WO2015025816A1 - Chymase inhibitor and medicine comprising chymase inhibitor - Google Patents

Abstract

Provided is a novel chymase inhibitor. The chymase inhibitor which comprises a compound represented by general formula (A) as an active ingredient displays selective and excellent inhibition of enzyme activity with respect to human chymase. The compound is preferably one type or more selected from the group consisting of quercitrin, isoquercitrin, hyperoside, kaempferol, and luteolin. A medicine comprising the chymase inhibitor as an active ingredient can suppress production of angiotensin II by suppressing activity of the human chymase, and thus is useful as a drug which is used to prevent and treat various diseases caused by angiotensin II. (In general formula (A), R¹ to R⁵ is each independently a hydroxyl group or methoxy group, and X¹ is a hydroxyl group or -OG group. G is a sugar chain.)

Description

Chymase inhibitor and drug containing chymase inhibitor

The present invention relates to a chymase inhibitor and a drug containing the chymase inhibitor.

The renin-angiotensin system is one of endocrine mechanisms that maintain homeostasis such as systemic blood pressure and body fluid volume. In the renin-angiotensin system, angiotensin I is produced from angiotensinogen by renin and further converted from angiotensin I to angiotensin II. This conversion is caused by angiotensin converting enzyme (ACE) in the circulating blood.
Angiotensin II is known to be a major causative factor in hypertensive diseases because of its strong vasoconstrictive action. Angiotensin II is a growth factor that promotes fibroblast proliferation. In cardiovascular diseases, cardiomyocyte hypertrophy, smooth muscle cell migration / proliferation, fibroblast intracellular matrix production stimulation, apoptosis It is widely involved in the regulation of cellular functions such as induction, and plays an important role in the formation of fibrosis, renal sclerosis, and arteriosclerotic lesions.
Thus, angiotensin II is the final active peptide of the renin-angiotensin system, and it can be expected to prevent or treat diseases caused by angiotensin II such as hypertension by suppressing the production of angiotensin II.

As a pathway for producing angiotensin II, a pathway produced from angiotensin I by angiotensin converting enzyme (ACE) is known. And the effectiveness of the ACE inhibitor which inhibits the activity of an angiotensin converting enzyme is clarified clinically as a prophylactic / therapeutic agent of the disease resulting from angiotensin II. However, ACE inhibitors are also known to have side effects such as dry cough, and ACE inhibitors are useful for the prevention and treatment of hypertension. There is hypertension (for example, hypertension caused by excessive salt intake) for which no effective antihypertensive effect is observed.

On the other hand, human chymase (hereinafter referred to as “chymase”) is an enzyme having the ability to produce angiotensin II in addition to ACE, and is a “non-ACE-dependent pathway that does not depend on ACE for the production of angiotensin II in human tissues. ”
The present inventors have isolated and extracted an enzyme that produces angiotensin II from angiotensin I from the human heart, and revealed that human chymase is responsible for the production of angiotensin II from the chemical structure and gene cloning of the enzyme (non- Patent Documents 1 to 3). Human chymase is produced in tissue-bound mast cells and stored in secretory granules, and has a physiological function in the tissue region after secretion from the secretory granules. In the blood vessel wall, normal, human chymase is more abundant in the outer membrane, and ACE is rather localized in the inner membrane, but once the blood vessel is damaged, chymase is involved in outer membrane remodeling and neointimal proliferation (Non-patent document 4). It has been reported that the non-ACE-dependent angiotensin II production pathway by this chymase contributes more in humans than other animal species, and the production of angiotensin II in the human heart is about 80%. There is also a report that it bears (Non-Patent Document 5).

Chymase is mainly present in the mast cell secretory granules of the heart, skin, blood vessel wall, intestinal tract, etc. When these tissues are damaged, they are secreted from the secretory granules and released extracellularly, and angiotensin I in the tissues is released. It is converted to angiotensin II (Non-patent Documents 3 and 6).
Angiotensin II is known to be deeply involved in cardiovascular hypertrophy and reconstruction, and it has been suggested that human chymase is deeply involved in cardiovascular lesions through the production of angiotensin II. Therefore, chymase inhibitors that inhibit the activity of human chymase prevent or treat diseases caused by angiotensin II by inhibiting local production of angiotensin II in human tissues, especially various heart diseases and vascular diseases. -It can be used as a therapeutic agent for kidney disease.

So far, several chymase inhibitors have been reported. Triazine derivatives (Patent Document 1), substituted aminopyrimidinylacetamide derivatives (Patent Document 2), benzimidazole derivatives (Patent Document 3) and the like have been reported as substances having a chymase inhibitory action.
In addition, the present inventors have found that chymase inhibitors are useful for the prevention and treatment of abnormal glucose tolerance (Patent Document 4) and diseases associated with abnormal vascular functions (Patent Document 5), and chymase inhibitors and ACE inhibitors. It has been reported that the combined use of these agents is useful for the prevention and treatment of cardiovascular diseases such as heart disease (Patent Document 6). A chymase inhibitor suppresses angiotensin II production by chymase released from each tissue mast cell by some kind of stimulation. Therefore, oral administration also prevents and improves both tissue lesions and systemic dysfunction (eg, hypertension). There is also an advantage that it is effective.
Furthermore, the present inventors have reported that inhibition of angiotensin converting enzyme (ACE) activity does not show an antihypertensive effect due to inhibition of chymase activity, and an antihypertensive effect is also observed against hypertension caused by excessive intake of salt. (Non-Patent Document 5).

JP-A-8-208654 JP 2003-104984 A Japanese Patent Laid-Open No. 2001-199983 International Publication No. 2005/018672 Pamphlet International Publication No. 2001/032214 Pamphlet International Publication No. 2003/018061 Pamphlet

On the other hand, the compounds that are active ingredients of chymase inhibitors that have been reported so far have an insufficient activity inhibiting action on human chymase, and many of them are difficult to obtain.
Under such circumstances, an object of the present invention is to provide a novel chymase inhibitor that has not been reported so far and a drug containing the chymase inhibitor as an active ingredient.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that the following inventions meet the above-mentioned object, and have reached the present invention.

That is, the present invention relates to the following inventions.
<1> A chymase inhibitor containing a compound represented by the following general formula (A) as an active ingredient.

(In the general formula (A), R ¹ to R ⁵ each independently represents a hydrogen atom, a hydroxyl group or a methoxy group, X ¹ represents a hydrogen atom, a hydroxyl group or a —OG group, and G represents a sugar chain.)
<2> The chymase inhibitor according to <1>, wherein X ¹ is a hydroxyl group, a rhamnose residue, a glucose residue, or a galactose residue.
<3> The chymase according to <1> or <2>, wherein the compound represented by the general formula (A) is at least one selected from the group consisting of quercitrin, isoquercitrin, hyperoside, kaempferol and luteolin. Inhibitor.
<4> The chymase inhibitor according to any one of <1> to <3>, comprising at least one selected from quercetin glycosides and quercetin derivatives.
<5> The chymase inhibitor according to any one of <1> to <4>, comprising a hyperoside.
<6> A drug containing the chymase inhibitor according to any one of <1> to <5>.
<7> The drug according to <6>, which is a non-ACE-dependent angiotensin II production inhibitor.
<8> The drug according to <6> or <7>, further containing a bioabsorption promoter.
<9> The drug according to any one of <6> to <8>, which is a preventive or therapeutic agent for cardiovascular disease.
<10> The drug according to <9>, wherein the cardiovascular disease is hypertension and / or hypertensive organ disorder.
<11> The drug according to <10>, which is a prophylactic or therapeutic agent for hypertension and / or hypertensive organ damage caused by excessive intake of salt.
<12> The drug according to any one of <6> to <8>, which is a prophylactic and / or therapeutic agent for arteriosclerotic diseases caused by lipid deposition in blood vessels.
<13> The drug according to any one of <6> to <8>, which is a prophylactic and / or therapeutic agent for a disease caused by impaired glucose tolerance.
<14> The drug according to <13>, wherein the disease caused by impaired glucose tolerance is diabetes and / or diabetic complications.
<15> The drug according to any one of <6> to <8>, which is one or more prophylactic and / or therapeutic agents selected from atopic dermatitis and psoriasis.
<16> The drug according to any one of <6> to <8>, which is one or more prophylactic and / or therapeutic agents selected from cirrhosis and pulmonary fibrosis.
<17> The drug according to any one of <6> to <8>, which is one or more prophylactic and / or therapeutic agents selected from Crohn's disease and ulcerative colitis.

The chymase inhibitor of the present invention exhibits selective and excellent enzyme activity inhibitory action against human chymase. Therefore, since the drug containing the chymase inhibitor as an active ingredient can suppress the production of angiotensin II by suppressing the activity of human chymase, it is useful as a medicine used for the prevention and treatment of various diseases caused by angiotensin II. It is.

It is the evaluation result (8 weeks) of the dose dependence of the hyperoside with respect to the systolic blood pressure (SBP) of the salt-dependent hypertensive mouse | mouth (six) which gave 2% saline as drinking water. It is the evaluation result (8 weeks) of the dose dependence of the hyperoside with respect to the diastolic blood pressure (DBP) of the salt-dependent hypertensive mouse | mouth (6 animals) which received 2% saline as drinking water.

Hereinafter, the present invention will be described in detail with reference to examples and the like, but the present invention is not limited to the following examples and the like, and can be arbitrarily modified and implemented without departing from the gist of the present invention.

The present invention relates to a chymase inhibitor containing as an active ingredient a compound represented by the following general formula (A) or a pharmacologically acceptable salt of the compound.
In addition, the compound represented by the general formula (A) has a flavonol skeleton or a flavonol-like skeleton, and is common in that it is substituted with a similar functional group.
Hereinafter, the compound represented by the general formula (A) may be referred to as “compound (A)”.

In the general formula (A), R ¹ to R ⁵ each independently represents a hydrogen atom, a hydroxyl group or a methoxy group, and X ¹ represents a hydrogen atom, a hydroxyl group or an —OG group. Here, G is a sugar chain, and details will be described later.

In the general formula (A), the number of hydroxyl groups or methoxy groups constituting R ¹ to R ⁵ is not limited, but the hydroxyl groups are 3 or more, more preferably 4 (all hydroxyl groups).

In the above general formula (A), X ¹ is a hydrogen atom, a hydroxyl group or an —OG group. When X ¹ is an —OG group, the sugar chain is glycosidically bonded through an oxygen atom.

There is no particular limitation on the structure of the sugar chain and the type of sugar constituting the sugar chain. The number of sugars constituting the sugar chain is not particularly limited, but is usually 1 to 4, preferably 1 or 2. In addition, when the number of sugar chains is 1, it is a so-called monosaccharide residue. The sugar constituting the sugar chain may be a pentose sugar or a hexose sugar.
The hydroxyl group contained in the sugar chain may be modified with other groups. For example, the hydroxyl group may be esterified as described later or may be alkoxylated.

Specific examples of the sugar constituting the sugar chain include ribose, glucose, mannose, galactose, rhamnose, xylose, arabinose, glucuronic acid, rutinose, sofolose, gentiobiose, zambubiose, latyrose, laminaribioose, gentiotriose, robinobiose, etc. Is mentioned.

The sugar may be a sugar (organic acid-bonded sugar) in which an organic acid is singly or plurally esterified. There is no particular limitation on the bonding position of the organic acid. Specific examples of the binding position of the organic acid include one or more of the 2-position, 3-position, and 6-position of the sugar. There is no limitation in particular in the kind and structure of the said organic acid. The organic acid may be an aromatic organic acid or an aliphatic organic acid.

In general formula (A), X ¹ is preferably a hydrogen atom, a hydroxyl group, a rhamnose residue, a glucose residue, or a galactose residue in terms of excellent human chymase inhibitory activity. In this case, the number of hydrogen atoms, hydroxyl groups or methoxy groups constituting R ¹ to R ⁵ is not limited, but hydroxyl groups of 3 or more are preferable.
In the chymase inhibitor of the present invention, as a compound represented by the general formula (A) (compound (A)), any one of quercetin, quercetin glycoside, and quercetin derivative having the following chemical formula is used as an active ingredient. It is preferable to contain, and quercetin glycosides and quercetin derivatives having particularly good absorbability in the body are preferable. Examples of quercetin glycosides and quercetin derivatives include quercitrin, isoquercitrin, and hyperoside.

In the chymase inhibitor of the present invention, examples of the compound (A) other than the quercetin, quercetin glycoside, and quercetin derivative include, for example, galangin, kaempferol, myricetin, luteolin, and luteolin. Is mentioned.

In the chymase inhibitor of the present invention, the compound (A) may be contained alone or in combination of two or more. Among the compounds (A), it is preferable to contain as an active ingredient at least one selected from the group consisting of quercitrin, isoquercitrin, hyperoside, kaempferol and luteolin having excellent chymase inhibitory activity.

In addition, quercitrin, isoquercitrin and hyperoside are excellent in chymase activity inhibitory action and have a functional group having a sugar chain in X ¹ , so that high bioabsorbability can be expected. Therefore, the chymase inhibitor of the present invention preferably contains at least one of quercitrin, isoquercitrin and hyperoside.
In particular, since hyperoside has a particularly excellent chymase activity inhibitory action and high bioabsorbability, the chymase inhibitor of the present invention preferably contains hyperoside.

Compound (A) may be artificially synthesized or derived from natural products. When a natural product is used as a raw material, an extract obtained from a raw material animal or plant by a known extraction method may be used as it is, or further separation and purification may be performed.

Examples of natural products containing quercetin, which is one of the compounds suitable as compound (A), include citrus fruits, onions, buckwheat, apples, green tea, broccoli, morohaya, and willow. Similarly, as a natural product containing a large amount of quercitrin, which is one of suitable compounds, there may be mentioned dokudami, ginkgo, and willow. Similarly, natural products containing a large amount of isoquercitrin, which is one of suitable compounds, include mango, dandelion, willow tree and the like.
In addition, examples of natural products that are rich in hyperoside (quercetin-3-O-galactoside) include moss, hypericum perforatum, and oak.
There is no restriction | limiting in particular as a site | part which can be used in these raw material plants, For example, an above-ground part, a root part, a fruit, etc. are mentioned.

These compounds may be contained as pharmacologically acceptable salts, and the form of the salt is not particularly limited. For example, alkali metal salts (sodium salt, potassium salt, lithium salt) Etc.), alkaline earth metal salts (calcium salt, magnesium salt, etc.), inorganic salts (acetate salt, ammonium salt, etc.), organic amine salts (dibenzylamine salt, glucosamine salt, ethylenediamine salt, diethylamine salt, triethylamine salt, dicyclohexyl) Amine salts, diethanolamine salts, tetramethylammonium salts, etc.), amino acid salts (glycine salts, lysine salts, arginine salts, ornithine salts, asparagine salts, etc.).

The chymase inhibitor of the present invention can suppress the production of angiotensin II and prevent and treat diseases caused by angiotensin II. Specific diseases will be described later.
The administration method of the chymase inhibitor of the present invention may be selected from oral administration and parenteral administration depending on the type of disease caused by angiotensin II.
It should be noted that the content of the compound (A) in the chymase inhibitor of the present invention is such that the necessary amount can be ingested in consideration of necessary activity inhibitory action and administration method for the disease to be prevented or treated. It is appropriately determined within a range.
The chymase inhibitor of the present invention can be administered once a day, but can also be administered in several divided portions at appropriate intervals. Furthermore, when used as an injection, it can be performed by stepwise administration by administration injection or continuous administration by infusion.

(Applied drug of chymase inhibitor)
The chymase inhibitor of the present invention inhibits the activity of human chymase, an enzyme that produces angiotensin II from angiotensin I independently of ACE. Therefore, the drug containing the chymase inhibitor of the present invention becomes a non-ACE-dependent angiotensin II production inhibitor, suppresses tissue angiotensin II production, and can prevent and treat diseases caused by angiotensin II.
Therefore, a drug containing the chymase inhibitor of the present invention (hereinafter sometimes referred to as “the drug of the present invention”) can be used as a prophylactic / therapeutic agent for various clinical pathologies associated with increased enzyme activity of human chymase. . In addition, although the chemical | medical agent containing the chymase inhibitor of this invention may contain the arbitrary components accept | permitted pharmacologically, you may use the chymase inhibitor of this invention alone as a chemical | medical agent of this invention.

Examples of diseases associated with increased enzyme activity of human chymase include many circulatory diseases and atherosclerotic diseases caused by lipid deposition in blood vessels.
Cardiovascular diseases include all hypertension and hypertensive organ disorders. That is, examples of the heart disease include coronary artery disease, heart failure, and arteriosclerotic valvular disease. Cerebral diseases include cerebrovascular lesions, and cerebrovascular dilatation is also a target. In addition, since all hypertensive arteriosclerotic diseases of systemic blood vessels are included, chronic kidney diseases including upper and lower limb obstructive arteriosclerosis, hypertensive nephropathy and hypertensive diabetic nephropathy are also targeted.
The agent of the present invention can be used as a prophylactic and / or therapeutic agent for these cardiovascular diseases. The dose of the drug of the present invention when the disease to be prevented or treated is appropriately selected in consideration of not only the type of disease but also individual differences such as the age and sex of the subject. Is done.

In particular, hypertension caused by excessive salt intake tends to be drug-resistant hypertension, so antihypertensive effects such as ACE inhibitors and angiotensin receptor antagonists (ARB) are attenuated, and the antihypertensive effect is insufficient. However, in the case of the drug of the present invention having chymase activity inhibition as a mechanism, an effective antihypertensive effect is observed even in hypertension caused by excessive intake of salt.
Therefore, the medicament of the present invention is particularly suitable as a prophylactic / therapeutic agent for hypertension itself due to excessive intake of salt or the aforementioned hypertensive organ disorder.
In addition, the amount of the drug of the present invention in the case where the disease to be prevented and treated is hypertension caused by excessive intake of salt is appropriately selected in consideration of individual differences such as age and sex of the subject. However, it is usually about 1 mg to 50 mg per day.

Arteriosclerotic diseases caused by lipid deposition in blood vessels are caused by abnormal blood vessel function caused by lipid deposition in blood vessels, and symptoms caused by abnormal blood vessel function caused by lipid deposition in blood vessels. Diseases that worsen, diseases that delay healing due to vascular lipid abnormalities caused by vascular lipid deposition, and the like. For example, arteriosclerosis, acute cardiac coronary syndromes such as unstable angina and acute myocardial infarction, restenosis after percutaneous coronary angioplasty, obstructive arteriosclerosis, occlusive thromboangiitis, atherosclerosis Cases of vascular dysfunction due to vascular lipid deposition in cerebral infarction, cerebral infarction, intermittent rupture, lower extremity gangrene, renovascular hypertension, renal aneurysm, renal infarction.
The agent of the present invention can be used as a prophylactic and / or therapeutic agent for diseases associated with vascular dysfunction involving lipid deposition of these blood vessels. In addition, non-alcoholic liver injury and improvement of vascular patency after bypass surgery (especially in the case of venous bypass) are also targeted.
The dose of the drug of the present invention when the disease to be prevented or treated is appropriately selected in consideration of not only the type of disease but also individual differences such as the age and sex of the subject. Is done.

Diseases resulting from impaired glucose tolerance include diabetes and / or diabetic complications. Diabetic complications include diabetic nephropathy, diabetic retinopathy, diabetic peripheral neuropathy, hyperinsulinemia, insulin resistance syndrome, arteriosclerosis, acute cardiac coronary syndrome, obstructive arteriosclerosis, vasculitis, Examples include cerebral infarction, hypertension, renal failure, nephropathy, renal aneurysm, renal infarction, or obesity.
It can be used as a glucose tolerance ameliorating agent containing the chymase inhibitor of the present invention or a prophylactic and / or therapeutic agent for diseases caused by impaired glucose tolerance (particularly diabetes and / or diabetic complications). The dose of the drug of the present invention when the disease to be prevented or treated is appropriately selected in consideration of not only the type of disease but also individual differences such as the age and sex of the subject. Is done.

In addition to the above diseases, diseases associated with increased enzyme activity of human chymase include skin diseases such as atopic dermatitis and psoriasis, fibrosis-related organ disorders such as arteriosclerosis, cirrhosis and pulmonary fibrosis, Crohn's disease and ulcerative colon Examples include digestive diseases such as flame, and the agent of the present invention can be used as a preventive agent and / or therapeutic agent for these diseases.
That is, the agent of the present invention can be used as one or more prophylactic and / or therapeutic agents selected from atopic dermatitis and psoriasis.
In addition, the drug of the present invention can be used as one or more prophylactic and / or therapeutic agents selected from cirrhosis and pulmonary fibrosis.
In addition, the drug of the present invention can be used as one or more prophylactic and / or therapeutic agents selected from Crohn's disease and ulcerative colitis.
The dose of the drug of the present invention when the disease to be prevented or treated is appropriately selected in consideration of not only the type of disease but also individual differences such as the age and sex of the subject. Is done.

(Form of chymase inhibitor)
An effective amount of the chymase inhibitor of the present invention can be blended with a pharmaceutically acceptable carrier, and can be administered orally or parenterally as a solid preparation or a liquid preparation. The dosage form may be any dosage form used for normal oral administration or parenteral administration.
Specific examples of dosage forms used for oral administration or parenteral administration include powders, granules, tablets, capsules, troches and the like as solid preparations. Examples of liquid preparations include internal solutions, external solutions, suspensions, emulsions, syrups, injection solutions, infusions, and the like, and these dosage forms and other dosage forms are appropriately selected according to the purpose.

In solid preparations, adjuvants such as excipients, binders, disintegrants, lubricants, flavoring agents and stabilizers may be used for the chymase inhibitor of the present invention which is the main agent. The ratio of the main agent and the auxiliary agent is appropriately selected according to the purpose.
Preferable examples of the excipient in the solid preparation include lactose, D-mannitol, starch and the like. Preferable examples of the binder include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropylcellulose and the like. Preferable examples of the disintegrant include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium and the like. Moreover, you may use formulation additives, such as antiseptic | preservative, antioxidant, a coloring agent, and a sweetener, as needed.

In addition, when used as a liquid preparation, a solvent having dispersibility of the above-mentioned compound as an active ingredient and having biological safety is selected. Preferable examples of the solvent include water for injection, ethanol, propylene glycol and the like.

The liquid preparation may contain auxiliary components such as a solubilizing agent, a suspending agent, an isotonic agent, a buffering agent and an antioxidant together with the chymase inhibitor of the present invention which is the main agent.
Preferable examples of the solubilizer include ethanol, polyethylene glycol, propylene glycol, benzyl benzoate, sodium carbonate, sodium citrate and the like. Preferable examples of the suspending agent include sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate, polyvinyl alcohol, polyvinyl pyrrolidone, cahydroxymethyl cellulose and the like. Preferable examples of the buffer include buffers such as phosphates, acetates and carbonates. Preferable examples of the antioxidant include sulfite and ascorbate.

In addition, the chymase inhibitor of the present invention may contain any known component as long as the activity inhibitory action is not suppressed. For example, a stabilizer, a wetting agent or an emulsifier can be added, or a salt can be used as an auxiliary agent as an osmotic pressure adjusting agent or a pH adjusting agent.

Moreover, the compound (A) which is an active ingredient of the chymase inhibitor of the present invention includes a compound having poor bioabsorbability. For example, quercetin is water soluble but not highly bioabsorbable. Since the compound (A) having poor bioabsorbability cannot be expected to have the above-mentioned medicinal effect based on chymase inhibitory action, it is preferable to take a bioabsorption accelerator in order to increase bioabsorbability. The bioabsorption enhancer may be contained as a component of the drug of the present invention, or the bioabsorption enhancer may be used in combination with the drug of the present invention.
Any bioabsorption promoter may be used as long as it has an action of enhancing the bioabsorbability of the compound (A), and examples thereof include fats and oils and citrate. Fats and oils are not particularly limited, but are vegetable oils obtained from cottonseed oil, soybean oil, palm oil, corn oil, sunflower oil, rapeseed oil, sesame oil, etc., and livestock products such as sardines, mackerel, fish, cows, pigs, etc. Animal fats and oils obtained from Examples of the citrate include sodium citrate, potassium citrate and iron citrate. One type of these bioabsorption promoters may be used, or two or more types may be mixed.

A general method for producing pharmaceuticals can be applied to the above-mentioned methods for producing solid preparations and liquid preparations.

Furthermore, the chymase inhibitor of the present invention can contain other pharmaceutically effective ingredients in addition to the above active ingredients as needed in the treatment of various diseases. For example, the drug of the present invention may be used in combination with an ACE inhibitor or an angiotensin II receptor antagonist when a synergistic preventive / therapeutic effect can be expected.
The agent of the present invention becomes a non-ACE-dependent angiotensin II production inhibitor due to chymase activity inhibitory action, but by using this in combination with an ACE inhibitor, the ACE inhibitor or chymase inhibitor is used alone. Compared to the above, it is possible to synergistically suppress the production of angiotensin II in the local tissue, maintain the action of ACE inhibitor such as bradykinin degradation, and suppress various physiological actions mediated by chymase. It is effective because of its therapeutic effect.

In addition, when it is desired to ingest the chymase inhibitor of the present invention by eating and drinking on a daily basis, it may be contained in the food or beverage as a functional food. The term “functional food” as used herein means foods and / or beverages ingested for the purpose of maintaining health, such as health foods, nutritional supplements, functional nutritional foods, nutrition insurance foods, in addition to general foods. Yes. In addition, when commercializing as a functional food, various additives used in food, specifically, coloring agents, preservatives, thickening stabilizers, antioxidant bleaching agents, antibacterial and antifungal agents, A sour agent, a seasoning, an emulsifier, a reinforcing agent, a manufacturing agent, a fragrance and the like may be added.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to the following examples unless the gist thereof is changed.

<Evaluation 1: Evaluation of chymase activity inhibitory action (in vitro)>
The compound (A) used for the evaluation of chymase activity inhibitory action is as follows.
"Compound (A)"
・ Quercetin (Wako Pure Chemical Industries, Ltd.)
・ Quercitrin (SIGMA)
・ Isoquercitrin (SIGMA)
・ Hyperoside (Wako Pure Chemical Industries, Ltd.)
・ Galangin (Wako Pure Chemical Industries, Ltd.)
・ Kaempferol (Wako Pure Chemical Industries, Ltd.)
・ Myricetin (Wako Pure Chemical Industries, Ltd.)
・ Luteolin (Funakoshi Co., Ltd.)

Further, as a comparative example, the chymase activity inhibitory action was also evaluated for compounds having the following flavanone skeleton, its similar skeleton, and flavanol-like skeleton.

・ Eriodictyol (Funakoshi Corporation)
・ (+)-Taxifolin (SIGMA)
・ (±) -Taxifolin ((±) -Taxifolin Hydrate) (SIGMA)
・ Catechin ((+)-Catechin) (SIGMA)
・ Epicatechin (Wako Pure Chemical Industries, Ltd.)

Evaluation of chymase inhibitory activity (in vitro) was performed by the method described below with a modification to the method described in Eur J Biochem 268 (22), 5985-93 (2001). This evaluation method is a simple method using a substrate in which angiotensin I, which is a substrate for human chymase, is Dnp / Nma-modified, and that the chymase cleaves this substrate to produce angiotensin II to produce fluorescence. The outline of the evaluation method is described below.
The incubation buffer is 20 mM phosphate buffer containing 100 mM NaCl and the total incubation solution volume is 100 μL. First, 5 μL of a food extract solution (assay sample) is added, adjusted to contain 0.0012 units of standard human chymase (SIGMA), and preincubated at room temperature for 30 minutes, followed by Dnp / Nma modified angiotensin as a substrate. Add I (prepared by Protein Research Institute) to a final concentration of 200 μM and incubate at 37 ° C. for 30 minutes. The incubation was terminated by adding 25 μL of 0.5M NaOH. The luminescence fluorescence (460 nm) of the produced Dnp angiotensin II was measured, and the production amount was calculated from a standard curve prepared by standard Dnp angiotensin II. Using a control without an inhibitor as a control, the human chymase inhibitory activity and inhibition rate of the assay sample were determined.

Compound (A) quercetin, quercitrin, isoquercitrin, hyperoside, galangin, kaempferol, myricetin and luteolin were added so as to have the concentrations shown in Table 1, respectively, and the human chymase inhibitory activity rate was evaluated by the method described above. did.
Further, eriodictyol, (+)-taxifolin, (±) -taxifolin, catechin and epicatechin, which are comparative examples, were added so as to have the concentrations shown in Table 1, and the human chymase inhibitory activity rate was increased by the method described above. evaluated.
The results are shown in Table 1.

From the above results, it was confirmed that the compound (A) has a chymase activity inhibitory action. In particular, it was found that quercitrin, isoquercitrin, hyperoside, kaempferol and luteonin, which are compounds (A), have an excellent chymase activity inhibitory action.

<Evaluation 2: Evaluation of the action to improve hypertension by chymase inhibitors>
Using the chymase inhibitor of the present invention, as a disease caused by angiotensin II, the effect of improving hypertension caused by excessive intake of salt was evaluated. Note that hyperoside was used as an active ingredient of the chymase inhibitor.

In the following 5 groups (control group, test groups 1 to 4), systolic blood pressure (SBP) and diastolic blood pressure (DBP) were evaluated.
As an experimental animal, a wild-type mouse (C57BL / 6J) obtained from Japan SLC was reared in a laboratory animal breeding room in which the temperature was 24 ± 1 ° C., the humidity was 50-60%, and the lighting was controlled by a 12-hour light / dark cycle. CE-2 (Nippon Kureha) was used as the feed, and 2% by weight saline was used as the drinking water.
As for the number of cases, the experiment was started with N = 6 for each group, and no mouse died during the experiment. In each group, there was no difference in body weight, food intake and water intake.

<Control group (saline solution only)>
Salt-dependent hypertensive mice in which 6 mice were freely fed CE-2 (Kureha, Japan) as feed, 2% saline was used as drinking water, and 2% saline solution was loaded for 3 months Was made. Thereafter, the salt-dependent hypertensive mice were reared under the same conditions, and the evaluation was performed every week for 8 weeks.

<Test group 1 (hyperoside administration: 20 mg / kg BW / day)>
Similar to the control group, 2% saline solution was given for 3 months to prepare salt-dependent hypertensive mice (6 mice). The salt-dependent hypertensive mice (6 mice) were given hyperoside at 20 mg / kg BW / day together with the feed and saline, and the evaluation was performed every week for 8 weeks.

<Test group 2 (hyperoside administration: 10 mg / kg BW / day)>
Similar to the control group, 2% saline solution was given for 3 months to prepare salt-dependent hypertensive mice (6 mice). The salt-dependent hypertensive mice (6 mice) were given hyperoside at 10 mg / kg BW / day together with the feed and saline, and the evaluation was performed every week for 8 weeks.

<Test group 3 (hyperoside administration: 5 mg / kg BW / day)>
Similarly to the control group, 2% saline solution was given for 3 months to prepare salt-dependent hypertensive mice (6 mice). The salt-dependent hypertensive mice (6 mice) were given hyperoside at 5 mg / kg BW / day together with the feed and saline, and the evaluation was performed every week for 8 weeks.

<Test group 4 (hyperoside administration: 2.5 mg / kg BW / day)>
Similar to the control group, 2% saline solution was given for 3 months to prepare salt-dependent hypertensive mice (6 mice). The salt-dependent hypertensive mice (6 mice) were given hyperoside at 2.5 mg / kg BW / day together with the feed and saline, and the evaluation was performed every week for 8 weeks.

1 to 3 show the evaluation results of systolic blood pressure (SBP) and diastolic blood pressure (DBP) in the control group and test groups 1 to 4. FIG. A two-way analysis of variance was performed, and Bonferroni was used for the post-test. The significance level was P <0.05.

As shown in FIGS. 1 to 3, in the control group, increases in systolic blood pressure and diastolic blood pressure were observed with the passage of the administration period for 3 months, and this state was maintained for the test period. That is, it is recognized that hypertension caused by excessive intake of salt was developed in the control group given only 2% by weight of saline. In the subsequent test period, increases in systolic blood pressure and diastolic blood pressure were observed.
On the other hand, in test groups 1 to 4 in which hyperoside was administered 3 months after administration of saline (after the onset of hypertension), systolic blood pressure and diastolic blood pressure were observed from the first week of hyperoside administration compared to the control group. It can be seen that all increases started to be suppressed, and each of them was significantly suppressed after the second week. And it was confirmed that the hypotensive action of systolic blood pressure and diastolic blood pressure was greater as the amount of hyperoside was increased. From this result, it was shown that hyperoside administration contributes to the improvement of salt-dependent hypertension. Moreover, salt-dependent hypertension becomes drug-resistant hypertension, and no antihypertensive effect is observed by inhibiting ACE. Therefore, it is considered that the antihypertensive action is expressed by suppressing the non-ACE-dependent angiotensin II production by the chymase activity inhibitory action by administration of hyperoside.

The chymase inhibitor of the present invention has an excellent inhibitory action on human chymase activity, and is effective as a medicament used for the prevention and treatment of various diseases caused by angiotensin II.

Claims (17)

1. A chymase inhibitor comprising a compound represented by the following general formula (A) as an active ingredient.
   

   

   
   (In the general formula (A), R ¹ to R ⁵ each independently represents a hydrogen atom, a hydroxyl group or a methoxy group, X ¹ represents a hydrogen atom, a hydroxyl group or a —OG group, and G represents a sugar chain.)
   
2. The chymase inhibitor according to claim 1, wherein X ¹ is a hydroxyl group, a rhamnose residue, a glucose residue, or a galactose residue.
3. The chymase inhibitor according to claim 1, wherein the compound represented by the general formula (A) is at least one selected from the group consisting of quercitrin, isoquercitrin, hyperoside, kaempferol and luteolin.
4. The chymase inhibitor according to claim 1, comprising at least one selected from quercetin glycosides and quercetin derivatives.
5. The chymase inhibitor according to claim 1, comprising a hyperoside.
6. A drug comprising the chymase inhibitor according to any one of claims 1 to 5.
7. The drug according to claim 6, which is a non-ACE-dependent angiotensin II production inhibitor.
8. The drug according to claim 6 or 7, further comprising a bioabsorption enhancer.
9. The drug according to claim 6, which is a preventive or therapeutic agent for cardiovascular disease.
10. The drug according to claim 9, wherein the cardiovascular disease is hypertension and / or hypertensive organ disorder.
11. The drug according to claim 10, which is a prophylactic or therapeutic agent for hypertension and / or hypertensive organ damage caused by excessive intake of salt.
12. The agent according to claim 6, which is a prophylactic and / or therapeutic agent for arteriosclerotic diseases caused by lipid deposition in blood vessels.
13. The drug according to claim 6, which is a preventive and / or therapeutic agent for a disease caused by impaired glucose tolerance.
14. The drug according to claim 13, wherein the disease caused by impaired glucose tolerance is diabetes and / or diabetic complications.
15. The drug according to claim 6, which is one or more prophylactic and / or therapeutic agents selected from atopic dermatitis and psoriasis.
16. The drug according to claim 6, which is one or more prophylactic and / or therapeutic agents selected from cirrhosis and pulmonary fibrosis.
17. The drug according to claim 6, which is one or more prophylactic and / or therapeutic agents selected from Crohn's disease and ulcerative colitis.

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