JPH05246870A - Human platelet aggregation-inhibiting agent - Google Patents

Abstract

PURPOSE:To provide the subject inhibiting agent containing the juice of the green leaves of a gramineous plant as an active ingredient, reduced in the toxicity and adverse actions, capable of being massively administered and excellent in the safety and the therapeutic effects. CONSTITUTION:The objective inhibiting agent contains as an active ingredient a supernatant component which is obtained by squeezing the green leaves of a gramineous plant such as a barley plant e.g. by a mechanical crashing means such as a mixer under a condition not causing any unseasonable thermal modification, subjecting the obtained green juice to a centrifugal separation, collecting the supernatant and subsequently subjecting the supernatant to a sterilizing filtration treatment using a sterilization filter, etc. The inhibiting agent is preferably administered at a dose of 100-500mg/day for an adult as the active ingredient.

Description

Detailed Description of the Invention

The present invention relates to a human platelet aggregation inhibitor, and more specifically, it is a substantially nontoxic and highly safe human platelet aggregation that contains, as an active ingredient, the squeezed component of green leaves of gramineous plants, particularly barley plants. Regarding inhibitors.

[0002] The present inventors have been focusing on the physiological action of juicing juice or its dry powder from young leaves of grasses, especially wheat plants before ripening, and have been researching it for many years. Preventive or curative action (Japanese Patent Publication No. 46-385)
48), anti-cancer action (Japanese Patent Publication No. 59-36890).
Japanese Patent Publication), blood pressure lowering action (Japanese Patent Publication No. 59-36888).
JP-B and JP-B-59-39410), blood glucose lowering action (see JP-A-62-226927), antiviral action (see JP-A-3-169823),
Antioxidant activity (see Japanese Patent Application No. 2-220398) has been found, and for some of them, isolation and confirmation of the active substance have been successful.

In the course of the above series of studies, the present inventors have
This time, it was revealed that the juice components of green leaves of grasses, especially wheat plants, have an inhibitory effect on human platelet aggregation, and the present invention has been completed.

Thus, according to the present invention, there is provided a human platelet aggregation inhibitor characterized by containing, as an active ingredient, the juice component of green leaves of a grass family plant.

The human platelet aggregation inhibitor of the present invention will be described in more detail below.

As a grass family plant which is a raw material of an active ingredient of the inhibitor of the present invention, a barley plant is suitable, and examples of the barley plant include barley, wheat, naked barley, oats, and oats.
Examples include corn, millet, and Italian diegrass, and barley, wheat, and barley are particularly suitable.

In the present invention, among these malted plants, fresh stem and / or leaf portions of young plants harvested before maturity (herein, stem and / or leaf portions are collectively referred to as "green leaf"). Is especially suitable.

First, the green leaves of grasses, especially barley plants,
The juice is squeezed by a mechanical crushing means such as a mixer or a juicer under a condition that does not cause undue heat denaturation, and a coarse solid content is removed by a means such as sieving or filtration (hereinafter, this is referred to as ""Aojiru") is prepared. The green juice thus obtained can be used as it is as the juice component of the present invention. In general, it is preferable to sterilize the green leaves with a sterilizing agent such as sodium hypochlorite prior to the above squeezing and then perform the squeezing treatment.

The green juice obtained as described above is preferably further centrifuged to collect a supernatant liquid, and in some cases, this is subjected to sterilization filtration treatment with a sterilizing filter or the like to obtain a supernatant component. Can also be used as the juice component of the present invention.

Further, the green juice or supernatant component obtained as described above is neutralized to a pH of about 5 to 9 according to the method described in, for example, Japanese Patent Publication No. 46-38548, and then spray-dried, freeze-dried, etc. It can be dried and powdered by an appropriate means that does not cause heat denaturation, to obtain green juice powder or supernatant powder. These green juice powder or supernatant powder can also be used as the juice component of the present invention.

Furthermore, a liquid obtained by redissolving the green juice powder or the supernatant powder in water, or a liquid obtained by removing insolubles from the redissolved aqueous solution can be used as the juice component of the present invention, or A liquid obtained by extracting green juice powder or supernatant powder with an aqueous alcohol, for example, 50% aqueous methanol, or a powdered product obtained by removing the aqueous alcohol therefrom can also be used as the juice component of the present invention.

A solution obtained by removing solids from the green juice or a solution obtained by re-dissolving green juice powder in water to remove solids, for example, the above-mentioned supernatant component is a reverse phase chromatograph, an ion exchange chromatograph, A fraction having human platelet aggregation inhibitory activity can also be collected by applying gel filtration or the like. Reversed-phase chromatography can be performed using, for example, Delta-Pak3
Using a 00Å 15 μm C ₁₈ column (manufactured by Waters),
Acetonitrile containing 0.01 to 0.5% trifluoroacetic acid and 0.01 to 0.5% trifluoroacetic acid can be used as an eluent to elute the active fraction. Further, as the ion exchange chromatography, for example, Protein-Pak DEAE-5P
W (Waters) containing 1-100 mM Tris-HCl buffer at pH 8.7 and 0.1-1 M NaCl 2
The active fraction can be obtained by performing chromatography with a 0-100% linear concentration gradient of 0 mM Tris-HCl buffer. Furthermore, gel filtration can be performed, for example, with Protein-
It can be performed using Pak 300SW (manufactured by Waters). The green juice or supernatant component obtained as described above can be orally administered as it is or by adding, for example, milk, skim milk, other colloidal protein-containing substances, sweeteners, etc., but generally, the quality is constant. It is preferable to use the green juice powder or supernatant powder, or the redissolved product or extract thereof, which is more stable and more stable.

The "squeezing component of green leaves of grasses" used as an active ingredient in the present invention means not only green juice obtained as described above from green leaves of grasses, especially barley plants, but It is intended to include all the treated components or fractions having a human platelet aggregation inhibitory action obtained by further treatment as described above.
The human platelet aggregation inhibitor of the present invention may consist essentially of only the above-mentioned squeezed components, but it is generally suitable for administration by incorporating various pharmaceutically acceptable additives. It is preferable to formulate into the above dosage form.

Examples of the additives that can be used include additives for freeze-drying or spray-drying, as well as optional additives for formulation which are usually used for formulation into a desired dosage form. Specifically, for example, ascorbic acid, biotin, calcium pantothenate, carotene, choline chloride, magnesium oxide, niacin, pyridoxine chloride, lipofurapine, sodium pantothenate, thiamine hydrochloride, tocopherol, vitamin A,
Nutrients such as vitamin B ₁₂ and vitamin D ₂ ; sodium metaphosphate, sodium phosphate (first, second, third
Salt), sodium pyrophosphate, sodium tripolyphosphate and the like; preservatives such as calcium sorbate, benzoic acid, methyl paraoxybenzoate, sodium benzoate and the like; arabia gum, tragacanth, sodium alginate, methylcellulose, carboxymethylcellulose. ,
Carriers or diluents such as calcium alginate, aluminum silicate, calcium silicate, mannitol, sorbitol, lactose, fructose, soluble starch, amino acids, glucose, sugar, honey, sucrose, fatty acid ester can be mentioned.

The human platelet aggregation inhibitor of the present invention can be administered orally or parenterally, and can be formulated into any dosage form suitable for each administration route.
Suitable dosage forms for oral administration such as granules, pellets or tablets, coated tablets, capsules, troches, solutions, syrups; non-injections, drops, suppositories, eye drops, nasal drops, sprays, etc. The dosage form may be suitable for oral administration.

Further, it is also possible to use an external preparation type such as an ointment, a cream, a tincture and a patch.

The dose of the human platelet aggregation inhibitor of the present invention depends on the weight, sex, age, and weight of the symptoms of the subject patient.
It can be varied within a wide range depending on the judgment of a doctor, etc., but as a rough guide, it is generally about 30 to about 1000 mg / day, preferably about 100 per adult as an active ingredient.
It can be exemplified within the range of about 500 mg / day. The above dose can be administered once or divided into several times a day. However, in the case of oral administration, the human platelet aggregation inhibitor of the present invention is substantially nontoxic and has no side effects as described below, and therefore, it can be administered in a large amount beyond the above range. The acute toxicity LD ₅₀ of green juice from green leaves of gramineous plants, which is an active ingredient of the human platelet aggregation inhibitor of the present invention, for example, green juice powder from young leaves of barley before ripening is 1
It is virtually nontoxic at 2,000 mg / kg (oral, mouse), and no toxicity or side effect is observed even from the results of the subacute toxicity test of 1000 mg / kg continuous administration (oral, mouse).

Therefore, in view of its large pharmacological effect and lack of toxicity and side effects, the juice component of green leaves of the Gramineae plant of the present invention is practically nontoxic and has a practical inhibitory effect on human platelet aggregation. It has been revealed that it becomes a very unique human platelet aggregation inhibitor that has both the ability to be administered in a large amount and.

The present invention will be described in more detail below with reference to examples.

[0020]

【Example】

Example 1: Preparation of green juice powder of young barley leaf 1 kg of young barley leaf before maturity was washed with water, crushed with a crusher, and squeezed using a squeezing machine to obtain green juice 1 of young barley leaf.
1 was obtained. Add calcium hydroxide to this green juice
Was adjusted to 7.0 and freeze-dried by a conventional method to obtain 40 g of green barley green juice powder (hereinafter referred to as BLE powder).

Example 2: Measurement of human platelet aggregation inhibitory activity Preparation of sample solution : 5 g of the BLE powder prepared in Example 1 was dissolved in 10 ml of distilled water, and a 0.22 μm sterilizing filter (Wa) was used.
(manufactured by ters) to obtain a sample solution.

Measurement method : 9 ml of blood of a healthy person was collected in a polypropylene tube containing 1 ml of a 3.13% sodium citrate aqueous solution and centrifuged at 2300 rpm for 4 minutes to separate the turbid supernatant. Platelet-rich plasma (hereinafter,
PRP) was obtained, on the other hand, a buffer solution was added to the remaining lower layer portion to make the total volume 10 ml, and then 1 at 2800 rpm.
After centrifugation for 0 minutes, the supernatant was separated to obtain platelet poor plasma (PPP).

Paybon dual-channel aggregometer with P
P was adjusted so that the optical density was 0% with RP and 100% with PPP, and P contained 1 mM CaCl ₂ in the measuring cell.
After pouring 40 μl of RP and allowing it to stand for 1 minute, the sample solution in the amount shown in Table 1 below was added, and after incubation at 37 ° C. for 2 minutes, a platelet aggregation promoter was added at the concentration shown in Table 1 and the optical density was read. This optical density corresponds to the human platelet aggregation inhibition rate.
The results are shown in Table 1.

[0024]

[Table 1] As shown in Table 1 above, BLE powder shows a significant inhibitory effect on human platelet aggregation.

Next, formulation examples using BLE powder produced in the same manner as in Example 1 are shown.

Example A: Tablets BLE powder 40.0 mg Lactose 82.4 mg Crystalline cellulose 70.0 mg Talc 5.0 mg Binder (carboxymethyl cellulose) 2.0 mg Magnesium stearate 0.6 mg 20.0 mg BLE powder, lactose, Crystalline cellulose, talc and binder are uniformly mixed and made into granules, and then magnesium stearate is added to the mixture to form a tablet of 200 mg.

Example B: Enteric coated tablet The tablets obtained in Example A were subjected to enteric coating having the following formulation to produce 1 tablet of 430 mg.

Methacrylic acid / ethyl acrylate copolymer 10.8% Polyethylene glycol 6000 1.6% Surfactant (Tween 80) 1.1% Talc 7.2% Purified water 79.3% 100% Example C: Granules BLE powder 1.0 mg Lactose 700.0 mg Starch 289.0 mg Gelatin 10.0 mg 100.0 mg BLE powder, lactose and starch are uniformly mixed, and a small amount of water is further mixed and kneaded and then granulated. And dry (particle size 0.8 mm columnar granules). Example D: Capsule Granules are prepared according to the following formulation, enteric coated, and filled in capsules.

Granules (particle diameter 0.8 mm, columnar granules) BLE powder 1.4 mg Lactose 140.0 mg Starch 56.6 mg Gelatin 2.0 mg 20.0 mg Enteric coating (coating amount 430 mg / g
Granules) The formulation is as in Example B above.

Capsule filling Gelatin capsule No. 2 is filled with 200 mg.

Example E: Lozenge BLE powder 1.0 mg white sugar 920.0 mg gum arabic 79.0 mg Purified water proper amount 100.0 mg BLE powder and white sugar were uniformly mixed, and gum arabic was dissolved in a small amount of purified water. In addition, the mixture was kneaded into granules, dried, and compressed into lozenges.

Example F: Suppository BLE powder 1.0 g Polyoxyethylene lauryl ether (21 E.O.) 30.0 g Polyoxyethylene sorbitan monostearate (6 E.O.) 100.0 g Lipophilic glyceryl monostearate 16 0.0 g polyethylene glycol 400 6.0 g polyethylene glycol 4000 proper amount polyoxyethylene lauryl ether, polyoxyethylene sorbitan monostearate, lipophilic glyceryl monostearate, polyethylene glycol 400 and polyethylene glycol 4000 were dissolved by heating at 60 ° C. After that, the mixture was cooled to 45 ° C., BLE powder was added thereto, and the mixture was mixed uniformly, and then molded into 2 g of a suppository with a suppository molding machine.

Example G: Microcapsules Lactose 740.0 g BLE powder 20.0 g Eudragit RS 50.0 g Magnesium stearate 10.0 g 100.0 g Lactose was put into a vacuum mixing and drying coater while being rotationally mixed. After heating to about 50 ° C., the inside of the tank was evacuated by a vacuum pump, and coating was performed with an aqueous solution of BLE powder. Eudragit RS after coating
Was coated under vacuum in the same manner with a methylene chloride solution (containing 1% magnesium stearate based on the total weight of the product) to obtain microcapsules of BLE powder.

Claims (2)

[Claims] 1. A human platelet aggregation inhibitor comprising a squeezed component of a green leaf of a grass family as an active ingredient. 2. The human platelet aggregation inhibitor according to claim 1, wherein the gramineous plant is a barley plant.