JP2003146883A - Preventing and treating agent for defect of memory - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a medicine, beverage and foods for preventing or treating Alzheimer type defect of memory. SOLUTION: This preventing and treating agent for the Alzheimer type defect of memory is provided by using sphingomyelin as an active ingredient of the agent. Sphingomyelin has an effect for suppressing the reduction of a protein kinase C activity caused by aging and it is suggested that the compound is effective for preventing and treating the Alzheimer type defect of memory.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an agent for preventing or treating Alzheimer's memory disorders, which contains sphingomyelin, which is one of phospholipids, as an active ingredient.

[0002]

2. Description of the Related Art In recent years, as the average life span has been extended, the number of elderly people has increased rapidly, and various problems have arisen as an aging society. Among them, the increase of senile dementia is a big problem. Senile dementia is roughly classified into cerebrovascular dementia and Alzheimer's dementia.Although Alzheimer's dementia is expected to increase rapidly in Japan in the future, its mechanism of onset and progress Active research is underway, but there is no effective treatment and it becomes a social problem, and the development of an effective therapeutic drug is desired.

In central degenerative diseases such as Alzheimer's dementia, it is a neurotransmitter due to degenerative loss of cholinergic neurons in the basal forebrain region (septal area, blocker diagonal zone and basal Meinert's nucleus). Acetylcholine is not produced, causing a significant decrease. Decreased acetylcholine is thought to be the cause of memory-learning impairment in Alzheimer-type dementia [Whiteahouse et al, Science, 21]
5, 1237-1239 (1982); Coyle et al, Science, 219, 118.
4-1190 (1983)]. Experimentally, it has been reported that destruction of cholinergic neurons in the basal forebrain of rats causes a decrease in the amount of acetylcholine and memory impairment [He.
lper et al, J. Neurosci., 5, 866-873 (1985)]. 1980
In the 1980s, the correlation between long-term potentiation of the hippocampus and learning / memory was noted, and it is considered that the increase in intracellular calcium activates the brain by inducing long-term potentiation via calmodulin-dependent kinase or protein kinase C. Nakagawa Hachiro, Metabolism, 26, Extra number, Brain metabolism and its abnormalities, 37-44 (1989)]. Also, protein kinase C
Has been reported to be decreased in the brain (temporal lobe tissue membrane fraction) of patients with Alzheimer's disease, and immunoreactivity is reduced in the hippocampal membrane fraction [Shun Shimohama et al., 35th Japan. Proceedings of the Society for Neurochemistry, 92 (1992); MaslishE. Et al,
J. Neurosci., 10, 2113-2124 (1990); Shun Shimohama, Saishin, 47, 602 (1992)]. It is considered that activation of protein kinase C, which is present in a high concentration in the brain, is effective in treating Alzheimer's dementia. From the above, it is considered that substances that increase protein kinase C activity in the brain have preventive and therapeutic actions on central degenerative diseases such as Alzheimer's dementia.

On the other hand, sphingomyelin is a kind of phospholipid, which is widely distributed in egg yolk and milk, and is distributed as a constituent component of cell membranes and serum lipoproteins. It is known that sphingomyelin in vivo influences cell proliferation and differentiation via an information transmission system. However, sphingomyelin is not known to be effective in preventing or treating Alzheimer's memory impairment.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a drug and food and drink for preventing or treating Alzheimer's memory disorder.

[0006]

[Means for Solving the Problems] The inventors of the present invention have conducted extensive studies to solve the above-mentioned problems. As a result, sphingomyelin was orally ingested to induce protein kinase C in the brain.
They have found that they have the activity of increasing the activity, and have completed the present invention. That is, the present invention relates to a prophylactic / therapeutic agent for Alzheimer-type memory disorders, which contains sphingomyelin as an active ingredient. In the present invention, sphingomyelin purified and purified may be used, or may be used in the form of a phospholipid containing sphingomyelin.

Sphingomyelin and sphingomyelin-containing phospholipids can be prepared by the following method. For example, milk-derived sphingomyelin-containing phospholipids obtained by a method of extracting milk or a dairy product such as whey protein concentrate (WPC) with ether or acetone (JP-A-3-47192) 28% content) can be used,
In addition, a butter curd obtained by heating and melting butter or an aqueous fraction containing butter serum can be used as a sphingomyelin-containing phospholipid (containing about 9% of sphingomyelin in phospholipids). The milk fat globule coating fraction contained in buttersarum can be used as a sphingomyelin-containing phospholipid (containing about 9% of sphingomyelin in phospholipids). In addition, animal brains and culture fluids of microorganisms capable of producing sphingomyelin can also be used as the sphingomyelin-containing phospholipid. Then, the sphingomyelin-containing phospholipids are purified by methods such as dialysis, ammonium sulfate fractionation, gel filtration, isoelectric point precipitation, ion exchange chromatography, and solvent fractionation to use sphingomyelin having a higher purity. Is also good. Further, these sphingomyelin and sphingomyelin-containing phospholipids may be used in a liquid state or a powder state.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention is a preventive and / or therapeutic agent for Alzheimer-type memory disorders containing sphingomyelin as an active ingredient. The dosage form of the preventive or therapeutic agent for Alzheimer-type memory disorders may be tablets, capsules, granules, powders, powders and the like. Further, these dosage forms can be produced by a conventionally known method, for example,
It may be molded by mixing with a carrier, an excipient, or the like that is allowed in the production of the preparation.

The present invention is also a food or drink for preventing or treating Alzheimer's memory disorders, which contains sphingomyelin as an active ingredient. This Alzheimer's type memory disorder prevention, therapeutic food and drink food and drink, milk, milk drink, coffee drink, juice, jelly, biscuits, bread, noodles, sausage and other food and drink, sphingomyelin,
Further, it is added to a nutritional composition or the like for the purpose of supplementing essential fatty acids and fat-soluble vitamins.

In the present invention, in order to exert the preventive and therapeutic effects on Alzheimer-type memory disorders, in the case of an adult, sphingomyelin is 10 to 2,500 mg per day.
The blending amount may be adjusted so that 50 to 2,500 mg can be ingested. Next, the present invention will be described more specifically with reference to Examples and Experimental Examples.

[0011]

Example 1 10% of whey protein concentrate (WPC)
After the reaction solution obtained by allowing the protease to act on the aqueous solution was extracted with a chloroform-methanol (2: 1) solution,
The mixture was concentrated and extracted with acetone to obtain a complex lipid fraction. Next, this complex lipid fraction was subjected to fluorosilyl column chromatography and stepwise extracted with a chloroform-methanol solution to obtain a phospholipid fraction. Then, this phospholipid fraction was subjected to silica gel column chromatography and stepwise extracted with a chloroform-methanol solution to obtain sphingomyelin. The resulting sphingomyelin was freeze-dried to give a prophylactic / therapeutic agent for Alzheimer-type memory disorders. The sphingomyelin content was 95.2% by weight when the sphingomyelin was subjected to thin-layer chromatography, developed with a Dittmer reagent and measured by a densitometry method. Therefore, if 11 mg or more of the preventive or therapeutic agent for Alzheimer's memory disorder is taken per day, it is effective for the prevention and treatment of Alzheimer's memory disorder.

[0012]

TEST EXAMPLE 1 Using the sphingomyelin obtained in Example 1, the preventive and therapeutic effects of Alzheimer's memory deficits were examined in animal experiments using mice. That is, 20-week-old senescence-accelerated mice (SAMP / 8 strain) were used as experimental animals in groups of 20 each. Then, in the experimental feed having the composition shown in Table 1, the amount of sphingomyelin added was 0.5%, and for the other compositions, the experimental feed according to the AIN-76 composition was fed for one month.

[0013]

[Table 1]

After the administration of the experimental feed, the blood was removed from the heart and the brain was collected. Immediately after that, 5 times the volume of the ice-cold homogenizing buffer solution (0.25 M
Sucrose, 10mM Hepes, 2mM EDTA, 5mM EGTA, 1mM P
After homogenizing in MSF, 10 mM β-mercaptoethanol, 10 μg / ml leupeptin, pH 7.5), 105,000 ×
The mixture was centrifuged at 4 ° C for 60 minutes for 60 minutes to obtain a supernatant. About this supernatant, Hepes buffer (10 mM Hepes, 20 mM NaCl, 0.2 mM
DERE-cellulose column equilibrated with EDTA, 0.5 mM EGTA, 1 mM PMSF, 10 mM β-mercaptoethanol, pH 7.4)
It was purified with (DE-52) to obtain a protein kinase C extract.

The protein kinase C activity of the protein kinase C extract thus obtained was evaluated by examining the incorporation of [γ-32P] ATP into histones. That is, 0.1 ml of reaction solution (20 mM Hepes pH7.5, 1 mM
Magnesium acetate, 50 μM [γ-32P] ATP (0.5 μCi), 500
After incubating μg / ml phosphatidylserine, 8 μg / ml 1-oleoyl-2-acetyl-sn-glycerol, the above protein kinase C extract) for 3 minutes at 30 ° C, add 0.1 ml of 40% trichloroacetic acid. The reaction was stopped by adding 0.1 ml each of 50 mM ATP and 5 mg / ml BSA, followed by centrifugation, and 32 P in the precipitate was measured with a liquid scintillation counter. Then, the protein kinase C activity was expressed as the amount of [γ- 32 P] ATP incorporated into histones per 1 minute of the protein of the protein kinase C extract. The result is shown in FIG.

According to this, the protein kinase C activity was significantly increased in the group of the present invention. Therefore, it was suggested that sphingomyelin has an effect of suppressing a decrease in protein kinase C activity due to aging, and is effective in preventing or treating Alzheimer's memory deficits.

[0017]

Example 2 A tablet having the composition shown in Table 2 was produced by a conventional method and used as a preventive or therapeutic agent for Alzheimer's memory disorder.
The amount of sphingomyelin contained in this tablet is 9.
It was 5% by weight. Therefore, this tablet is
Ingestion of more than mg is effective for prevention and treatment of Alzheimer's memory disorder.

[0018]

[Table 2]

[0019]

Example 3 A phospholipid fraction was prepared by the same method as that described in Example 1, and this phospholipid fraction was used as a sphingomyelin-containing phospholipid. The resulting sphingomyelin-containing phospholipid was lyophilized to give a prophylactic / therapeutic agent for Alzheimer's memory disorder. In addition,
The sphingomyelin content in this sphingomyelin-containing phospholipid was 25.0% by weight. Therefore, ingestion of 40 mg or more of the prophylactic / therapeutic agent for Alzheimer's memory disorder per day is effective for the prevention and treatment of Alzheimer's memory disorder.

[0020]

Example 4 The sphingomyelin-containing phospholipid obtained in Example 3 was blended to produce a milk drink for the prevention and treatment of Alzheimer-type memory disorders. That is, after mixing 20 kg of milk, 21.6 kg of skim milk powder and 56 kg of water to prepare a milk base, 1 kg of granulated sugar and 0.37 kg of phospholipids containing sphingomyelin are added to this milk base, and a homogenizer gives a homogeneous pressure of 200 kg / cm2. Emulsified as. And the filling temperature
Fill it in a 200 ml can at 90 ° C, tighten it, and then
Retort sterilization was performed at 10 ° C for 10 minutes, followed by cooling to produce a canned milk drink. In addition, 185 mg of sphingomyelin was contained in 1 can (200 ml) of this canned milk drink. Further, this canned milk drink had almost no browning and had a milk flavor.

[0021]

Example 4 The sphingomyelin-containing phospholipid obtained in Example 3 was blended to produce a tube / oral nutritional diet for the prevention and treatment of Alzheimer's memory disorders. Ie, 75
Dextrin 632g, sodium caseinate 174g, skim milk powder 33g, sucrose fatty acid ester in 1.5 l of water heated to ℃
6 g, palm oil fractionated oil 62 g and sphingomyelin-containing phospholipid 45 g mixed fat and oil mixture 107 g and potassium triphosphate 7 g dissolved in 50 ml of water were added and mixed, and further calcium chloride 4 g / 160 ml solution, Magnesium sulfate
A solution of 7 g / 160 ml, a solution of vitamin mixture 4 g / 53 ml and a solution of mineral mixture 6 g / 53 ml were added and mixed. Next, this mixture was pre-emulsified with a colloid mill and then homogenized with a homogenizer at a uniform pressure of 400 kg / cm 2. And 121
After heat sterilization at 15 ° C. for 15 seconds, spray drying was performed to obtain about 1 kg of tube / oral nutrition food, and 92 g of nitrogen foil was packed in each aluminum foil container to produce tube / oral nutrition food. The sphingomyelin content contained in this nutritional composition was 1.1% by weight. Therefore, ingestion of one tube (92 g) of this tube / oral nutritional food per day can supply about 1 kcal of energy, and it is also effective for the prevention and treatment of Alzheimer's memory disorder.

[0022]

Example 5 A cream having a fat content adjusted to 50% was washed with 3 times the amount of water, allowed to stand at 4 ° C. overnight and then treated with butter churn to separate butter granules and butter milk. Next, the buttermilk was subjected to ammonium sulfate fractionation, centrifugation and dialysis to prepare a milk fat globule coating fraction, and this milk fat globule coating fraction was used as a sphingomyelin-containing phospholipid.
The resulting sphingomyelin-containing phospholipid was lyophilized to give a prophylactic / therapeutic agent for Alzheimer's memory disorder. The sphingomyelin content in this sphingomyelin-containing phospholipid was 10.4% by weight. Therefore, if you take 100mg or more of the prophylactic / therapeutic agent for Alzheimer's memory disorder per day,
It is effective for the prevention and treatment of Alzheimer-type memory disorders.

[0023]

Example 6 The sphingomyelin-containing phospholipid obtained in Example 5 was blended to produce a nutritional composition for preventing / ameliorating / treating Alzheimer's memory disorder. That is, in 60 kg of dissolved water, 3.4 kg of milk protein and soy protein
Disperse 1.14 kg, dissolve by heating to 70 ° C, add 12.4 kg of dextrin, and further add 2.4 kg of fats and oils that mixed soybean oil, palm oil and purified fish oil, and 0.2 kg of phospholipid containing sphingomyelin. Was added. Next, after preliminarily emulsifying with a TK homomixer, it was concentrated and spray-dried to prepare a raw material powder. And, to this raw material powder, an appropriate amount of minerals,
Vitamins, flavors and the like were added and mixed to prepare a powdery nutritional composition for the prevention and treatment of Alzheimer's type memory disorders. The sphingomyelin content contained in this nutritional composition was 0.3% by weight. Therefore, ingestion of 3.5 g or more of the present nutritional composition per day is effective for prevention and treatment of Alzheimer's memory disorder.

[0024]

The prophylactic and therapeutic agent for Alzheimer's type memory disorders containing sphingomyelin of the present invention as an active ingredient has an action of increasing protein kinase C activity in the brain by oral ingestion. It can be advantageously used as a preventive or therapeutic agent.

[Brief description of drawings]

FIG. 1 shows the protein kinase C activity in each experimental group of Test Example 1.

Claims (2)

[Claims] 1. A preventive and / or therapeutic agent for Alzheimer-type memory disorders, which contains sphingomyelin as an active ingredient. 2. A food or drink for preventing and / or treating Alzheimer-type memory disorders, which contains sphingomyelin as an active ingredient.