JPH08198754A - Agent for improving cerebral function - Google Patents

Abstract

PURPOSE: To obtain a cerebral function improving agent containing phosphatidyl- L-serine originated from the brain of an animal except a cattle, capable of being massively produced at a low cost, without a problem on the aspects of the cost and supply, and capable of improving the cerebral functions. CONSTITUTION: This improving agent contains phosphatidyl-L-serine originated from the brain of an animal (e.g. pig, sheep, chicken) except cattle or its salt as an active ingredient. The salt of the phosphatidyl-L-serine is preferably Na salt or K salt. The phosphatidyl-L-serine is obtained e.g. by extracting a swine brain or an ovine brain with acetone, filtering the filtrate, extracting the residue with petroleum ether, vacuum-drying the extract, dissolving the dried product in ethyl ether, adding ethyl alcohol to the solution to produce precipitates, dissolving the precipitates in chloroform, and subsequently purifying the solution with silica gel column chromatography.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for improving brain function, for example, a composition for improving brain function, which is a composition effective for preventing or treating dementia such as Alzheimer's disease and Parkinson's disease.

[0002]

2. Description of the Prior Art Bruni et al. Have reported that when phosphatidylserine extracted from bovine brain is injected into the tail vein of mice, the glucose concentration in the brain is increased by about 4 times that in the control group. (Nature, vol.260, p.331,1976). It has also been reported that the oral administration of this phosphatidylserine extracted from bovine brain for 12 weeks improves the behavior of aged rats showing a decrease in memory.
(A.Zanotti et al., Psychopharmacology Berl., Vol.99,
p316, 1989).

Further, in a human clinical study, it was reported in a double-blind-placebo study that bovine brain phosphatidylserine is effective for Alzheimer's disease and memory loss in old age (PJ Delwaide et al., Acta Neurol. .Scand., Vol.7
3, p. 136, 1986; RR Engel et al., Eur. Neuropsychophar
macol., vol.2, p.123, 1993).

As described above, bovine brain phosphatidylserine showing an increase in brain glucose concentration shows an effect of improving brain function in rats and humans. Therefore, an increase in glucose concentration in the brain is considered to be an index in selecting a substance having an effect on improving brain function, while on the other hand, according to the article by Bruni et al. Mentioned earlier, phosphatidylserine extracted from soybean has such an effect. It has been reported that the fatty acid composition in phosphatidylserine is important for the expression of the effect of improving brain function.

That is, the phosphatidylserine extracted from bovine brain has a very specific fatty acid chain structure in which the constituent fatty acid chains are mostly stearic acid chain at the 1-position and oleic acid chain at the 2-position. Therefore, it was considered that having this unique constituent fatty acid chain is necessary for the expression of the improving action of brain function.

[0006] On the other hand, recently, a phosphatidylserine having a specific fatty acid composition obtained by a synthetic method is expected to be used for senile dementia based on the activating action of protein kinase C isozyme. However, it has not been confirmed to be effective in vivo (Japanese Patent Laid-Open No. 6-279311).

On the other hand, with respect to lyso-type phosphatidylserine, it has been reported that lysophosphatidyl-L-serine, which is derived from bovine brain phosphatidyl-L-serine, exhibits an action of increasing glucose in the brain or blood.
(HW Chang et al., Br.J.Pharmacol., Vol.93, p.611,198
8).

[0008] As described above, it has been conventionally considered that phosphatidylserine extracted from bovine brain exhibits an action of increasing glucose in the brain based on its characteristic composition of constituent fatty acids.

[0009]

As described above, in the conventional literatures and the like, only the phosphatidylserine extracted from bovine brain or its lyso form has been known to have an action of increasing glucose in the brain. However, since only about 1 g of phosphatidylserine extracted from bovine brain can be obtained from the brain of one bovine, it is obvious that there are significant restrictions in terms of cost and quantitative supply.

As a result of earnest studies, the present inventors have found that, in addition to phosphatidylserine derived from bovine brain, phosphatidyl-L derived from brains of animals other than bovine, meat or fish meat, viscera, plants, or microorganisms. The present invention has been completed by discovering that serine also has a brain function improving effect (memory disorder recovery effect), and clarified that phosphatidylserine derived from sources other than bovine brain can also be used as an anti-dementia substance.

That is, an object of the present invention is to obtain a brain function improving agent which has no problem in terms of cost and supply and can improve brain function.

[0012]

In the brain function improving agent according to the invention described in claim 1, phosphatidyl-L-serine or a salt thereof derived from the brain of an animal other than bovine is used as an active ingredient. .

In the brain function improving agent according to the invention described in claim 2, the animal other than the bovine described in claim 1,
They are pigs, sheep and chickens.

In the brain function improving agent according to the invention described in claim 3, phosphatidyl-L-serine derived from cottonseed or a salt thereof is used as an active ingredient.

In the brain function improving agent according to the invention described in claim 4, yeast-derived phosphatidyl-L-serine or a salt thereof is used as an active ingredient.

In the brain function improving agent according to the present invention as defined in claim 5, phosphatidyl-L-serine or a salt thereof derived from any one of chicken, chicken internal organs, fish body and fish meat is used as an active ingredient.

[0017]

The brain function-improving agent of the present invention contains phosphatidyl-L-serine or a salt thereof derived from the brain of an animal other than bovine as an active ingredient, and therefore can increase the brain glucose concentration. Thus, it has the effect of improving brain function in the administered subject. Specific animals other than cattle include pigs, sheep and chickens.

In another brain function improving agent of the present invention, phosphatidyl-L-serine derived from cottonseed or its salt is used as an active ingredient, and phosphatidyl-L-serine derived from yeast or its salt is used as an active ingredient. In addition, since phosphatidyl-L-serine or a salt thereof derived from any one of chicken, chicken internal organs, fish bodies, and fish meat is used as an active ingredient, the glucose concentration in the brain can be similarly increased. Thus, it has the effect of improving brain function in the administered subject.

Specifically, the chicken internal organs include chicken liver. Examples of fish bodies and meat include fish bodies such as sardines, tuna, mackerel, etc., fish meat, mixed meat, squeezed lees of fish oil, viscera of fish, and residues obtained by squeezing liver oil from viscera of fish.

As described above, in the brain function improving agent according to the present invention, phosphatidyl-L-serine derived from the brain of animals other than bovine, meat or fish, fish meat, viscera, plant origin, or microorganism origin, or a derivative thereof. It uses salt as an active ingredient. This phosphatidyl-L-serine other than bovine brain has a fatty acid chain composition described later, and the relationship between the fatty acid chain composition and the brain function improving effect (effect of increasing brain glucose concentration) is not always clear. At least within the range where the effect was confirmed in the examples described later, it was confirmed to be effective.

The phosphatidyl-L-serine salt other than bovine brain of the present invention may be used in the form of a pharmaceutically acceptable salt. Specifically, there are sodium salt, potassium salt, magnesium salt, ammonium salt, phosphate, hydrochloride, sulfate and the like, but sodium salt and potassium salt are preferable.

Furthermore, the administration of the phosphatidyl-L-serine of the present invention other than bovine brain or a salt thereof is effective both intravenously and orally. Further, other excipients such as lipids, sugars, proteins and the like may be mixed and processed into capsules or granules with improved easiness of handling and preservability. Furthermore, since there is no problem in terms of safety, it can be incorporated into foods and drinks that are ingested daily and used for reducing or preventing brain dysfunction.

The phosphatidyl-L-serine other than bovine brain or a salt thereof according to the present invention is phosphatidyl-L-serine or a salt thereof by a purification operation using a conventional organic solvent from various animal tissues as raw materials, cottonseed lecithin, yeast and the like. L-serine can be produced. Various animal tissues used as raw materials,
Cottonseed lecithin, yeast, etc. can be provided in a much larger amount and at a lower cost than bovine brain, and there are few quantitative problems in terms of supply.

It is desirable that various animal tissues used, cottonseed lecithin, and yeast-derived phosphatidyl-L-serine are subjected to an appropriate purification treatment step to remove impurities, but do not cause problems or effects in administration. As long as there is no problem that hinders it, it may be used while containing the impurities derived from the raw materials or impurities in the production process.

[0025]

【Example】

Example 1. Purification of Phosphatidyl-L-serine from Animal Tissue Purification of phosphatidyl-L-serine from animal tissue is
"Shinsei Chemistry Laboratory 4 Lipid II Phospholipid (Tokyo Kagaku Dojin, 1991) 8.3 Phosphatidylserine (Hiroyoshi Arai)" was used as a reference. 200 minced animal tissues (porcine brain, sheep brain, chicken, chicken liver, sardine fish, mackerel blood)
60 ml of acetone was added per g and the mixture was homogenized with a Waring blender. To this, 200 ml of acetone was added, and the supernatant was removed by suction filtration.
After washing with l of ethanol, the lipid was extracted with 800 ml of petroleum ether with stirring overnight. The extract is dried under reduced pressure,
It was dissolved in 10 ml of ethyl ether, 100 ml of ethyl alcohol was gradually added thereto, and the precipitate was collected by suction filtration. The recovered precipitate was dried under reduced pressure, and 5 g of the precipitate was dissolved by adding chloroform to the column (φ32, packed with silica gel (Silica gel 60, MERCK)).
Fractions containing phosphatidyl-L-serine (mm × 300 mm) were collected. As a result of analysis by silica gel thin layer chromatography, the phosphatidyl-L-serine content of each purified product was 90% or more.

Example 2. Purification of phosphatidyl-L-serine from cottonseed lecithin Purification of phosphatidyl-L-serine from cottonseed is also described in "Shinsei Chemistry Laboratory Course 4 Lipid II Phospholipids (Tokyo Kagaku Dojin,
1991) 8.3 Phosphatidylserine (Hiroyuki Arai) ”. Chloroform was added to 5 g of cotton seed lecithin and dissolved to give silica gel (Silica gel 60, MERCK
Purification was carried out using a column (φ32 mm × 300 mm) packed with a product (manufactured by K.K.) and a fraction containing sfatidyl-L-serine was collected. As a result of silica gel thin layer chromatography analysis, the content of sfatidyl-L-serine in each purified product was 90%.
That was all.

Embodiment 3 FIG. Composition of fatty acid chain of various phosphatidyl-L-serine The composition of fatty acid chain of various phosphatidyl-L-serine obtained in Examples 1 and 2 was analyzed. Specifically, the methyl esterified sample was analyzed by capillary GLC according to "Biochemistry Experimental Method 9 Introduction to Lipid Analysis Method (Yasuhiko Fujino, Academic Society Publishing Center)". The results are shown in Table 1 below. In the table, "PS" is phosphatidyl-L-serine, "16: 0" is palmitic acid, "18: 0" is stearic acid, "18: 1" is oleic acid, and "18: 2" is linole. acid,
“20: 4” indicates arachidonic acid.

[0028]

[Table 1]

As shown in Table 1, most of the fatty acids of phosphatidyl-L-serine extracted from bovine brain are stearic acid and oleic acid. On the other hand, the fatty acids of phosphatidyl-L-serine derived from pig brain and sheep brain have the same constitution, and the constitution in which stearic acid and oleic acid account for the majority constitutes the fatty acid peculiar to the animal brain itself. Was inferred.

However, the fatty acids of phosphatidyl-L-serine derived from chicken, chicken liver, mixed mackerel meat, and cottonseed yeast are:
It has been found that the fatty acid composition of phosphatidyl-L-serine from bovine and other animal brains and from yeast is quite different.

Phosphatidyl-L derived from chicken and liver
In the fatty acids of serine, a large amount of linoleic acid is used, and in the fatty acids of phosphatidyl-L-serine derived from mackerel hemp meat, fatty acids other than palmitic acid, stearic acid, oleic acid, linoleic acid, and arachidonic acid account for the majority. In addition, the fatty acids of phosphatidyl-L-serine derived from cottonseed are similar to those derived from chicken liver, and most of them are linoleic acid and palmitic acid. Furthermore, the fatty acids of yeast-derived phosphatidyl-L-serine are mostly palmitic acid and oleic acid.

Example 4. Improvement of scopolamine-induced memory impairment Each group of 10 male rats (SD strain, body weight about 300g) was given 1.0ml / kg of scopolamine solution (3.0mg / ml buffer) and various PS solutions (60mg / ml buffer). It was administered intraperitoneally, and 20 minutes after the administration, it was placed in the light room of a rat step-through cage (Muromachi Kikai Co., Ltd.), and after about 10 seconds, the door separating the light room and the dark room was opened and the rat entered the dark room. Immediately after that, electrical stimulation (4 mA, 100 V, direct current) for 2 seconds was given. Then, 24 hours after the administration, the rat was placed in the light room again, and the time until the extremities entered the dark room (reaction latency) was measured up to 5 minutes. It is judged that the longer this reaction latency is, the better the memory of the experience of receiving the electric stimulation.

[0033]

[Table 2]

As shown in Table 2, in the control group (administered the buffer solution), the reaction latency of all individuals was 5 minutes or more, whereas in the scopolamine-only administration group, 9 out of 10 animals were detected. The individual entered the dark room within 5 minutes, and the induction of memory impairment was confirmed. On the other hand, in the case where both scopolamine and PS were administered, all the individuals remained in the dark room, and PS extracted from other than bovine brain also had an anti-dementia effect (improvement effect on scopolamine-induced memory impairment) similar to bovine brain PS. It was confirmed. The same anti-dementia effect was also confirmed in these PS lysoforms.

As described above, the finding of the present invention that other than phosphatidyl-L-serine extracted from bovine brain has an effect of improving brain function, (1) phosphatidyl-L-serine effective for improving brain function is obtained. Since it can be taken orally, continuous ingestion has become possible without pain. (2) Phosphatidyl-L-serine, which is effective in improving brain function, can now be manufactured in large quantities at low cost from raw materials that are inexpensively available.

[0036]

Industrial Applicability As described above, the present invention uses phosphatidyl-L-serine or a salt thereof derived from the brains of animals other than bovine as an active ingredient, so that the glucose concentration in the brain can be increased. Thus, it has the effect of improving brain function in the administered subject. Specific animals other than cattle include pigs, sheep and chickens.

In another brain function improving agent of the present invention, phosphatidyl-L-serine derived from cottonseed or its salt is used as an active ingredient, and phosphatidyl-L-serine derived from yeast or its salt is used as an active ingredient. Furthermore, since the active ingredient is phosphatidyl-L-serine or its salt derived from any one of chicken, chicken internal organs, fish bodies, and fish meat, the glucose concentration in the brain can be similarly increased. Thus, it has the effect of improving brain function in the administered subject.

As described above, in the brain function-improving agent of the present invention, phosphatidyl-L-serine other than bovine brain can be provided in a much larger amount and at lower cost than bovine brain, and can be supplied. There are few quantitative problems.

Claims (5)

[Claims] 1. A brain function improving agent comprising phosphatidyl-L-serine or a salt thereof derived from the brains of animals other than bovine as an active ingredient. 2. The brain function improving agent according to claim 1, wherein the animal other than the bovine is pig, sheep or chicken. 3. A brain function improving agent, which comprises phosphatidyl-L-serine derived from cottonseed or a salt thereof as an active ingredient. 4. A brain function improving agent comprising yeast-derived phosphatidyl-L-serine or a salt thereof as an active ingredient. 5. A brain function improving agent comprising phosphatidyl-L-serine or a salt thereof derived from any one of chicken meat, chicken internal organs, fish bodies and fish meat as an active ingredient.