JP2016199470A - Human chymase activity inhibitor and preventive/therapeutic agent of hypertension - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide novel human chymase activity inhibitors.SOLUTION: A human chymase activity inhibitor contains the compound represented by formula (A) and/or a piririniumu body thereof as an active ingredient. (In the general formula (A), Rto Rrepresent each independently a hydroxyl group or a methoxy group; Xrepresents a hydroxyl group or a -OG group. In addition, G represents a sugar chain.) The compound represented by the general formula (A) is quercetin, quercitrin, isoquercitrin, or idaein.SELECTED DRAWING: None

Description

  The present invention relates to an agent that inhibits the activity of human chymase.

In recent years, a non-angiotensin-dependent pathway has been found in the production of angiotensin II in the local area of human tissue, and in particular, attention has been focused on the angiotensin II production pathway by human mast cell human chymase (human chymase) (see, for example, Non-Patent Document 1).
Human chymase is mainly present in the heart, skin, blood vessel wall, intestinal tract, etc., and is released from the heart or the like by some stimulus, and converts angiotensin I into angiotensin II. 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, a human chymase activity inhibitor that inhibits the activity of human chymase has the potential to be used as a therapeutic agent for various heart diseases by inhibiting local production of angiotensin II in the heart.
In addition, human chymase is considered to be involved in hypertension, organ damage, arteriosclerosis and the like, and human chymase activity inhibitors are also expected to have preventive and therapeutic effects on such diseases.

  So far, several human chymase activity inhibitors have been reported. Triazine derivatives (Patent Document 1), substituted aminopyrimidinyl acetamide derivatives (Patent Document 2), benzimidazole derivatives (Patent Document 3) and the like have been reported as substances having an inhibitory action on human chymase activity.

JP-A-8-208654 JP 2003-104984 A Japanese Patent Laid-Open No. 2001-199983

J. et al. Biol. Chem 1990; 265: 222348-22357, J. MoI. Biol. Chem. 1991; 266: 17173-17179

On the other hand, the human chymase inhibitor compounds disclosed so far are often inactive or unstable.
Under such circumstances, an object of the present invention is to provide a novel inhibitor of human chymase activity.

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

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

(In general formula (A), R ^{1 to} R ⁴ each independently represents a hydroxyl group or a methoxy group, X ¹ represents a hydroxyl group or —OG group, and G represents a sugar chain.)
<2> The human chymase activity inhibitor according to <1>, wherein X ¹ is a hydroxyl group, a rhamnose residue, or a glucose residue.
<3> The human chymase activity inhibitor according to <1> or <2>, wherein the compound represented by the general formula (A) is quercetin, quercitrin, or isoquercitrin.
<4> A human chymase activity inhibitor containing a compound represented by the following general formula (B) as an active ingredient.

(In General Formula (B), R ^{5 to} R ⁸ each independently represent a hydroxyl group or a methoxy group, X ² represents a hydroxyl group or —OG group, and G represents a sugar chain.)
<5> The human chymase activity inhibitor according to <4>, wherein X ² is a hydroxyl group or a galactose residue.
<6> The human chymase activity inhibitor according to <4> or <5>, wherein the compound represented by the general formula (B) is idein.
<7> A prophylactic / therapeutic agent for hypertension comprising the human chymase activity inhibitor according to any one of <1> to <6>.

  The human chymase activity inhibitor of the present invention is selective and effective against human chymase, and is effective as a pharmaceutical used for the prevention and treatment of various diseases caused by angiotensin II.

  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 human chymase activity inhibitor containing as an active ingredient a compound represented by the following general formula (A) and / or a compound represented by the general formula (B).
Note that the compound represented by the general formula (A) and the compound represented by the general formula (B) have a flavanone skeleton or a flavanone-like skeleton, and are substituted with the same functional group at an equivalent position of the skeleton. In common.

R ¹ to R ⁴ in the general formula (A) are each independently a hydroxyl group or a methoxy group, X ¹ is a hydroxyl group, or a -OG group. Here, G is a sugar chain, and details will be described later.

In the general formula (B), R ^{5 to} R ⁸ each independently represent a hydroxyl group or a methoxy group, and X ² represents 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 ⁴ is not limited, but the hydroxyl groups are 3 or more, more preferably 4 (all hydroxyl groups).
In the general formula (B), the number of hydroxyl groups or methoxy groups constituting R ^{5 to} R ⁸ is not limited, but the hydroxyl groups are 3 or more, more preferably 4 (all hydroxyl groups).

The general formula (A) and the general formula ^{(B), X 1, X} 2 is a hydroxyl group or -OG group. When X ¹ and X ² are 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 the general formula (A), X ¹ is preferably a hydroxyl group, a rhamnose residue, or a glucose residue in terms of excellent human chymase inhibitory activity. In this case, there is no limit to the number of hydroxyl or methoxy group representing R ¹ to R ^4, a hydroxyl group 3 or more.
That is, the human chymase activity inhibitor of the present invention preferably contains as an active ingredient any one of quercetin, quercetin glycoside, and quercetin derivative, which has the following chemical formula. Among these, it is particularly preferable to contain any one of quercetin, quercitrin and isoquercitrin as an active ingredient.

The compound represented by the general formula (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 for the compound represented by the general formula (A), include citrus fruits, onions, buckwheat, apples, green tea, broccoli, Morohayana 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.
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.

In the compound represented by the general formula (B), X ² is preferably a hydroxyl group or a galactose residue, and particularly preferably idein (cyanidin 3-O-galactoside) represented by the following chemical formula.

The compound of the general formula (B) may be artificially synthesized or derived from a natural product. 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 idein, which is one of the compounds suitable for the compound represented by the general formula (B), include various autumnal plants, and specific examples include penicitane, kogoumetsugi, nutsaki, azalea, and willow. 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.

Here, the willow tree or its cultivar is one of suitable raw materials because it contains various compounds having the human chymase activity represented by the general formula (A) or the general formula (B). As the willow tree or its cultivar, the willow tree, which is also commonly referred to as matade or honde, preferably, the red cultivar (purple red), azabutade, hosobatade or the like is used, and particularly preferred is the red seed.
Therefore, the willow tree can be suitably used as the human chymase activity inhibitor of the present invention as it is or after separation and purification by a known method (for example, Japanese Patent No. 5186628).

  Tadeonal (polycodial) represented by the following chemical formula, which is a component contained in Willow tree, also has a human chymase activity inhibitory action.

In the human chymase activity inhibitor of the present invention, the compound having the human chymase activity represented by the general formula (A) or the general formula (B), which is an active ingredient, may be contained alone or in combination of two or more.
Among the compounds having human chymase activity, quercitrin, isoquercitrin and idein have particularly large chymase activity inhibitory action. Therefore, the human chymase activity inhibitor of the present invention preferably contains at least one of quercitrin, isoquercitrin and idein.

  These compounds may be contained as medically 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, dicyclohexylamine) Salt, diethanolamine salt, tetramethylammonium salt, etc.), amino acid salts (glycine salt, lysine salt, arginine salt, ornithine salt, asparagine salt, etc.) and the like.

The human chymase activity inhibitor of the present invention can be administered orally. Further, it may be administered parenterally by injection or the like.
The concentration of the above-mentioned compound, which is an active ingredient contained in the human chymase activity inhibitor of the present invention, is not particularly limited as long as it exhibits the human chymase activity inhibitory action, taking into account the type of the target compound and the necessary activity inhibitory action. Thus, the amount used is appropriately determined.
The human chymase inhibitor of the present invention can be administered once a day, but can also be administered by dividing it at appropriate intervals in 2 to 3 times. Furthermore, when used as an injection, it can be performed by stepwise administration by administration injection or continuous administration by infusion.

(Applied drug of human chymase activity inhibitor)
The human chymase activity inhibitor of the present invention is derived from the enzyme activity of human chymase by inhibiting the activity of human chymase, which is an enzyme that produces angiotensin II (AII) from angiotensin I (AI) independently of an angiotensin converting enzyme. The disease can be prevented and treated. Diseases associated with increased enzyme activity of human chymase include hypertension and related organ disorders, diabetes and related organ disorders, arteriosclerotic vascular diseases, heart failure, myocardial remodeling after myocardial infarction, postoperative scar formation, inflammatory bowel disease, etc. Is mentioned.
Therefore, the human chymase activity inhibitor of the present invention can be used as a prophylactic / therapeutic agent for the various clinical conditions described above, and a prophylactic / therapeutic agent for hypertension is one of its preferred applications.

(Form of human chymase activity inhibitor)
The human chymase activity inhibitor of the present invention can be administered orally or parenterally as a solid or liquid preparation by blending an effective amount thereof together with a pharmaceutically acceptable carrier. 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, corrigents and stabilizers may be used for the human chymase activity 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, hydroxypropyl cellulose 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.

  Moreover, when using as a liquid formulation, what has the dispersibility of the said compound which is an active ingredient, and a biosafety is selected as a solvent. 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, a tonicity agent, a buffering agent, and an antioxidant together with the human chymase activity inhibitor of the present invention as 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.

  Moreover, the human chymase activity 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.

  Furthermore, the human chymase inhibitor of the present invention can contain other pharmaceutically effective ingredients in addition to the above active ingredients as necessary in the treatment of various diseases.

  The manufacturing method of the said solid formulation and liquid formulation can apply the manufacturing method of a general pharmaceutical.

  Moreover, when it is desired to ingest the human chymase inhibitor of the present invention by eating and drinking on a daily basis, it may be contained in the food or beverage to obtain 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.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited to a following example, unless the summary is changed.

The compounds used for evaluation are as follows.
・ Quercetin (Wako Pure Chemical Industries)
・ Kercitrin (SIGMA)
・ Isoquercitrin (SIGMA)
・ Ideine (SIGMA)
・ Tadeonal (Wako Pure Chemical Industries)

<Method for evaluating human chymase inhibitory activity>
The evaluation of human chymase inhibitory activity (in vitro) was carried out by the method described below with some modifications and modifications to the method described in Eur J Biochem 268 (22), 5585-93 (2001). In this method, for experiments (in vitro) on suppression of human chymase activity, a substrate obtained by Dnp / Nma modification of angiotensin I, which is a substrate for human chymase, is used, and when chymase cleaves this substrate to produce angiotensin II, fluorescence is developed. Method. The outline of the measurement 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.

Quercetin, quercitrin, isoquercitrin and idein, and tadeonal (reference example) were added to the concentrations shown in Table 1, respectively, and the human chymase inhibitory activity rate was evaluated by the method described above.
As shown in Table 1, it was found that quercetin, quercitrin, isoquercitrin, idein and tadeonal have an action of selectively inhibiting human chymase activity. In particular, quercetin, isoquercitrin, and idein were found to be excellent in human chymase activity inhibitory action.

  The human chymase activity inhibitor of the present invention is effective as a medicament used for the prevention and treatment of various diseases caused by angiotensin II.

Claims (7)

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

(In general formula (A), R ^{1 to} R ⁴ each independently represents a hydroxyl group or a methoxy group, X ¹ represents a hydroxyl group or —OG group, and G represents a sugar chain.) The human chymase activity inhibitor according to claim 1, wherein X ¹ is a hydroxyl group, a rhamnose residue or a glucose residue.   The human chymase activity inhibitor according to claim 1 or 2, wherein the compound represented by the general formula (A) is quercetin, quercitrin, or isoquercitrin. A human chymase activity inhibitor comprising a compound represented by the following general formula (B) as an active ingredient.

(In General Formula (B), R ^{5 to} R ⁸ each independently represent a hydroxyl group or a methoxy group, X ² represents a hydroxyl group or —OG group, and G represents a sugar chain.) The human chymase activity inhibitor according to claim 4, wherein X ² is a hydroxyl group or a galactose residue.   6. The human chymase activity inhibitor according to claim 4 or 5, wherein the compound represented by the general formula (B) is idein.   A prophylactic / therapeutic agent for hypertension, comprising the human chymase activity inhibitor according to any one of claims 1 to 6.