JPS59128328A - Agent for lowering cholesterol level - Google Patents

Abstract

PURPOSE:To provide an agent for lowering the cholesterol level, containing phosphatidylethanolamine as an active component, free of side effects, and effective in a low dose. CONSTITUTION:The objective agent for lowering the cholesterol level is composed of a phospholipid containing >=60% of phosphatidylethanolamine of formula. Phospholipid exsiting in vegetables and animals is effective to lower the cholesterol level in the living body. Within the components of phospholipid, the phosphatidylethanolamine of formula is contained e.g. in egg yolk, fishoil, soybean, etc. Phospholipid containing >=80% of phosphatidylethanolamine can be prepared, e.g. by extracting phospholipid from edd yolk with ethanol, etc., and adding a phospholipase to the obtained phospholipid to substitute the choline segment of phosphatidylcholine with ethanolamine hydrochloride by base exchange reaction.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a cholesterol-lowering agent, and more particularly to a cholesterol-lowering agent containing phosphatidylethanolamine as an active ingredient.

Cholesterol plays an important physiological role in the body, but on the other hand, it is well known that when a large amount of cholesterol accumulates in the body, it causes arteriosclerosis. For this reason, traditionally,
Synthetic agents such as phenylethyl acetic acid and triparanol, and natural products such as konjac mannan and hemi-1 cellulose have been developed as cholesterol-lowering agents.

However, conventional cholesterol-lowering agents have the disadvantage that when used during the prenatal period, side effects such as diarrhea occur, and they are not effective unless administered in large amounts.

In order to provide a natural cholesterol-lowering agent that does not cause side effects and is effective in small amounts, the present inventors have conducted various studies and found that phosphatidylethanolamine, a type of phospholipid, is found in phospholipids. They came to the knowledge that it has a specific cholesterol-lowering effect, and based on this knowledge, they completed the present invention.

That is, the present invention aims to provide a cholesterol-lowering agent that is effective in a small amount without causing side effects, and which uses phosphatidylethanolamine as an active ingredient as a characteristic of the present invention. It has long been known that phospholipids, which are found in animals and animals, have the effect of lowering cholesterol in living organisms. ), phosphatidylinositol, sphingomilliene, and lysophosphatidylcholine, and it is a mixture of several bases such as phosphatidylinositol, sphingomilliene, and lysophosphatidylcholine.As it was not clear which of these bases contributed to the cholesterol-lowering effect, phospholipids were used as cholesterol-lowering agents. However, in the past, it was necessary to administer large amounts of phospholipids.

Among the phospholipid components, the cholesterol-lowering agent used in the present invention is phosphatidylethanolamine ()'E)
β9, its chemical structure is CH, -U-cog□, and is also called cephalin.

The phosphatidylethanolamine used in the present invention can be found, for example, in animal organs such as egg yolks, fish oil, and brain, and in soybeans and
It is included in plants such as cottonseed, corn, and Rakukasei.

The content of phosphatidylethanolamine is approximately 1.2% in egg yolk (occupies lθ~15X in egg yolk phospholipids)
In the case of soybeans, it is about 0.5% (accounting for about 20% of soybean phospholipids), and even natural products contain phosphatidyl. It is necessary to obtain a product with a high amount of ethanolamine.

In order to obtain a product with a higher content, a known method may be used. For example, when using egg yolk (I) as a raw material, 1 l of phosphorus Ba is extracted from the egg yolk using ethanol or acetone,
Phospholibase'f-ti was added to the obtained phospholipid, and phosphatidylcholine (accounting for about 80% of egg yolk phospholipid) was obtained by a total base exchange reaction with ethanolamine hydrochloride.
By substituting the choline moiety of , a phosphatidylethanolamine-containing jft 80 X or higher can be obtained. As another method, instead of the above-mentioned enzymatic method, there is a method in which phospholipids are fractionated by a common medium fractionation method or a column fractionation method.

However, the issue here is the content (purity) of phos-7 atidylethanolamine, which is close to 100% as this substance has a specific cholesterol-lowering effect among other lipids. is desirable. However, although it is technically possible to produce 100% pure phospholipids, since phospholipids have similar properties,
It is difficult in terms of cost because it is not possible to efficiently create a purity amplifier with simple operations.

Therefore, the cost of purification and the i
It: Considering both , If it were made into a powder of such purity, it would be possible to make fats and oils and milk for preventing arteriosclerosis by simply mixing it with fats and oils or milk.

The cholesterol-lowering agent produced as described above can be used orally for health purposes to prevent arteriosclerosis, for example, by adding 0.1 to 2.0 melamine to oil-based foods such as salad oil, margarine, and butter-Q milk. Foods can be manufactured. Furthermore, it can be encapsulated in gelatin or the like to be used as a product for arteriosclerosis patients. Furthermore, it can be emulsified with glucose and water to form oil droplets with a diameter of 1 micron or less to produce an intravenous injection for arteriosclerosis patients.

The reason why phos-7 atidylethanolamine used in the present invention exhibits a cholesterol-lowering effect in vivo is thought to be due to the action of ethanolamine, which is a constituent base.

This is because even in animal test M using ethanolamine, it shows a cholesterol-lowering effect. It should be noted that it is not preferable to use ethanolamine as it is as a cholesterol-lowering agent, since negative phase administration of ethanolamine may cause a side effect of causing gastrointestinal disorders.

As mentioned above, the cholesterol-lowering agent of the present invention is
Natural 'Pl! Since it is made from L as a raw material, there are no side effects even when administered for a long period of time, and it has systemic effects even in small doses, so it is effective as an agent for preventing arteriosclerosis or as a therapeutic agent for arteriosclerosis patients.

The experimental animals were Wistar male rats with a body voltage of approximately 1301, which were preliminarily fed with a standard diet for 7 days, and then divided into 3 groups of 6 rats per group, and the experimental diets shown in Table 1 were administered to each group. and reared for 14 days.

Place one rat at a time in a stainless steel gauge and keep at room temperature 22
The animals were reared in a back room at 1°C, and food and water were provided to Mede.

(Experimental feed) The values in the table indicate feed m%. In addition, soybean oil does not contain any phospholipids that have been bleached and stripped, so it does not contain any phospholipids.
A (4001U/100y+N shout), vitamin y l
) (200iU/1uOr@fee) and vitamin E
(,10nv/100r Copperaceae) has been analyzed by TAS6, and lecithin is a pure phosphatidylcholine [95
X, PE is the purity of phosphatidylethanolamine 8
0% 4 was used.

Test Results After the experiment, rats were weighed, blood was collected, and laparotomy was performed, but no differences were observed between the groups in intake, body weight gain, organ weight, and blood biochemical tests.

Then, blood cholesterol and liver cholesterol were measured by conventional methods, and the results shown in Table 2 were obtained.

As is clear from Table 2, blood talesterol levels and liver cholesterol levels were lower in the test group compared to control 1 #11 and control #2. It was found that the effect was excellent.

Example 1 Phospholyberase prepared from cabbage was added to egg yolk lecithin (70% phospholipids), and the choline moiety of phos-7-acylcholine was directly converted by a base conversion reaction with ethanolamine salt, resulting in phosphatidylethanolamine ( 80% phosphatidylethanolamine in phospholipids)
I got it.

One part of this phosphatidylethanolamine was thoroughly mixed with 50 parts of commercially available milk to prepare milk for preventing arteriosclerosis. .

Example 2 Soybean lecithin (95% phospholipids) was separated by 90% ethanol swamp fractionation to obtain phos-7-acytidylethanolamine (65% phosphatidylethanolamine in phospholipids). 1. 1 part of this phosphatidylethanolamine and After mixing 1.5 m of Kozuekama rice oil, the mixture was encapsulated in gelatin by a conventional method to prepare a therapeutic agent for arteriosclerosis (5 m in diameter/rnA).

Exclusion Example 3 Egg yolk lecithin (phospholipid'x98%) was subjected to bath medium fractionation according to Exclusion Example 2, and then a phosphatidylethanolamine fraction was obtained using an alumina column. The purity of this fraction was 95%.

This phosphatidylethanolamine 1 (5 l for injection)
Emulsify by pressurizing it in 100% Pudoku sugar solution to obtain an emulsion consisting of 0.5 micron particles.The obtained emulsion was treated aseptically and a small portion was sealed in a pile for 20mg% intravenous injection. ,
An injection for patients with dynamic brittle sclerosis was prepared.

Applicant: Kewpie Co., Ltd. Procedural Amendment Omega 5 February 4, 1980 Director-General of the Patent Office Kazuo Wakasugi 1, Indication of the case 1985 Patent Application No. 485 2, Name of the invention Cholesterol-lowering agent 3, Amendment (142) Kewpie Co., Ltd. 4, Agent: 150 [Telephone (Tokyo) 486-3342] Address: 1-4-13 Shibuya, Shibuya-ku, Tokyo Internal corrections within Kewpie Co., Ltd. Statement 1: Invention Name: Cholesterol-lowering agent 2, claims (1) Cholesterol-lowering agent L0 characterized by containing phosphatidylethanolamine as an active ingredient (2) The proportion of phosphatidylethanolamine in phospholipids is 60% The scope of the claims which is the above (
1) The cholesterol-lowering agent described above.

3. Detailed Description of the Invention The present invention relates to a cholesterol-lowering agent, and more particularly to a cholesterol-lowering agent containing phos-7 atidylethanolamine as an active ingredient.

Cholesterol plays an important physiological role in the body, but on the other hand, it is known that when a large amount of cholesterol accumulates in the body, it causes arteriosclerosis.

For this reason, synthetic agents such as phenylethyl acetic acid, tol and lanol, and natural products such as konjac mannaya hemicellulose have been developed as cholesterol-lowering agents.

However, conventional cholesterol-lowering agents have the disadvantage that when used for a long period of time, side effects such as diarrhea occur, and that they are not effective unless administered in large amounts.

In order to provide a natural cholesterol-lowering agent that does not cause side effects and is effective in small amounts, the present inventors have conducted various studies and discovered that phos-7 atidylethanolamine, one of the phospholipids, is a phospholipid. Among them, it was discovered that it has a specific cholesterol-lowering effect, and based on this knowledge, the present invention was completed.

That is, the present invention aims to provide a cholesterol-lowering agent that is effective in a small amount without causing side effects, and is characterized by containing 7-osphatidylethanolamine as an active ingredient.

It has long been known that phospholipids contained in plants and animals have the effect of lowering plasma cholesterol. It is composed of a mixture of several bases such as diethanolamine (pg), phosphananolinositol, sphingomilliene, and lysophosphanadylcholine, and it is unclear which of these bases contributes to the cholesterol-lowering effect. Therefore, even if phospholipids were to be used as cholesterol-lowering agents, it was necessary to administer multiple doses of phospholipids.

Among the phospholipid components, the cholesterol-lowering agent used in the present invention includes phosphonadylethanolamine (PK).
), the chemical structure is CH2-0-C0R1.

The phosphonadylethanolamine used in the present invention is
For example, it is found in animal matter such as egg yolk, fish oil, and brain, and in plant seeds such as soybeans, cottonseed, corn, and peanuts.

The content of phosphonadylethanolamine is 1.2% in egg yolk (accounting for 10-15% of egg yolk phospholipid), and 0.5% in soybean (approximately 20% in soybean phospholipid).
%) Since it is a trace ingredient even in natural products,
In order to provide for the present invention, by methods such as extraction and fractionation,
It is necessary to obtain a product with a high content of phosphonadylethanolamine.

In order to obtain a product with a high content, it is sufficient to use a known method. For example, when egg yolk is used as a raw material, the syphospholipids are removed from the egg yolk using ethanol or acetone, and the resulting phospholipids are converted into phospholipids. By adding lipase and replacing ethanolamine hydrochloride with the choline moiety of 7-osphatylcholine (accounting for about 80% of egg yolk phospholipids) by base exchange reaction, the phosphatylethanolamine content is 80%. You can get more than just that.

As another method, instead of the enzyme method described above, there is also a method of fractionating phospholipids by a solvent fractionation method or a column fractionation method.

However, the issue here is the content (purity) of phosphoanadylethanolamine, but since this substance has a specific cholesterol-lowering effect among phospholipids, it must be 100% pure. Preferably close. However, although it is technically possible to produce 100% pure phospholipids, since phospholipids have similar properties,
Purity cannot be efficiently increased with simple operations, making it difficult in terms of cost.

Therefore, considering both the cost of purification and the cholesterol-lowering effect, the phospholipid should be purified so that the proportion (content) of phosphoanadylethanolamine in the phospholipid is 60% or more, preferably 80% or more. This is desirable. If it is made with such purity, it will be possible to produce fats and oils and milk for preventing arteriosclerosis by simply adding a small amount of it to fats and oils or milk.

The cholesterol-lowering agent prepared as described above can be used orally, for example, by adding 0.1 to 2. (A health food for preventing arteriosclerosis can be produced by adding about 1.
It can also be encapsulated in gelatin capsules and used as a drug for arteriosclerosis patients. Furthermore, it can be emulsified with an aqueous glucose solution to form oil droplets with a diameter of 1 micron or less to produce an intravenous injection for arteriosclerosis patients.

The fact that the 7-osphatidylethanolamine used in the present invention exhibits a cholesterol-lowering effect in vivo is thought to be due to the action of ethanolamine, which is a constituent base.

This is because even in animal tests using ethanolamine, it shows a cholesterol-lowering effect. -In addition,
Since long-term administration of ethanolamine may cause side effects such as inducing gastrointestinal disorders, it is not preferable to use ethanolamine directly as a cholesterol-lowering agent.

As mentioned above, the cholesterol-lowering agent of the present invention is
Since it is made from a natural product, there are no side effects even when administered for a long period of time, and it is effective even in small doses, so it is effective as an agent for preventing arteriosclerosis or as a therapeutic agent for patients with arteriosclerosis.

Test Example The experimental animals were Wistar detachment rats weighing 130.9 kg, which were preliminarily fed with a standard diet for 7 days, and then divided into 3 groups of 6 animals per group, each of which was administered the experimental diet shown in Table 1, and kept for 14 days. .

Rats were placed one by one in stainless steel darts and kept at room temperature 22
The animals were reared in a breeding room at ±1°C and given free access to food and water.

In addition, the numerical values in the table indicate weight factors. In addition, soybean oil is bleached and deodorized and does not contain any phospholipids.
200IU/100.9 feed) and vitamin 7E (107
9/100.9 feed) is added. Furthermore, the lecithin used was phosphonadylcholine with a purity of 95% Ni, and the PE used was phosphatidylethanolamine with a purity of 80%.

Test Results After the experiment, rats were weighed, blood sampled, and laparotomy was performed, but no differences were observed between the groups in intake, body weight gain, organ weight, and blood biochemical tests. .

Then, plasma cholesterol and liver cholesterol were measured using conventional methods, and the results shown in Table 2 were obtained.

As is clear from Table 2, it was observed that the plasma cholesterol level and liver cholesterol level were significantly lowered in the test group compared to control group 1 and control group 2.

Example 1 Phospholipase prepared from cabbage was added to egg yolk lecithin (70% phospholipids), and the choline moiety of 7-o-7-atedylcholine was replaced by a base exchange reaction with ethanolamine hydrochloride to form phos-7-atidylethanolamine. (7 osphatylethanolamine 8 in phospholipids)
0%) was obtained.

50 seconds of commercially available milk was thoroughly mixed with 21 parts of this phosphonadylethanolamine to prepare milk for preventing arteriosclerosis.

Example 2 Soybean lecithin (95% phospholipid) was separated into phosphatidylethanolamine (
7 osphatylethanolamine (65% in phospholipid)
I got it.

After mixing 1 part of this phosphonanolethanolamine and 1.5 parts of refined rice oil, the mixture was encapsulated in wide gelatin by a conventional method to prepare a therapeutic agent for arteriosclerosis having a particle size of 5 m/m.

Example Egg yolk lecithin (98 phospholipids) was subjected to solvent fractionation according to Example 2, and then subjected to an alumina column to obtain a 7 male 7 atedyl tanolamine fraction. The purity of this fraction was 95%.

1 to 0 g of this 7-mole 7-acylated tanolamine was emulsified in 100 g of a 5-glucose solution for injection.
An emulsion consisting of 5 micron particles was obtained. 20ml of the obtained emulsion! A small portion was sealed in a pile for intravenous injection and treated aseptically to prepare an injection for arteriosclerosis patients.

Claims (2)

[Claims] (1) A cholesterol-lowering agent characterized by containing phosphatidylethanolamine as an active ingredient. (2) Claim (1) in which the proportion of phosphatidylethanolamine in the phospholipid is 60% or more
Cholesterol lowering agent as described.