者へ投与する工程を含む、腎間質線維化の抑制方法または予防方法。 A method for suppressing or preventing renal interstitial fibrosis, comprising a step of administering a compound capable of suppressing the functional expression of CRTH2 protein to a patient with renal disease.
  7. 腎間質線維化の抑制または予防に使用するための、CRTH2タンパク質の機能発現を抑制できる化合物。 A compound that can suppress the functional expression of CRTH2 protein for use in the suppression or prevention of renal interstitial fibrosis.
  8. CRTH2タンパク質の機能発現を抑制できる化合物の、腎間質線維化の抑制剤または予防剤を製造するための使用。 Use of a compound capable of suppressing the functional expression of CRTH2 protein for producing an inhibitor or preventive agent for renal interstitial fibrosis.
PCT/JP2012/059411 2011-04-05 2012-04-05 Agent for inhibiting or preventing renal interstitial fibrosis WO2012137885A1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017104728A1 (en) * 2015-12-16 2017-06-22 国立大学法人東京大学 Medicine for treating food allergy

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
AIHUA ZHANG, ZHENG DONG, AND TIANXIN YANG: "Prostaglandin D2 inhibits TGF-beta1-induced epithelial-to-mesenchymal transition in MDCK cells", AM.J.PHYSIOL.RENAL PHYSIOL., vol. 291, 2006, pages F1332 - F1342 *
NANAE NAGATA, ET AL.: "De novo synthesis, uptake and proteolytic processing of lipocalin-type prostaglandin D synthase, beta-trace, in the kidneys", FEBS J., vol. 276, no. 23, December 2009 (2009-12-01), pages 7146 - 7158 *
RUFINA SCHULIGOI, ET AL.: "Prostaglandin H2 induces the migration of human eosinophils through the chemoattractant receptor homologous molecule of Th2 cells, CRTH2", J.LEUKOC.BIOL., vol. 85, no. 1, January 2009 (2009-01-01), pages 136 - 145 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017104728A1 (en) * 2015-12-16 2017-06-22 国立大学法人東京大学 Medicine for treating food allergy

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