JP2003231640A - Agent for treatment of atopic dermatitis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an agent for the treatment of atopic dermatitis, containing a vegetable-originated sphingoglycolipid effective for quickly supplying ceramide to the ceramide-deficient skin of a patient of atopic dermatitis and expected to mitigate and cure the symptom. <P>SOLUTION: The agent for the treatment of atopic dermatitis contains the sphingoglycolipid extracted from a vegetable, preferably fine Konjak (tuber of devil's tongue) powder with an organic solvent as an active component. The agent is administered by externally applying to the skin or by oral administration. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for atopic dermatitis containing a glycosphingolipid extracted from potatoes.

[0002]

BACKGROUND ART Atopic dermatitis is a kind of allergic dermatitis, but the number of patients has increased in recent years, and 20% of children under the age of 5 years and 10% of adults have atopic dermatitis. There is a report. The symptoms of atopic dermatitis are eczema with strong itching, and in severely ill patients, chronic drowsiness continues. The pain experienced by a person in this state is immeasurable. At present, research is being conducted on various methods for treating atopic dermatitis, but most of the treatment methods used are symptomatic steroids and antihistamines. Such treatment temporarily improves the symptoms, but there are many cases where the symptoms worsen as soon as the treatment is stopped.

Recently, it has been revealed that atopic dermatitis patients have an extremely small amount of ceramide in the stratum corneum as compared with healthy persons, and the barrier function of the skin is deteriorated because ceramide is not sufficiently produced. It was It is known that IL-1α production from keratinocytes is enhanced in the skin in which the barrier is destroyed. Therefore, it has been found that supplementing ceramide-depleted skin with ceramide is effective in improving the skin barrier function and alleviating the symptoms of atopic dermatitis.

Heretofore, there have been many cases in which a therapeutic drug for atopic dermatitis contains ceramide extracted from bovine or porcine brain, chemically synthesized ceramide or a pseudo-ceramide having a structure similar to ceramide. It was

[0005]

However, animal-derived glycosphingolipids have a possibility of virus infection. Moreover, since the occurrence of mad cow disease in 1986, the supply amount has drastically decreased due to the possibility of infection to humans. Further, chemically synthesized ceramides and pseudo-ceramides have structural differences from natural products and are obtained as racemates, so the effect is questioned, and plant-derived ceramides have been demanded.

As plant-derived ceramides, those extracted from wheat, rice bran, soybean, etc. are known so far, and they are used as a raw material for cosmetics because of their excellent moisturizing effect. As an example of orally ingesting glucosylceramide extracted from wheat and rice bran as a beauty food, Fragrance Journal, 23 (1), 81 (19)
95), etc., and it has been reported that oral administration of 20 mg of a ceramide-containing substance per day is effective for moisturizing the skin. However, these plant-derived ceramides are difficult to be highly purified, and high-purity products are very expensive, so although they are used for cosmetics and food applications,
It has not been used aggressively in the treatment of atopic dermatitis.

It is an object of the present invention to provide a therapeutic agent for atopic dermatitis containing a plant-derived glycosphingolipid, which is considered to be effective for alleviating and treating atopic dermatitis.

[0008]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that atopic dermatitis can be applied orally ingested by applying an extract extracted from a plant to the affected part of the atopic dermatitis. The present inventors have found that the symptoms of dermatitis can be alleviated or cured and have reached the present invention.

That is, the present invention is directed to a therapeutic agent for atopic dermatitis, which comprises a glycosphingolipid extracted from a plant with an organic solvent as an active ingredient, and its application method is skin. It is used by applying to or to be taken orally. In addition, the above-mentioned therapeutic agent for atopic dermatitis is characterized in that a preferable extraction raw material is potatoes.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below. The glycosphingolipid, which is an active ingredient in the therapeutic agent for atopic dermatitis of the present invention, contains glucosylceramide as a main component, and may contain other components. Glycosphingolipids are metabolized in the body when percutaneously or orally absorbed and reach the epidermal tissue in the form of ceramide, which suppresses the evaporation of water from the inside of the skin and prevents the entry of harmful substances from the outside. It constitutes a layer.

The glycosphingolipid, which is an active ingredient in the present invention, is obtained by extracting a plant with an organic solvent, and specifically, it can be obtained by the following method. The plant used as the extraction raw material is not particularly limited. Specifically, for example, almond, aosa, aonori, acacia, acacia, akane, red grape, red pine (including pine tar, amber, copal. The same applies to the following pine species), agaricus, akinogoshi, akebi, asagao, azalea, hydrangea, ashitaba. , Adzuki bean, asparagus, acerola, ashen yak, anise, avocado, amatcha, amaccha vine, amaryllis, altea, arnica, aloe, angelica, apricot, ansopkou, rush, izayoi rose, yew, fig, ginkgo, ylang ylang, corn, fennel. , Scots mallow, Nepenthes, Udo, Ume, Vladimir, Wenzhou mandarin orange, Ages, Shallot, Eleuthero, Enishida, Enokitake, Elderflower, Pea, Orchid, Psyllium, Plantain Zami, (including orange peel) barley, Atractylodes, osmanthus, St. John's wort, dead nettle, Onidokoro, olives, oregano, orange, carnation, cocoa, oysters,
Oysterfish, cuckoo, oak, anemone, pumpkin,
Chamomile, camu camu, chamomile, crow cucumber, larch, karin, garcinia, cardamom, raspberry, kiwi, kyokyo, cabbage (including kale), caraway, cucumber, kumquat, ginnan, guava, wolfberry, kudzu, gardenia, cumin, cranberry, Walnuts, grapefruit, cloves, black pine, black beans, chlorella, ketsumeishi, ginger ginger, cowberry, pepper, cosmos, burdock, wheat (including wheat germ), sesame, komatsuna, rice (including rice bran), coriander, konjac potato (konjaktobi) (Including powder), kelp, salmon berry, cypress, pomegranate, sweet potato, sweet potato,
Sugar cane, sugar beet, saffron, pomelo, hawthorn, hawthorn, shiitake, cyclamen, perilla, shimeji, potato, peony, jasmine, juzdama,
Shungyk, ginger, ginger, white oak, ginkgo, cinnamon, watermelon, water pie, horsetail, star anise, star apple, sudachi, stevia, plum, sage (salvia), mallow, celery, senkyu, senburi, buckwheat, broad bean, radish, soybean (Including okara), daidai, thyme, bamboo shoots, onion, tarragon, taro, tanjin, dandelion, chicory, gypsophila, horsetail, camellia, camellia, clover, clover, tuna, tuna, tsubaki, dill, amanita (geranium), touga. , Capsicum, Capsicum, Astragalus, Corn, Dodami, Turmeric, Eucommia, Ash, Naga, Nasuna, Nutmeg, Nanten, Nigauri, Ningyogi, Chinese chive, Carrots, Garlic, Green onion, Yarrow Saw palmetto, Nobile, verbena, palm, pineapple, hibiscus, chickweed,
Basil, parsley, naked mulberry, mint, pearl barley, banana, banaba, vanilla, paprika, hamamelis, beet,
Bell pepper, hempflower, hisb, hijiki, pistachio, hissop, willow, daisy, cypress, hiba, castor, sunflower, loquat, phalaenopsis,
Fenugreek, Fukinoto, blackberries, plums, blueberries (including bilberries), prunes, loofah, safflower, belladonna, bergamot, spinach, spinach, physalis, bodice, button, hops, jojoba, maitake, maou, maca, macadamian nuts, Matatabi, marigold, mango, honeywort, mimosa, myoga, myrrh, purple, mace, melissa, meliloto, melon, men (including cottonseed meal), moyashi, yam yam, yam yam, yam yogi, eucalyptus, yukinoshita, yuzu, Lily, yokinin, yomena (aster), mugwort, lime, rye,
Lilac, raspberry, peanut, lacquer, apple (including apple fiber), gentian, litchi, lettuce, lemon, lotus root, lotus root, rosehip, rosemary, bay leaf, radish, horseradish (including horseradish), etc. Potatoes such as sweet potato, potato, sato, yama, konjac, and naga are preferable because they have a high content of glycosphingolipids and have a high healing effect on atopic dermatitis, and konjac is more preferable. It is preferable to use konjaktobi powder because konjak potato can be obtained at low cost. Konnyakutobi flour has a peculiar astringent taste and irritating odor as a by-product during the production of konnyaku, which is made from konjac yam, but it has a peculiar astringent taste and irritating odor. Therefore, it is used as a thickener for some fertilizers and concrete. However, it is a resource that has never been used as food. These plant raw materials may be used as they are, or may be processed by operations such as drying, mashing, and heating.

Any organic solvent can be used as the extraction solvent in the present invention as long as it does not impair the effects of the present invention by reacting with the raw material and the glycosphingolipid during extraction. In addition, one kind of solvent may be used alone or a plurality of solvents may be mixed and used. Examples of the organic solvent include alcohols such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, and tert-butanol, hexane, pentane, diethyl ether, chloroform, dichloromethane, acetone, acetonitrile, Examples include ethyl acetate. Of these, preferred examples include methanol, ethanol, hexane, and acetone, and particularly preferred examples include ethanol.
In addition, when extracting with these organic solvents, additives such as water and surfactants can be added within a range that does not impair the effects of the present invention in order to improve extraction efficiency.

The amount of the organic solvent used for the extraction is preferably about 1 to 30 times, more preferably about 1 to 10 times the amount of the starting plant. If the amount of the solvent used is less than this range, the solvent may not spread to the entire raw material, and extraction may be insufficient.Therefore, adding an amount of solvent exceeding this range no longer affects the amount of extraction, Only the burden of solvent removal work in the subsequent concentration step increases.

The extraction temperature depends on the boiling point of the solvent used, but is preferably 0 to 80 ° C, more preferably about room temperature to 60 ° C. If the extraction temperature is below this range, the extraction efficiency will decrease, and even if a temperature above this range is applied, the extraction efficiency will not be significantly affected, and the amount of energy used will only unnecessarily increase.

The extraction time is preferably 1 to 48 hours, more preferably 2 to 20 hours. When the extraction time is shorter than this range, the extraction is not sufficiently performed, and even if the extraction is performed for a long time without exceeding this range, the extraction amount can no longer be expected to increase.

The extraction operation is not limited to one batch operation. It is also possible to add a fresh solvent to the residue after extraction again to perform an extraction operation, or to bring the extraction solvent into contact with the extraction raw material a plurality of times. That is, as the extraction operation, any of batch operation, semi-continuous operation, and countercurrent multistage contact operation can be used. Also, a known extraction method such as Soxhlet extraction may be used.

Next, the extraction residue is separated and removed. The method of separation is not particularly limited, and known methods such as suction filtration, filter press, cylinder press, decanter, centrifuge, and filter centrifuge can be used.

The extract thus obtained is sent to the concentration step. The concentration method is not particularly limited, and for example, it can be concentrated by a vacuum concentration device such as an evaporator or a solvent removal by heating.

The extract for the treatment of atopic dermatitis of the present invention can be used as it is, but since the concentration of glycosphingolipid contained in the extract is as low as 10.0% by mass or less, impurities are continuously extracted. It is more preferable to remove the above to further improve the purity. Any method may be used to improve the purity. For example, washing with water or an organic solvent, washing with an alkaline solution, purification by column chromatography, a method of passing activated carbon or the like, distribution with a solvent having different polarities, a recrystallization method and the like are included. When it is necessary to obtain a particularly high-purity glycosphingolipid, after treating with an alkaline solution, partition with chloroform, diethyl ether, etc., separate and concentrate the organic layer, and further use a resin column or silica gel column chromatography. It is preferred to separate the glycosphingolipids by. These can be performed by a known method.

Next, regarding the method for analyzing the obtained mixture containing glycosphingolipid, the simplest analysis method is thin layer chromatography. Since glucosylceramide derived from human or bovine is commercially available in the standard, it is used and developed using a suitable solvent system such as chloroform-methanol system using a silica gel thin layer plate,
It can be easily analyzed by coloring with concentrated sulfuric acid or anthrone reagent. In addition, it is also possible to perform analysis by known methods such as high performance liquid chromatography and various chromatograph-mass spectrometry methods.

The therapeutic agent for atopic dermatitis of the present invention comprises dry skin caused by atopic dermatitis, cracks, erythema, abrasions,
It relieves various skin symptoms such as exfoliation and pruritus and pain. The prescription may be a method of externally applying the therapeutic agent for atopic dermatitis of the present invention, or a method of orally ingesting, the application method is, for example, a cream,
The dosage form may be a lotion, an ointment or the like, and in the method of oral ingestion, it can be ingested after being processed into powder, tablets, capsules, gels, soft capsules, drinks and the like.

The therapeutic agent for atopic dermatitis of the present invention is highly safe since it contains a plant-derived glycosphingolipid, and the content of the glycosphingolipid is not particularly limited. The preferable content of the plant-derived glycosphingolipid for causing expression is 0.001 to 95% by mass if it is applied. When the content is less than this, the effect of the present invention tends to be delayed or reduced, and when the content is more than this, the effect is not further enhanced and the usability is only deteriorated. If taken orally, the daily intake of glycosphingolipid is 1 μg to 100 g, and the frequency of ingestion is not limited, but it is continued at a frequency of once every 3 days to 10 times a day. It is preferable to ingest. If the amount of glycosphingolipid taken per day is less than 1 μg, the effect may not be sufficiently exhibited, and even if the amount of 100 g or more is taken, the effect is no longer improved.

The therapeutic agent for atopic dermatitis of the present invention is not limited to those intended for humans. In recent years, 7 of dog skin diseases
It is said that atopic dermatitis accounts for 0% or more, and atopic dermatitis of pets is becoming a big problem. Therefore, it is prescribed for domestic animals such as dogs, cats and other pets, cattle, pigs, sheep, etc. May be. In the case of prescribing to such an animal, the one to be applied may be formulated in the above-mentioned form, and the one to be orally ingested may be formulated by adding it to pet foods, beverages, livestock feed and the like.

The therapeutic agent for atopic dermatitis of the present invention is a lotion, emulsion, moisture cream, sunscreen, sunscreen cosmetic, pack, foundation, which is applied to the skin.
It can also be used by adding to white powder, blusher, eye make-up, perfume, cologne, lip balm, lipstick, etc., and it is also a skin care product for patients with atopic dermatitis, face cleansing cream, face soap, shampoo, rinse, treatment, and May be added to a bath agent or the like.
When taken orally, it may be taken by adding it to ordinary foods. Common foods include, for example, bread, udon, soba, foods such as rice, cheese, wiener, sausage, ham, food products such as processed seafood, cookies, cakes, jellies, puddings, candies, chewing gum,
Examples include confectionery such as yogurt, soft drinks, alcoholic beverages, nutritional drinks, coffee, tea, milk and other beverages.

The therapeutic agent for atopic dermatitis of the present invention has a vitamin, collagen, squalane, soybean lecithin, plant-derived sterols, hyaluronic acid, sorbitol, chitin, chitosan, glycerin, in order to promote its effect.
Butylene glycol, propylene glycol, ethylene glycol, polyethylene glycol, petrolatum, silicone, waxes, xanthan gum, guar gum, gum arabic, pectin, quince seed, ascorbic acid,
Sodium lactate solution, carboxyvinyl polymer, polyvinylpyrrolidone, polyvinyl alcohol, niacin,
Niacinamide, ingredients of various plant extracts such as olive extract and ginkgo biloba extract, antihistamines, antileukotriene drugs, naloxone hydrochloride, cyclosporine drugs,
It can also be used in combination with medicines such as steroidal preparations, protopic ointment, neoral, and macrolide immunosuppressants.

The effects of using the therapeutic agent for atopic dermatitis of the present invention include dry skin caused by atopic dermatitis,
It relieves various symptoms of the skin such as cracks, erythema, abrasions and exfoliation of the skin and symptoms such as itching and pain.
The improved state of the barrier ability of the skin and the increase of the water content of the skin can be known using conventionally known measurement methods such as the measurement of transepidermal water loss (TEWL) and the measurement of keratin water content. TEWL
For example, an Evaporimeter (Servo Med, Sweden), Tewameter (Courage + Khazaka, Germany), etc. can be used for the measurement. For measuring the water content of the keratin, for example, Corneometer (Courage + Khazaka, Germany), Skikon-2
For example, 00 (IBS Co., Ltd.) can be used.
In addition, the healing state of atopic dermatitis can be grasped by measuring the IgE value.

[0027]

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited thereto. First, the measuring device and measuring method used in the following examples will be described. (1) Qualitative method of glycosphingolipids Silica gel thin layer chromatography (TLC) was used for the qualification of glycosphingolipids. Silica gel plate (Sillicagel 60F254 type manufactured by Merck & Co., Inc.)
It was applied to a developing tank of chloroform: methanol: water = 87: 13: 2 (volume ratio) and developed. After the development, the silica gel plate was dried with a drier or the like, sprayed with sulfuric acid and heated to develop the color. (2) Method for quantifying glycosphingolipids High performance liquid chromatography (HPLC) was used for quantifying glycosphingolipids. Using Waters LC Module 1,
As the column, GL Science Inertsil SIL 100A was used. Solvent is chloroform: methanol = 9: 1 (volume ratio)
Was used at a flow rate of 1.0 ml / min at 25 ° C. A light scattering detector (500 ELSD manufactured by ALLTECH) was used for detection.
As a standard substance for glycosphingolipids, 2 times amount of ethanol was added to konjac millet flour, and the extract extracted by stirring for 2 hours at room temperature was purified by silica gel column chromatography and PLC plate, and then the above liquid chromatography was performed. The konjac-derived glucosylceramide, which was confirmed to have no other peak under the conditions of, was used.

Production Example 1 1 kg of konjac millet powder was placed in a stirring tank, 2 L of ethanol was added thereto, and the mixture was stirred at room temperature for 2 hours. Then, the extract and the residue were separated by filtration. The extract was concentrated by an evaporator to obtain 10.7 g of a brownish waxy concentrate. When this was measured based on the above qualitative and quantitative methods, it was found that spots of glycosphingolipid were detected by TLC, and 0.55 g of glycosphingolipid was contained by HPLC. The purity of the glycosphingolipid in the Tobi powder extract was 5.1% by mass.

20.7 g of 10.7 g of this soybean flour extract
Was dissolved in ethanol and the solution was introduced into 500 ml of an aqueous methanol solution of 0.4 mol / L potassium hydroxide while stirring. After stirring for 2 hours as it is, 500 m of water
1, chloroform (500 ml) were introduced, and the mixture was vigorously stirred and then allowed to stand to recover the chloroform layer. Chloroform was distilled off with an evaporator, and a glycosphingolipid-containing product was obtained.
6 g was obtained. The content of glycosphingolipid measured by HPLC was 0.57 g. The purity of the glycosphingolipid after the alkali treatment was improved to 8.6% by mass.

Then, a glass empty column having an inner diameter of 5 cm and a length of 100 cm was wet-filled with HP-Cellulofine manufactured by Seikagaku Kogyo Co., Ltd., which had been sufficiently swollen in methanol, to a height of 60 cm. After flowing 2 L of methanol, when the liquid surface of methanol reached the upper end of the resin, 6.6 g of the above-mentioned glycosphingolipid-containing material was dissolved in methanol to give 20
What was made into ml was introduced. Subsequently, methanol was added at a flow rate of
Flowed at 0.0 ml / min, and the effluent was collected in 40 ml portions. As a result of analyzing the collected effluent by thin layer chromatography, nothing was contained in 500 ml from the beginning, and then glycosphingolipid was observed in 160 ml in a dark spot. When 160 ml of this was collected and the solvent was distilled off, 0.61 g of a brownish pasty substance was obtained. The glycosphingolipid content was determined by HPLC to be 0.32 g, and the glycosphingolipid purity of the obtained extract was 52% by mass.

Next, a glass empty column having an inner diameter of 2 cm and a length of 100 cm was filled with 60 cm of silica gel (silica gel 60, 70-230 mesh manufactured by Nacalai Tesque, Inc.).
Filled up to the height of ethyl acetate: methanol = 95: 5
2 L of the mixed solvent of was poured. When the surface of the developing solvent reached the upper end of the resin, 0.61 g of the brownish paste-like substance obtained by the above method was dissolved in methanol to make 2 ml. Subsequently, the same developing solvent was added at a flow rate of 5.0 ml /
It was run for a minute, and the effluent was collected in 20 ml portions. As a result of analyzing the collected effluent by thin layer chromatography, nothing was contained in 120 ml from the beginning, and there was no spot of glycosphingolipid in 180 ml after that, but other impurities flowed out. 120 ml after that
The glycosphingolipid was observed in the dark spot. When 120 ml was collected and the solvent was distilled off, 0.31
g white solid was obtained. When the glycosphingolipid content was determined by HPLC, it was 0.31 g, and the purity of the glycosphingolipid of the obtained treated product was 100% by mass.

Example 1 (Cream) 1.0 g of the konjac potato-derived sphingoglycolipid obtained in Production Example 1 was mixed with 15 g of stearic acid and 2 parts of cetanol.
g, liquid paraffin 1 g, octyldodecanol 1 g,
Composition A was prepared by stirring and dissolving with 0.05 g of propylparaben at 70 ° C. On the other hand, in another container, 12 g of glycerin, 0.5 g of potassium hydroxide and 0.2 g of methylparaben were dissolved in 67.25 g of purified water at 70 ° C. to obtain a composition B. The composition A was introduced while the composition B was strongly stirred, and the mixture was stirred under reduced pressure for 5 hours to obtain 100 g of a cream for treating atopic dermatitis.

Example 2 (Lotion) 1.0 g of the konjac potato-derived sphingoglycolipid obtained in Production Example 1 was mixed with 5 g of propylene glycol, 3 g of 70% sorbit solution, and polyoxyethylene sorbitol lauryl ether (20 E.O.) 0 0.1 g, ethanol 10
g, methylparaben 0.1 g, sodium citrate 0.
A composition A was prepared by stirring together with 2 g, cetanol 2 g, liquid paraffin 1 g, octyldodecanol 1 g, and propylparaben 0.05 g at 70 ° C. for dissolution. On the other hand, in another container, 12 g of glycerin and 0.5 of potassium hydroxide.
g and 0.2 g of methylparaben were introduced into 76.55 g of purified water to obtain a composition B. Mixing Compositions A and B at 60 ° C,
Lotion 1 for treating atopic dermatitis by vigorously stirring for 3 hours with an emulsifying machine "Polytron PT10-35"
00g was obtained.

Production Example 2 10 kg of wheat flour was placed in a stirring tank, and ethanol 20 was added thereto.
L was added and the mixture was stirred at room temperature for 2 hours. Then, the extract and the residue were separated by filtration. The extract was concentrated by an evaporator to obtain 68.2 g of a brownish waxy concentrate. When this was measured based on the above qualitative and quantitative methods, TL
A spot of glycosphingolipid was detected by C, and HPL
C revealed that the glycosphingolipid contained 0.50 g. The purity of the glycosphingolipid in the wheat flour extract was 0.7% by mass.

68.2 g of this wheat flour extract was dissolved in 200 g of ethanol and introduced into 5 L of 0.4 mol / L potassium hydroxide aqueous methanol solution while stirring. After stirring as it is for 2 hours, 5 L of water and 5 mL of chloroform are added.
L was introduced, and the mixture was vigorously stirred and then allowed to stand, and the chloroform layer was recovered. Chloroform was distilled off with an evaporator to obtain 26.8 g of a glycosphingolipid-containing substance. The content of glycosphingolipid measured by HPLC was 0.48 g.
The purity of the glycosphingolipid after the alkali treatment was 1.8% by mass.

Next, a glass empty column having an inner diameter of 5 cm and a length of 100 cm was wet-filled with HP-Cellulofine manufactured by Seikagaku Kogyo Co., Ltd., which had been sufficiently swollen in methanol, to a height of 60 cm. After flowing 2 L of methanol, when the liquid surface of methanol reached the upper end of the resin, 6.7 g of the above-mentioned glycosphingolipid-containing material was dissolved in methanol to give 20
What was made into ml was introduced. Subsequently, methanol was added at a flow rate of
Flowed at 0.0 ml / min, and the effluent was collected in 40 ml portions. As a result of analyzing the collected effluent by thin layer chromatography, nothing was contained in 500 ml from the beginning, and then glycosphingolipid was observed in 100 ml in a dark spot. When 160 ml of this was collected and the solvent was distilled off, 0.27 g of a yellow paste-like substance was obtained. This operation was repeated 3 times, and 26.8 g of the glycosphingolipid-containing material was treated to obtain 1.09 g of the purified product. The glycosphingolipid content was determined by HPLC to be 0.44 g, and the glycosphingolipid purity of the obtained extract was 40% by mass.

Next, 60 cm of silica gel (silica gel 60, 70-230 mesh, manufactured by Nacalai Tesque, Inc.) is placed in a glass empty column having an inner diameter of 2 cm and a length of 100 cm.
Filled up to the height of ethyl acetate: methanol = 95: 5
2 L of the mixed solvent of was poured. When the liquid level of the developing solvent reached the upper end of the resin, 1.09 g of the yellow paste-like substance obtained by the above method was dissolved in methanol to make 3 ml. Subsequently, the same developing solvent was added at a flow rate of 5.0 ml /
It was run for a minute, and the effluent was collected in 20 ml portions. As a result of analyzing the collected effluent by thin layer chromatography, 120 ml did not contain anything from the beginning, and then 220 ml had no spots of glycosphingolipid, but other impurities flowed out. After that 160 ml
The glycosphingolipid was observed in the dark spot. When this 160 ml was collected and the solvent was distilled off, 0.35
g white solid was obtained. When the glycosphingolipid content was determined by HPLC, it was 0.35 g, and the purity of the glycosphingolipid of the obtained treated product was 100% by mass.

Example 3 (Cream) Wheat-derived glycosphingolipid obtained in Production Example 2
Composition A was dissolved by stirring 1.0 g together with 15 g of stearic acid, 2 g of cetanol, 1 g of liquid paraffin, 1 g of octyldodecanol, and 0.05 g of propylparaben by stirring at 70 ° C. On the other hand, in another container, 12 g of glycerin,
70 g of 0.5 g of potassium hydroxide and 0.2 g of methylparaben
It was dissolved in 67.25 g of purified water at 0 ° C. to obtain a composition B. The composition A was introduced while the composition B was strongly stirred, and the mixture was stirred under reduced pressure for 5 hours to obtain 100 g of a cream for treating atopic dermatitis.

Comparative Example 1 In Example 1, 1.08 g of ceramide 3 (purity: 92.3%) manufactured by Cosmo Farm Co., Ltd., a synthetic ceramide, was added in place of the konjac potato-derived sphingoglycolipid obtained in Production Example 1. Other than that was manufactured similarly, and 100 g of creams not containing glycosphingolipid were obtained.

Comparative Example 2 In Example 1, the sphingoglycolipid derived from konjac potato obtained in Production Example 1 was not added, and instead of 1.
The same production was carried out except that 0 g of purified water was added to obtain 100 g of a cream containing no glycosphingolipid.

Test Example 1 <Atopic dermatitis therapeutic agent application test> Since the patient had atopic dermatitis symptoms and had been treated by applying a steroid agent, and the symptoms were improving, application of a steroid agent was recommended. We had 25 men and women in their 20s cooperate one week after they stopped working together. TEWL of the affected area
Measure the values and make sure that the averages are almost the same, 5 for each man and woman
Five groups are formed for each person, and the cream of Example 1, the lotion of Example 2, the cream of Example 3, the cream of Comparative Example 1, and the cream of Comparative Example 2 are applied to the affected area twice a day in each group. I got it applied. Therefore, the skin condition and itchiness were examined after 1 week and 2 weeks. Each item was evaluated by a questionnaire, and "completely cured" was given 3 points, "improved" was given 2 points, "no change" was given 1 point, and "deteriorated" was given 0 point. The total value is shown. The results are shown in Table 1.

[0042]

[Table 1]

As described above, the group in which the cream or lotion containing the glycosphingolipid of the present invention was prescribed showed a clear improvement, but the group containing the synthetic ceramide of Comparative Example 1 and Comparative Example 2 were found. A group formulated with a cream containing no glycosphingolipid of the present invention is
Symptoms tended to worsen with the cessation of steroids.

Example 4 (drinking agent) 1.0 g of the konjac potato-derived sphingoglycolipid obtained in Production Example 1 was dissolved in 5 g of soybean lecithin ("SLP-paste" manufactured by Trulecithin Industry Co., Ltd.) at 70 ° C. Then, it was introduced into 99 g of strongly stirred drinking water at 70 ° C. As-is emulsifier "Polytron PT10-3
The mixture was vigorously stirred at 30 ° C. for 5 hours at 30 ° C., stirring was stopped, and the mixture was gradually cooled to obtain 100 g of a drink for treating atopic dermatitis. The obtained drink was homogeneous and no precipitation occurred even after 1 month.

Example 5 (Soft Capsule) 1.0 g of the konjac potato-derived sphingoglycolipid obtained in Production Example 1 was dissolved in 5 g of soybean lecithin (“SLP-paste” manufactured by Trulecithin Industry Co., Ltd.) at 70 ° C. . 0.6 g of this solution was enclosed in a film composed of gelatin, glycerin and water to prepare a soft capsule for treating atopic dermatitis.

Example 6 (drink) Wheat-derived glycosphingolipid obtained in Production Example 2
1.0 g was dissolved in 5 g of soybean lecithin (“SLP-paste” manufactured by Trulecithin Industry Co., Ltd.) at 70 ° C., and this was introduced into 99 g of strongly stirred drinking water at 70 ° C.
As it is, at an emulsifying machine “Polytron PT10-35” at 30 ° C.
After stirring vigorously for 3 hours, the stirring was stopped and gradually cooled to obtain 100 g of a drink for treating atopic dermatitis. The obtained drink was homogeneous and no precipitation occurred even after 1 month.

Comparative Example 3 The procedure of Example 4 was repeated except that 1.0 g of edible collagen was added instead of the konjac potato-derived sphingoglycolipid obtained in Preparation Example 1, and no sphingoglycolipid was contained. 100 g of a drink was obtained.

Comparative Example 4 In Example 4, the sphingoglycolipid derived from konjac potato obtained in Production Example 1 was not added, and instead of 1.
The same production was carried out except that 0 g of purified water was added to obtain 100 g of a drink formulation containing no glycosphingolipid.

Test Example 2 <Ingestion test for therapeutic agent for atopic dermatitis> Forty 40-week-old male NC mice were bred in cages of five mice each. After cutting the abdomen of these mice, 5
Sensitization treatment was performed by continuously applying 0.1 ml of a 0.2 vol% dinitrofluorobenzene ethanol solution to the abdominal skin for consecutive days. Ten days after the last treatment, 0.02 ml of 0.1% by volume dinitrofluorobenzeneacetone solution was applied to induce an allergic reaction. 10 of these mice
Each of the animals was divided into 4 groups, and 100 μl of each of them was orally administered once a day for 3 weeks in a gastric tube to each of the drinks of Example 4, Example 6, Comparative Example 3, and Comparative Example 4. One day later,
The number of scratches per 10 minutes and the serum IgE value of each group after 1 week and 2 weeks were measured, and the average value for each group is shown in Table 2.

[0050]

[Table 2]

As described above, the number of scratches in the group orally ingested with glycosphingolipid was remarkably reduced, and the serum IgE value was also rapidly reduced.

[0052]

INDUSTRIAL APPLICABILITY According to the present invention, by application or ingestion, ceramide which is deficient in the skin of patients with atopic dermatitis can be promptly supplied, and it has an excellent effect of alleviating or healing symptoms. It is possible to provide a therapeutic agent for atopic dermatitis. Furthermore, the therapeutic agent for atopic dermatitis of the present invention can be easily produced at low cost by using an extract extracted from potatoes.

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Kazuko Shizama             23 Uji Kozakura, Uji City, Kyoto Prefecture Unitika Ltd.             Shikisha Central Research Institute (72) Inventor Katsuyuki Mukai             23 Uji Kozakura, Uji City, Kyoto Prefecture Unitika Ltd.             Shikisha Central Research Institute F-term (reference) 4C084 AA17 MA17 MA28 MA37 MA52                       MA63 ZA892 ZB132                 4C088 AB80 AB84 BA10 CA06 MA17                       MA28 MA37 MA52 MA63 ZA89                       ZB13

Claims (4)

[Claims] 1. A therapeutic agent for atopic dermatitis, which comprises a glycosphingolipid extracted from a plant with an organic solvent as an active ingredient. 2. The therapeutic agent for atopic dermatitis according to claim 1, which is used by being applied to the skin. 3. The therapeutic agent for atopic dermatitis according to claim 1, which is taken orally. 4. The therapeutic agent for atopic dermatitis according to claim 1, wherein the plant is a potato.