JP5259601B2 - Collagen production promoter - Google Patents

Description

The present invention relates to medicines blended with it and collagen production accelerator, in particular, those that exhibit the collagen production promoting effect by oral ingestion.

  Skin aging (wrinkles, sagging, etc.) and rough skin have become major cosmetic concerns. There are various causes of skin aging, rough skin, etc., but the underlying phenomenon is caused by a decrease in collagen production activity of dermal fibroblasts, a decrease in hyaluronic acid synthesis activity, an increase in collagenase activity due to UV rays, UV rays and the environment. It is considered that the skin moisturizing function is lowered due to the damage caused by the active oxygen generated, and the skin components are deteriorated, denatured or reduced. If the skin's moisturizing function decreases due to external factors such as the environment, meals, luxury items, etc., and aging, the skin becomes dry and loses its elasticity, causing dry skin and wrinkles. It is thought to lead to the onset of atopic dermatitis.

  The epidermis and dermis of the skin are composed of epidermal cells and fibroblasts, and extracellular matrices (collagen, elastin, hyaluronic acid, etc.) that are structural supports outside these cells. These skin tissues are newly reborn on a daily basis through a turnover cycle, and in addition to maintaining moisture and maintaining flexibility and elasticity of the skin, they are effective in preventing the entry of external enemies. However, the above external and internal factors and aging promote the decrease in production, degradation, and denaturation of collagen, which is a major component of the extracellular matrix, resulting in a decrease in skin moisture and flexibility. It loses its elasticity and causes aging such as rough skin and wrinkle formation.

  Collagen is the main component of connective tissue and occupies 25-30% of the protein constituting the body. It exists especially in skin, bone, cartilage, tendon, ligament, and plays an important role in maintaining their structure and function. Collagen is said to take a long time to turn over compared to a normal protein, and its cycle is slowed down with aging. When the cycle is lowered, the collagen itself is also denatured (aging), and it is thought that the function of each organ is lowered. In the case of skin, when collagen ages, the flexibility and elasticity of the skin decrease. Furthermore, as collagen ages, its function as a structural support declines, preventing the growth, differentiation, and migration of fibroblasts present in the skin base, creating a vicious cycle in which the skin turnover cycle is further delayed. It is believed that.

  Based on this idea, attempts have been made to promote collagen production. For example, reduction of collagen includes supplementation with collagen (Patent Documents 1 and 2), promotion of collagen metabolic activity (Patent Documents 3 and 4), and promotion of collagen production (Patent Documents 5 and 6).

  In recent years, the search for plant-derived collagen-like substances and collagen production-promoting substances and development of products containing them have been actively carried out by reducing the image of animal-derived raw materials (Patent Documents 4 to 9).

  Phospholipids exist specifically in cell membrane-like structural sites, and are known as major components of biological membranes along with proteins. It is contained in many parts such as brain, nerves, internal organs, blood, eggs, and seeds, and plays many functions for life support. Based on the difference in structure, it is classified into glycerophospholipid and sphingophospholipid, and various types and functions are known (Non-patent Documents 1 and 2).

  Lecithin is widely recognized as a mixture containing various glycerophospholipids, and is widely used as a surfactant for food, industrial, pharmaceutical and cosmetic purposes. Furthermore, the use for the purpose of the selective transport to the action site | part by forming the closed vesicle (liposome) which consists of a lipid bilayer membrane, and enclosing an active ingredient is also made | formed (nonpatent literature 2).

  In addition, the functionality of various glycerophospholipids has recently become apparent. For example, phosphatidylcholine (PC), which is a main component of lecithin, has an action of obtaining a whitening effect (Patent Documents 10 and 11), an action of suppressing collagen production induced by inflammatory stimulation (Non-Patent Document 3), and damaged skin. Has been reported to regulate the recovery by suppressing the contraction of hair (Non-patent Document 4), and the action of activating protein kinase C (PKC) to phosphatidic acid (PA) to promote hair regeneration (Patent Document 12) An action that improves the membrane fluidity of tumor cells and changes the multidrug resistance (Patent Document 13), a cell proliferation action for lysophosphatidic acid (LPA), and a cell growth inhibitory activity for cyclic lysophosphatidic acid (cPA) ( Non-patent document 5) has been reported. In addition, phosphatidylserine (PS) has a brain function improving effect (Non-patent document 6) and neurite outgrowth activity (Non-patent document 7), and phosphatidylethanolamine (PE) has a neurotrophic effect (Non-patent document 8). Has been reported. Furthermore, there is a report of a collagen synthesis promoting action using cultured cells through a lysophosphatidic acid or phosphatidic acid nitric oxide production promoting action (Patent Document 14).

However, it has never been known that collagen production is promoted by ingesting these glycerophospholipids orally.
JP 2002-51734 A JP 2003-238597 A Japanese Patent Application Laid-Open No. 07-2699 Japanese Patent No. 3696464 Japanese Patent Laid-Open No. 2001-278783 JP 2003-121748 A JP 2003-206226 A JP 2003-277286 A JP 2006-8571 A Biochemistry Experiment Course 3, Lipid Chemistry, 251-341 Emulsifiers for food-basics and applications-, 67-91 J. et al. Lab. Clin. Med. 139 (2002), 202-210 J. et al. Invest. Surg. 17 (2004), 15-22 Protein Nucleic Acid Enzyme, Vol. 44, no. 8 (1999), 1118-1125 FOOD Style 21, Vol. 6, no. 11 (2002), 108-116 Japan Agricultural Chemistry 2004 Conference, 3A19p23 “Effect of yolk-derived phosphatidylserine on neurite outgrowth” J. et al. Lipid Research, 47 (2006), 1434-1443. JP 2004-59496 A JP 2005-272444 A JP 2006-76967 A JP 2006-143744 A JP 2005-200370 A

  However, many of the plant-derived collagen-like substances and collagen production-promoting substances described above are derived from herbal medicines, and are generally expensive, and have a limited capacity range that exhibits side effects and proper effects due to the original herbal medicine. There was a problem. In addition, there are many things that contain irritants in the components, and various considerations are necessary when using such as a strong action (toxicity) on skin fibroblasts.

The present invention is inexpensive and safe collagen production promoter, as well as to provide the medicines like blended therewith, in particular, it aims to provide a collagen production promoting agent exhibiting collagen production promoting effect by oral ingestion To do.

  As a result of intensive studies to solve the above problems, the present inventors have found that glycerophospholipids, particularly glycerophospholipids containing unsaturated fatty acids, may have a high collagen production promoting action. Reached.

  That is, the first aspect of the present invention is a collagen production promoter characterized by comprising glycerophospholipid as an active ingredient, and preferably the glycerophospholipid is phosphatidylcholine, lysophosphatidylcholine, phosphatidylserine, lysozyme. One or more selected from the group consisting of phosphatidylserine, phosphatidylethanolamine, lysophosphatidylethanolamine, phosphatidylinositol, lysophosphatidylinositol, phosphatidic acid, lysophosphatidic acid, phosphatidylglycerol and lysophosphatidylglycerol, and preferably , Wherein at least one of the fatty acids constituting the glycerophospholipid is an unsaturated fatty acid, more preferably, the collagen production described above Susumuzai is intended to demonstrate the collagen production promoting action by oral ingestion.

A second aspect of the present invention is to summarized as to that and Drug characterized by blending the collagen production promoter.

  Furthermore, the present invention provides the use of the glycerophospholipid for producing a collagen production promoter, a method for increasing the collagen content in the skin of a subject, including administering the glycerophospholipid to the subject, and administering the glycerophospholipid to the subject. A method for increasing the collagen content in the tendon and / or bone of the subject, including increasing the collagen content in the skin, tendon and / or bone of the subject including orally administering the glycerophospholipid to the subject It is also possible to provide a method.

According to the present invention, inexpensive and safe to provide collagen production promoter and medicines like blended with it, in particular, collagen enhancing agent exhibiting collagen production promoting effect by oral ingestion can be provided.

  Hereinafter, the present invention will be described in detail.

  The collagen production promoter of the present invention uses glycerophospholipids alone or in combination of two or more. As glycerophospholipids, the presence of ester type (monoacyl type, diacyl type), ether type (alkyl type, alkylacyl type, alkenylacyl type, dialkyl type) and phosphono type (CP compound) are known.

  Any glycerophospholipid used in the present invention may be used as long as the effects of the present invention are not impaired. For example, it is also possible to use glycerophospholipids derived from plant materials, animal materials, fungi and bacteria, and glycerophospholipids obtained by purifying or chemically / enzymatically treating these, chemically synthesized products, and enzyme synthesized products.

  The glycerophospholipid used in the present invention is preferably an ester glycerophospholipid, and more specifically, phosphatidylcholine, lysophosphatidylcholine, phosphatidylserine, lysophosphatidylserine, phosphatidylethanolamine, lysophosphatidylethanolamine, phosphatidylinositol, lysophosphatidylinositol, phosphatidine Examples include acids, lysophosphatidic acid, phosphatidylglycerol, lysophosphatidylglycerol, and the like.

  At least one of the fatty acids constituting the ester-type glycerophospholipid used in the present invention is preferably an unsaturated fatty acid, and the unsaturated fatty acid has an unsaturation degree of 1 or more and a carbon number of 4 or more. Is preferable from the viewpoint of collagen production promoting activity. More specifically, butenoic acid (C4: 1, such as crotonic acid and isocrotonic acid), pentenoic acid (C5: 1), hexenoic acid (C6: 1), heptenoic acid (C7: 1), octenoic acid (C8 1), nonenoic acid (C9: 1), decenoic acid (C10: 1), undecenoic acid (C11: 1), dodecenoic acid (C12: 1 such as lauroleic acid), tridecenoic acid (C13: 1), tetradecene Acids (C14: 1, such as myristoleic acid, myristaleic acid, etc.), pentadecenoic acid (C15: 1), hexadecenoic acid (C16: 1, such as palmitoleic acid, palmitelaidic acid, etc.), heptadecenoic acid (C17: 1) ), Octadecenoic acid (C18: 1, such as petroselinic acid, petroselinic acid, oleic acid, elaidic acid, vaccenic acid, etc.), nonadecene (C19: 1), eicosenoic acid (C20: 1, such as gadoleic acid, gondrenic acid, etc.), docosenoic acid (C22: 1, such as erucic acid, brassicic acid, setoleic acid, etc.), tetracosenoic acid (C24: 1, such as nervone, etc.) Acid), hexacosenoic acid (C26: 1), octacosenoic acid (C28: 1), triacontenoic acid (C30: 1), pentadienoic acid (C5: 2), hexadienoic acid (C6: 2, such as sorbic acid), Peptadienoic acid (C7: 2), octadienoic acid (C8: 2), nonadienoic acid (C9: 2), decadienoic acid (C10: 2), undecadienoic acid (C11: 2), dodecadienoic acid (C12: 2), tridecadienoic acid (C13: 2), tetradecadienoic acid (C14: 2), pentadecadienoic acid (C15: 2), hexadecadi Acid (C16: 2), Heptadecadienoic acid (C17: 2), Octadecadienoic acid (C18: 2, such as linoleic acid, linoelaidic acid, etc.), Eicosadienoic acid (C20: 2), Docosadienoic acid (C22: 2) Tetracosadenoic acid (C24: 2), Hexacosadienoic acid (C26: 2), Octacosadienoic acid (C28: 2), Triacatadienoic acid (C30: 2), Hexadecatrienoic acid (C16: 3), Octadecatrienoic acid ( C18: 3, such as α-linolenic acid, γ-linolenic acid, etc.), eicosatrienoic acid (C20: 3, such as dihomo-γ-linolenic acid, mead acid, etc.), docosatrienoic acid (C22: 3), tetracosa Trienoic acid (C24: 3), hexacosatrienoic acid (C26: 3), octacosatrienoic acid (C28: 3), tria Contatrienoic acid (C30: 3), octadecatetraenoic acid (C18: 4, such as stearidonic acid), eicosatetraenoic acid (C20: 4, such as arachidonic acid), docosatetraenoic acid (C22: 4, For example, adrenic acid etc.), tetracosatetraenoic acid (C24: 4), hexacosatetraenoic acid (C26: 4), octacosatetraenoic acid (C28: 4), triacontatetraenoic acid (C30: 4), Eicosapentaenoic acid (C20: 5), docosapentaenoic acid (C22: 5, such as crupadnonic acid), tetracosapentaenoic acid (C24: 5), docosahexaenoic acid (C22: 6), tetracosahexaenoic acid (C24) : 6, such as nisic acid), and the like.

  In addition to the linear unsaturated fatty acids shown above, lumenic acid (C18: 2), calendic acid (C18: 3), jacaric acid (C18: 3), eleostearic acid (C18: 3), catalpinic acid (C18) : 3), conjugated fatty acids such as punicic acid (C18: 3), luminenic acid (C18: 3), ricinoleic acid (C18: 1), ricinaleic acid (C18: 1), dimorphecolic acid (C18: 2) Hydroxyl unsaturated fatty acids such as epoxy fatty acids such as bemolic acid (C18: 1), furanoid fatty acids such as urofuranic acid, high molecular weight branched unsaturated fatty acids such as mycolic acid, other methoxy unsaturated fatty acids, A cyclic unsaturated fatty acid may be used, and the structure and type are not particularly limited as long as the degree of unsaturation is 1 or more and the number of carbon atoms is 4 or more.

  The glycerophospholipid used in the present invention can be obtained by extracting from a plant material with water or an organic solvent. The plant material to be used is not particularly limited, but it is particularly desirable to use cereals, seeds, potatoes, fruit vegetables, leaf vegetables, stem vegetables, root vegetables, flower vegetables and the like containing a large amount of glycerophospholipid. For example, almond, Aosa, Aonori, Akaza, Acacia, Akane, red grape, red pine (including pine ani, cocoon, copal; the same applies to pine species), agaricus, akinonogeshi, akebi, morning glory, azalea, hydrangea, ashitaba, azuki bean , Asparagus, Acerola, Asenyaku, Anise, Abragiri, Avocado, Ama, Achacha, Achacharu, Amaryllis, Altea, Arnica, Aloe, Angelica, Apricot, Angusok, Rabbit, Izayoi Rose, Ichii, Fig, Ginkgo, Iran, Iran , Turmeric, euglena, moray, prickly pear, udo, ume, vulture, wenzhou mandarin, age, sesame, shallot, elephant, staghorn, enokitake, elderflower, pea, orchid, Obaco, Great White Thistle, Barley, Ochera, Osmanthus, Hypericum, Odoricho, Onidokoro, Olive, Oregano, Orange (including Orange Peel), Carnation, Cacao, Oyster, Kakidooshi, Cuckoo, Oak, Katakuri, Pumpkin, Chamomile, Camomile, Camo Raven, larch, quince, garcinia, cardamom, raspberry, kiwi, kikyo, cabbage (including kale), caraway, cucumber, kumquat, ginnan, guava, wolfberry, kudzu, gardenia, cumin, cranberry, walnut, grapefruit, clove, Black pine, black bean, chlorella, tsutsumeishi, genosho, berry, coconut, pepper, cosmos, burdock, wheat (including wheat germ), sesame, komatsuna, rice (including rice bran) ), Coriander, konnyaku rice cake (including konjac powder), kombu, salmon berry, cypress, pomegranate, sweet potato, sugarcane, sugarcane, sugar beet, saffron, pomelo, hawthorn, salamander, shiitake mushroom, cyclamen, perilla, shimeji , Potato moth, peonies, jasmine, juzudama, shungiku, ginger, ginger, ginger, ginkgo, cinnamon, watermelon, sweet pea, horsetail, star anise, star apple, sudachi, stevia, plum, sage (salvia), mallow, celery, senkyu , Assembly, buckwheat, broad bean, radish, soybean (including okara), Daidai, thyme, bamboo shoot, onion, tarragon, taro, tanjin, dandelion, chicory, tea, camellia, tsukushi, camellia, camellia, tsutsu Mexa, Tsuruxa, Tsuruna, Tsuwabuki, Dill, Tengekuaoi (Geranium), Toga, Pepper, Toki, Red pepper, Corn, Dokudami, Tokon, Tochu, Toneriko, Nagaimo, Nazuna, Rapeseed, Nutmeg, Nigiri, Nigori Carrot, garlic, leek, yarrow, saw palmetto, nobil, verbena, palm (nuclear), pineapple, hibiscus, chickweed, basil, parsley, peppercorn, mint, pearl barley, banana, vanaba, vanilla, paprika, hamamelis, beet, bell pepper , Hinoki, hinoki, pistachio, hyssop (willow mint), daisies, daisies, cypress, hiba, castor, sunflower, loquat, phalaenopsis, phenegrek, fukinotou, grape (including seeds), buoy Cucurrant, blackberry, plum, blueberry (including bilberry), prunes, loofah, safflower, belladonna, bergamot, spinach, spinach, physalis, bodaiju, button, hop, jojoba, borage, maitake, maou, maca, macadamian nut, Matatabi, Marigold, Mango, Honey Bee, Mimosa, Myoga, Myrrh, Murasaki, Mace, Melissa, Merryroth, Melon, Men (including cottonseed oil lees), Palms, Cornflower, Yamamata, Yamayuri, Yamamomogi, Eucalyptus, Yukinoshita, Yuzu, Lily, Yokuinin, Yomena (Aster), Artemisia, Lime, Rye, Lilac, Raspberry, Peanut, Pepper, Apple (including Apple Fiber), Gentian, Ganoderma, Lettuce, Lemon, Forsythia Lotus root, rose hips, rosemary, bay leaf, onion, (including horseradish) wasabi, and the like.

  As long as it does not impair the effect of this invention as an organic solvent used in order to obtain said plant raw material-derived glycerophospholipid, what kind of thing may be used. For example, alcohols such as methanol, ethanol, propanol and butanol, polyhydric alcohols such as ethylene glycol, propylene glycol and glycerin, ketones such as acetone and methyl ethyl ketone, esters such as methyl acetate and ethyl acetate, tetrahydrofuran and diethyl ether Ethers such as dichloromethane, halogenated hydrocarbons such as dichloromethane, dichloroethane and chloroform, aliphatic hydrocarbons such as hexane and pentane, aromatic hydrocarbons such as toluene, polyethers such as polyethylene glycol, pyridines, etc. Can be mentioned. One of these solvents may be used alone, or a plurality of solvents may be used.

  Among them, when used for food, ethanol and hexane are desirable, and water, enzymes, various surfactants, etc. can be used within a range not impairing the effects of the present invention in order to increase extraction efficiency. is there. Further, in addition to using an organic solvent as described above, it is also possible to use a supercritical extraction method which has been attracting attention in recent years.

  The glycerophospholipid-containing extract thus obtained may be used as it is as the collagen production promoter of the present invention, and is concentrated, decolored, desalted, distributed, powdered as long as the effects of the present invention are not impaired. You may use what performed processing, such as conversion. For example, the solvent may be removed by concentration under reduced pressure to increase the solid content, or the colored component may be removed by activated carbon treatment, and water-soluble by liquid / liquid distribution between the aqueous layer and the organic solvent layer. What removed the component may be used. Moreover, you may refine | purify by various chromatography of a normal phase type | system | group or a reverse phase type | system | group. Further, those extracts or processed products may be powdered by adding excipients such as dextrin and lactose. The content of glycerophospholipid is preferably 0.01% by mass to 50% by mass and more preferably 0.1% by mass to 10% by mass with respect to the solution or powder. If it is less than 0.01% by mass, the effect is low, and if it exceeds 50% by mass, it is not desirable from the viewpoint of production cost and handling.

  Since the glycerophospholipid (including the extract) used in the present invention exhibits an excellent collagen production promoting action particularly by oral ingestion, the collagen production promoter of the present invention comprising this as an active ingredient is taken orally. There will be improvements in skin flexibility and elasticity, bone and tendon strength.

  The subject of the present invention is a warm-blooded animal, preferably a mammal, most preferably a human.

  Various pharmaceutically acceptable carriers can be added to the formulation. For example, excipients, binders, disintegrants, lubricants, flavoring agents, coloring agents, sweeteners, corrigents, solubilizers, suspending agents, emulsifiers, coating agents can be included. It is not limited to. The collagen production promoter of the present invention may be sustained or sustained release.

  The active ingredient of the collagen production promoter of the present invention is an extremely safe substance that is glycerophospholipid and has sufficient food experience. From this point, it is considered that the dose of the collagen production promoter of the present invention is not strictly limited, but in general, the lower limit is the minimum amount that can exert an effect according to the purpose of prevention or treatment, the upper limit is From the viewpoint of ease of taking, economical efficiency, etc., the actual amount is used as a standard, and usually about 5 mg to about 50 g, preferably about 50 mg to about 5 g per day, in terms of glycerophospholipid may be taken. . Of course, it can be changed according to the age, weight, symptom, duration of treatment, course of treatment, etc. of the person taking the medicine. The daily dose can be administered in several divided doses. It can also be taken in combination with other collagen production promoters.

When the collagen production promoter of the present invention is added to food or drink , the content of the collagen production promoter of the present invention is not considered to be strictly limited, but generally the lower limit depends on the purpose of prevention or treatment. The upper limit is based on the practical amount from the viewpoint of ease of intake, economics, etc., and is usually about 5 mg to about 50 g per day for an adult in terms of glycerophospholipid, Preferably about 50 mg to about 5 g may be taken. Of course, it can be changed according to the age, weight, symptom, intake period, course of treatment, etc. of the person who takes it. The daily dose can be taken in several divided doses. It can also be taken in combination with other collagen production promoters.

The collagen production promoter can and formulation child so that the above amounts in food and drink of the raw material of the present invention. In is al, can be added iron, inorganic components such as calcium, various vitamins, oligosaccharides, dietary fibers such as chitosan, proteins, such as soybean extract, lipids, sugars sucrose and lactose, and the like.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

Example 1 (Preparation 1 of collagen production promoter)
A collagen production promoter was prepared by adding 1 mL of ethanol to 10 mg of the following unsaturated fatty acid ester type glycerophospholipid, stirring and dissolving.

(Unsaturated fatty acid ester type glycerolysophospholipid)
<Collagen production promoter 1> 1-Oleoyl-sn-glycero-3-phosphate sodium salt (O-LPA; manufactured by Sigma)
<Collagen production promoter 2> 1-Oleoyl-sn-glycero-3-phosphocholine (O-LPC; manufactured by Sigma)

(Unsaturated fatty acid ester type glycerophospholipid)
<Collagen production promoter 3> 1,2-Dioleyl-sn-glycero-3-phosphate sodium salt (DOPA; manufactured by Sigma)
<Collagen production promoter 4> 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC; manufactured by Sigma)
<Collagen production promoter 5> 1-Oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine (OPPC; manufactured by Sigma)
<Collagen production promoter 6> 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC; manufactured by Sigma)
<Collagen production promoter 7> 1-Palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC; manufactured by Sigma)
<Collagen production promoter 8> 1,2-Dilinoleoyl-sn-glycero-3-phosphocholine (DLPC; manufactured by Sigma)
<Collagen production promoter 9> 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE; manufactured by Sigma)
<Collagen production promoter 10> 1,2-Dioleoyl-sn-glycero-3-phosphoserine sodium salt (DOPS; manufactured by Sigma)

Example 2 (Preparation 2 of collagen production promoter)
10 kg of konjac fly flour was charged into a stirring bowl, 20 L of ethanol was added thereto, stirred at room temperature for 2 hours, and the extract and the residue were separated by filtration. The extracted solution was concentrated by an evaporator to obtain about 100 g of a brownish waxy concentrate. 24 g of the waxy concentrate was applied to a silica gel column, eluted with chloroform and acetone, and then eluted with methanol to obtain 4 g of an extract containing konjac and powdered phospholipids. From the NMR and HPLC analysis results, it was confirmed that the extract was a glycerophospholipid-rich extract containing an unsaturated fatty acid such as linoleic acid. A collagen production promoter was prepared by adding 10 mL of ethanol to 100 mg of this glycerophospholipid-rich extract and stirring and dissolving (collagen production promoter 11).

Example 3 (Preparation 3 of collagen production promoter)
A collagen production promoter was prepared by adding 10 mL of ethanol to 100 mg of soybean-derived lecithin, stirring and dissolving (collagen production promoter 12).

Example 4 (Evaluation of collagen production promoting effect; cell experiment 1)
Normal human skin fibroblasts (NHDF: manufactured by Kurabo Industries) are seeded in a 24 well plate at 5 × 10 ⁴ cells / well, and contain 2% fetal bovine serum (FBS) in the presence of 37 ° C. and 5% carbon dioxide gas. The cells were cultured in a medium (low serum medium for human skin fibroblast proliferation containing a low serum growth additive: Cascade, Cascade). 24 hours later, the cells were washed twice with the above-mentioned dedicated medium not containing FBS, and collagen production promoters 1 to 10 containing the unsaturated fatty acid ester type glycerolysophospholipid and the unsaturated fatty acid ester type glycerophospholipid prepared in Example 1 were used. 500 μl of the same medium mixed with (prepared to 20 μM) was added, and the culture was continued. After 24 hours, the type I procollagen-I carboxylate peptide (PIP) contained in the culture supernatant was measured using Procollagen Type-I Peptide EIA Kit (manufactured by Takara Bio Inc.). PIP is a propeptide cleaved from procollagen, which is a type I collagen precursor, and is an index reflecting the amount of collagen synthesis. About the production promotion effect of lysophospholipid, the PIP production amount of the comparative control group (blank) was evaluated as 100%.

  As is clear from Table 1, the unsaturated fatty acid ester type glycerolysophospholipid and the unsaturated fatty acid ester type glycerophospholipid of the present invention exhibited collagen production promoting activity. It was shown that the production promoting activity was strong in the order of phosphatidic acid> phosphatidylcholine> phosphatidylethanolamine> phosphatidylserine, and became stronger as the degree of unsaturation of the constituent fatty acids and the number of unsaturated fatty acids increased.

Example 5 (Preparation 4 of collagen production promoter)
A collagen production promoter was prepared by adding 1 mL of ethanol to 10 mg of the following saturated fatty acid ester type glycerophospholipid, stirring and dissolving.
(Saturated fatty acid ester type glycerophospholipid)
<Collagen production promoter 13> 1,2-Dipalmitoyyl-sn-glycero-3-phosphate sodium salt (DPPA; Sigma)
<Collagen-Producing Agent 14> 1,2-Dipalmitoyyl-sn-glycero-3-phosphocholine (DPPC; Sigma)

Example 6 (Evaluation of collagen production promoting action; cell experiment 2)
As in Example 4, normal human dermal fibroblasts were seeded in a 24-well plate and cultured in a dedicated medium containing 2% fetal bovine serum (FBS) at 37 ° C. in the presence of 5% carbon dioxide gas. After 24 hours, the cells were washed twice with the above-mentioned dedicated medium containing no FBS, and 500 μl of the same medium (prepared to 20 μM) mixed with the saturated fatty acid ester-type phospholipid prepared in Example 5 was added to continue the culture. did. After 24 hours, PIP contained in the culture supernatant was measured by the same method as in Example 4. About the production promotion effect of phospholipid, the PIP production amount of the comparative control group (blank) was evaluated as 100%.

  As is apparent from Table 2, those containing saturated fatty acid ester type phospholipids exhibited collagen production promoting activity, although their activities were lower than those containing at least one unsaturated fatty acid shown in Table 1.

Example 7 (Evaluation of collagen production promoting action of plant material-derived glycerophospholipid; cell experiment 3)
As in Example 4, normal human dermal fibroblasts were seeded in a 24-well plate and cultured in a dedicated medium containing 2% fetal bovine serum (FBS) at 37 ° C. in the presence of 5% carbon dioxide gas. After 24 hours, the cells were washed twice with the above-described dedicated medium not containing FBS, and 500 μl of the same medium (prepared to 10 mg / ml) mixed with collagen production promoters 11 and 12 prepared in Examples 2 and 3 was added. The culture was continued. After 24 hours, PIP contained in the culture supernatant was measured by the same method as in Example 4. About the production promotion effect of phospholipid, the PIP production amount of the comparative control group (blank) was evaluated as 100%.

  As is apparent from Table 3, the plant material-derived glycerophospholipid-containing collagen production promoter of the present invention exhibits a collagen production-promoting activity, and in particular, konjac fly flour-derived phospholipid has a high collagen production-promoting activity. It was.

Example 8 (Evaluation of collagen production promoting effect by oral ingestion; animal experiment)
Wistar rats (male, 4 weeks old, 5 animals / group) were bred for 3 weeks on a diet containing 6% protein to obtain pseudo-aging model rats. Collagen production promoter 8 (DLPC) prepared in Example 1 was suspended in tap water so as to be 1 mg / ml, and 1 mL was orally administered once a day for 5 weeks for 100 g body weight of the pseudo-aged model rat. Ingested (equivalent to 10 mg / day / kg-BW). Similarly, the collagen production promoters 11 and 12 prepared in Examples 2 and 3 are suspended in tap water so as to be 1 mg / ml, and 1 mL is once a day for 100 g body weight of the simulated aging model rat. Orally ingested weekly (equivalent to 10 mg / day / kg-BW).

  After the administration was completed, the hair on the back of the rat was removed, and the skin was excised into a 2 × 5 cm rectangle together with the subcutaneous tissue. After weighing, 10 mL of distilled water cooled with ice was added to the extracted skin and homogenized sufficiently, and the precipitate was collected by centrifugation (7000 rpm × 20 min). 10 mL of ice-cooled 0.1N sodium hydroxide was added to the precipitate, shaken overnight under refrigeration (6 ° C.), the precipitate was recovered by centrifugation, and the same operation was performed again. The precipitate recovered by centrifugation is washed with ice-cooled distilled water, and 15 mL of ice-cooled 0.5 M acetic acid is added to the precipitate recovered again after centrifugation, and collagen extraction is performed overnight under refrigeration (6 ° C.). Extraction supernatant was obtained by centrifugation. Soluble collagen in the extracted solution was quantified using Sircol Collagen Assay Kit (Funakoshi).

  As is apparent from Table 4, it was shown that the collagen production promoter of the present invention promotes the production of collagen in the skin by increasing the skin weight and the amount of skin soluble collagen and taking it orally.

Although the present invention has been described in detail and with reference to specific embodiments, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the spirit and scope of the invention.
This application is based on a Japanese patent application filed on August 28, 2007 (Japanese Patent Application No. 2007-221384), the contents of which are incorporated herein by reference.

  INDUSTRIAL APPLICABILITY The present invention can provide a collagen production promoter, a food / beverage product containing the same, and a pharmaceutical product.

Claims (5)

  A collagen production promoter comprising a glycerophospholipid as an active ingredient, wherein at least one of the fatty acids constituting the glycerophospholipid is an unsaturated fatty acid.   The glycerophospholipid is composed of phosphatidylcholine, lysophosphatidylcholine, phosphatidylserine, lysophosphatidylserine, phosphatidylethanolamine, lysophosphatidylethanolamine, phosphatidylinositol, lysophosphatidylinositol, phosphatidic acid, lysophosphatidic acid, phosphatidylglycerol, and lysophosphatidylglycerol. The collagen production promoter according to claim 1, which is one or more selected.   The collagen production promoter according to claim 1 or 2, which exhibits a collagen production promoting effect by oral ingestion.   The collagen production promoter according to any one of claims 1 to 3, wherein the glycerophospholipid is phosphatidic acid. The pharmaceutical containing the collagen production promoter of any one of Claims 1-4 .