JPH08225453A - Lipoprotein (a) depressor, cholesterol depressor and medicine containing the same - Google Patents

Abstract

PURPOSE: To provide a depressor exhibiting exceeding serum lipoprotein (a) level depressing action, having high safety and useful for curing or prevention of ischemic heart disease or arterial sclerosis, etc., by including proanthocyanin as an active component. CONSTITUTION: This depressor contains proanthocyanin as an active component. As an extracting raw material of proanthocyanin, a strained draff secondary produced in a wine-squeezing process in producing of red wine or a strained draff in producing of white wine or grape juice, etc., is preferable. Proanthocyanin may be obtained by dissolving 1wt. of an extracting raw material into 2-1000wt. times of solvent, extracting in a range from room temperature to boiling point of the solvent under normal pressure for 2 hours-2 weeks and purifying resultant extract in a method such as an adsorbent treating or a membrane separation, etc. Dose of the proanthocyanin is preferably 0.5-150mg/ day in a dry weight.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a lipoprotein (a) lowering agent and a cholesterol lowering agent containing proanthocyanidin as an active ingredient, and a medicine containing these as an active ingredient. The drug of the present invention is ischemic heart disease, arteriosclerosis,
It is useful for the treatment and prevention of cerebrovascular dementia, diabetes and angiopathic Parkinson's disease, or for reducing the total blood cholesterol level without affecting HDL-cholesterol.

[0002]

2. Description of the Related Art Serum lipoprotein (a) (hereinafter referred to as "L
p (a) ”is rich in cholesterol, easily aggregated in the presence of calcium, and easily binds to connective tissues such as glycosaminoglucan, and thus it is suggested that lipid deposition on the arterial wall may occur ( "Arteriosclerosis", 17th
Vol. 639-658, (1989)). Also,
It is considered that Lp (a) bound to glycosaminoglucan is easily taken up by macrophages and may promote foam cell formation.

Furthermore, apoprotein (a), which is an apoprotein peculiar to Lp (a), has high homology with plasminogen (Biochem.
Biophys. Acta., 960, 91-97, 1988), which is known to suppress the fibrinolytic system (Biochemistry, 28, 2370-23).
74, 1988; Nature, 339, 303-305, 1989).

From these, it is considered that Lp (a) is involved in the onset and progress of arteriosclerosis. In fact, Lp (a) is associated with ischemic heart disease, cerebral infarction, carotid atherosclerosis,
It is considered to be a risk factor different from conventional low-density lipoprotein cholesterol and platelet aggregation for cerebrovascular dementia, diabetes and vascular disorder Parkinson's disease ("arteriosclerosis", Vol. 17, 639-). Pp. 658, (1989
Year)).

By the way, conventional therapeutic agents for hyperlipidemia are agents developed with a focus on the action of lowering low-density lipoprotein cholesterol and triglyceride among plasma lipids, and their action on Lp (a). Is hardly considered. For example, probucol (Atherosclerosis, 79, 267 to 269, 1989) and clofibrate (Metabolism, 24, 1047 to 1054, 1975), which are known as therapeutic agents for hyperlipidemia,
And cholestyramine (Atherosclerosis, 73, 135-141,
1988) have no Lp (a) lowering action and even show an increasing tendency. Only a slight reduction in serum Lp (a) was observed in nicotinic acid and its derivatives (At
herosclerosis, 57, 293 ~ 301, 1985; Current Therapu
tic Res. 54, 550-552, 1993).

[0006] Further, stanozolol (Biochem. Biophys. Acta., 795, 293 to 296, 1984) which is a steroid, and neomycin (Atherosclerosis, 57, 293) which is an antibiotic.
, 301, 1985), a serum Lp (a) lowering effect is recognized, but there is a problem in terms of side effects.

Regarding food, palm oil and fish liver oil (Cli
n. Res., 38, 250A, 1990) has only been reported to reduce serum Lp (a). However, regarding fish liver oil, there is also a report (Thrombosis Res., 58, 667 to 668, 1990) that there is no Lp (a) lowering effect, and conflicting results have been obtained.

[0008] Moreover, if the dose is increased to enhance the effectiveness of fish liver oil or palm oil, there is a risk of excess calories due to fats and fatty acids. Moreover, in the case of fish liver oil, there is a risk of excessive intake of vitamins A and D, and the incidence of side effects such as nausea, vomiting, diarrhea, and abdominal pain is high (La
ncet, No.8658, 177-181, 1989).

On the other hand, a state in which blood cholesterol level is high (so-called hypercholesterolemia) is recognized as a risk factor in cardiovascular disease. Epidemiologic studies have demonstrated that populations consuming large amounts of saturated fatty acids and cholesterol almost always have relatively high levels of serum cholesterol and high mortality from coronary heart disease.

Conventionally, as cholesterol lowering agents for lowering blood cholesterol levels, exogenous cholesterol absorption inhibitors, cholesterol excretion and excretion of bile acids into catabolism, cholesterol biosynthesis inhibitors, etc. have been used. It has been.

Although various kinds of these drugs are known, many of them are not sufficiently satisfactory, and some of them are chemical synthetic products which cannot always guarantee safety in long-term administration. It was a situation in which we couldn't get what we wanted. In particular, when used in combination with foods and drinks, there has been a demand for safer cholesterol-lowering agents of natural origin.

[0012]

As described above, as described above,
Drugs and foods that are known to lower serum Lp (a) levels and blood cholesterol levels have problems in terms of safety and harmful effects when ingested in large amounts. Therefore, the development of Lp (a) -lowering agents and cholesterol-lowering agents that have sufficiently strong effects and do not cause any problems in terms of safety for the human body has been left as an issue.

[0013]

[Means for Solving the Problems] Although the amount of fat intake is correlated with the incidence of heart disease, the incidence of heart disease is low in France despite the large amount of fat intake, and is called the French paradox. On the other hand, red wine is a brewed liquor that has been widely consumed mainly in Europe since ancient times, but its consumption in France is particularly large, and in recent years, the relationship between the French paradox and red wine has attracted attention (Lance.
t, 339, 1523-1526, 1992).

The inventors of the present invention have conducted intensive studies focusing on this French paradox. As a result, the proanthocyanidins contained in the grape extract have the effect of lowering the serum Lp (a) level and the blood cholesterol level. The inventors have found that it has a reducing effect and have completed the present invention.

That is, an object of the present invention is to use grape extract, particularly Lp containing proanthocyanidin as an active ingredient.
(A) To provide a lowering agent, a cholesterol lowering agent, and a medicine containing these as an active ingredient.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION Proanthocyanidin, which is the active ingredient of the Lp (a) lowering agent and cholesterol lowering agent of the present invention (hereinafter, both are collectively referred to as "lipid etc. lowering agent"), is present in plants. A condensed tannin, that is, a compound in which flavan-3-ol or flavan-3,4-diol is bonded as a unit by condensation or polymerization, and is propellalgonidine (Properalgonidin), procyanidin (Procyanidin), prodelphinidin (Prodefinidine).
lfinidin), Proguibourtinidi
n), Profisetinidin, Prorobinetinidin, Proteracacidin, Promeracacidin
in), proapigeninidin, proluteolinidin, and other proanthocyanidins, and stereoisomers thereof.

In obtaining the proanthocyanidins of the present invention, the extraction raw material is not limited at all, and various fruits and plants such as trees can be used, but grape fruits are preferable, and among them, red wine is produced. Pomace produced as a by-product in the sake brewing process and pomace produced during the production of white wine, grape juice and the like are preferably used.

The solvent used for extracting the plant is not limited, but may be any solvent selected from the group consisting of water, lower alcohols, acetone, alkyl ketones, ethyl acetate and the like, or any two or more solvents. Mixed solvent of.

In extracting the plant, 2 to 1000 times by weight of the solvent is added to 1 dry or wet weight of the plant, and the temperature is within the range of the boiling point of the solvent from room temperature to normal pressure for 2 hours to It is preferable to extract for 2 weeks. The extract is concentrated under reduced pressure by a conventional method, and if necessary, freeze-dried. The plant extract thus obtained is in powder form, and exhibits a remarkable serum Lp (a) level lowering action and cholesterol lowering action even at the stage of a crude extract having a relatively low content of proanthocyanidins.

The plant extract obtained as described above is further subjected to a purification method such as an adsorbent treatment method, a membrane separation method or a solvent fractionation method to obtain a highly pure proanthocyanidin.

For example, a highly pure proanthocyanidin can be obtained by subjecting the plant extract to adsorption column chromatography and extracting with one or more kinds of solvents.

In order to carry out adsorption distribution by molecular weight by subjecting the plant extract to adsorption column chromatography, for example, after dissolving this in about 10 times the amount of water, Sephadex LH-20 (manufactured by Pharmacia, USA) is used. ), Diaion HP20 (manufactured by Mitsubishi Kasei), SepaBeads HP1MG
(Mitsubishi Kasei Kogyo), Toyopearl HW40F (Toyo Soda Kogyo), etc. are subjected to column chromatography for adsorption, and the column is washed with water and then extracted with an appropriate solvent. A column fraction containing anthocyanidins can be obtained.

The organic solvent used for elution is not particularly limited, and examples thereof include lower alcohol, acetone, alkyl ketone, ethyl acetate and n-hexane.
Particularly, lower alcohol and acetone are preferable. Two or more kinds of these solvents may be mixed and used, and the elution operation is preferably performed at room temperature.

To prepare the lipid etc. lowering agent of the present invention using the proanthocyanidin obtained as described above,
This may be formulated as it is or with a known pharmaceutical carrier.

The lipid etc. lowering agent of the present invention can be formulated together with commonly used additives, carriers, auxiliaries and the like, and can be used as an oral or parenteral product in the field of pharmaceuticals according to a conventional method. You can Oral preparations include tablets, capsules, granules, fine granules, syrups, drinks, etc., and parenteral preparations include ointments, creams, external preparations such as liquid preparations, sterile solution preparations, suspension preparations, etc. Injections and the like.

The lipid lowering agent of the present invention is formulated into a required unit dose form together with a pharmaceutical vehicle, carrier, excipient, integrator, preservative, stabilizer, flavoring agent and the like. It is done by

In formulating the lipid etc. lowering agent of the present invention, the following may be mentioned as adjuvants to be mixed in tablets, capsules and the like. That is, gum arabic, corn starch, integrators such as gelatin, lubricants such as magnesium stearate, excipients such as microcrystalline cellulose, gelatinized starch, puffing agents such as alginic acid, sucrose, lactose, Examples include sweeteners such as saccharin, peppermint, and flavors such as cherry. Further, in the case of a capsule, a liquid carrier such as oil and fat may be contained in addition to the above materials.

In addition, other materials may be included as coatings or to alter the physical form of the formulation,
For instance, tablets may be coated with shrack, sugar and syrup or elixir as the active compound,
Sucrose can be included as a sweetener, methylparaben or propylparaben as a preservative, and peppermint or orange flavor can be included as a flavor, respectively.

Sterile compositions for injection are prepared by dissolving or suspending proanthocyanidins in water for injection, natural vegetable oils such as coconut oil, sesame oil, cottonseed oil or the like, or synthetic fat vehicles such as ethyl oleate. It can be prescribed by the method. In this injection, antiseptics, antioxidants, buffers and the like can be added if necessary.

When used as an external preparation, petrolatum, hydrophilic ointment, paraffin, lanolin, oils and fats, macrogol and the like can be used as the base.

Among the lipid lowering agents of the present invention, Lp (a)
The lowering agent can be used as a medicine for treating ischemic heart disease, arteriosclerosis, cerebrovascular dementia, diabetes, vascular disorder Parkinson's disease and the like. On the other hand, the cholesterol lowering agent can be used as a medicine for lowering the total blood cholesterol without affecting the HDL-cholesterol.

When the Lp (a) lowering agent is used for the above-mentioned purpose, the dose of proanthocyanidin is 0.5 to 50 mg as a dry weight to be administered once or several times a day, that is, at a dose of 0.5. The dose may be about 5 to 150 mg / day. On the other hand, when it is used as a cholesterol lowering agent, the dry weight of proanthocyanidins may be administered at about 50 to 1,500 mg / day.

The lipid-lowering agent of the present invention is a serum Lp
(A) The dosage form may be added to various foods for the purpose of lowering the level and the total amount of cholesterol in blood. Examples of foods to which the lipid etc. lowering agent of the present invention can be added include tea drinks, juices, coffee drinks, carbonated drinks,
Examples include chewing gum, candy, caramel, chocolate, ice cream and the like.

Since the main component of the lipid lowering agent of the present invention is proanthocyanidin which is also a grape component, there is no problem in terms of safety, but when the lipid lowering agent of the present invention is added to food, From the point of view of color tone, 0.001-5.0
It is desirable that the concentration range be%.

When the food containing the Lp (a) lowering agent of the present invention is ingested as a health food or a functional food for preventing diseases and maintaining health, it is divided into several times a day, and 0.5-150m as anthocyanidin conversion
It is desirable to take g. In addition, when the food containing the cholesterol lowering agent of the present invention is ingested as a health food or a functional food for reducing the total cholesterol level in blood, it is divided into several times a day, and as a proanthocyanidin conversion per day. It is desirable to take 50-1,500 mg. In addition, as the compounding amount in the food,
It is desirable to set it to about 1 to 5%.

[0036]

INDUSTRIAL APPLICABILITY The Lp (a) lowering agent of the present invention shows a remarkable serum Lp (a) level lowering effect and is highly safe. Therefore, ischemic heart disease, arteriosclerosis, cerebrovascular dementia, diabetes and It can be used for treatment and prevention of vascular disorder Parkinson's disease and the like. The Lp (a) lowering agent can be added to foods and used for prevention of the above diseases.

On the other hand, the cholesterol lowering agent of the present invention is
Since it does not affect HDL-cholesterol and can lower the level of total cholesterol in the blood, it can be used to lower the total cholesterol in hypercholesterolemic patients. In addition, when it is added to foods that are ingested daily, it is possible to prevent spontaneous increase in total cholesterol level.

The Lp (a) level lowering agent and cholesterol lowering agent of the present invention can be used not only for humans but also for animals. In particular, for pet animals bred in the city, obesity due to lack of exercise is a concern, but for these animals, by providing a feed containing proanthocyanidins, preferably 1 to 5%, It is possible to reduce Lp (a) and cholesterol that cause obesity.

[0039]

[Examples] Next, production examples of the grape extract of the present invention, test examples of its pharmacological action, formulation examples and the like will be given as examples, but the present invention is not limited to these examples.

Example 1 Production Example (1) of Grape Extract: To 43 g wet weight of pomace produced as a by-product in the squeezing process during the production of red wine, 200 ml of 80% by volume acetone water was added and left overnight at room temperature. . The liquid obtained by filtering with a glass filter was concentrated to dryness under reduced pressure,
0.23 g of a brown powder was obtained. This powder is about 30
% Proanthocyanidins. The quantification of proanthocyanidins was performed by the method of R. Jambunathan et al. (J. Agric. Food Chem., 34, 425-429 (1986)).
Went by.

Example 2 Production Example (2) of Grape Extract: 500 g of red grape fruit
To the above, 2,000 ml of 50% ethanol was added and left at room temperature for one week. The liquid obtained by filtering with a glass filter was concentrated to dryness under reduced pressure to obtain 30 g of a brown powder. This powder contained about 3% proanthocyanidins.

Example 3 Effect on Serum Lp (a) Level: The red grape fruit extract obtained in Example 2 was examined for Lp (a) lowering effect. First, the serum Lp (a) level is 25 mg / d
150mg / of red grape fruit extract for 5 males over 1
Oral administration daily, before and 2 weeks after administration to collect blood and serum L
The p (a) value was measured by enzyme immunoassay. Table 1 shows the results.

[0043]

As is clear from the above results, significant Lp was observed in all the cases by continuous administration of red grape fruit extract for 2 weeks.
(A) A lowering effect was recognized. The test was based on the paired t-test.
The risk factor was less than 0.001.

Example 4 Production Example of Proanthocyanidin: 25 kg of Red Grape Fruit
As a raw material, 1.5 kg of extract was obtained in the same manner as in Example 2. This was dissolved in 10 liters of 50% ethanol, adsorbed on a column (φ30 cm × 32 cm) of Diaion HP-20 (manufactured by Mitsubishi Kasei Co., Ltd.), washed with 100 liters of water, and then extracted with 100 liters of ethanol. 105 g of a proanthocyanidin fraction was obtained. This fraction contained 97% proanthocyanidins.

Example 5 Effect on Serum Cholesterol: The effect of the proanthocyanidin fraction obtained in Example 4 on serum lipid was examined as follows. 10 male subjects (33-
(Age 37) was orally administered with the proanthocyanidin fraction at 0.5 g / day for 2 weeks. The intakes of calories and nutrition of all the subjects were controlled by ingesting a standard diet (manufactured by Taihei Co., Ltd .; Tokyo) from 2 weeks before administration until the end of the test. Total cholesterol and H of these subjects
The DL-cholesterol level was measured 2 weeks before the administration, on the day after the administration and 2 weeks after the administration, and the action of the proanthocyanidin fraction on blood cholesterol was examined from the changes in these values. The results are shown in Table 2.

[0047] (Numerical values are mean ± standard deviation; ^* indicates p <0.05 with respect to the value on the day of administration)

As is clear from the above results, the administration of the proanthocyanidin fraction for 2 consecutive weeks did not affect HDL-cholesterol but significantly reduced serum total cholesterol.

Example 6 Capsules and tablets: (Composition) (wt%) Proanth 2 cyanidin fraction of Example 4 Lactose 83 Magnesium stearate 15 (Preparation) A capsule or tablet was prepared according to the method.

Example 7 Powder and granules: (Composition) (wt%) Proanth 5 of Example 4 Cyanidine fraction Starch 55 Lactose 40 (Production method) After uniformly mixing the components, the usual method was used. It was made into powder or granules according to.

Example 8 Injection: (Composition) (wt%) Proanth 2 cyanidin fraction of Example 4 Surfactant 8 Physiological saline 90 (Production method) Heat-mixed each component and sterilized It was an injection.

Example 9 Tea Drink: Oulon tea leaves were soaked in 10 times volume of boiling water at about 90 ° C. for 3 to 5 minutes to remove tea leaves from the infusion, and 1% each was applied to a nylon mesh filtered solution. Example 2 extract and L-
Sodium ascorbate was added to each to make a tea beverage. that's all

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A23L 1/30 A23L 1/30 B A61K 31/35 ABN A61K 31/35 ABN // C07D 311/62 C07D 311/62 (72) Inventor Hirofumi Tanahashi 1-1-1, Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka Suntory Ltd. Wine Research Institute (72) Inventor Kazuaki Hosoda 4-1-1 Kioicho, Chiyoda-ku, Tokyo Santo Lee Co., Ltd. Healthcare Business Development Department

Claims (11)

[Claims] 1. A lipoprotein (a) lowering agent containing proanthocyanidins as an active ingredient. 2. The lipoprotein (a) lowering agent according to claim 1, wherein proanthocyanidin is contained in the grape extract. 3. A medicine comprising the lipoprotein (a) lowering agent according to claim 1 or 2 as an active ingredient. 4. The lipoprotein (a) lowering agent according to claim 1, which is a food additive. 5. A cholesterol lowering agent containing proanthocyanidins as an active ingredient. 6. The cholesterol lowering agent according to claim 5, wherein the proanthocyanidin is contained in the grape extract. 7. A medicine comprising the cholesterol lowering agent according to claim 5 or 6 as an active ingredient. 8. The cholesterol lowering agent according to claim 5, which is a food additive. 9. A food containing 1 to 5% by weight of proanthocyanidins. 10. 1-5% by weight of proanthocyanidins
A method for lowering an animal lipoprotein (a) level, which comprises feeding the animal the feed contained therein. 11. 1-5% by weight of proanthocyanidins
A method for lowering a cholesterol level in an animal, which comprises feeding the animal the feed contained therein.