JP2009155337A - Treating agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a treating agent comprising as an effective ingredient α-GPC (L-α-glycerylphosphorylcholine), which has been utilized, for example, as an ingredient of a skin care preparation for external use or as an ingredient of a treating agent for chronic obstructive pulmonary diseases, for other diseases than the above. <P>SOLUTION: An anti-osteoporosis agent, anti-depression agent, anti-stress agent, antihypertensive agent, antidiabetic agent, antiallergic agent, anti-fatigue agent, hair-growing agent and hair-restoring agent, each comprising as an effective ingredient α-GPC (L-α-glycerylphosphorylcholine), are provided. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a therapeutic agent useful for various diseases containing alpha-GPC as an active ingredient.

Alpha-GPC is used, for example, as a component of a skin external preparation (Patent Document 1) and as a component of a therapeutic agent for chronic obstructive pulmonary disease (Patent Document 2).
However, there is no view that alpha-GPC is effective with a significant effect on the various diseases applied in the present invention described below.

Special table 2004-536838 gazette Special Table 2002-532520

  An object of the present invention is to provide various therapeutic agents that are effective with a particularly remarkable effect on various diseases applied in the present invention described below.

The invention according to claim 1 is an anti-osteoporosis agent containing alpha-GPC as an active ingredient.
The invention according to claim 2 is an antidepressant / antistress agent containing alpha-GPC as an active ingredient.
The invention according to claim 3 is an antihypertensive agent containing alpha-GPC as an active ingredient.
The invention according to claim 4 is an antidiabetic agent comprising alpha-GPC as an active ingredient.
The invention according to claim 5 is an antiallergic agent containing alpha-GPC as an active ingredient.
The invention according to claim 6 is an anti-fatigue agent containing alpha-GPC.
The invention according to claim 7 is a hair nourishing and hair restorer containing alpha-GPC.

  According to the present invention, there are provided various therapeutic agents that are particularly effective for various diseases applied in the present invention described below with remarkable effects.

  Hereinafter, the present invention will be described in more detail.

Alpha-GPC (L-alpha-glyceryl phosphorylcholine; sn-glycero (3) phosphocholine) is a widely known compound as described in Patent Documents 1 and 2 above.
Alpha-GPC can be produced by a known chemical synthesis method, and commercially available products can also be used.

  The dose of alpha-GPC varies depending on the age, weight, indication symptoms, etc. of the patient. For example, adults are administered once to several times a day, 0.001 mg to 1 g, preferably about 0.1 mg to 500 mg per day. It is good to do.

  The therapeutic agent of the present invention can be appropriately prepared by a known method in the form of a solid or solution (hereinafter, also referred to as a preparation) such as a tablet, pill, capsule, granule, powder, powder, or liquid. That is, the solid preparation or liquid preparation useful in the present invention is produced by using a well-known preparation method that has been well established. Examples of additives include excipients, pH adjusting agents, cooling agents, suspending agents, diluents, antifoaming agents, thickeners, solubilizers, disintegrating agents, binders, lubricants, antioxidants. Agents, coating agents, coloring agents, flavoring agents, surfactants, plasticizers or fragrances.

  The therapeutic agent of the present invention can be taken as various health foods and functional foods. Examples of these include various products, but for the production of health foods and functional foods, in addition to commonly used food materials and food additives, excipients, extenders, binders, It can be used in the form of food and drink preparations using adjuvants such as disintegrants, lubricants, dispersants, preservatives, wetting agents, solubilizers, preservatives, stabilizers, capsule bases and the like. Specific examples of the adjuvant include lactose, fructose, glucose, starch, gelatin, magnesium carbonate, synthetic magnesium silicate, talc, magnesium stearate, calcium carbonate, methylcellulose, carboxymethylcellulose, or a salt thereof, gum arabic , Polyethylene glycol, syrup, petrolatum, glycerin, ethanol, propylene glycol, citric acid, sodium chloride, sodium sulfite, sodium phosphate, pullulan, carrageenan, dextrin, reduced palatinose, sorbitol, xylitol, stevia, synthetic sweetener, citric acid Ascorbic acid, acidulant, baking soda, sucrose ester, hardened vegetable oil, potassium chloride, safflower oil, beeswax, soybean lecithin, flavor, and the like can be blended. Regarding the production of such health foods and functional foods, reference books for pharmaceutical preparations such as “Japanese Pharmacopoeia Manual (General Rules for Preparations)” (Yodogawa Shoten) can be referred to.

  In addition to the above, the therapeutic agent of the present invention can be taken as a food or drink. Specifically, natto, thick fried, tofu, konjac, dumplings, pickles, boiled, croquettes, sandwiches, pizzas, hamburgers, dumplings, shumai, salads, etc., various powders (beef, pork, chicken and other livestock products) Marine products such as shrimp, scallops, sea bream, kelp, vegetables / fruits, plants, yeast, algae, etc.), pudding, cookies, crackers, bread, cakes, chocolate, potato chips, biscuits, donuts, jellies and other confectionery, Japanese rice crackers such as rice crackers, sheep crab, daifuku, ohagi, other buns, Japanese confectionery such as castella, frozen confectionery (rice cake etc.), bread and confectionery such as chewing gum, noodles such as udon, buckwheat and kishimen, kamaboko, ham, fish sausage, etc. Fish paste products, meat products such as ham, sausage, hamburger, corn beef, salt, pepper, miso, soy sauce, sauce, dressy Seasonings such as gyu, mayonnaise, ketchup, sweeteners, spices, teppanyaki foods such as Akashi-yaki, takoyaki, monja-yaki, okonomiyaki, yakisoba, fried udon, dairy products such as cheese, hard-type yogurt, and fats and oils Various foods such as powdered solids of flavors and fragrances (vanilla, citrus fruits, bonito etc.) and powdered foods and drinks (instant coffee, instant tea, instant milk, instant soup, miso soup, etc.) These are not particularly limited.

  Furthermore, in the present invention, for example, royal jelly, propolis, vitamins (A, C, D, E, K, folic acid, pantothenic acid, biotin, derivatives thereof, etc.), minerals (iron, magnesium, calcium, zinc, etc.), Selenium, lecithin, carotenoids (lycopene, astaxanthin, zeaxanthin, lutein, etc.), saponins (gymnemic acid, soybean saponin, ginseng saponins, etc.), fatty acids, proteins (collagen, elastin, etc.), oligosaccharides (isomalto-oligosaccharides, cyclic oligosaccharides, etc.) ), Phospholipids and derivatives thereof (sphingomyelin, ceramide, etc.), sulfur-containing compounds (allyin, sepaene, taurine, glutathione, methylsulfonylmethane, etc.), sugar alcohols, lignans (sesamin, etc.), animal and plant extracts containing these , Root vegetables (turmeric, show Etc.), it can be used in combination, and the like.

In addition, when alpha-GPC is used as an external preparation for skin, known materials that are usually used as external preparations for skin, such as pigments, fragrances, preservatives, surfactants, pigments, antioxidants, humectants, ultraviolet absorbers Etc. can be appropriately blended.
The external preparation for skin of the present invention can be used in a known form such as cream, milky lotion, lotion, pack and the like. The blending ratio of alpha-GPC is not particularly limited, but is, for example, 0.0001% by mass to 5% by mass, and preferably 0.001% by mass to 1% by mass in the external preparation for skin.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further, this invention is not limited to these. The therapeutic agent of the present invention is extremely useful as an anti-osteoporosis agent, an antidepressant / anti-stress agent, an antihypertensive agent, an anti-diabetic agent, an anti-allergic agent, an anti-fatigue agent, an external preparation for skin, and a hair restorer / hair restorer. Hereinafter, the various medicinal effects will be described with examples.

Example 1 (Anti-osteoporosis effect)
Osteoporosis Improvement Effect Test SD rat (22 weeks old) Female ovaries were surgically removed to create osteoporosis model rats. Ovariectomized rats were divided into 6 groups of 7 animals, and orally administered a liquid dissolved in physiological saline so that alpha-GPC was 40 mg / kg every other day (total 17 times) during the 35-day test period. did. The feed was the solid feed CRF-1 for mice, rats and hamsters from Oriental Yeast Co., Ltd. There was no difference in food intake between groups during the study period. On the 35th day after the start of the test, the weight of the rat was measured, and then the femur was removed. The femur was used for analysis with the adhesive tissue and muscle removed. After measuring the volume of the femur, it was washed three times with ethanol, then washed three times with acetone, dried overnight, and then weighed to determine the dry weight of the femur. From the volume and dry weight, the bone density (dry weight g / volume mm ³ ) was measured. In addition, as a control experiment, an example (comparative example) in which the above experiment was repeated was performed except that saline containing no alpha-GPC was administered to the rat. As a result, the bone density of the comparative example was 1.011 mg / mm ³ , whereas the bone density of Example 1 was 1.098 mg / mm ³ .

Example 2 (Antidepressant / Anti-stress effect)
The therapeutic effect of alpha-GPC was examined.
Evaluation of tranquilizing effect by forced mouse swimming test The therapeutic agent of the present invention was evaluated by the forced mouse swimming test developed by Porsolt in 1977. This is one of the most frequently used animal model experiments for depression. In this test, the mouse is forced to swim in a limited space to cause “immobility”. This immobility state is thought to reflect a kind of “despair state” in which stressed animals abandon their escape from water, and is associated with depression and stress in humans. In fact, antidepressants have been found to specifically reduce the duration of immobility in this situation, and this shortening has been found to have a significant correlation with clinical titer.

The test method is as follows.
Mice are forced to swim in a plastic cylinder containing 25 ° C. water to a depth of 15 cm. After forced swimming for 5 minutes, dry in a dryer at 30 ° C. for 15 minutes and return to the home cage. On the next day, the test sample is intraperitoneally administered to the mouse, one hour later, forced swimming for 5 minutes is imposed again, and the duration of the immobility that appears is measured using a stopwatch. A state in which the mouse floats on the water and is stationary is determined as an immobile state. For the duration of stationary state, a significant difference test is performed, and a statistically significant difference is tested. For the experiment, male ddY mice are used, and there are 6 mice per group. All tests are conducted between 1pm and 6pm. A test using imipramine, an antidepressant, as a positive control will also be conducted.

  As a result, the duration of immobility in mice administered with 30 mg / kg of alpha-GPC was 178.8 ± 2.8 seconds. The control (saline only) was 209.0 ± 3.2 seconds. The duration of immobility in the positive control (30 mg / kg dose) mice was 180.1 ± 5.6 seconds. The duration of immobility in this example and the positive control is significantly different at a risk rate of 1%.

Example 3 (blood pressure lowering effect)
Alpha-GPC was added to a general commercial feed (Funabashi Farm, Funabashi SP), and the changes in systolic blood pressure and body weight were compared using stroke-prone spontaneously hypertensive rats (SHR-SP). As a control group, a general sample to which no alpha-GPC was added was used. The A group is the control group and the B group is the present invention group, and each group of 6-week-old male SHR-SPs is bred for 7 weeks, and changes in blood pressure and body weight when they reach 12 weeks of age. Investigated about. As shown in Table 1, in the change of blood pressure, significant suppression of blood pressure increase was observed in the present invention group. In the present invention group, the concentration of alpha-GPC in the feed was adjusted so that the daily intake of alpha-GPC was 50 mg / kg body weight.

Example 4 (anti-diabetic effect)
Diabetes was induced by administering streptozotocin once to the tail vein of 6-week-old male SD rats (6 rats per group).
The dose of alpha-GPC was 30 mg / kg, and it was orally administered 1 hour before the administration of streptozotocin (STZ). From the next day, it was orally administered once a day for 13 consecutive days. The day after the final administration, 50% glucose aqueous solution (10 ml / kg) was orally administered, and blood glucose level (mg / dl) was measured over time (o-toluidine / boric acid).

  In addition, as a normal control group, a group in which only sterile water was administered without administering STZ, a group in which STZ was administered and sterilized water was administered as a disease state control group, and STZ was administered as a positive control group and nicotinamide ( 50 mg / kg) was provided. Nicotinamide has been reported to be effective against STZ diabetes models (Animal utilization for new drug development, 419-422, R & D planning, 1985).

  Diabetes is a disease in which the ability to metabolize glucose is reduced and hyperglycemia occurs. In this example, a peak of blood glucose level was observed 1 hour after glucose administration, but the maximum blood glucose level was 338 mg / dl in the pathological control group, and the maximum blood glucose level was 165 mg / dl in the normal control group. The maximum blood glucose level of the pathological condition control group was about twice that of the normal control group, and a decrease in glucose metabolism ability was observed in the pathological condition control group.

  With respect to the activity of alpha-GPC, the inhibition rate (%) with respect to the blood glucose level of the pathological condition control group was calculated by Formula 1.

(Formula 1)
Inhibition rate (%) = [1- (high blood sugar level of alpha-GPC administration group or positive control group−high blood sugar level of normal control group) / (high blood sugar level of disease state control group−high blood sugar level of normal control group)] ] × 100

  As a result, the inhibition rate in the alpha-GPC administration group was 55.5%. The suppression rate of the positive control group was 43.0%. Therefore, in the alpha-GPC administration group, an excellent decrease in blood glucose level was observed and the glucose metabolism ability was improved as compared with the disease state control group.

Example 5 (anti-allergic effect)
Twenty patients (10 males and 10 females aged 20-22 years) with food allergen positive chronic urticaria by the RAST method were administered alpha-GPC at a dose of 500 mg for 1 month. The results are shown in Table 2 below.

Example 6
20 patients with atopic dermatitis by RAST method (10 men and 10 women aged 20 to 22 years) were administered 500 mg of alpha-GPC together with 1 meal for 1 month. The results are shown in Table 3 below.

Example 7 (anti-fatigue effect)
STD DDY male mice (5 weeks old: each group n = 3 to 4) were orally ingested with alpha-GPC. The intake is 50 mg / kg body weight. Ingestion was performed using a solution in which alpha-GPC was dissolved in pure water. In addition, the test group mice were fed with pure water only.

Thirty minutes after the ingestion, the mouse was placed in an 80 cm deep water tank, and the time until immobility was measured. As a mean value of time until immobilization of mice in each test group (each group n = 3 to 4), the control group was about 112 seconds, whereas the alpha-GPC administration group was about 250 seconds. Met.
From the above, a high anti-fatigue effect was confirmed for alpha-GPC.

Reference example 1 (melanin suppression effect)
As cells producing melanin, cultured B16 melanoma cells derived from mice were cultured in Eagle's MEM medium supplemented with fetal bovine serum to a final concentration of 10%, and the cells were cultured at a concentration of 3 × 10 ³ cells / ml. Inoculate 6 ml of each well of a 6-well plate, and after culturing in a CO ₂ incubator for 5 days, replace with a medium supplemented with alpha-GPC, and further culture under the same conditions for 3 days. After washing the cells, the cells are peeled off by a scraper treatment, solubilized with sodium dodecyl sulfate (SDS), and the absorbance at 475 nm and 260 nm is measured to obtain S ₄₇₅ and S ₂₆₀ . The melanin inhibition rate was calculated by Equation 1 with the absorbance at 475 nm and 260 nm of cells cultured in a medium not added with the test sample as C ₄₇₅ and C ₂₆₀ . As a positive control, kojic acid was used.

  As a result, the melanin inhibition rate of the positive control (addition of 3 mM kojic acid in the medium) was about 56%, and the melanin inhibition rate of the test sample to which alpha-GPC was added (500 μg / ml of alpha-GPC was added to the medium). Was about 56%.

Reference example 2 (wrinkle formation inhibitory effect)
A wrinkle formation model was prepared by irradiating UVB (long wavelength ultraviolet rays) for 10 weeks on the back of two groups of hairless mice, each of 5 mice. Thereafter, one group of this wrinkle formation model contains a liquid obtained by dissolving alpha-GPC in a 10% ethanol aqueous solution at a ratio of 0.5% by mass (Example 1), and the other group contains only a 10% ethanol aqueous solution ( Comparative Example), the coating was continued once a day on the back where wrinkles were formed at a rate of 5 days per week for 4 weeks. After the application period of 4 weeks, replicas were collected from the back of each model, and wrinkle scores were assigned to each of the obtained replicas according to the following criteria.
(Wrinkle score standard)
0: Directional structure is not recognized.
1: A fibrous thin structure is recognized with directionality.
2: A thick rod-like structure is recognized together with a fibrous thin structure having directionality.
3: A thick rod-like structure with directionality is observed.

  The evaluation using the wrinkle score is an evaluation by defining a linear structure with directionality on a replica as a wrinkle. Therefore, according to the wrinkle score standard, the higher the score, the better the skin state where wrinkle formation has progressed. When the average value of each group was computed about the wrinkle score obtained above, the reference example 2 was 0 and the comparative example was 1.0. When the wrinkle score of both groups was subjected to a significant difference test by Mann-Whitney test, the wrinkle score of Example 9 was significantly smaller than the wrinkle score of the comparative example at a risk rate of 0.05 or less.

Reference example 3
A cream having the following composition containing alpha-GPC was prepared by a conventional method.
Alpha-GPC 10 parts by weight Glycerol sorbitan fatty acid ester 60 parts by weight Microcrystalline wax 10 parts by weight Olive oil 30 parts by weight Liquid paraffin 180 parts by weight Magnesium stearate 10 parts by weight Propylene glycol 37 parts by weight Magnesium sulfate 7 parts by weight Purified water 655 parts by weight Part

Reference example 4
About the cream of the said reference example 3, the wrinkle formation inhibitory effect was investigated. That is, using a paneler worried about wrinkles, the cream of Reference Example 4 was applied to the right side of the face twice a day in the morning and evening for 1 month every day, and the improvement of the right side wrinkles with respect to the left side was improved. +: Obviously improved, ±: Slightly improved,-: Evaluation was made based on no improvement. As a result, all the panelists evaluated it as very improved.

Example 8 (hair growth and hair growth effect)
According to the experimental procedure described in Japanese Patent No. 4041451, the effect of alpha-GPC was verified.
(1) Mice test C3H mice (8 weeks old, male, average body weight 35 g) dorsal skin (2 cm × 4 cm) were cut with an electric clipper and a shaver, and each sample of the examples was applied to the test subject skin twice a day in the morning and evening. Then, 0.2 mL per animal was applied continuously for 2 weeks. A group of 10 animals was used for one sample. On the 22nd day from the start of application, the skin of the test area of each sample was photographed with a video camera, and the areas of the hair cutting part and the hair growth part were measured with an image analyzer. The determination of the hair nourishing effect was carried out by calculating the hair growth rate (%) shown below, and calculating the average hair growth rate of each group in the examples for comparison.
Hair growth rate (%) = (Area of hair growth part) / (Area of hair cutting part) × 100

(2) Human test At 10 cm above the ears of 10 subjects with male pattern baldness, the hair was shaved into a circular shape with a diameter of 5 mm at two locations on the left and right, and each sample of the example was placed on the entire left hair. About 3 mL was applied twice daily in the morning and evening, and the right and left sides were compared without applying anything on the right side. In the determination of the effect of the hair growth rate, the hair growth rate was evaluated with the value obtained by the following formula after removing 20 test part hairs at the left and right shaved sites 1 month after the start of the test.
Hair growth rate = (average length of 20 left hairs) ÷ (average length of 20 right hairs)

  In addition, the hair growth effect, the itch prevention effect, the hair loss effect, and the anti-dandruff effect were determined for each item 3 months after the start of the test, compared to the test site where nothing was applied on the right side. It was shown by the number of respondents who responded that they became bristle or increased their vellus hair, “doesn't feel itchy much”, “has lost hair loss”, and “had less dandruff”.

  A hair nourishing and hair restorer with the formulation shown in Table 6 was prepared according to a conventional method, and the above tests were conducted for evaluation. The concentration of alpha-GPC in the sample is as shown in Table 6. The results are also shown in Table 6.

  It should be noted that no abnormalities were observed in the skin condition even during and after the long-term use of the hair nourishing / hair-growing agent of the present invention.

Claims (7)

  Anti-osteoporosis agent containing alpha-GPC as active ingredient   An antidepressant / anti-stress agent containing alpha-GPC as an active ingredient.   An antihypertensive agent comprising alpha-GPC as an active ingredient.   An anti-diabetic agent containing alpha-GPC as an active ingredient.   An antiallergic agent containing alpha-GPC as an active ingredient.   Anti-fatigue agent containing alpha-GPC.   A hair nourishing agent containing alpha-GPC.