JP2008239505A - Neuroblast growth promoter and neurite extender - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new neuroblast growth promoter or neurite extender which promotes the neuroblast growth or the neuroblast neurite extension, promotes the change of neuroblast to nerve cell even in a state that the neuroblast is not damaged, thereby effectively prevents the ageing of brain, and further improves the functions of the brain. <P>SOLUTION: This neuroblast growth promoter is characterized by containing an extract of a Cistanche plant or a phenylethanoid glycoside as an active ingredient. It is preferable to contain at least one of echinacoside and acteoside or both the echinacoside and the acteoside as the phenylethanoid glycoside. The neuroblast growth promoter and the neurite extender can be used as medicines and external preparations for skins for mammals including human being, foods, drinks, and feeds for mammals. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a neuroblast proliferation promoter and neurite extension agent using a novel component. The present invention is widely used for medicines for mammals including humans, external preparations for skin, foods and the like.

  Conventionally, it has been known that fresh stalks of herbs belonging to the family Cistanche are effective in treating infertility, impotence, constipation, and the like (see Patent Document 1). In addition, preparations obtained from fresh perennial herb stems provide blood and kidney nutrition. These parasites and perennial herbs are widely cultivated in the northwestern region of China and are known in some regions as “desert ginseng”. The most cultivated species of Cistanche are all parasitic plants [Cistanche tubulosa].

  Japanese scientists have systematically studied the chemical constituents and pharmacological activities of Cistanche, and found that phenylethanoid glycosides are the main active ingredients in these perennial herbs (eg, Non-patent documents 1 to 3). Such active ingredients are effective antioxidants, metabolism promoters, memory promoters, libido promoters and the like. The medical properties of various phenylethanoid glycoside compounds have been studied by many researchers.

Further, according to Patent Document 1, it is reported that when a phenylethanoid glycoside is administered to a mouse or a rat that has caused some kind of disorder in the brain, the brain function is recovered.
Furthermore, it has been reported that when a phenylethanoid glycoside is added to a neuroblast that has been damaged by the addition of TNFα, the neurite outgrowth returns to normal (ie, almost before the cell is damaged). (Non-Patent Document 4).

JP 2004-250449 A Sato T., et al. Yakugaku Zasshi, 1985, 105 (12): 1131 Jimenez C., et al. Nat Prod Rep, 1994, 11 (6): 591 Cometa F., et al. Fitoterapia, 1993, 64 (3): 195 Deng Min et al. Acta Pharmacoe Sin, 2004, 25 (10) 1276-1284

However, Patent Document 1 has found that brain function is restored by administering a phenylethanoid glycoside in the event of a brain function disorder, but brain function disorder occurs. It has never been described before that brain function is improved.
Further, according to Non-Patent Document 4 described above, there is a description that neurites are extended and brain function is restored by adding phenylethanoid glycoside after causing damage to neuroblasts. However, when phenylethanoid glycosides are added in a state where the neuroblasts are not damaged, it is known that the neuroblasts proliferate and promote the neurite outgrowth of the neuroblasts. It is not done.
Under such a background, the present inventors added a phenylethanoid glycoside compound or an extract of a plant of the genus Agrocyceae containing it to an uninjured neuroblast (SK-N-SH). It has been found that the proliferation of neuroblasts is promoted, and further, the extension of neurites of neuroblasts is promoted, and the present invention has been completed.
That is, the present invention promotes the proliferation of neuroblasts and promotes neurite outgrowth of neuroblasts even when the neuroblasts are not damaged. It promotes the change to nerve cells, and even when nerve cell degeneration occurs, it effectively prevents the decrease of nerve cells, thereby effectively preventing aging by the brain, Another object of the present invention is to provide a novel neuroblast proliferation promoter and neurite extension agent that can further improve brain function.

In order to solve the above-mentioned problems, the neuroblast growth promoter of the first invention is characterized by comprising phenylethanoid glycoside as an active ingredient.
The phenylethanoid glycoside preferably contains at least one of echinacoside and acteoside.
Further, the phenylethanoid glycoside preferably contains both echinacoside and acteoside.
The neuroblast growth-promoting agent of the second invention is characterized by containing an extract of a plant of the genus Oleaceae as an active ingredient.
Moreover, it is preferable that the said extract contains a phenylethanoid glycoside as an active ingredient.
Further, the extract preferably contains at least one of echinacoside and acteoside as the phenylethanoid glycoside.
Moreover, it is more preferable that the extract contains both echinacoside and acteoside as the phenylethanoid glycoside.
The neurite extender of the third invention comprises phenylethanoid glycoside as an active ingredient.
The neurite extender preferably contains at least one of echinacoside and acteoside as the phenylethanoid glycoside.
Furthermore, it is more preferable that the phenylethanoid glycoside contains both echinacoside and acteoside.
The neurite extender of the fourth invention is characterized by comprising an extract of a plant of the genus Oleaceae as an active ingredient.
Moreover, it is preferable that the said extract contains a phenylethanoid glycoside as an active ingredient.
Further, the extract preferably contains at least one of echinacoside and acteoside as the phenylethanoid glycoside.
Moreover, it is more preferable that the extract contains both echinacoside and acteoside as the phenylethanoid glycoside.

The neuroblast proliferation-promoting agent of the first and second inventions promotes the proliferation of neuroblasts even in a state where no damage has occurred in the neuroblasts. Can be promoted. Thereby, functions, such as memory power, calculation power, and judgment power, can further be improved. In addition, even when nerve cells degenerate due to aging or the like, brain functions can be maintained by nerve cells that have changed from neuroblasts. Thereby, cerebral disorders such as aging of the brain and dementia can be effectively prevented.
In addition, the neurite extender of the third and fourth inventions promotes neurite extension even when the neuroblast is not damaged. Thereby, since more synapses can be constructed with respect to a normal brain, the function of the brain can be further improved. In addition, even when neuronal degeneration occurs due to aging or the like, it is possible to quickly construct a synapse for supplementing the function, so that the function of the brain can be maintained. Thereby, cerebral disorders such as aging of the brain and dementia can be effectively prevented.

Hereinafter, the present invention will be described in detail.
The “neuroblast proliferation promoter and neurite extension agent” (hereinafter also simply referred to as “neuroblast proliferation promoter etc.”) of the present invention is characterized by comprising phenylethanoid glycoside as an active ingredient. To do.
In the present specification, “neuroblast proliferation promoter and the like” mean both “neuroblast proliferation promoter” and “neurite extension agent”.

Here, the above “phenylethanoid glycoside” refers to a compound represented by the following chemical formula (1).

  The phenylethanoid glycoside is not particularly limited. For example, echinacoside, acteoside, 2′-acetylacetoside; campneoside I; campneoside II; (cistantubuloside) A, B1, B2, C1, C2; crenatoside; decaffeoylacteoside; isoacteoside; rhodioloside; syringalide A; 3 ' -Α-L-rhamnopyranoside, tubuloside A and the like. These may be used alone or in combination of two or more.

The neuroblast proliferation promoter or the like preferably contains at least one of echinacoside and acteoside as a phenylethanoid glycoside. This is because it has higher neuroblast proliferation action and neurite extension action.
Furthermore, particularly in neurite extenders, it is more preferable to contain at least echinacoside as an active ingredient. This is because it has a better neurite extension.

The method for obtaining the phenylethanoid glycoside is not particularly limited, and may be obtained by synthesis or may be obtained by extraction from a plant.
Moreover, when extracting from a plant, the plant used as a raw material is not specifically limited, For example, the ground medicine which is a crude drug, the rhododendron plant chorogi, the pepper family pepper genus, the olive fruit, the herbaceous plant herbaceous plant etc. are mentioned. These may be used alone or in combination of two or more. Of these, it is most preferable to use plants of the family Amaranthaceae. In addition to echinacoside and acteoside as phenylethanoid glycosides, 2'-acetylacteoside; campneoside I; campneoside II; cistantubuloside A, B1, B2, C1, C2 Decaffeoylacteoside; isoacteoside; rhodioloside; syringalide A; 3′-α-L-rhamnopyranoside, tubuloside A, etc. This is because it has higher neuroblast proliferation action and neurite extension action.
Here, the plant of the Amaranthaceae used as a raw material is not particularly limited. However, it is not limited to these. These may be used alone or in combination of two or more. Of these, the whole parasitic plant [Cistanche tubulosa: kankanikujuyou] is particularly preferable.

  The relationship between the above-described components of the phenylethanoid glycoside and the chemical formula (1) is shown below. However, the structural formula of crenatoside is shown in the following chemical formula (2). Among the above components, most of the components other than echinacoside and acteoside are contained in a small amount or a trace amount in the preparation.

All the compounds shown in Table 1 can be confirmed by high performance liquid chromatography (HPLC). Under the present circumstances, the conditions of high performance liquid chromatography are as follows. The stationary phase is C18 alkylsilane silicone, the mobile phase is acetonitrile-0.05M phosphoric acid aqueous solution (elution gradient 4: 96 → 15: 85), the flow rate is 1 ml / min, and the detection wavelength is 330 nm. It is.

  In addition, another neuroblast proliferation promoter of the present invention is characterized by using an extract extracted from a plant of the genus Oleaceae as an active ingredient.

Here, the plant of the Amaranthaceae used as a raw material is not particularly limited. rossica) and the like, but is not limited thereto. These may be used alone or in combination of two or more.
Further, it is particularly preferable to use a total parasitic plant [Cistanche tubulosa: kankanikujuyo] among the plants of the genus Oleaceae.

Furthermore, the “extract” preferably contains a phenylethanoid glycoside. Here, the “phenylethanoid glycoside” is not particularly limited, but echinacoside, acteoside, 2′-acetylacetoside; campneoside I; campneoside II; cis Cistantubuloside A, B1, B2, C1, C2; crenatoside; decaffeoylacteoside; isoacteoside; rhodioloside; syringalide A 3′-α-L-rhamnopyranoside, tubuloside and the like. In this case, the above components may be included alone or in the form of a mixture of two or more.
Among these, it is preferable to contain at least one of echinacoside and acteoside.
In particular, when used as a neurite extender, it is preferable that at least echinacoside is contained in the extract. This is because it has a more excellent neurite extension action.

At this time, the content of echinacoside is not particularly limited, but 10 to 70 when the total mass of the extract of the plant of the family Crestaceae (for example, the total parasitic plant [Cistanche tubulosa]) is 100% by mass. It can be made into mass%, Preferably it is 15-50 mass%, More preferably, it is 20-40 mass%, More preferably, it can be 23-38 mass%.
Further, the content of acteoside is not particularly limited. However, when the total mass of the extract of the plant of the family Amaranthaceae is 100% by mass, it is 1 to 40% by mass, preferably 5 to 30% by mass, more preferably 7 to 20%. It can be made into the mass%, More preferably, it is 8-10 mass%.

  Here, the part of the plant of the Amaranthaceae extracted is not particularly limited, and may be any of leaves, roots, stems, and the like as long as a desired effect is obtained. Preferably, it is extracted from the stems of the genus Oleaceae, in particular fresh stems.

  The method for obtaining the extract from the plant of the genus Oleaceae is not particularly limited, and examples thereof include a solvent extraction method and a supercritical extraction.

Moreover, when extracting by a solvent extraction method, the solvent to be used is not specifically limited, However, It is preferable to use a polar solvent. Moreover, although the said polar solvent is not specifically limited, For example, water, methanol, ethanol, isopropanol, acetone, 1, 3- butylene glycol, ethylene glycol, propylene glycol, glycerol, acetic acid, ethyl acetate, ether etc. are mentioned. These may be used alone or in combination of two or more.
This is because it is possible to obtain an extract containing a high concentration of phenylethanoid glycoside, which is preferably subjected to further treatments such as dilution, concentration, drying and purification.
Examples of the purification method include activated carbon treatment, resin adsorption treatment, ion exchange resin, liquid-liquid countercurrent distribution, and the like.

One embodiment of a specific method for producing the extract is described in detail below. In addition, the manufacturing method of an extract is not limited to the following.
The manufacturing method of the said extract consists of two steps, extraction and refinement | purification. In the first stage, the stems of the genus Oleaceae (for example, the total parasitic plant [Cistanche tubulosa]), more preferably the fresh stems of the plant are cut into flakes, or Grind into powder. Next, the flakes, fine particles or powder thus obtained are immersed in a solvent such as water, a lower aliphatic alcohol such as ethanol or methanol, or a mixture thereof. At this time, the extraction is performed at room temperature. Next, the mixed solution is filtered, and the filtrate is concentrated under reduced pressure or in vacuo to obtain an extract. Next, as a second step, the extract is heated in water and then transferred to an adsorption column packed with D-101 type or AB-8 type macroporous adsorption resin to purify the extract. . At this time, the column is eluted using water, methanol, ethanol, a mixed solution of water and methanol, or a mixed solution of water and ethanol as an elution solvent. The elution may be carried out by using an elution solvent as described above in a solution having a constant concentration or by adding a concentration gradient to the solution. The eluate is collected and concentrated, and then dried by a known drying method. When the eluate has dried, the extract is obtained. The extract thus obtained contains a phenylethanoid glycoside.

  The neuroblast proliferation promoter of the present invention can be used as a material for various foods and drinks. Examples of food and drink include edible oil (salad oil), confectionery (gum, candy, caramel, chocolate, cookie, snack, jelly, gummy, tablet confection etc.), noodles (soba, udon, ramen etc.), dairy products ( Milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, tea, tea, carbonated drinks, sports drinks, etc.) and health foods (tablets , Capsules, etc.) and nutritional supplements (nutrient drinks, etc.). The neuroblast proliferation promoter of the present invention and the like may be appropriately blended with these foods and drinks.

These foods and drinks can be blended with various ingredients depending on the type, for example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid. Acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic, Food materials such as carrageenan, casein, gelatin, pectin, agar, vitamin Bs, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, pigments, fragrances and preservatives can be used. In addition, this neuroblast proliferation promoter with health maintenance function includes other antioxidants and health food ingredients such as antioxidants, reduced ascorbic acid (vitamin C), vitamin E Reduced glutatin, tocotrienol, vitamin A derivative, lycopene, β-cryptoxanthin, astaxanthin, zeaxanthin, fucoxanthin, uric acid, ubiquinone, coenzyme Q10, folic acid, garlic extract, allicin, sezamin, lignans, catechin, isoflavone, chalcone, Tannins, flavonoids, coumarin, isocoumarins, blueberry extract, arbutin, tannin, anthocyanin, apple polyphenol, grape seed extract, ellagic acid, kojic acid, surge extract health food material, V. (vitamin) A, V.B1 , V.B2, V.B6, V.B12, VC, VD VE, VP, choline, niacin, pantothenic acid, calcium folate, EPA, oligosaccharide, dietary fiber, squalene, soy lecithin, taurine, donariella, protein, octacosanol, DHA, egg yolk lecithin, linoleic acid, lactoferrin, magnesium, zinc, chromium , Selenium, potassium, heme iron, oyster meat extract, chitosan, chitin oligosaccharide, collagen, chondroitin, turmeric, licorice, kokushi, keihi, hawthorn, ginger, ganoderma, rainbow trout extract, suppon, licorice, kokushi, keihi, hawthorn, Ginger, reishi, plantain, chamomile, chamomile, dandelion, hibiscus, honey, boren, royal jelly, lime, lavender, rosehip, rosemary, sage, bifidobacteria, faecalis, lacris, wheat germ oil, go Oil, perilla oil, soybean oil, medium chain fatty acid, agaricus, ginkgo biloba extract, turmeric, chondroitin, brown rice germ extract, litchi, onion, DHA, EPA, DPA, green tea, cordyceps, garlic, bee, papaya, puer, Propolis, Meguri Tree, Yabushake, Royal Jelly, Saw Palmetto, Hyaluronic Acid, Collagen, Gabba, Harp Seal Oil, Shark Cartilage, Glucosamine, Lecithin, Phosphatidylserine, Seven Carrots, Mulberry Leaf, Soybean Extract, Echinacea, Ezoukogi, Barley Extract, Olive leaf, Olive fruit, Gymnema, Banaba, Salacia, Garcinia, Chitosan, St. John's wort, Jujube, Carrot, Passion flower, Broccoli, Placenta, Barley, Grape seed, Peanut seed coat, Bilberry, Black cohosh, Ma Rear thistle, laurel, sage, rosemary, raffma, black vinegar, bitter gourd, maca, safflower, flax, oolong tea, flower thorn, caffeine, capsaicin, xylooligosaccharide, glucosamine, buckwheat, citrus, dietary fiber, protein, prunes, spirulina, Barley young leaves, nucleic acids, yeast, shiitake mushrooms, plum meat, amino acids, deep sea potato extract, noni, oyster meat, suppon, champignon, plantain, acerola, pineapple, banana, peach, apricot, melon, strawberry, raspberry, orange, fucoidan, Meshimakobu, cranberry, chondroitin sulfate, zinc, iron, ceramide, silk peptide, glycine, niacin, chesttree, ceramide, L-cysteine, L-carnitine, red wine leaf, millet, horsetail, biotin, centella asiatica, hascup, Picnoji Enol, Japanese cypress, rhubarb, clove, rosemary, catechin, puer, citric acid, brewer's yeast, merirot, black zinger, ginger, gadget, nattokinase, benicouji, tocotrienol, lactoferrin, cinnamon, buckwheat, cocoa, yuzu seed extract, perilla Berry extract, lychee seed extract, evening primrose extract, black rice extract, α-lipoic acid, gabba, fresh coffee bean extract, buffalo extract, kiwi seed extract, Unshu mandarin orange extract, red ginger extract, astaxanthin) and the like.

  As a specific production method, the neuroblast growth promoter of the present invention is spray-dried or freeze-dried as it is or together with powdered dextrin, and this is easily converted into a powder, granule, tablet, or solution. Food) and the like. Further, if necessary, it can be mixed with a binder such as gum arabic to form a powder or granules, and added to a solid food. Moreover, it is also possible to add to a beverage as it is or after being dispersed and dissolved in water, ethanol, glycerin or a mixture thereof.

  The neuroblast proliferation promoter and the like of the present invention may be used as a raw material for drugs (including pharmaceuticals and quasi drugs). It can be produced by appropriately blending the neuroblast proliferation promoter of the present invention with the raw material for the pharmaceutical preparation. In addition, the said chemical | medical agent may be used for a human and may be used for mammals other than a human. Examples of the pharmaceutical raw material that can be blended in the neuroblast proliferation promoter of the present invention include, for example, excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hydrogenated vegetable oil, Kaolin, talc, etc.), binder (distilled water, physiological saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrant (sodium alginate, agar) Sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose, gum arabic powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (Quaternary ammonium base, sodium lauryl sulfate Um, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearates, polyethylene glycol, and the like) and the like.

  The administration method of the neuroblast proliferation promoter and the like according to the present invention can be generally administered orally in the form of tablets, pills, soft / hard capsules, fine granules, powders, granules and the like. The water-soluble preparation can be administered orally as a liquid. Furthermore, parenteral administration may be used. When administered as a parenteral agent, the neuroblast growth promoter of the present invention is dispersed in a suitable solubilizer such as ethanol or water, and then a poultice, lotion, ointment, tincture, cream, etc. Can be applied in any dosage form. In addition, water-soluble preparations such as this neuroblast proliferation promoter, etc., as it is or with a dispersant, suspension, stabilizer, etc. added, poultices, lotions, ointments, tinctures, creams, etc. Can be applied in any dosage form.

  The dosage may vary depending on the administration method, medical condition, age of the patient, etc., but for adults, it can usually be administered as an active ingredient per day from 5 to 400 mg, and for children, usually from about 0.5 to 200 mg.

  The compounding ratio when using the neuroblast proliferation promoter of the present invention as a drug can be appropriately changed depending on the dosage form, but is usually about 0.01 when administered orally or by mucosal absorption. In the case of parenteral administration, about 0.01 to 20 wt% is recommended. Since the dose varies depending on various conditions, a dose smaller than the above dose may be sufficient, or the dose may need to be administered beyond the range. The pharmaceutical composition may include, in addition to the neuroblast proliferation promoter, etc., other known compounds commonly used in the pharmaceutical field, and compounds necessary for molding into a form suitable for oral administration. . Examples of such compounds include lactose, starch, hydroxypropyl cellulose, kaolin, talc, calcium carbonate and the like.

In addition, the neuroblast proliferation promoter and the like of the present invention can be contained in mammalian animal feed. The animal feed can be contained in the same manner as the food and drink. The animal feed is not particularly limited. For example, it is used for domestic animals such as cattle and pigs, companion animals such as dogs, cats and hamsters (animals kept as pets). can do.
For example, flour, meat, etc. can be used as companion animal feed. At this time, examples of the flour include wheat flour, rice flour, rye flour, corn flour, millet flour, bubble flour, corn flour, and soybean flour, and these flours may be used in combination of two or more. By using flour, nutrients such as carbohydrates can be supplied to companion animals. Among the above flours, it is most preferable to use flour, and as flour, strong flour, medium flour, thin flour can be used alone or in combination, and even when such flour and other flour are used in combination Good. Furthermore, in order to adjust the elasticity of the animal feed after the heat treatment, wheat flour, wheat gluten, soybean protein and the like may be combined. In addition, the network structure derived from gluten contained in wheat flour can constitute a puffed tissue structure when heat-treated, which contributes to improving the texture.

  At this time, the meat to be used is not particularly limited, and chicken, pork, beef, mutton, goat meat, crab meat, turkey meat, horse meat, etc. can be used, but chicken is preferably used in terms of flavor. The The above meat is obtained by slaughtering and demolishing livestock in the usual way. In addition, since the deterioration of the quality of a product with intermediate moisture or low moisture is mainly caused by oxidation of fat, the meat used is preferably lean meat with little fat content or fat removed. In addition, the coexistence of meat can improve the palatability of companion animals as well as strengthening high quality animal proteins.

The companion animal feed can be prepared by various methods. As a preferred method, a kneaded raw material mixture (hereinafter referred to as “dough”) containing flour (preferably wheat flour) and meat is prepared and molded. Thereafter, a heat treatment method can be exemplified. The composition of flour and meat in the dough is not particularly limited, but is usually about 5 to 60% flour, preferably about 10 to 50%, about 5 to 80% meat, preferably about 20 to 50%, and the required amount. It is adjusted to consist of water. Moreover, when using Aw regulator, the said Aw regulator is added so that it may become about 5-30%, Preferably it is about 10-20%. In addition, what is necessary is just to adjust the usage-amount of water suitably according to usage-amounts, such as flour, meat, and Aw regulator, so that dough can be knead | mixed and shape | molded.
The method for preparing the dough is not particularly limited, but preferably the meat is first ground with a silent cutter, chopper, or the like. At this time, it is preferable to chop the chopped meat so that bubbles are sufficiently contained therein. Then, the flour-containing dough can be prepared by adding flour, water and, if necessary, an Aw adjusting agent to the finely ground meat and sufficiently kneading to contain bubbles. In preparing the dough, a foaming agent may be added. It is preferable to use a foaming agent, particularly when flour other than wheat flour is used as the flour. By adding the foaming agent, fine bubbles can be uniformly contained in the dough. Various foaming agents can be used as the foaming agent, but it is preferable to use a soy protein foaming agent and / or an enzyme-decomposed soybean protein foaming agent from the viewpoint of the stability of air bubbles.

The dough prepared in this manner is molded and heat-treated, whereby the companion animal feed of the present invention is obtained. The dough may be formed into an appropriate shape according to the ease of eating when the companion animal eats the companion animal feed of the present invention, the handling of the owner, etc., for example, plate shape, stick shape, Examples include a disk shape, a donut shape, and a heart shape. Also, from doughs prepared from the same formulation, dye them with pigments of different colors, or mix vegetables or fruits to prepare multiple doughs with different appearances, and combine them in a multilayer or concentric form You can also
The heating means for the molded dough is not particularly limited, and examples thereof include oven heating and microwave heating. These heating methods are known, and heat treatment may be performed according to a conventional method. The water content of the feed after the heat treatment is usually about 20 to 40%. By the above heat treatment, the dough swells due to the evaporation of water and the expansion of bubbles, and the water evaporates in a short time, so Aw is lowered and the storage stability is improved. Moreover, when wheat flour is used as the flour, the mesh structure derived from gluten contained in the wheat flour is fixed by heat treatment, and the texture is improved. In the case of oven heating, there is an advantage that a unique color tone (amber color) and aroma can be generated in the feed. On the other hand, in the case of microwave heating, it can be heated from inside the dough. There is an advantage that it can be uniformly expanded and a feed having uniform air bubbles can be obtained. In the above heat treatment, it is preferable to adjust so that the Aw of the obtained feed is in the range of 0.6 to 0.9. As described above, the storage stability of the feed can be remarkably improved by adjusting Aw to this range.

The above-mentioned companion animal feed thus obtained is a feed having a bread-like property and has a soft texture and moderate flexibility and elasticity, so that it is a companion with weak teeth such as a young dog, an old dog or a cat. Suitable as animal feed, snacks, etc. Of course, it can be used as a feed or a snack for healthy adult dogs and cats. The animal feed is produced by storing an appropriate amount in a packaging container and sealing it. As the packaging container, it is preferable to use an oxygen gas-impermeable packaging material. Examples of the packaging form include vacuum packaging, inert gas filling packaging, and the like, but it is preferable to carry out inert gas filling packaging together with an oxygen scavenger (for example, Ageless ^TM ). According to such a packaging form, it is possible to prevent quality deterioration and microorganism growth due to oxygen during the storage period.

The neuroblast proliferation-promoting agent and the like of the present invention can be expected to have a neurite extension action even when used as a skin external preparation (including cosmetics, pharmaceuticals and quasi drugs). In addition, the said skin external preparation may be used for a human and may be used for mammals other than a human.
Examples of the external preparation for skin to which the neuroblast proliferation promoter of the present invention can be formulated include, for example, emulsion, soap, facial cleanser, bath preparation, cream, emulsion, lotion, eau de cologne, shave cream, shave Lotion, cosmetic oil, suntan / sunscreen lotion, powder, foundation, perfume, pack, nail cream, enamel, enamel remover, eyebrow, blusher, eye cream, eye shadow, mascara, eyeliner, lipstick, lip balm, Shampoos, rinses, hair dyes, dispersions, cleaning agents and the like can be mentioned.
Examples of the form of a pharmaceutical or quasi-drug that can contain the neuroblast proliferation promoter of the present invention include ointments, creams, and liquids for external use.

In addition to the neuroblast proliferation promoter according to the present invention and the like in the form of the above-mentioned external preparation for skin, the ingredients blended in the external preparation for skin such as cosmetics and quasi-drugs as long as the neurite extension action is not impaired, Oils, higher alcohols, fatty acids, UV absorbers, powders, pigments, surfactants, polyhydric alcohols / sugars, polymers, physiologically active ingredients, solvents, antioxidants, fragrances, preservatives, etc. can be blended. .
Examples are listed below, but the present invention is not limited to these examples.

(1) Examples of oil components Ester-based oil phase components: glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearic acid Isocetyl, butyl stearate, butyl myristate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoaralkyl neopentanoate, glyceryl tri (capryl / caprate), trimethylolpropane tri-2-ethylhexanoate, trimethylol triisostearate Propane, pentaerythritol tetra-2-ethylhexanoate, cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isolaurate Stearyl, isotridecyl myristate, isocetyl myristate, isostearyl myristate, isocetyl palmitate, isostearyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl linoleate, octyldodecyl isoleate, isopropyl isostearate Cetostearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethyleneglycol dioctanoate , Ethylene glycol dioleate, propylene glycol dicaprate, propylene glycol di (capryl / capric acid), propylene glycol dicaprylate, neopentyl glycol dicaprate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, triisopalmitine Glyceryl acid, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyl decyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleic acid Ester, polyglycerol isostearate, dipropyl carbonate, charcoal Dialkyl (C12-18), triisocetyl citrate, triisoaralkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyl decyl lactate, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, Trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, olein Cholesteryl acid, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, 12-stearoyl hydroxystearate Cetyl, 12-stearoyl-hydroxystearic acid stearyl 12-stearoyl-hydroxystearic acid isostearate, and the like.
Hydrocarbon oil phase components: squalane, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, petrolatum and the like.
Animal and vegetable oils and their hydrogenated oils, and naturally occurring waxes: beef tallow, hydrogenated beef tallow, lard, hydrogenated tallow, horse oil, hydrogenated horse oil, mink oil, orange luffy oil, fish oil, hydrogenated fish oil, egg yolk oil and other animal oils and Its hardened oil, avocado oil, almond oil, olive oil, cacao butter, kiwi seed oil, apricot oil, kukui nut oil, sesame oil, wheat germ oil, rice germ oil, rice bran oil, safflower oil, shea butter, soybean oil, evening primrose oil , Perilla oil, tea seed oil, camellia oil, corn oil, rapeseed oil, hydrogenated rapeseed oil, palm kernel oil, hydrogenated palm kernel oil, palm oil, hydrogenated palm oil, peanut oil, hydrogenated peanut oil, castor oil, hydrogenated castor oil , Vegetable oils such as sunflower oil, grape seed oil, jojoba oil, hardened jojoba oil, macadamia nut oil, medhome oil, cottonseed oil, hardened cottonseed oil, coconut oil, hardened coconut oil and its hardened , Beeswax, high acid value beeswax, lanolin, reduced lanolin, hydrogenated lanolin, liquid lanolin, carnauba wax and montan wax.
Silicone oil phase components: dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, methylhydrogenpolysiloxane, polyether-modified organopolysiloxane, dimethyl Siloxane / methylcetyloxysiloxane copolymer, dimethylsiloxane / methylstearoxysiloxane copolymer, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, amino-modified silicone oil, amino-modified organopolysiloxane, dimethiconol, silicone gel, acrylic Examples include silicone, trimethylsiloxysilicic acid, and silicone RTV rubber.
Fluorine oil phase components: perfluoropolyether, fluorine-modified organopolysiloxane, fluorinated pitch, fluorocarbon, fluoroalcohol, fluoroalkyl / polyoxyalkylene co-modified organopolysiloxane, and the like.

(2) Examples of higher alcohols Lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, behenyl alcohol, 2-ethylhexanol, hexadecyl alcohol, octyldodecanol and the like can be mentioned.

(3) Examples of fatty acids Caprylic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, arachidonic acid, behenic acid , Erucic acid, 2-ethylhexanoic acid and the like.

(4) Examples of UV absorbers Paraaminobenzoic acid, amyl paraaminobenzoate, ethyldihydroxypropyl paraaminobenzoate, glyceryl paraaminobenzoate, ethyl paraaminobenzoate, octyl paraaminobenzoate, octyldimethyl paraaminobenzoate, ethylene glycol salicylate, salicylic acid Octyl, triethanolamine salicylate, phenyl salicylate, butylphenyl salicylate, benzyl salicylate, homomenthyl salicylate, benzyl cinnamate, octyl paramethoxycinnamate, 2-ethylhexyl paramethoxycinnamate, mono 2-diparamethoxycinnamate Glyceryl ethylhexanoate, isopropyl paramethoxycinnamate, diethanolamine salt of paramethoxyhydrocinnamic acid, diisopropyl diisopropylcinnamic acid Stealth mixture, urocanic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone sodium disulfonate, dihydroxybenzophenone, dihydroxydimethoxybenzophenone, hydroxyoctoxybenzophenone, tetrahydroxybenzophenone, Butylmethoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2-ethylhexyl-1-oxy) -1,3,5-triazine, 2- (2-hydroxy-5-methylphenyl) benzotriazole , Methyl-O-aminobenzoate, 2-ethylhexyl-2-cyano-3, 3-diphenyl acrylate, phenylbenzimidazole sulfate, 3- (4-methylbenzylidene) can Examples include full, isopropyldibenzoylmethane, 4- (3,4-dimethoxyphenylmethylene) -2, 5-dioxo-1-imidazolidinepropionate 2-ethylhexyl, and the like, polymer derivatives, silane derivatives, and the like.

(5) Examples of powders and pigments Red 104, Red 201, Yellow 4, Blue 1, Black 401 and other dyes, Yellow 4 AL lake, Yellow 203 BA lake and other lake dyes, nylon powder , Silk powder, urethane powder, Teflon (registered trademark) powder, silicone powder, polymethyl methacrylate powder, cellulose powder, starch, silicone elastomer spherical powder, polyethylene powder, yellow iron oxide, red iron oxide, black Colored pigments such as iron oxide, chromium oxide, carbon black, ultramarine and bitumen, white pigments such as zinc oxide, titanium oxide and cerium oxide, extender pigments such as talc, mica, sericite, kaolin and barium sulfate plate, titanium mica Pearl pigments such as barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate, metal salts such as magnesium silicate, Inorganic powders such as Rica and alumina, metal soaps such as aluminum stearate, magnesium stearate, zinc palmitate, zinc myristate, magnesium myristate, zinc laurate, zinc undecylenate, bentonite, smectite, boron nitride, etc. It is done. There are no particular restrictions on the shape of these powders (spherical, rod-like, needle-like, plate-like, irregular shape, flake-like, spindle-like, etc.) and particle diameter. These powders are conventionally known surface treatments such as fluorine compound treatment, silicone treatment, silicone resin treatment, pendant treatment, silane coupling agent treatment, titanium coupling agent treatment, oil agent treatment, N-acylated lysine treatment, The surface treatment may or may not be performed in advance by polyacrylic acid treatment, metal soap treatment, amino acid treatment, lecithin treatment, inorganic compound treatment, plasma treatment, mechanochemical treatment or the like.

(6) Examples of surfactants Anionic surfactants: fatty acid soap, α-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate , Alkylamide sulfate, alkyl phosphate, POE alkyl phosphate, alkylamide phosphate, alkyloylalkyl taurine salt, N-acyl amino acid salt, POE alkyl ether carboxylate, alkyl sulfosuccinate, alkyl sulfoacetic acid Sodium, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate, etc. are mentioned.
Cationic surfactant: alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride , Behenic acid amidopropyldimethylhydroxypropylammonium chloride, stearic acid diethylaminoethylamide, stearic acid dimethylaminopropylamide, lanolin derivative quaternary ammonium salts, and the like.
Amphoteric surfactants: carboxybetaine type, amide betaine type, sulfobetaine type, hydroxysulfobetaine type, amide sulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amidoamine type and the like.
Nonionic surfactant: propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE sorbitol fatty acid ester, POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE cured castor Oil, POE castor oil, POE / POP copolymer, POE / POP alkyl ether, polyether-modified silicone lauric acid alkanolamide, alkylamine oxide, hydrogenated soybean phospholipid, and the like.
Natural surfactant: lecithin, saponin, sugar surfactant and the like.

(7) Examples of polyhydric alcohols and sugars Ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, diglycerin, polyglycerin, 3-methyl-1,3-butanediol, 1, 3 -Butylene glycol, sorbitol, mannitol, raffinose, erythritol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, trehalose, alkylated trehalose, mixed isomerized sugar, sulfated trehalose, pullulan and the like. These chemically modified compounds can also be used.

(8) Examples of polymers Acrylic ester / methacrylic acid ester copolymer (plus size, manufactured by Kyoyo Chemical), vinyl acetate / crotonic acid copolymer (resin 28-1310, manufactured by NSC), vinyl acetate / croton Acid / vinyl neodecanate copolymer (28-2930, manufactured by NSC), methyl vinyl ether maleic acid half ester (Gantrez ES, manufactured by ISP), T-butyl acrylate / ethyl acrylate / methacrylic acid copolymer (rubymer) , BASF), vinyl pyrrolidone / vinyl acetate / vinyl propionate copolymer (Rubicol VAP, BASF), vinyl acetate / crotonic acid copolymer (Rubiset CA, BASF), vinyl acetate / croton Acid / vinyl pyrrolidone copolymer (Rubiset CAP, manufactured by BASF), vinyl pyrrolidone / acrylate copolymer (Rubiflex, BA SF), acrylate / acrylamide copolymer (Ultrahold, BASF), vinyl acetate / butyl maleate / isobornyl acrylate copolymer (Advantage, ISP), carboxy vinyl polymer (Carbopol, BF Goodrich), anionic polymer compounds such as acrylic acid / alkyl methacrylate copolymers (Pemulene, BF Goodrich) and acetic acid amphoteric compounds of dialkylaminoethyl methacrylate polymers (Yukaformer, Mitsubishi Chemical) Amphoteric polymer compounds such as octyl acrylate acrylate / hydroxypropyl acrylate / butyl aminoethyl methacrylate copolymer (AMPHOMER, NSC), quaternized vinylpyrrolidone / dimethylaminoethyl methacrylate (GAFQUAT, ISP) , Methyl vinyl imidazolium chloride Cationic polymer compounds such as vinyl / vinylpyrrolidone copolymer (rubicoat, manufactured by BASF), polyvinylpyrrolidone (rubiscol K, manufactured by BASF), vinylpyrrolidone / vinyl acetate copolymer (rubiscol VA, manufactured by BASF) ), Nonionic polymer compounds such as vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (copolymer 937, manufactured by ISP), vinylcaprolactam / vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (copolymer VC713, manufactured by ISP), etc. There is. Cellulose or derivatives thereof, keratin and collagen or derivatives thereof, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharide, xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabino Naturally derived polymer compounds such as galactan, gum karaya, gum tragacanth, alginic acid, albumin, casein, curdlan, gellan gum and dextran can also be suitably used.

(9) Examples of physiologically active ingredients Examples of physiologically active ingredients include substances that impart some physiological activity to the skin when applied to the skin. For example, whitening ingredients, immunostimulants, anti-aging agents, UV protection agents, slimming agents, tanning agents, antioxidants, hair growth agents, hair restorers, moisturizers, blood circulation promoters, antibacterial agents, bactericides, desiccants, A cooling sensation agent, a warming sensation agent, vitamins, an amino acid, a wound healing promoter, an irritation relaxation agent, an analgesic agent, a cell activator, an enzyme component, etc. are mentioned. Examples of these suitable ingredients include, for example, ashitaba extract, avocado extract, amateur extract, altea extract, arnica extract, aloe extract, apricot extract, apricot kernel extract, ginkgo biloba extract, fennel extract, turmeric extract, oolong tea extract, ages Extract, Echinacea leaf extract, Ogon extract, Oat extract, Oen extract, Barley extract, Hypericum extract, Oyster extract, Dutch mustard extract, Orange extract, Seawater dried product, Seaweed extract, Hydrolyzed elastin, Hydrolyzed wheat powder, Hydrolyzed silk , Chamomile extract, Carrot extract, Kawara mugwort extract, Licorice extract, Calcade extract, Oyster extract, Kina extract, Cucumber extract, Guanosine, Gardenia extract, Kumazasa extract, Clarae Kiss, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentian extract, tea extract, yeast extract, burdock extract, fermented rice bran extract, rice germ oil, comfrey extract, collagen, bilberry extract, saicin extract, psycho extract Extract Kizuta extract, hawthorn extract, elderberry extract, yarrow extract, mint extract, sage extract, mallow extract, senki Sue extract, assembly extract, soybean extract, tiso extract, thyme extract, tea extract, clove extract, chigaya extract, chimpi extract, touki extract, toshinsen extract, tonin extract, spruce extract, dokudami extract, tomato extract, natto extract, carrot extract, garlic extract, Novara extract, hibiscus extract, bacmond extract, parsley extract, honey, hamamelis extract, parietalia extract, toad extract, bisabolol, loquat extract, dandelion extract, burdock extract, bukuro extract, butcher bloom extract, grape extract, propolis, loofah extract, Safflower extract, peppermint extract, bodaige extract, button extract, hop extract, pine extract, marronnier extract, mizubasho Kiss, Mukuroji extract, Melissa extract, Peach extract, Cornflower extract, Eucalyptus extract, Yukinoshita extract, Yokuinin extract, Artemisia extract, Lavender extract, Apple extract, Lettuce extract, Lemon extract, Lotus root extract, Rose extract, Rosemary extract, Roman chamomile extract And royal jelly extract.
Biopolymers such as deoxyribonucleic acid, mucopolysaccharide, sodium hyaluronate, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, hydrolyzed eggshell membranes, amino acids, hydrolyzed peptides, sodium lactate, urea, sodium pyrrolidonecarboxylate , Moisturizing ingredients such as betaine, whey, trimethylglycine, oily ingredients such as sphingolipid, ceramide, phytosphingosine, cholesterol, cholesterol derivatives, phospholipids, ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, guaiazulene, Immunostimulants such as hydrocortisone, vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, vitamin C ester Vitamins, active ingredients such as allantoin, diisopropylamine dichloroacetate, 4-aminomethylcyclohexanecarboxylic acid, antioxidants such as tocopherol, carotenoid, flavonoid, tannin, lignan, saponin, α-hydroxy acid, β-hydroxy acid, etc. Cell activators, blood circulation promoters such as γ-oryzanol and vitamin E derivatives, wound healing agents such as retinol and retinol derivatives, arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, glutathione, etc. Whitening agent, cephalanthin, licorice extract, red pepper tincture, hinokitiol, garlic iodide extract, pyridoxine hydrochloride, DL-α-tocopherol, DL-α-tocopherol acetate, nicotinic acid, nicotinic acid derivative, calcium pantothenate, D-pan Tothenyl alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropylmethylphenol, estradiol, ethinyl estradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, takanal, camphor, salicylic acid, nonyl acid vanillylamide, nonanoic acid vanillylamide, pyroctone Olamine, glyceryl pentadecanoate, L-menthol, mononitroguaiacol, resorcin, γ-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormone, cantalis tincture, cyclosporine, zinc pyrithione, hydrocortisone, minoxidil, monostearic acid Examples include hair growth agents such as polyoxyethylene sorbitan, mint oil, and Sasanishiki extract.

(10) Examples of antioxidants: sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tolyl biguanide, nordihydroguaiaretic acid, parahydroxyanisole, butylhydroxyanisole, dibutylhydroxy Examples include plant extracts having an antioxidant effect such as toluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoid, flavonoid, tannin, lignan, saponin, apple extract and clove extract.

(11) Examples of solvents Purified water, ethanol, lower alcohol, ethers, LPG, fluorocarbon, N-methylpyrrolidone, fluoroalcohol, volatile linear silicone, next-generation chlorofluorocarbon and the like.

Examples of the present invention will be described, but the present invention is not limited to these examples.
Examples Preparation of Neuroblast Growth Promoter, etc. of the Present Invention Example 1; Extract of Cistanche tubulosa (hereinafter simply referred to as “kankanikujuyou”) <Extraction process>
A fresh stalk flake of Kankaniku Juyo was immersed in 8 times the amount of water of the flake. The flakes were soaked in water for 1 hour and then decocted with water for 2 hours. The mixture brewed in this way was filtered to obtain a first filtrate. Next, the residue of the decocted mixture was mixed with 40% ethanol of 4 times the residue, decocted for 4 hours, and the decocted mixture was filtered to obtain a second filtrate. About the residue of the 2nd filtrate, the same operation as the above was repeated twice, and the 3rd and 4th filtrate was obtained. These four kinds of filtrates were combined and concentrated in vacuo so that the specific gravity was 1.05 (50 ° C.), thereby obtaining a final extract. The final extract thus obtained weighed 6.2 kg.

<Purification process>
6 kg of the final extract was dissolved in half the amount of water of the final extract while heating. The extract solution was then placed on an adsorption column packed with pre-treated macroporous adsorption resin of D-101 type. The column is first eluted with water to obtain twice the amount of fresh water eluate, and the column is further eluted with 20% ethanol to double the amount of fresh stem first 20% ethanol eluate. Got. The water eluate was subjected to the adsorption-desorption operation again to obtain a second ethanol eluate. These two 20% ethanol eluates were combined, concentrated and dried to obtain a preparation containing phenylethanoid glycoside (Example 1). The weight of the preparation thus obtained was 0.865 kg.

  The contents of echinacoside and acteoside were measured by high performance liquid chromatography (HPLC). At this time, the HPLC conditions are as follows. The stationary phase was C18 alkylsilane silicone, the mobile phase was methanol-0.15% acetic acid aqueous solution (30:70), the flow rate was 1 ml / min, and the detection wavelength was 330 nm.

  The echinacoside and acteoside dried in vacuum at 60 ° C. for 24 hours were measured and dissolved in 50% methanol to prepare a reference solution (each 1 ml solution contains 0.1 mg of solute).

  The test solution was prepared by dissolving a preparation containing 50 mg of phenylethanoid glycoside in an appropriate amount of 50% methanol in a 25 ml graduated container while sonicating. Then, 50% methanol was added to the solution up to a 25 ml scale. Thereafter, about 1 ml of this solution was accurately taken and transferred to a container with a 10 ml scale, and 50% methanol was added to the 10 ml scale. Thereafter, this solution was filtered through a 0.45 μm membrane to obtain a test solution.

  5 μl of each of the reference solution and the test solution was taken and injected into a liquid chromatography column, and the peak areas of echinacoside and acteoside were measured. The content was calculated using these peak areas. As a result, the content of echinacoside was 29.3% by mass of the preparation (Example 1), and the content of acteoside was 10.0% of the preparation (Example 1).

Example 2 and Example 3
Example 2 used pure echinacoside, and Example 3 used pure acteoside (both manufactured by Kyokusei Industrial Co., Ltd.).

Test Example 1: Evaluation Test Method of Neuroblast Proliferation Action Human neuroblasts (SK-N-SH) were suspended in MEM medium (containing 10% FCS, 100 units / mL penicillin, 100 μg / mL streptomycin) (1 × 10 ^Four
cells / mL) and seeded 5 mL each in a 6 mm petri dish. Then, the citrus extract of Example 1 prepared in various concentrations was added and cultured for 5 days, and the degree of cell proliferation of the cells on the 5th day of culture was evaluated by the MTT assay. The result is shown in FIG. Example 2 and Example 3 were tested in the same manner. The result is shown in FIG.

Results and Effects of Examples in Test Example 1 According to FIG. 1, when the degree of cell proliferation was measured using an MTT assay, a neuroblast proliferation action was observed in a concentration-dependent manner in the kankanikusuyo extract. (Figure 1). Cell proliferation was 7% at 10 μg / mL and 16% at 30 μg / mL. Thereby, it was confirmed that the extract of Kanka has a proliferation action of human neuroblasts (SK-N-SH) even in a state where no damage has occurred in the neuroblasts.
Furthermore, according to FIG. 2, when the degree of cell proliferation was measured using the MTT assay, neuroblast proliferation action was observed in a concentration-dependent manner in echinacoside and acteoside. In particular, a significant difference was observed between echinacide 1 μg / mL and 10 μg / mL. This confirms that phenylethanoid glycosides, in particular echinacoside and acteoside, have a proliferative effect on human neuroblasts (SK-N-SH) even when neuroblasts are not damaged. It was done.

Test Example 2: Evaluation of neurite outgrowth action Human neuroblasts (SK-N-SH) were suspended in MEM medium (containing 10% FCS, 100 units / mL penicillin, 100 μg / mL streptomycin) (1 × 10 ⁴
cells / mL) and seeded 5 mL each in a 6 mm petri dish. And the kankanikujou extract of Example 1 prepared in various density | concentrations was added, the time-dependent change of the neurite was observed with the microscope, and photography was performed. The result is shown in FIG. In the same manner, tests were performed on Example 2 and Example 3. The results are shown in FIGS. 4 and 5, respectively.

  According to FIG. 3, although cell growth of the control was observed, the neurite outgrowth was not observed much, whereas more neurite outgrowth was observed in the kankanikuyuyo extract. (Refer to the solid circle in FIG. 3). In particular, network formation with adjacent cells was confirmed at a concentration of 30 μg / mL. Thus, it was confirmed that the kankanikuyu extract of this example has a neurite extension action in a concentration-dependent manner even in a state in which no damage has occurred in neuroblasts.

  In FIGS. 4 and 5, control cell proliferation was observed, but neurite outgrowth was not observed much, whereas echinacoside (FIG. 4) and acteoside (FIG. 5) showed more Neurite outgrowth was observed (see the dashed circles in FIGS. 4 and 5). Moreover, in echinaccoside, formation of a network with adjacent cells at a low concentration was remarkably confirmed (see the broken line circle in FIG. 5). As a result, it was confirmed that phenylethanoid glycosides, particularly echinacoside, have an action of promoting neurite outgrowth in a concentration-dependent manner even when neuroblasts are not damaged.

Although the compounding examples, such as a neuroblast proliferation promoter of this invention, are given to the following, the following compounding example does not limit this invention.
Formulation Example 1: Chewing gum
53.0wt% sugar
Gum base 20.0
Glucose 10.0
Minamata 16.0
Fragrance 0.5
Neuroblast proliferation promoter, etc.0.5
100.0wt%

Formulation Example 2: Gummy
Reduced water tank 40.0wt%
Granulated sugar 20.0
Glucose 20.0
Gelatin 4.7
Water 9.68
Grape juice 4.0
Grape flavor 0.6
Dye 0.02
Neuroblast proliferation promoter, etc. 1.0
100.0wt%

Formulation Example 3: Candy
50.0 wt% sugar
Minamata 33.0
Water 14.4
Organic acid 2.0
Fragrance 0.2
Neuroblast proliferation promoter, etc.0.4
100.0wt%

Formulation Example 4: Yogurt (hard / soft)
Milk 41.5wt%
Nonfat dry milk 5.8
Sugar 8.0
Agar 0.15
Gelatin 0.1
Lactic acid bacteria 0.005
Neuroblast proliferation promoter, etc.0.4
Perfume
Water residue
100.0wt%

Formulation Example 5: Soft drink
Fructose glucose liquid sugar 30.0wt%
Emulsifier 0.5
Neuroblast proliferation promoter, etc.0.05
Perfume
Purified water residue
100.0wt%

Formulation Example 6: Tablets
76.4 wt% sugar
Glucose 19.0
Sucrose fatty acid ester 0.2
Neuroblast proliferation promoter, etc.0.5
Purified water 3.9
100.0wt%

Formulation Example 7: Soft capsule
Grape seed oil 87.0wt%
Emulsifier 12.0
Neuroblast proliferation promoter, etc. 1.0
100.0wt%

Formulation Example 8: Tablet
Lactose 54.0wt%
Crystalline cellulose 30.0
Starch degradation product 10.0
Glycerin fatty acid ester 5.0
Neuroblast proliferation promoter, etc. 1.0
100.0wt%

Formulation Example 9: Oral granules (pharmaceuticals)
Neuroblast proliferation promoter, etc. 1.0 wt%
Lactose 30.0
Cornstarch 60.0
Crystalline cellulose 8.0
Polyvinylpyrrolidone 1.0
100.0wt%

Formulation Example 10: Cosmetic cream
Squalane 20.0wt%
Beeswax 5.0
Refined jojoba oil 5.0
Glycerin 5.0
Glycerol monostearate 2.0
Polyoxyethylene (20) sorbitan-
Monostearate 2.0
Neuroblast growth promoter 2.0
Preservative appropriate amount
Perfume
Purified water residue
100.0wt%

Formulation Example 11: Lotion
Ethanol 5.0wt%
Glycerin 2.0
1,3-butylene glycol 2.0
Polyethylene oleyl ether 0.5
Sodium citrate 0.1
Citric acid 0.1
Neuroblast growth promoter 0.1
Purified water residue
100.0wt%

Formulation Example 12: Body gel
Macadamia nut oil 2.0wt%
Octyldodecyl myristate 10.0
Methylphenylpolysiloxane 5.0
Behenyl alcohol 3.0
Stearic acid 3.0
Batyl alcohol 1.0
Glyceryl monostearate 1.0
Tetraoleic acid polyoxyethylene sorbit 2.0
Hydrogenated soybean phospholipid 1.0
Ceramide 0.1
Retinol palmitate 0.1
Preservative appropriate amount
Centella extract 1.0
Neuroblast proliferation promoter, etc. 1.0
1,3-butylene glycol 5.0
Purified water residue
100.0wt%

Formulation Example 13: Latex
Squalane 4.0wt%
Vaseline 2.5
Cetanol 2.0
Glycerin 2.0
Lipophilic glyceryl monostearate 1.0
Stearic acid 1.0
L-Arginine 1.0
Neuroblast proliferation promoter, etc.0.5
Potassium hydroxide 0.1
Perfume
Purified water residue
100.0wt%

Formulation Example 14: Bath agent (liquid)
Propylene glycol 50.0wt%
Ethanol 20.0
Sodium sulfate 5.0
Neuroblast proliferation promoter, etc.0.5
Lanolin 0.5
Avocado oil 0.5
Dye 1.5
Fragrance 22.0
100.0wt%

Formulation Example 15: Cat food
Corn 34.0wt%
Flour 35.0
Meat meal 15.0
Beef tallow 8.9
Salt 1.0
Skipjack extract 4.0
Neuroblast proliferation promoter, etc. 1.0
Taurine 0.1
Vitamins 0.5
Minerals 0.5
100.0wt%

Formulation Example 16: Dog food
Corn 30.0wt%
Meat (chicken) 15.0
Defatted soybean 10.0
Flour 25.0
Mosses 5.0
Neuroblast proliferation promoter, etc.5.0
Animal fats and oils 8.9
Oligosaccharide 0.1
Vitamin 0.5
Mineral 0.5
100.0wt%

  As described above, the present invention promotes the proliferation of neuroblasts and promotes the extension of neurites of the neuroblast even when the neuroblast is not damaged. , Promoting the transformation of neuroblasts into neurons, effectively preventing the decrease of neurons even when neuronal degeneration occurs, thereby effectively aging the brain Thus, it is possible to provide a novel neuroblast proliferation promoter and neurite extension agent that can prevent and further improve brain function.

It is a graph which shows the neuroblast proliferation effect | action of a kankanikujuyo extract. It is a graph which shows the neuroblast proliferation effect | action of an echinacoside and an acteoside. It is the photograph which image | photographed the extension state of the neurite by a kankanikujuyo extract. It is the photograph which image | photographed the extension state of the neurite by echinacoside. It is the photograph which image | photographed the extension state of the neurite by acteoside.

Claims (18)

A neuroblast growth promoter comprising phenylethanoid glycoside as an active ingredient. The neuroblast proliferation promoter according to claim 1, wherein the phenylethanoid glycoside includes at least one of echinacoside and acteoside. The neuroblast proliferation promoter according to claim 2, wherein the phenylethanoid glycoside includes echinacoside and acteoside. A neuroblast growth-promoting agent comprising an extract of a plant of the clover family as an active ingredient. The neuroblast proliferation promoter according to claim 4, wherein the extract contains phenylethanoid glycoside as an active ingredient. 6. The neuroblast proliferation promoter according to claim 5, wherein the extract contains at least one of echinacoside and acteoside as the phenylethanoid glycoside. The neuroblast proliferation promoter according to claim 6, wherein the extract contains echinacoside and acteoside as the phenylethanoid glycoside. A neurite extension agent containing phenylethanoid glycoside as an active ingredient. The neurite extender according to claim 8, wherein the phenylethanoid glycoside contains at least one of echinacoside and acteoside. The neurite extender according to claim 9, comprising echinacoside and acteoside as the phenylethanoid glycoside. A neurite extender that contains an extract of a plant of the genus Crestaceae as an active ingredient. The neurite extender according to claim 11, wherein the extract contains a phenylethanoid glycoside as an active ingredient. The neurite extender according to claim 12, wherein the extract contains at least one of echinacoside and acteoside as the phenylethanoid glycoside. The neurite extender according to claim 13, wherein the extract contains echinacoside and acteoside as the phenylethanoid glycoside. A medicinal product for mammals including humans, comprising as an active ingredient the neuroblast proliferation promoter according to any one of claims 1 to 6 or the neurite extender according to any one of claims 7 to 14. The topical skin for mammals including humans comprising the neuroblast proliferation promoter according to any one of claims 1 to 6 or the neurite extender according to any one of claims 7 to 14 as an active ingredient. Agent. A food or drink comprising as an active ingredient the neuroblast proliferation promoter according to any one of claims 1 to 6 or the neurite extender according to any one of claims 7 to 14. A feed for mammals, comprising as an active ingredient the neuroblast growth promoter according to any one of claims 1 to 6 or the neurite extender according to any one of claims 7 to 14.