JPH07223962A - Thrombogenesis suppressing agent and composition containing the same - Google Patents

Abstract

PURPOSE:To obtain a thrombogenesis suppressing agent composed of a specific 1,2-dihydroxy-4-(2-hydroxyethyl)benzene derivative, having thrombogenesis suppressing action and high safety and useful as pharmaceuticals, foods, etc., for the prevention and treatment of thrombosis. CONSTITUTION:This thrombogenesis suppressing agent is composed of a compound of formula I [R1, R2 and R3 each is H or group of formula II (R7 and R8 each is H or a lower alkyl); R4 and R5 each is R7; R5 is H or OH; Rham is rhamnose residue; Ara is arabinose residue] or its salt, e.g. 1-{2-[3-(2- arabinosylrhamnosyl)-4-(3,4-dihydroxycinnamoyl)glycosyl]ethyl}-3,4- dihydrobenzene. The compound of formula I can be extracted e.g. from Stachys riederi (a kind of plant of the family Labiatae). The daily administration rate of the agent is preferably 1-1,000mg for adult.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a thrombus inhibitor and a composition such as a drug, a food or the like having a thrombus inhibitory action.

[0002]

2. Description of the Related Art In recent years, as the lifestyle has become westernized, the patterns of disease incidence and causes of death have changed in Japan. That is, infectious diseases, which were the leading cause of death, have been significantly reduced, and instead, cardiovascular diseases such as hypertension and stroke and cancer have been greatly increased.

Of these, various drugs such as lauolphia type drugs, node blockers, and hydralazine type drugs have been developed for hypertension, but for thrombosis such as stroke,
Only the dicoumarol drug warfarin potassium is known. However, it has been reported that this warfarin potassium has side effects such as gin machine, fever, vomiting, nausea, chills, and dermatitis, and also induces congenital abnormalities, and still has sufficient efficacy and safety. The current situation is that a drug for treating thrombosis that combines the above is not available.

On the other hand, 1,2-dihydroxy-4- (2-
The hydroxyethyl) benzene derivative is a compound known to be contained in a large amount in the extract of sesame, a plant of the Labiatae family. This sesame seed is a plant that grows naturally in Japan and can be found in most parts of Japan, and the above components are present in extracts from almost any part of the plant.

However, the sesame extract or 1,2-
It is not known that the dihydroxy-4- (2-hydroxyethyl) benzene derivative has a thrombosis-inhibiting effect,
In addition, no attempts have been reported to prevent or treat thrombosis using these substances.

[0006]

SUMMARY OF THE INVENTION The present invention has been made from the above viewpoint, provides a thrombosis inhibitor having a sufficient thrombus-inhibiting effect and excellent in safety, and further provides safe thrombosis. An object of the present invention is to provide a composition such as a drug or food capable of preventing or treating the above.

[0007]

In order to solve the above problems, the present inventor screened various plant extracts and their purified products in consideration of safety, using a platelet aggregation inhibitory action as an index of thrombosis inhibition. As a result of repeating the above, Inukoma (Stachys Rieder)
It was found that the hot water extract of i)) contained a component having an inhibitory effect on platelet aggregation, and as a result of further purification and further investigation, 1,2-dihydroxy-4-
The inventors have found that the (2-hydroxyethyl) benzene derivative has a strong platelet-suppressing action, and completed the present invention.

That is, the present invention is a thrombus inhibitor comprising a 1,2-dihydroxy-4- (2-hydroxyethyl) benzene derivative represented by the following general formula (I) and / or a salt thereof, and a composition containing the same. It is a thing.

[0009]

[Chemical 9]

However, in the chemical formula 9, R ₁ , R ₂ and R ₃ are each independently a hydrogen atom or a group represented by the chemical formula 10,
R ₄ and R ₅ are each independently a hydrogen atom or a short chain length alkyl group, R ₆ is a hydrogen atom or a hydroxyl group, and —R
am represents a rhamnose residue and -Ara represents an arabinose residue.

[0011]

[Chemical 10]

However, in the chemical formula 10, R ₇ and R ₈ each independently represent a hydrogen atom or a short chain length alkyl group. Less than,
The present invention will be described in detail.

<1> Thrombus Inhibitor of the Present Invention The thrombus inhibitor of the present invention is 1,2-dihydroxy-4- (2-hydroxyethyl) represented by the above general formula (I).
It consists of a benzene derivative and / or its salt.

In the compound represented by the general formula (I), R ₄ , R ₅ and R ₇ , R ₈ each independently represent a hydrogen atom or a short chain length alkyl group. The number is preferably 1 to 10, more preferably 1 to 6, and further preferably 1 to 4.

In the general formula (I), the bond between rhamnose and glucose is at the 1-position of rhamnose, and the bond between rhamnose and arabinose is a glucosidic bond,
2nd, 3rd or 4th of rhamnose and 1 of arabinose
It is preferable that it is performed between positions. Further, among these, those in which rhamnose and arabinose are bound at the 2-position of rhamnose are more preferable. The glucosidic bond may be in either α or β form.

Further, the above 1,2-dihydroxy-4-
Of the (2-hydroxyethyl) benzene derivatives,
1- (2- (3- (2-arabinosyl rhamnosyl) -4- (3,4-dihydroxycinnamoyl) represented by 1
Glycosyl) ethyl) -3,4-dihydroxybenzene, 1- (2- (3- (2-arabinosylrhamnosyl) -6- (3,4-dihydroxycinnamoyl) glycosyl) represented by Chemical formula 12 Ethyl) -3,4-dihydroxybenzene, 1- (2- (3- (2-arabinosyl rhamnosyl) -6- (4-hydroxy-3-) represented by Chemical formula 13
Methoxycinnamoyl) glycosyl) ethyl) -3-hydroxy-4-methoxybenzene, represented by the formula:
1- (2- (3- (2-arabinosyl rhamnosyl) -4
-(4-Hydroxy-3-methoxycinnamoyl) glycosyl) ethyl) -3,4-dihydroxybenzene, 1- (2- (3- (2-arabinosylrhamnosyl) -6) represented by Chemical formula 15 -(3,4-Dihydroxycinnamoyl) glycosyl) -1-hydroxyethyl) -3,4-
Dihydroxybenzene, 1- (2-
(3- (2-arabinosyl rhamnosyl) -4- (3,4
-Dihydroxycinnamoyl) glycosyl) -1-hydroxyethyl) -3,4-dihydroxybenzene is mentioned as a compound preferably used according to the invention.

[0017]

[Chemical 11]

[0018]

[Chemical 12]

[0019]

[Chemical 13]

[0020]

[Chemical 14]

[0021]

[Chemical 15]

[0022]

[Chemical 16]

The above 1,2-dihydroxy-4- (2-hydroxyethyl) benzene derivatives are well known for their structural formulas and the like as compounds extracted from sesame seeds. As a method of obtaining from sesame seeds, for example, whole grass of dried or sesame seeds is extracted with a solvent, insoluble matter is removed from the extract by filtration or the like, and the solvent is distilled off from the obtained filtrate, followed by column chromatography. Examples of the method include purification by a usual means such as chromatography.

In this case, the extraction may be carried out in a batch system or a continuous system, and a preferable extraction method is to soak whole sesame seeds or a dried whole plant in a solvent for several days at room temperature. Examples of the method include a method of placing it and a method of soaking it at a temperature near the boiling point for several hours.

The solvent used for the extraction is preferably a polar solvent, and examples thereof include alcohols such as methanol and ethanol, ketones such as acetone and methyl ethyl ketone, and water. One of these solvents is used alone. Alternatively, two or more kinds may be mixed and used.

In the thrombotic inhibitor of the present invention, it is possible to use a salt of this compound in addition to the 1,2-dihydroxy-4- (2-hydroxyethyl) benzene derivative obtained as described above, 1,2-dihydroxy-4- (2-
One of the hydroxyethyl) benzene derivative and its salt may be used alone, or two or more thereof may be mixed and used.

Examples of such salts include sodium obtained by reacting a 1,2-dihydroxy-4- (2-hydroxyethyl) benzene derivative with an inorganic or organic base in equal amounts.
Salts with alkali metals such as potassium, salts with alkaline earth metals such as calcium and magnesium, salts with amines such as ammonia, triethylamine, triethanolamine, salts with basic amino acids such as arginine, lysine, etc. Can be illustrated.

<2> Composition Containing Thrombosis Inhibitor of the Present Invention The composition of the present invention is a composition prepared by mixing one or more of the above thrombosis inhibitors according to a conventional method.
Examples include pharmaceutical products.

When the above thrombus suppressor is used in food, as a general food, the required amount is added to each food material,
It can be used by being processed and manufactured by an ordinary manufacturing method.

The dosage form of the pharmaceutical composition of the present invention is not particularly limited, but is generally one of generally harmless pharmaceutically acceptable vehicles, or several kinds of vehicles, carriers, excipients, binders, disintegrants, dispersants, and lubricants. With agents, coating agents, flavoring agents, coloring agents, etc.
Tablets, capsules, granules, powders, drenches, suppositories, injections and the like can be formulated using conventionally known techniques. Examples of administration routes of injections include intraarterial administration, intravenous administration, subcutaneous administration, intramuscular administration and the like.

The amount of the above thrombotic inhibitor in these compositions is usually the daily intake per adult (though this varies depending on the patient's symptoms, weight, age, sex, etc.).
However, it is preferable that the amount of the thrombosis inhibitor is 1 to 1000 mg.

[0032]

EXAMPLES Examples of the present invention will be described below. First, examples of the thrombus inhibitor of the present invention will be described.

[0033]

Example 1 10 kg of dried sesame seeds were added to 5-10 mm
The mixture was cut into small pieces, to which 5 l of ethanol and 5 l of purified water were added, and the mixture was heated under reflux for 3 hours with stirring. Then, the extract was cooled, filtered to remove insoluble matter, and the filtrate was concentrated under reduced pressure to remove the solvent. The obtained extract was dissolved in purified water and poured into a column filled with Diaion HP-20 (manufactured by Mitsubishi Kasei Co., Ltd.), and further 2 l of purified water was flowed to wash it.

Thereafter, the column adsorbate was eluted with 2 l of ethanol, concentrated under reduced pressure, and then subjected to preparative high performance liquid chromatography (manufactured by Tosoh Corp., HLC837, ODS).
Column, elution solvent: 5% → 50% acetonitrile aqueous solution, UV 220 nm), and purified as a thrombosis inhibitor of the present invention by the following 6 types of 1,2-dihydroxy-
The 4- (2-hydroxyethyl) benzene derivative was obtained in the following yields.

(Compound 1) 1- (2- (3- (2-arabinosylrhamnosyl) -4- (3,4-dihydroxycinnamoyl) glycosyl) ethyl) -3,4-dihydroxybenzene: 145 mg

(Compound 2) 1- (2- (3- (2-arabinosylrhamnosyl) -6- (3,4-dihydroxycinnamoyl) glycosyl) ethyl) -3,4-dihydroxybenzene: 91 mg

(Compound 3) 1- (2- (3- (2-arabinosylrhamnosyl) -6- (4-hydroxy-3-methoxycinnamoyl) glycosyl) ethyl) -3-hydroxy-4- Methoxybenzene: 83mg

(Compound 4) 1- (2- (3- (2-arabinosylrhamnosyl) -4- (4-hydroxy-3-methoxycinnamoyl) glycosyl) ethyl) -3,4-
Dihydroxybenzene: 121 mg

(Compound 5) 1- (2- (3- (2-arabinosylrhamnosyl) -6- (3,4-dihydroxycinnamoyl) glycosyl) -1-hydroxyethyl)-
3,4-dihydroxybenzene: 144 mg

(Compound 6) 1- (2- (3- (2-arabinosylrhamnosyl) -4- (3,4-dihydroxycinnamoyl) glycosyl) -1-hydroxyethyl)-
3,4-dihydroxybenzene: 107 mg

<Evaluation of Thrombosis Inhibitors of the Present Invention> With respect to the thrombosis inhibitors of Compounds 1 to 6 obtained in Example 1 above, an acute toxicity test and a test for thrombosis inhibitory action were conducted.

(1) Acute toxicity test 6 groups of 6 ICR mice (male, body weight 25-35)
g), each thrombosis inhibitor (compounds 1 to 1 obtained in Example 1).
6) as a 10% physiological saline solution, 100
It was injected intraperitoneally at a rate of 0 mg / kg. Life or death was determined 14 days after administration, but no death was observed. From this, the thrombotic inhibitor of the present invention has an LD ₅₀ value of 1000 mg.
From the above, it can be seen that the safety is high.

(2) Thrombus Inhibitory Action The thrombus inhibitory action was evaluated by measuring the platelet aggregation inhibitory rate by the following method, using this as an index.

9 ml of blood was collected from a rabbit, 1 ml of a 3.8 wt% sodium citrate aqueous solution was added thereto, and the mixture was stirred well and then centrifuged at 950 rpm for 10 minutes. Platelet-rich plasma obtained from this supernatant was placed in 5 test tubes in 0.2 ml aliquots,
At 37 ° C., each thrombosis inhibitor obtained in Example 1 (Compound 1
0.01 ml of 1% aqueous solution of ~ 5) was added and stirred for 3 minutes. As a control, 0.2 ml of the above platelet-rich plasma
Prepared in the same manner as above using 0.01 ml of purified water.

2 μM adenosine 5′2 phosphate monosodium salt (ADP: Boehringer Mannheim Japan) was added to each of these as a platelet aggregation inducer in an amount of 0.02 m.
After adding 1 liter each and stirring for 7 minutes, the degree of platelet aggregation was measured using a platelet aggregometry device (Hematracer 601, manufactured by Nikko Bioscience). In addition, the same operation and measurement were performed using a 2 μg / ml collagen solution (hormone Chemie Munich) as a platelet aggregation inducer.

The platelet aggregation inhibitory rate, which is an index of the thrombotic inhibitory effect, was calculated by dividing the platelet aggregation of the specimen by the control platelet aggregation and multiplying by 100 to subtract from 100. The results are shown in Table 1.

[0047]

[Table 1]

From this, it is clear that the thrombus inhibitor of the present invention shows a strong platelet inhibitory rate and has an excellent thrombus inhibitory action. Next, examples of the composition of the present invention containing the thrombosis inhibitor obtained in the above examples will be shown. In addition, all compounding amounts used below are parts by weight.

[0049]

Examples 2 to 7 Injections The thrombosis inhibitor of Compound 1 obtained in Example 1 was dissolved in purified water to a concentration of 1%, sterile filtered, and aseptically filled in vials, freeze-dried, sealed. Then, a vial for injection of Example 2 was obtained. Similarly, the injections of Examples 3 to 7 containing the thrombosis inhibitors of Compounds 2 to 6 were produced.

[0050]

Examples 8 to 9 Injection The thrombosis inhibitor of Compound 1 obtained in Example 1 was dissolved in purified water to a concentration of 1%. Separately, the thrombosis inhibitor of Compound 5 obtained in Example 1 was dissolved in purified water to a concentration of 1% and mixed with the above solution in an equal amount. Thereafter, this was subjected to aseptic filtration, and aseptically filled into a vial, lyophilized, and sealed to obtain a vial for injection of Example 8. Similarly, the injection of Example 9 containing the thrombus suppressors of Compound 3 and Compound 4 was obtained.

[0051]

Example 10 Granules The formulation components shown in Table 2 were put in a Numal Melizer (manufactured by Fuji Paudal Co., Ltd.), granulated while moistening, and dried by blowing air to obtain granules.

[0052]

[Table 2]

[0053]

Example 11 Enteric coated granules A component shown in Table 3 was moistened in a new mulmelizer,
After granulating and drying, a solution obtained by dissolving the component B in ethanol and methylene chloride was gradually sprayed and coated. This was blow-dried and sized to obtain an enteric-coated granule.

[0054]

[Table 3]

[0055]

Example 12 Sustained Release Granules The granules of Example 10 and the enteric coated granules of Example 11 were mixed in equal amounts to obtain sustained release granules.

[0056]

Examples 13 to 20 Candy Component A in Table 4 was heated and dissolved at 150 ° C., then cooled to 120 ° C., and Component B was added thereto. Stir it evenly,
It was molded to obtain a candy.

[0057]

[Table 4]

[0058]

Example 21 Gummy A component of Table 5 was dissolved by heating at 110 ° C., B component separately swollen and dissolved was added thereto, and further C component was added and poured into a mold, and left for one day and night, then from the mold. Removed and got a gummy.

[0059]

[Table 5]

[0060]

The thrombus inhibitor of the present invention has a sufficient thrombus inhibitory action and is excellent in safety. In addition, the composition of the present invention can effectively and safely prevent and treat thrombosis such as cerebral infarction and stroke by including the thrombosis inhibitor.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Shin Yamamoto 560 Pola Kasei Co., Ltd., Kashio-cho, Totsuka-ku, Yokohama-shi, Kanagawa Prefecture Totsuka Laboratory Co., Ltd. (72) Inventor Toshio Miyase 4-chome, Nakada, Shizuoka-shi 5-29

Claims (3)

[Claims] 1. A thrombotic inhibitor comprising a 1,2-dihydroxy-4- (2-hydroxyethyl) benzene derivative represented by the following general formula (I) and / or a salt thereof. [Chemical 1] However, in Chemical formula ₁ , R ₁ , R ₂ and R ₃ are each independently a hydrogen atom or a group represented by Chemical formula 2, and R ₄ and R ₅ are each independently a hydrogen atom or a short chain alkyl group. R ₆ is
A hydrogen atom or a hydroxyl group, -Ram represents a rhamnose residue, and -Ara represents an arabinose residue. [Chemical 2] However, in Chemical formula 2, R ₇ and R ₈ each independently represent a hydrogen atom or a short chain length alkyl group. 2. The 1,2-dihydroxy-4- (2-
A hydroxyethyl) benzene derivative is represented by Chemical Formula 1- (2- (3- (2-arabinosylrhamnosyl) -4)
-(3,4-dihydroxycinnamoyl) glycosyl)
Ethyl) -3,4-dihydroxybenzene, 1- (2- (3- (2-arabinosyl rhamnosyl) represented by Chemical formula 4
-6- (3,4-Dihydroxycinnamoyl) glycosyl) ethyl) -3,4-dihydroxybenzene, 1- (2- (3- (2-arabinosylrhamnosyl) -6) represented by Chemical formula 5 -(4-hydroxy-3-methoxycinnamoyl) glycosyl) ethyl) -3-hydroxy-4-methoxybenzene, 1- (2- (3- (2-
Arabinosyl rhamnosyl) -4- (4-hydroxy-3)
-Methoxycinnamoyl) glycosyl) ethyl) -3,
4-dihydroxybenzene, 1- (2-
(3- (2-arabinosyl rhamnosyl) -6- (3,4
-Dihydroxycinnamoyl) glycosyl) -1-hydroxyethyl) -3,4-dihydroxybenzene,
From 1- (2- (3- (2-arabinosylrhamnosyl) -4- (3,4-dihydroxycinnamoyl) glycosyl) -1-hydroxyethyl) -3,4-dihydroxybenzene represented by The selected one is characterized in that it is selected.
The thrombus suppressor described. [Chemical 3] [Chemical 4] [Chemical 5] [Chemical 6] [Chemical 7] [Chemical 8] 3. A composition containing the thrombosis inhibitor according to claim 1 or 2.