JP2016135757A - Cognitive function maintaining or improving agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel cognitive function maintaining or improving agent which can be administered more easily, and is capable of maintaining or improving cognitive function.SOLUTION: The present invention relates to a maintaining or improving agent for cognitive function which is administered orally and contains a swine liver degradation product by protease. The swine liver collected from a sugar pork (R) is suitably used. Taking the cognitive function maintaining or improving agent of the invention enables prevention of dementia, or maintenance or improvement of symptoms thereof.SELECTED DRAWING: None

Description

  The present invention relates to an agent for maintaining or improving cognitive function. It relates to an improving agent.

Dementia is known as one of the diseases associated with aging.
Dementia is a condition in which the intellectual ability (cognitive function) acquired so far has been lost as a result of damage to the developed brain. As specific symptoms (cognitive dysfunction) of dementia, in addition to memory impairment such as forgetfulness, declines in judgment, calculation, understanding, learning, etc. appear. For this reason, those who have become dementia will have problems in social life and interpersonal relationships.

Major diseases that cause dementia include degenerative diseases such as Alzheimer's disease and Parkinson's disease, cerebrovascular disorders, normal pressure hydrocephalus, metabolic nutrition disorders such as vitamins, and hypothyroidism. Dementia is diagnosed when cognitive impairment appears to interfere.
The above-mentioned diseases tend to increase with age, so in Japan, with the progress of aging, the number of people diagnosed with dementia is also increasing.
Furthermore, in 2015, the “baby boom generation” will reach an older age, and the number of elderly people will increase further in the future. Therefore, it is considered that more people are diagnosed with dementia.

Under such circumstances, there is a demand for an agent for maintaining or improving cognitive function.
For example, as one of them, there has been proposed a pharmaceutical composition for treating dementia comprising a mammalian liver extract that is insoluble in acetone and soluble in water (Patent Document 1).
The liver extract is obtained by adding a large excess of acetone to the raw material (liver) and mixing, separating the resulting acetone insoluble fraction from acetone, and further dissolving the obtained acetone insoluble fraction in water. be able to. In addition, the therapeutic pharmaceutical composition of Patent Document 1 is administered to a subject by injection such as intramuscular injection.

Japanese Patent Laid-Open No. 5-255096

  As described above, the therapeutic pharmaceutical composition described in Patent Document 1 is administered to a subject by injection. However, taking into consideration that the burden of care is further increased as the number of elderly people increases, a dosage form that can be used more easily, for example, can be administered by a person with dementia, is preferable.

  The present invention has been made based on such circumstances, and an object thereof is to provide a novel agent for maintaining or improving cognitive function that can be used more easily and can maintain or improve cognitive function.

  The present inventor has conceived an oral administration form as an agent for maintaining or improving cognitive function that can be used more easily, and has conducted extensive research. As a result, the present inventors have found that when a porcine liver degradation product by proteolytic enzyme is orally administered to a subject, the cognitive function of the subject can be maintained or improved, and the present invention has been completed.

That is, the gist of the present invention is as follows.
[1] An agent for maintaining or improving cognitive function, wherein the agent is administered orally and contains a proteolytic enzyme porcine liver degradation product.
[2] The agent for maintaining or improving cognitive function according to [1], wherein the porcine liver degradation product is orally administered in an amount to be taken at least 266 mg per day.
[3] The agent for maintaining or improving cognitive function according to [1] or [2], wherein the porcine liver degradation product is obtained from the liver of a pig free of specific pathogens.
[4] The cognitive function maintenance or improvement agent according to [3], wherein the specific pathogen-free pig is sugar pork (registered trademark, hereinafter the same).

An object of the present invention is to provide a novel agent for maintaining or improving cognitive function that can be used more easily and can maintain or improve cognitive function.
That is, cognitive function can be maintained or improved by taking the cognitive function maintenance or improvement agent of the present invention containing porcine liver degradation products by proteolytic enzymes, so that dementia can be prevented or symptoms can be prevented. It can be maintained or improved. Moreover, the cognitive function maintenance or amelioration agent of the present invention can be used more easily than the conventional one because it can be taken by oral administration, for example, by the subject itself.

It is a graph which shows the relationship between the proliferation of the rat astroglial cell (RCR-1) cell, and the added component in connection with Test Example 1. FIG. 4 is a graph showing the relationship between the proliferation of mouse microglia cells (MG6) and the added components in Test Example 1. FIG. It is a graph which shows the relationship between the proliferation of the neuroblast (IMR-32) cell, and the added component in connection with Test Example 1. It is a graph which shows the score value of the Hasegawa type simple cognitive evaluation scale (HDR-S) before and after administration in connection with Test Example 2.

  The present embodiment relates to an agent for maintaining or improving cognitive function, characterized by containing a porcine liver degradation product by a protease (protease). Hereinafter, the cognitive function maintenance or improvement agent of the present embodiment will be described in detail.

The mechanism of maintaining or improving cognitive function by orally ingesting the cognitive function maintaining or improving agent of the present embodiment is not necessarily clear at present, but it is not always clear, such as peptides and phospholipids derived from pig liver It is considered to be a complex action.
The invention disclosed in Patent Document 1 described above has water-soluble peptides as active ingredients, and completely removes water-insoluble fractions including functional lipids such as phospholipids during the production process. Has been. Moreover, since the liver degradation product marketed is also manufactured using solid-liquid separation methods, such as ultrafiltration, the component which does not melt | dissolve in water, such as a lipid, is not contained.
On the other hand, the porcine liver degradation product according to the present embodiment contains abundant peptide fractions derived from porcine liver, and a wide range of components including phospholipids remain as they are.

The kind, breed, and the like of the pig from which the pig liver according to this embodiment is obtained are not particularly limited, and can be appropriately set by those skilled in the art.
On the other hand, it is preferable that the pig which concerns on this embodiment is a specific pathogen free pig (Specific Pathogen Free pig, SPF pig).
A specific pathogen-free pig is a progeny of a pig removed from a mother pig at the end of pregnancy by cesarean section, a disease that greatly affects the growth of the pig, specifically toxoplasma infection, mycoplasma pneumonia, atrophic rhinitis, OS This refers to pigs not suffering from key disease or swine dysentery.

Furthermore, in this embodiment, it is more preferable that said specific pathogen-free pig is a sugar pork.
Sugar pork is a specific pathogen-free pig that has been sanitized over three generations. Specifically, just before the parent pigs are delivered, a cesarean section is taken out to take out the piglets, making them safe native species that do not have the five pathogens mentioned above, and then the children born from this pig to normal delivery Furthermore, the pig born from this pig becomes the parent pork of the sugar pork.
Sugar pork is bred only on plant-based feed, based on thorough hygiene management, and grown without administration of antibiotics, antibacterial agents, chemicals, and the like.

  The liver degradation product according to the present embodiment can be obtained by, for example, a method of filtering a treatment liquid after enzymatic treatment of pig liver such as sugar pork.

An example of the manufacturing method will be described more specifically.
First, the liver raw material is pulverized using a homogenizer, a minced cutter or the like, and water is added to the obtained pulverized product to obtain a reaction solution. The amount of water to be added is not particularly limited and can be appropriately set by those skilled in the art.
In addition, the form of the pig liver which concerns on this embodiment is not specifically limited, If it is the liver obtained from the pig, the raw | natural thing may be processed, such as freezing. In addition, the reaction liquid preparation process is not limited to the above-mentioned process. For example, a pig liver that has been previously subjected to processing such as pulverization is obtained and suspended in water to prepare a reaction liquid. Also good.

Next, a proteolytic enzyme is added to the reaction solution to perform an enzyme treatment.
In the present embodiment, the proteolytic enzyme to be used can be appropriately selected, and may be either an endopeptidase or an exopeptidase, or a combination thereof.
Endopeptidases include: Neutase (Novozymes), Alcalase (Novozymes), Nucleicin (Hichiai), Sumiteam MP (Shin Nihon Chemical Industry), Bromelain F (Amano Pharmaceutical), Orientase 20A (Hichiai), Morcine F (Kikkoman), Newase F (Amano Pharmaceutical), Sumiteam AP (Shin Nippon Chemical Industry), etc. are mentioned.
Examples of exopeptidases include flavorzyme (Novozymes), Sumiteam FP (Nippon Chemical Co., Ltd.), actinase (Kaken Pharmaceutical), cochlase P (Genencore) and the like. Although reaction conditions are not specifically limited, For example, it can carry out as reaction time of about 12 to 24 hours at the temperature of 50-55 degreeC. Moreover, although the usage-amount of an enzyme can also be set suitably, it can be set as about 0.1-0.5 mass% of a liver weight, for example.

  Next, the proteolytic enzyme contained in the reaction solution subjected to the enzyme treatment is inactivated. The conditions for the deactivation treatment are also not particularly limited. For example, the deactivation treatment can be performed by heating the reaction solution at 90 ° C. for about 1 hour.

  Subsequently, the reaction solution in which the proteolytic enzyme is inactivated is subjected to filtration to remove undegraded products, thereby obtaining the porcine liver degraded product of this embodiment. The obtained porcine liver degradation product can be dried and used as a powder, for example, or can be used in a liquid state. Further, for example, an excipient such as dextrin may be added by adding to a liquid liver degradation product (filtrate) before the drying treatment.

The agent for maintaining or improving the cognitive function of the present embodiment contains, for example, a porcine liver degradation product obtained as described above. The content thereof is not particularly limited and can be appropriately set by those skilled in the art. For example, the content is 30% by mass or more, more preferably 30% by mass or more and 80% by mass or less per cognitive function maintenance or improvement agent.
In addition to porcine liver degradation products, it may contain other components in an amount of 100% by mass, and includes, for example, the above-mentioned excipients, bases, lubricants, fragrances, and the like. May be.

The form (drug form) of the cognitive function maintenance or improvement agent of the present embodiment is not particularly limited, and can be produced as a pharmaceutical, a quasi-drug, a food, or the like. It can be set as the form which disperse | distributed to the form filled with the liver degradation product, or the above-mentioned pig liver degradation product in a liquid component.
In addition, the amount of the cognitive function maintenance or improvement agent of the present embodiment to be taken is not particularly limited, and can be appropriately set in consideration of the form thereof, for example, a porcine liver degradation product per hard gelatin capsule It can be made to contain 100-500 mg.
Regarding the daily dose, for example, from the viewpoint of maintaining or improving the cognitive function, the pig liver degradation product according to this embodiment is taken at 266 mg or more, preferably 790 mg or more, more preferably 1580 mg or more. can do. Further, the upper limit of the daily dose is not particularly limited, but since there is no difference in the effect even if it is taken more than this, it can be taken in an amount of 2660 mg or less.

As described above, according to the present embodiment, although there are individual differences, an agent for maintaining or improving cognitive function that can maintain or improve cognitive function by oral administration can be provided.
By taking the cognitive function maintenance or improvement agent of this embodiment containing porcine liver degradation products, it is possible to maintain or improve cognitive function to prevent dementia or maintain or improve symptoms. is there.

  In addition, since the cognitive function maintenance or improvement agent of this embodiment can maintain or improve cognitive function by oral administration, it can be administered to a subject more easily than conventional ones. It is also possible to do.

  Moreover, the cognitive function maintenance or improving agent of the present embodiment can be prepared without using an organic solvent such as acetone. Therefore, it is excellent in safety compared with conventional ones and can be taken in the form of food or the like.

  Hereinafter, the cognitive function maintenance or improvement agent of the present invention will be described in more detail by way of examples. Note that the present invention is not limited to this.

[Preparation of cognitive function maintenance or improvement agent]
750 g of liver raw material (frozen) collected from sugar pork was pulverized with a homogenizer or mince cutter until it became a paste, and 8.0 L of ion-exchanged water was added as a reaction solution.
To the 5 L reaction tank, 3.75 g of the obtained reaction solution and proteolytic enzyme (Neutrase (Novothymus)) were added, and the mixture was stirred for 15 hours at 50 ° C. and 300 rpm to carry out the enzyme reaction.
After completion of the reaction, heat treatment was performed at 90 ° C. for 1 hour to inactivate the enzyme. Next, the reaction solution was filtered to remove undecomposed matter.
Subsequently, 200 g of dextrin is added to the filtrate as an excipient, followed by spray drying (conditions: atomizer rotational speed 10,000 rpm, hot air inlet temperature 185 ° C., exhaust outlet temperature 100 ° C.), and powdered porcine liver degradation product And a vehicle mixture (609 g, pig liver degradation product: 409 g).

  Next, the obtained powdery mixture (396 mg) was filled into gelatin hard capsules, and used as an agent for maintaining or improving the cognitive function of Example 1 (the ratio of porcine liver degradation product was 266 mg / capsule).

[Reference test: Effect of porcine liver degradation products on nerve activation]
A mixture of powdered porcine liver degradation product and excipient was prepared in the same manner as in Example 1, and this mixture was used as an agent for maintaining or improving the cognitive function of the reference example (hereinafter referred to as the PLD of the reference example). Also called).
Using PLD as a reference example, the effect on nervous system cells was examined. Further, phosphatidylcholine (PC: manufactured by Wako Pure Chemical Industries) and choline chloride (C: manufactured by Wako Pure Chemical Industries) were also examined for reference. In the test for examination, the growth promotion or growth inhibitory effect of each nervous system cell was used as an index. PLD and PC were suspended or dissolved in dimethyl sulfoxide (DMSO; Nacalai Tesque), and C was suspended or dissolved in phosphate buffered saline (PBS; Wako Pure Chemical Industries, Ltd.), and adjusted to the concentrations shown in FIGS.
Rat astroglial cells (RCR-1), mouse microglia cells (MG6) and neuroblasts (IMR-32) in culture medium at 6 × 10 ⁴ cells / mL (for IMR-32, 1 × 10 ⁵ cells / mL) Then, 0.1 mL each was added to a 96-well microplate (Tlue line) and cultured in a CO ₂ incubator for 24 hours. Thereafter, PLD, PC and C prepared at the respective concentrations were added and cultured for 24 hours. After aspirating the medium, add 0.1 mL of medium supplemented with cell growth test reagent (Alamar Blue reagent, Thermo scientific) at 10%. After 2 hours, the fluorescence intensity at 530/590 nm (Ex / Em) was measured using a multiplate reader. (Day 1) measured by (BioTech). After measuring the fluorescence luminance, the reaction reagent was removed, fresh medium was added to each well, and the cells were cultured again for 24 hours. Thereafter, 0.1 mL of a cell proliferation test reagent-containing medium was added, and fluorescence intensity at 530/590 nm was measured after 2 hours (Day 2).

The results are shown in FIGS. PC and C promoted the proliferation of RCR-1 cells, but the proliferation of RCR-1 cells was suppressed in PLD. On the other hand, in MG6 cells, PLD and C promoted cell proliferation. No significant effect was observed in IMR-32 cells. These results were more prominent on Day 2 than on Day 1.
From the above results, it was suggested that the action of the PLD of the reference example differs depending on the type of nerve cell. In particular, the fact that PLD promoted the growth of MG-6 cells is expected to have effects such as nutritional supplementation to neurons by microglia cells and elimination of deposits. This was considered to be expected to improve the cognitive function of PLD.
That is, PLD does not simply show cell growth promotion or cell growth suppression but acts on the intracellular signal transmission system to regulate cell growth. In addition, it was considered that the fact that a remarkable effect was observed in Day 2 compared to Day 1 suggests the possibility of acting on the intracellular signal transmission system.

[Evaluation when porcine liver degradation product is ingested]
Evaluation in humans was performed using the cognitive function maintenance or improvement agent of Example 1 (hereinafter also referred to as PLD of Example 1). The test was conducted with the approval of the Japan Pharmaceutical University Ethics Committee. Specifically, the cognitive function of 8 persons (75.0 ± 11.2 years old) who orally administered 6 capsules of PLD daily and 5 persons (80.8 ± 8.9 years) who orally administered 3 capsules of PLD daily was evaluated.
Evaluation of cognitive function was carried out by implementing the “Hasegawa Simple Cognitive Evaluation Scale (HDR-S)”, and HDR-S score values 2 weeks and 4 weeks after the start of administration were used as an evaluation index. The collaborators in this study are those who have visited a medical institution for the sequelae of hypertension, diabetes, and stroke.
The result is shown in FIG. In the experimental group in which PLD was orally administered 6 capsules daily, the HDR-S score value before taking PLD was 25.3 ± 2.1, and an increase in score value was observed at 28.0 ± 2.1 two weeks after administration. In addition, an increase in score value of 28.0 ± 1.9 was observed even after 4 weeks. As a result of Dunnett's test, there was a statistically significant difference in the improvement of HDR-S after 2 weeks and 4 weeks after oral administration of 6 capsules of PLD daily, with a risk rate of 1% or less. In oral administration of 6 capsules daily, 6 out of 8 patients (75%) showed improvement of 2 points or more in HDR-S after 2 weeks and 4 weeks. On the other hand, when PLD was orally administered 3 capsules daily, 24.4 ± 3.8 before administration, 25.8 ± 3.0 after 2 weeks, and 24.6 ± 4.2 after 4 weeks, a statistically significant improvement in HDR-S Was not recognized. However, 2 out of 5 HDR-S improvements were observed in 3 out of 5 (60%) after 2 weeks and 1 out of 5 (20%) after 4 weeks.
That is, in Test Example 2, although there are individual differences, the cognitive function improving effect was confirmed by oral administration of the cognitive function maintaining or improving agent of Examples.

Claims (4)

  An agent for maintaining or improving cognitive function, wherein the agent is administered orally and contains a proteolytic enzyme porcine liver degradation product.   The agent for maintaining or improving cognitive function according to claim 1, wherein the porcine liver degradation product is orally administered in an amount to be taken at least 266 mg per day.   The agent for maintaining or improving cognitive function according to claim 1 or 2, wherein the porcine liver degradation product is obtained from the liver of a pig free of specific pathogens.   4. The agent for maintaining or improving cognitive function according to claim 3, wherein the specific pathogen-free pig is Sugar Pork (registered trademark).