JP6274714B2 - Sebum production inhibitor - Google Patents

Description

  The present invention relates to a sebum production inhibitor that suppresses the production of sebum, and cosmetics, foods and drinks, pharmaceuticals, feeds, and pet foods containing the agent.

  Sebum secreted from the sebaceous glands, which are the appendages of the skin, forms physiologically important functions such as buffering, bactericidal, and excretion of endogenous substances by forming a fat film in the epidermis. (For example, refer nonpatent literatures 1 and 2). Thus, sebum line dysfunction, that is, excessive secretion and decreased secretion of sebum, is closely related to the onset of skin diseases such as acne vulgaris, seborrheic rash and xeroderma (dry skin), respectively. Especially, acne caused by increased sebum secretion is a skin disease accompanied by mental stress not only in adolescence but also in adult men and women because it develops mainly on the face, and lowers the quality of life of patients (for example, Non-patent documents 1 to 4).

  Production and secretion of sebum has been reported to involve male hormones such as 5α-dihydrotestosterone and insulin (see, for example, Non-Patent Documents 5 to 8). In addition, it has been reported that perilipin, which is one of phosphorylated proteins and is involved not only in the formation of intracellular lipid droplets but also in lipid synthesis / degradation regulation, is expressed in hamster sebaceous glands (for example, non-patented) References 9 and 10).

  Acne is a non-inflammatory acne that causes comedones due to follicular occlusion and sebum retention, and inflammation with an inflammatory response due to the growth and immune response to Propionibacteriumacne (P. acne), an obligate anaerobic Gram-positive gonococci It is classified as acne (see, for example, Non-Patent Documents 11 and 12). Since both are pathologically different, it is considered that a medical strategy based on the two aspects of acne treatment is essential (see, for example, Non-Patent Document 13). For non-inflammatory acne, 13-cis Retinoic acid (isotretinoin, hereafter abbreviated as “13cisRA”) is a retinoic acid preparation to improve abnormal skin keratinization, while P. acne is targeted for inflammatory acne. An oral antibacterial agent is prescribed (see, for example, Non-Patent Document 14). 13cisRA can also directly suppress abnormal increase in sebum production, which is the root cause of acne (see, for example, Non-Patent Documents 15 to 17). Is limited (see, for example, Non-Patent Document 14). Therefore, the present situation is that development of a safe and effective pharmaceutical that can directly suppress sebum production is desired.

  In view of these situations, there are a plurality of sebum production inhibitors such as inhibitors using extracts from natural herbal medicines (see, for example, Patent Documents 1 and 2).

  However, the known sebum production inhibitors as described above have problems in safety and the like, and none have satisfactory effects. Under these circumstances, there has been a demand for a product having an effect of suppressing sebum production that is safe even when used in a small amount for a long period of time.

  Carotenoids are compounds having a conjugated double bond and a methyl group side difference at the center, and are classified into xanthophylls containing carotenes and oxygen atoms, which are composed only of hydrocarbons. Carotenes are also classified into α-carotene and β-carotene having a cyclic hydrocarbon at both ends of the molecular chain, and lycopene having no cyclic hydrocarbon at both ends of the molecular chain. There are many types of these carotenoids depending on the conjugated double bond chain and the side chain added to the cyclic hydrocarbon, and there are more than 750 types of carotenoids whose structures are determined. Animals have no action to synthesize carotenoids, and plants and some microorganisms can synthesize carotenoids.

  Of the carotenoids, α-carotene, β-carotene, lutein / zeaxanthin, lycopene, and cryptoxanthin are known as carotenoids that have particularly high transferability into the blood, and are also known to accumulate in organs and the like. . In addition, epidemiological studies and the like have been reported to have a negative correlation with the occurrence of cardiovascular disease (for example, see Non-Patent Document 18) and to have effects such as prevention of macular degeneration (for example, see Non-Patent Document 19). .

  However, it has not been known that carotenoids and / or derivatives thereof act on sebaceous glands and suppress sebum production.

JP 2003-212789 A JP-A-2005-206588

Standard Dermatology (Masahiro Sasakawa, Satoshi Tomita, Takashi Hashimoto) (2007) Medical School, 25-27, 339-341. Thiboutot, D. (2004) J. Invest. Dermatol. 123, 1-12 Farrar, M. D. and Ingham, E. (2004) Clin. Dermatol. 22, 380-384. Zouboulis, C. C., Eady, A., Philpott, M., Goldsmith, L. A., Orfanos, C., Cunliffe, W. C., and Rossenfield, R. (2005) Exp. Dermatol. 14, 143-152. Plewig, G. and Luderschmidt, C. (1977) J. Invest. Dermatol. 68, 171-176. Akamatsu, H., Zouboulis, C. C., and Orfanos, C. E. (1992) J. Invest. Dermatol. 99, 509-511. Deplewski, D. and Rosenfield, R. L. (1999) Endocrinology 140, 4089-4094. Sato, T., Imai, N., Akimoto, N., Sakiguchi, T., Kitamura, K., and Ito, A. (2001) J. Invest. Dermatol. 117, 965-970. Akimoto, N., Sato, T., Iwata, C., Koshizuka, M., Shibata, F., Nagai, A., Sumida, M., and Ito, A. (2005) J. Invest. Dermatol. 124 , 1127-1133. Kimmel, A. R., Brasaemle, D. L., McAndrews-Hill, M., Sztalryd, C., and Londos, C. (2010) J. Lipid Res. 51, 468-471. Bojar, R. A. and Holland, K. T. (2004) Clin. Dermatol. 22, 357-379. Pawin, H., Beylot, C., Chivot, M., Poli, F., Revuz, J., and Dreno, B. (2004) Eur. J. Dermatol. 14, 4-12. Dermatological practice 18 Techniques for acne treatment (Fukumi Yoshikawa, Yoshiki Miyaji, Masahiro Sasakawa) (2005) Bunkodo, 48-52. 171-175. Thiboutot, D., Gollnick, H., Bettoli, V., Dreno, B., Kang, S., Leyden, JJ, Shalita, AR, Lozada, VT, Berson, D., Finlay, A., Goh, CL , Herane, MI, Kaminsky, A., Kubba, R., Layton, A., Miyachi, Y., Perez, M., Martin, JP, Ramos-E-Silva, M., See, JA, Shear, N ., Wolf, J. Jr. (2009) J. Am. Acad. Dermatol. 60, S1-S50. Tsukada, M., Schroder, M., Roos, T. C., Chandraratna, R. A., Reichert, U., Merk, H. F., Orfanos, C. E., and Zouboulis, C. C. (2000) J. Invest. Dermatol. 115, 321-327. Zouboulis, C. C. (2006) J. Invest. Dermatol. 126, 2154-2156. Sato, T., Takahashi, A., Kojima, M., Akimoto, N., Yano, M., and Ito, A. (2007) J. Invest. Dermatol. 127, 2740-2748. Treatment Vol. 84 Fruits and lifestyle-related diseases (Min Sugiura) (2002) No. 84 4 142-144 D Max Snodderly (1995) Am J Clin Nutr. 62,1448S-61S

  The object of the present invention is to solve the above-mentioned problems, exhibit effects in a small amount, and prevent sebum production even when used for a long period of time, and cosmetics, foods and drinks, and pharmaceuticals containing the sebum production inhibitor To provide feed, pet food and the like.

  As a result of intensive studies to solve the above problems, the present inventors have unexpectedly found a sebum production inhibitory effect on carotenoids and / or derivatives thereof, particularly cryptoxanthin and / or derivatives thereof, and have reached the present invention.

That is, the present invention is as follows.
(1) A sebum production inhibitor comprising a carotenoid and / or a derivative thereof.
(2) A sebum production inhibitor, wherein the carotenoid and / or its derivative is cryptoxanthin and / or its derivative.
(3) The sebum production inhibitor according to (2), wherein cryptoxanthin and / or a derivative thereof is derived from citrus.
(4) The sebum production inhibitor according to (3), wherein the citrus fruit is Wenzhou mandarin orange.

It is the graph which compared the inhibitory effect by (beta) -cryptoxanthin and 13cisRA with respect to the triacylglycerol (TG) production of a hamster sebocyte of non-insulin induction. It is the graph which compared the inhibitory effect by (beta) -cryptoxanthin and 13cisRA with respect to the triacylglycerol (TG) production of an insulin induction hamster sebocyte. It is the graph which compared the time-dependent inhibitory effect by (beta) -cryptoxanthin and 13cisRA with respect to the triacylglycerol (TG) production of an insulin induction hamster sebocyte. It is the graph which compared the intracellular lipid droplet formation inhibitory effect by (beta) -cryptoxanthin and 13cisRA in the insulin acyl-inducible ([A]) or insulin-induced ([B]) hamster sebaceous gland triacylglycerol (TG) production. .

  Hereinafter, the present invention will be described in detail.

  The carotenoid in the present invention is not particularly limited, but α-cryptoxanthin, β-cryptoxanthin, isocryptoxanthin, actinoerythrol, astaxanthin, astaxin, adonixanthin, phenicoxanthine, β-apo-8′-carotenal, β-apo-8′-carotenol, alloxanthin, anteraxanthin, isoagelaxanthin, isoseasatintin, isoreniellaten, β-isorenieraten, idoxanthin, equinone, 3′-epilutein, erythroxanthin sulfate, Okenone, osylaxanthin, capsanthin, cabsorbin, caroxanthin, α-carotene, β-carotene, β-carotene, δ-carotene, ε-carotene, ε, ε-carotene-3.3'-diol, η-carotene, ζ -Carotene, Kanta Santine, guilloxanthine, guaaxanthin, crystaltaxanthin, crocoxanthin, crocetin, chloroxanthine, chlorobactene, ketomyxocoxanthin, ketomixol fucoside, geliodesxanthine, zynoxanthine, thermozeaxanthin, diazinoxanthine, diatoxanthine Diaponeurosporene, diapophytoene, 4.4'-diapolycopene dial, 2,2'-diketospiriloxanthine, 3,4-dehydrorhodopine, dehydrolycopene, citranaxanthin, citranaxanthin , Siphonaxanthin, staphyloxanthine, spiroxanthin, spheroidenone, sphenoidene, zeaxanthin, taraxanthin, tunaxanthin, deoxyflexoxanthin, decaprenoxanthin, Thiaten, demethylspheroidene, paplioerythrin, trikentriorhodin, tolralozin, tolylene, neurospoxanthine, neurosporen, neoxanthine, nostoxanthin, bacteriorubixanthin, bacteriovelin, bastaxanthin, paplioerythrinone , Paplioerythrin, parasiloxanetin, halacinthiaxanthine, violaxanthin, fizzarien, phytoene, phytofluene, philipsiaxanthine, phenicoxanthine, fucoxanthin, prasinoxanthine, flavuxanthin, frituraxanthin, plectiniaxanthin, hetero Xanthine, peridinin, helenien, bokelia xanthine, myxoxanthine, mytiloxanthin, mutoxanthine, mutochrome Meso-zeaxanthin, monads xanthine, easy Tsuka xanthine, lycopene, lutein, ruby xanthine, rodoxanthin, Rodopinaru, Rodopinoru, Rodobiburin, Rodopin, and the like Lolo xanthine.

  The carotenoid derivative in the present invention is not particularly limited, and examples thereof include fatty acid ester, sulfate ester, hydroxyl adduct, dehydroxylated and glycoside of the carotenoid.

  Examples of the fatty acid ester include fatty acid esters such as lauric acid (C12), myristic acid (C14), palmitic acid (C16), and stearic acid (C18).

  Among the carotenoids and / or derivatives thereof, β-cryptoxanthin, α-carotene, β-carotene, lycopene, lutein / zeaxanthin, and / or derivatives thereof are used when taken orally from the viewpoint of the sebum production inhibitory effect. Β-cryptoxanthin and / or its derivatives are particularly preferable.

  In the case of transdermal absorption, β-cryptoxanthin and / or a derivative thereof, astaxanthin and / or a derivative thereof, fucoxanthin and / or a derivative thereof are preferable from the viewpoint of sebum production inhibitory effect, and β-cryptoxanthin and / or a derivative thereof are preferable. Its derivatives are particularly preferred.

  The sebum production inhibitor of the present invention has eight carotenoids (β-cryptoxanthin, α-carotene, β-carotene, lycopene, lutein / zeaxanthin, astaxanthin, fucoxanthin) with respect to total carotenoids from the viewpoint of sebum production inhibitory effect and / The total proportion of carotenoids of these derivatives is preferably 50% by mass or more, more preferably 70% by mass or more, and even more preferably 90% by mass or more.

  The dosage form of the sebum production inhibitor of the present invention may be any of solid, semi-solid, and liquid, and is appropriately set according to the form of the product to be blended. In the product in which the sebum production inhibitor of the present invention is blended, water, fats and oils, waxes, hydrocarbons, fatty acids, higher alcohols, and the like within the range that does not impair the effects of the present invention, depending on the form and the like. , Esters, plant extracts, water-soluble polymers, surfactants, metal soaps, alcohols, polyhydric alcohols, pH adjusters, antioxidants, UV absorbers, preservatives, fragrances, powders, thickening You may contain additives, such as an agent, a pigment | dye, and a chelating agent. Moreover, the product in which the sebum production inhibitor of the present invention is blended may contain a functional component in a range that does not impair the effects of the present invention, depending on the form and use. Examples of the functional component include vitamin C, collagen, squalane, niacin, niacinamide, hyaluronic acid, placenta extract, sorbitol, chitin, chitosan, various plant extracts, and the like. About these compounding quantities, unless the effect of this invention is impaired, it is not limited.

  The sebum production inhibitor in the present invention is preferably ingested as a carotenoid so that the daily intake per adult is 0.01 to 100 g, more preferably 0.1 to 50 g, and even more preferably 0.1 to 30 g. preferable. In addition, when the intake amount per day for an adult is less than 0.01 g, the sebum production inhibitory effect may be lowered. On the other hand, if the intake amount per day for an adult exceeds 100 g, the raw material cost increases for the resulting sebum production inhibitory effect, which is not preferable.

  Since the sebum production inhibitor in the present invention can be used in any administration form such as oral, enteral, diatomical mucosa, injection, etc., it is used by being blended in products such as cosmetics, foods and drinks, pharmaceuticals, pet foods, and feeds. .

  When the sebum production inhibitor of the present invention is used in cosmetics, the sebum production inhibitor of the present invention is prepared in a desired form in combination with a cosmetically acceptable base material and additive ingredients, and used for suppressing sebum production. Offered as a cosmetic. The form of such cosmetics is not particularly limited, and specific examples include emulsions, creams, lotions, packs, cosmetic liquids, cleaning agents, makeup cosmetics, and the like.

  Further, when the sebum production inhibitor of the present invention is used in foods and drinks, the sebum production inhibitor of the present invention is prepared alone or in combination with other food materials and additive ingredients into a desired form, for sebum production suppression. Provided as a food and drink. Examples of such foods and drinks include foods for specified health use, dietary supplements, functional foods, foods for the sick, and the like in addition to general foods and drinks. Although it does not restrict | limit especially as a form of these food / beverage products, Specifically, Main dishes, such as breads and noodles; Side dishes, such as cheese, ham, winner, and processed seafood; Fruit juice drink, carbonated drink, milk drink, etc. Beverages; cookies, cakes, jelly, pudding, candy, yogurt and other favorite products; tablets, granules, powders, capsules, soft capsules, nutritional drink supplements, etc. In addition, about the food for sick people, it can be used as a meal for the patient of the disease from whom improvement of a symptom is anticipated by sebum production suppression.

  Furthermore, when using the sebum production inhibitor of this invention for food-drinks, the sebum production inhibitor of this invention can also be provided as a food additive for sebum production suppression individually or in combination with another component.

  In addition, when the sebum production inhibitor of the present invention is used in pharmaceuticals, the sebum production inhibitor of the present invention is desired alone or in combination with other pharmacologically active ingredients, pharmaceutically acceptable base materials, additive ingredients, etc. And is provided as a pharmaceutical product for suppressing sebum production. Although it does not restrict | limit especially as a form of such a pharmaceutical, Specifically, oral administration formulation, such as a tablet, a granule, a powder agent, a capsule, a soft capsule, a syrup, external preparation, an inhalant, a suppository, etc. Transdermal or transmucosal preparations; injections and the like.

  In addition, when the sebum production inhibitor of the present invention is used for feed or pet food, the sebum production inhibitor of the present invention is prepared alone or in combination with other feed ingredients into a desired form, and used for suppressing sebum production. Provided as feed or pet food. Examples of feed ingredients used in the feed or pet food include cereals such as corn, wheat, barley and rye; brans such as bran and rice bran; moss such as corn gluten meal and corn jam meal; skim milk powder; Animal feeds such as whey, fish meal and bone meal; yeasts such as beer yeast; calciums such as calcium phosphate and calcium carbonate; vitamins; fats and oils; amino acids;

  Regarding the administration or intake of the sebum production inhibitor when the sebum production inhibitor of the present invention is used as cosmetics, foods / drinks, pharmaceuticals, feeds and pet foods, the dosage form of the agent, the age of the subject of administration, and expectation The amount effective for the suppression of sebum production may be appropriately set according to the effect to be achieved. For example, the administration or intake per day as a carotenoid is usually set to about 0.01 to 100 g, preferably about 0.1 to 50 g, and more preferably about 0.1 to 30 g. By satisfying such administration or intake, the effect of suppressing sebum production can be effectively achieved in vivo.

  In the sebum production inhibitor of the present invention, cosmetics, foods and drinks, pharmaceuticals, feeds and pet foods containing the same, various functional ingredients may be blended as necessary within a range not impairing the effects of the present invention. I can do it. Examples of functional ingredients include other sebum production inhibitors, vitamin C, collagen, squalane, niacin, niacinamide, hyaluronic acid, placenta extract, sorbitol, chitin, chitosan, and various other plant extracts. Thing etc. are mentioned. About these compounding quantities, unless the effect of this invention is impaired, it is not limited.

  The method for producing the sebum production inhibitor of the present invention will be described below with examples.

  The carotenoid and / or derivative thereof of the present invention can be obtained by a method of extracting from animals or plants or bacteria using a solvent or the like. Further, the carotenoid extract is preferably concentrated by concentrating to dryness by a conventional method.

  When obtaining carotenoids derived from animals and plants, the animals and plants to be used are not particularly limited, but grains such as brown rice, corn, sweet potatoes, sweet potatoes, sweet potatoes such as sweet potatoes, and processed products thereof, azuki bean, green beans, sweet potatoes, sweet potatoes, broad beans , Daizu, Chickpea, Benibanengen, Ramame, Ryoto, Lentil and other beans, Almond, Asa, Sesame, Cashew nut, Kaya, Ginkgo, Chestnut, Walnut, Coconut, Sesame, Shi, Watermelon, tochi, lotus, rhinoceros, pistachio, sunflower, brazil nut, hazelnut, macadamia nut, pine, peanut and other seeds, artichoke, morning glory, tomorrow, asparagus, udon, funny, hill hijiki, okura , Turnip, pumpkin, mustard, cauliflower, kanpyo, chrysanthemum, cabbage, cucumber, Japanese garlic, Kyosai, Mushrooms, Kinsai, Watercress, Hakui, Kale, Kohlrabi, Garbage, Burdock, Komatsuna, Zhasai, Santo Usagi, Shisogarashi, Shiso, Jukusai, Shungiku, Junsai, Ginger, Shiro Zucchini, crispy, celery, mainspring, taasai, daikon, daisai, takana, bamboo shoot, onion, tuna, chicory, chingensai, boar, vine, tsurumura saki, hot pepper, chili, Winter potato, tomatoes, trevis, tonbutsu, nagasaki haikusai, eggplants, nazuna, nigari, leeks, carrots, garlic, green onions, nozawana, nobori, napa cabbage, pakchoi, basil, parsley, sardine konnyaku, Hayatouri, beet , Peppers, hinona, hiroshimana, wipes, buns, block Lee, Spinach, Horseradish, Makomo, Mizukake, Trefoil, Myouga, Mago, Mameha, Sprouts, Morohaya, Yamagobo, Yuri, Yosai, Yomagi, Peanuts, Rakikyo, Riki, Rhubarb, Vegetables such as lettuce, lotus root, shrimp, wasabi, bracken, acerola, avocado, apricot, strawberry, fig, yokan, plum, unshu mandarin orange, olive, orange, oroblanco, persimmon, pumpkin, karin, kiwifruit, kiwano, kumquat , Guava, gooseberry, gummy, grapefruit, gourmet, cherry, sanbokan, shikuwasha, watermelon, sudachi, plum, daikon, tangor, danzero, cherimoya, durian, summer mandarin, natsume, jujube, pineapple, hascup , Hassle, passion flu Tsu, banana, papaya, hyuga natsu, loquat, grape, blueberry, buntan, white support, ponkan, makiwauri, quince, mango, melon, peach, yamomo, yuzu, raspberry, apple, lemon , Aonori, Amanori, Arame, Iwanori, Enogori, Ogori, Kawari, Constricted, Kombu, Suizenji, Amakusa, Tosakari, Hijiki, Hiigashi, Funori, Matsumo, Mudori , Seaweeds such as mozuku, sea bream, sea bream, sea bream, eel, caviar, kochi, salmon / masal, sea bream, sea bream, fish, fish, scallop, sea bream, Abalone, Itayagai, Oyster, Sazae, Shijimi, Tani, Mushroom, Fist, Top shell, Garage, Ribbon, Clam, Scallop , Shellfish such as sea bream, potato, shellfish such as shrimp and crab, cephalopods such as squid and octopus, sea urchin, jellyfish, jellyfish, sea bream, sea cucumber, sea bream, wild boar, etc. , Pigs, rabbits, cows, horses, whales, deer, pigs, noodles, goats, Aigamo, poultry and other meats, eggs, quail eggs, chicken eggs, peas and other eggs, milk such as milk And processed products.

  Among the above-mentioned animals and plants, those derived from citrus are preferable. Citrus in the present invention can include plants belonging to the citrus family and the like. More specifically, Wenzhou orange, Iyokan, Summer orange, Orange, Cabos, Kawabata, Kishumikan, Kiyomi, Kumquat, Grapefruit, Gecki, Sanpokan, Shikuwasa, Jabara, Sweety, Sudachi, Daidai, Tachibana, Dekopon, Natsudaidai, Hassaku, Navel orange, Valencia orange, Evening white cocoon, Hyuganatsu, Bungtan, Ponkan, Mandarin orange, Yatsushiro, Yuzu, Lime, Lemon, Karatachi (including varieties equivalent or similar to these) and the like. Among them, Unshu mandarin is desirable because it has a high content of cryptoxanthin and / or its derivatives.

  When obtaining a carotenoid derived from a microorganism, the microorganism to be used is not particularly limited, and examples thereof include microorganisms selected from algae such as Dunaliella and Haematococcus having carotenoid production ability, yeasts such as Phaffia and Rhodotorula. Among these, from the viewpoint of carotenoid production ability, bacteria belonging to the genus Paracoccus, Sphingomonas, Brerevundimonas or Erythrobacter are preferred, and bacteria belonging to the genus Paracoccus are more preferred.

  Among the bacteria belonging to the genus Paracoccus, Paracoccus carotinifaciens, Paracoccus marcusii, Paracoccushaeundaensis and Paracoccus zeaxanthinifaciens are preferable, and Paracoccus carotinifaciens is particularly preferable. Specific examples of bacteria belonging to the genus Paracoccus include Paracoccus carotinifaciens E-396 strain (FERM BP-4283) and Paracoccus genus bacteria A-581-1 strain (FERM BP-4671), and these mutant strains are also included. It is preferably used in the present invention.

  When extracting from animals and plants and / or microorganisms with a solvent, a polar solvent such as water or an organic solvent can be used. The organic solvent is not particularly limited, but 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, methyl acetate, ethyl acetate and the like Esters, tetrahydrofuran, diethyl ether and other ethers, dichloromethane, dichloroethane, chloroform and other halogenated hydrocarbons, hexane, pentane and other aliphatic hydrocarbons, toluene and other aromatic hydrocarbons, polyethylene glycol, etc. Examples include polyethers and pyridines. These may be used alone or in combination of two or more.

  In particular, when the sebum production inhibitor of the present invention is contained in food, it is preferable to use ethanol or hexane among the above organic solvents from the viewpoint of safety. In order to increase the extraction efficiency, water, enzymes, various surfactants, and the like can be used as long as the effects of the present invention are not impaired.

  In the extraction with a solvent, the extraction can be performed a plurality of times. It is also possible to use a supercritical extraction method that has been attracting attention in recent years.

  The extract thus obtained is preferably made into a concentrate by a concentration operation. Examples of the concentration operation include removal of the solvent by using a vacuum concentration device such as an evaporator or heating. Then, the concentrate is purified by a known purification means to obtain carotenoids having high purity and / or derivatives thereof. Specifically, for example, the carotenoid is obtained by purifying the extract several times by silica gel column chromatography, or by removing impurities by alkali treatment or solvent fractionation and then purifying by silica gel column chromatography. Can be obtained.

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

<Production inhibitory effect of triacylglycerol (hereinafter abbreviated as “TG”) by β-cryptoxanthin>
Example 1
(Experimental items and cell culture method)
In order to evaluate the sebum production inhibitory effect of β-cryptoxanthin on hamster sebaceous cells, the following confirmation tests (1) to (4) were carried out.
(1) Lipid synthesis is not induced by insulin (may be abbreviated as “insulin not induced”). For hamster sebaceous cells, β-cryptoxanthin is added at 1.1 μM, 3.3 μM, 10 μM, or 13 cisRA. 0.01 μM, 0.1 μM, and 1 μM were added, and the amount of accumulated TG was compared and evaluated.
(2) Lipid synthesis was induced with 10 nM insulin (may be abbreviated as “insulin induction”). For hamster sebocytes, β-cryptoxanthin was added at 1.1 μM, 3.3 μM, 10 μM, or 13 cis RA. 0.01 μM, 0.1 μM, and 1 μM were added, and the amount of accumulated TG was compared and evaluated.
(3) Addition of β-cryptoxanthin (10 μM lanes 2 and 5) or 13 cisRA (1 μM lanes 3 and 6) to hamster sebocytes that are not induced with insulin (lanes 1 to 3) and insulin (lanes 4 to 6) The amount of TG accumulated on the 3rd, 6th, and 8th days of culture was compared and evaluated.
(4) The effects of β-cryptoxanthin (10 μM) and 13 cisRA (1 μM) on lipid droplets formed in hamster sebaceous gland cells without insulin induction ([A]) and insulin induction ([B]) were evaluated.

[Standard operation]
Standard operations in the above evaluation are shown below.
The cells were seeded in a 35 mm dish (manufactured by Falcon) to 2.7 × 10 ⁵ cells / dish and cultured for 1 day, and then in the presence and / or absence of 10 nM insulin (manufactured by Sigma Chemical), Various concentrations of β-cryptoxanthin (trade name: β-Cryptoxanthin (for experimental research), product code: SY-001, manufactured by Shikoku Yasshu Co., Ltd., sometimes abbreviated as “β-cpx”) or 13cisRA (Sigma Chemical) After being added to Sebocyte-growth medium (sometimes abbreviated as “SG medium”), the cells were cultured for a certain period of time. In addition, what added neither beta-cryptoxanthin nor 13cisRA was set as the solvent control (it may be abbreviated as "C").
The composition of SG medium is shown below.
SG medium:
6 v / v% fetal bovine serum (manufactured by Thermo Fisher Scientific), 2 v / v% human serum (manufactured by MP Biomedicals), 0.68 mM L-glutamine (manufactured by Wako Pure Chemical Industries), 100 unit / ml penicillin G ( MP Biomedicals) and Dulbecco's modified Eagle's medium / Ham's F-12 medium (1: 1) containing 100 μg / ml sulfate streptomycin (Meiji Seika) (Invitrogen)

After the above culture, the following operation was performed in order to quantify the amount of TG accumulated in the cells.
[Measurement of TG amount]
The cultured cells were washed twice with a phosphate buffer, and the cells were used with a phosphate buffer (1 ml) containing 0.25% trypsin (DIFCO Laboratries) and 0.02 wt / v% EDTA (Sigma Chemicals). Was recovered. The obtained cell suspension was pulverized by ice chilling using an ultrasonic pulverizer (Bioruptor, manufactured by Cosmo Bio Inc.) 10 times at 200 W for 10 seconds. The amount of TG in the cells was measured using a Liquitec TGII kit (Roche Diagnostics) according to the attached operation method.
That is, treatment solution 1 [1.65 IU / ml glycerol kinase / 6 IU / ml glycerol-3-phosphate oxidase / catalase / 2.95 mM Disodiumadenosine-5'-triphosphate / 0.65 mM sodium N- (3,5-dimethoxyphenyl) ) -N′-succinylethylenediamine] was added and reacted at 37 ° C. for 10 minutes to remove free glycerol. Immediately after, the attached treatment solution 2 [0.55 IU / ml lipoprotein lipase / peroxidase / 0.65 mM 4-aminoantipyrin] was added, and further reacted at 37 ° C. for 10 minutes. After completion of the reaction, the absorbance at 590 nm was measured with a microplate reader (Infinite F200, manufactured by Tecan Japan), and the amount of TG obtained based on the absorbance of the triolein standard solution (60 mg / ml) was calculated.

  The results of the confirmation tests (1), (2), and (3) described above are shown in FIGS.

Furthermore, intracellular lipid droplet formation was analyzed by the nile red staining method shown below.
[Detection method of lipid droplet formation]
A mixture of dimethyl sulfoxide (DMSO) (Sigma Chemicals) solution containing 0.1v / v% nile red (Sigma Chemicals) and purified water at a ratio of 1: 1000 (volume ratio) was used as the nile red staining solution. . Hamster sebaceous cells were seeded in 96-well multiplates (Falcon) and cultured for 24 hours, and insulin was absent ([A]) or present ([B]) (10 nM), b-cryptoxanthin ([A] Lane 2 and [B] Lane 3 3 10 μM) and 13 cisRA ([A] Lane 3 and [B] Lane 4 1 μM) were cultured for 8 days in SG medium. In addition, Lane 1 of [A] and [B] was used as a solvent control.
After completion of the culture treatment, the cells were washed with a phosphate buffer, and then a phosphate buffer containing 4 wt / v% paraformaldehyde (manufactured by Wako Pure Chemical Industries, Ltd.) was added and fixed at room temperature for 15 minutes. Thereafter, the cells were washed with a phosphate buffer, and nile red staining solution was added, and the cells were protected from light and stained at room temperature for 15 minutes. After staining, the cells were washed with PBS (−), and the fluorescence intensity (excitation wavelength: 535 nm, fluorescence wavelength: 595 nm) of nile red incorporated into the lipid droplets was measured with a microplate reader.

  The result of the confirmation test (4) is shown in FIG.

As is clear from FIGS. 1 to 3, TG accumulation was suppressed in a dose-dependent and time-dependent manner by adding β-cryptoxanthin both when insulin was not induced and when induced. In addition, FIG. 4 shows that β-cryptoxanthin suppresses the formation of lipid droplets regardless of the presence or absence of insulin induction. From the above, it has been clarified that β-cryptoxanthin suppresses the accumulation of TG through the inhibition of the formation of lipid droplets accumulated in the sebaceous cells.

Claims (3)

A sebum production inhibitor containing cryptoxanthin and / or a fatty acid ester thereof (excluding those containing crust or a crust extract) .   The sebum production inhibitor according to claim 1, wherein the cryptoxanthin and / or fatty acid ester thereof is derived from citrus.   The sebum production inhibitor according to claim 2, wherein the citrus fruit is Wenzhou mandarin orange.