JPH09188629A - Medicine for inhibiting antibody-producing cell and composition containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject highly safe medicine capable of inhibiting the antibody-producing cells of a spleen, etc., and useful for treating and preventing autoimmune diseases such as atopic dermatitis and systemic lupus erythematous by utilizing a specific natural polymer. SOLUTION: This medicine for inhibiting antibody-producing cells contains a silk-related polymer as an active ingredient. The silk-related polymer is either of silk, a silk extract and the enzymatic degradation product of the silk. The antibody-producing cell-inhibiting medicine is high in the effect for preventing autoimmune diseases, especially atopic dermatitis appearing in dependence on seasons, because the antibody-producing cell-inhibiting medicine can be administered in good timing, before the immune reaction is flared up. One or more kinds of the antibody-producing cell-inhibiting medicines can be added to a medicine composition, a food composition, a skin preparation for external use, a cosmetic, etc. The content of the antibody-producing cell-inhibiting medicine is approximately 0.1-60wt.% in the medicine composition, approximately 0.1-80wt.% in the food composition, and approximately 0.05-30wt.% in the skin preparation for external use or the cosmetic.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an antibody-producing cell inhibitor useful for treating or preventing autoimmune diseases.

[0002]

2. Description of the Related Art In recent years, autoimmune diseases such as atopic dermatitis, systemic lupus erythematosus, bronchial asthma, and allergic rhinitis are greatly affected by sudden changes in living environment and eating habits. The number is increasing. For these autoimmune diseases, at present, inflammation is only symptomatically suppressed by administration of a non-steroidal or steroidal anti-inflammatory drug.

In such autoimmune diseases, various autoantibodies and autoantigen-sensitized lymphocytes are known to exist, and autoantibodies alone, complement-dependent, phagocytic antibody-dependent, killer cell-dependent. It has also been experimentally confirmed that sexually induced tissue damage occurs. However, its specific mechanism has not been elucidated, and therefore, attempts to reduce autoimmune ability often cause immunodeficiency, so that there is currently no radical treatment method. That is, there has been a demand for a drug that suppresses cells that produce autoantibodies and antibody-producing cells such as the spleen.

On the other hand, it has not been known at all that silk-related macromolecules are useful in suppressing antibody-producing cells and preventing or treating autoimmune diseases.

[0005]

The present invention has been made in view of such circumstances, and an object thereof is to provide a substance that suppresses antibody-producing cells such as the spleen.

[0006]

In view of such a situation,
The present inventors have conducted intensive studies in search of a substance that suppresses antibody-producing cells, and as a result, found that silk-related polymers have such an action and completed the invention. Hereinafter, the present invention will be described in detail.

(1) The antibody-producing cell inhibitor of the present invention The antibody-producing cell inhibitor of the present invention comprises a silk-related polymer. The silk-related polymer means a part or all of proteins or peptides constituting silk, and specifically, silk or cocoon itself, silk or cocoon processed by chopping or crushing, Solvent extract of silk or cocoon, pepsin, trypsin, chymotrypsin, cathepsin, papain, promelin, ficin and other enzymatic decomposition products of silk and cocoon, hydrolyzate of silk and cocoon with acid or base, fractional purification of these Examples thereof include things. Among these, preferred are enzymatic decomposition products of silk or cocoons, solvent extracts, solvent extracts of silk or cocoons and fractions thereof, and more preferred are enzymatic decomposition products of silk and fractions thereof. is there. These can be manufactured as follows.

(Material) (1-1) Silk or cocoon Silk or cocoon may be taken as it is or after heat sterilization, and may or may not be specially processed. The processing method may be, for example, fine cutting or crushing with a mill or the like. By processing, it can be expected that oral administration will be easier and the absorption of the active ingredient will be improved.

(1-2) Enzymatic degradation product Silk or cocoon is dispersed in a solvent such as water, and pepsin,
Trypsin, chymotrypsin, cathepsin, papain,
Proteases such as promelin and ficin may be added and the protease may be decomposed at a temperature around 40 ° C. for 2 to 72 hours. Microbial proteases such as yeast-derived proteases may be used. The pH may be the optimum pH for the protease. The preferred group of these enzymes is the readily available pepsin. After enzymatic decomposition, the protease may be inactivated by heating or the like to evaporate the solvent and then used.

(1-3) Hydrolyzate The hydrolyzate is prepared by dispersing the raw material in water or the like, adding a catalyst such as an acid or a base, heating and stirring at a temperature near the boiling point for 1 to 24 hours, and concentrating. Can be obtained. Examples of the catalyst include acids such as hydrochloric acid, sulfuric acid and nitric acid, examples of the base include caustic soda and caustic potash, and examples of the acid include hydrochloric acid, sulfuric acid and nitric acid. Among these, hydrochloric acid is preferable because it is easy to carry out post-treatment such as neutralization.

(1-4) Solvent Extract The solvent extract is prepared by adding a solvent to the above-mentioned drug substance, enzymatic decomposition product, and hydrolyzate, and soaking it for several hours at a temperature near the boiling point and for several days at room temperature. The solvent may be distilled off. The solvent is preferably a polar solvent, for example, water, alcohols such as ethanol and methanol, ketones such as acetone and methyl ethyl ketone, ethers such as diethyl ether and tetrahydrofuran, halogenated hydrocarbons such as chloroform and carbon tetrachloride, Nitriles such as acetonitrile can be mentioned. These may be used alone or in combination of two or more. A particularly preferable solvent is a mixed liquid of water and alcohol, which is excellent in safety and extraction efficiency.

(1-5) Fractionated product As the fractionated product, the above-mentioned drug substance, enzymatic degradation product, hydrolysis product,
The solvent extract was converted to diethyl ether-water, chloroform-
Liquid-liquid extraction with a biphasic solvent system such as water, hexane-hydrous alcohol, water-butanol, column chromatography using a carrier such as silica gel, ODS, ion exchange resin, fractionated product by gel filtration, dialysis membrane The fractionated product and the like can be exemplified. Of these, preferred are ion exchange resin column chromatography and gel filtration.

(2) Antibody-Producing Cell Inhibitor of the Present Invention The antibody-producing cell inhibitor of the present invention comprises the above silk-related polymer. The antibody-producing cell inhibitor referred to in the present invention has the action of suppressing the antibody-producing activity of antibody-producing cells and decreasing the amount of antibody produced. Since the silk-related polymer is excellent in the action of suppressing antibody-producing cells such as spleen and decreasing the amount of antibody production as shown in Examples below, it is excellent in the treatment and prevention of autoimmune diseases, especially, atopic dermatitis. . In particular, for atopic dermatitis that develops depending on the season, the antibody-producing cell inhibitor can be administered in good timing before the immune reaction flares up, so a high preventive effect can be expected.

(3) Composition of the present invention The composition of the present invention is characterized by containing one or more of the above antibody-producing cell inhibitors. The types of the composition of the present invention include, for example, pharmaceutical compositions for treating or preventing autoimmune diseases, food compositions for treating or preventing autoimmune diseases, and skins such as atopic dermatitis and systemic lupus erythematosus. Examples include external preparations for skin and cosmetics for autoimmune diseases. In addition to the antibody-producing cell inhibitor of the present invention, these compositions may contain optional components usually used in these compositions. Such optional ingredients include, in pharmaceutical compositions, excipients, binders, coating agents,
Lubricants, sugar coatings, disintegrants, bulking agents, flavoring agents, emulsifying agents
Examples include solubilizing / dispersing agents, stabilizers, pH adjusters, isotonic agents, and the like. Examples of foods include colorants, thickeners, flavoring agents, stabilizers, emulsifiers, preservatives, and oxidation stabilizers. In skin external preparations and cosmetics, hydrocarbons such as petrolatum and microcrystalline wax, esters such as jojoba oil and gayleu, triglycerides such as beef tallow and olive oil, high-grade alcohol such as cetanol and oleyl alcohol. Alcohols, stearic acid, fatty acids such as oleic acid,
Polyhydric alcohols such as glycerin and 1,3-butanediol, nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, thickeners such as ethanol and carbopol, preservatives, and ultraviolet rays Absorbents, antioxidants,
Examples thereof include dyes and powders. Furthermore, it is possible to include components preferable for treatment or prevention of autoimmune relaxation, for example, herbal components such as perilla, steroid hormones such as dexamethasone, and anti-inflammatory agents such as indomethacin. These compositions of the present invention can be manufactured by a conventional method.
The content of the preferable antibody-producing cell inhibitor of the present invention in these compositions is 0.1 to 60% by weight in the pharmaceutical composition.
, More preferably 1 to 40% by weight, still more preferably 5 to 30% by weight. Further, in the food composition, it is preferably 0.1 to 80% by weight, 1 to 70% by weight
Is more preferable, and 5 to 50% by weight is further preferable. Further, for external skin preparations and cosmetics, 0.05 to 30% by weight is preferable, 0.1 to 20% by weight is more preferable, and 0.5 to 10% by weight.
% By weight is more preferred. The application amount of the composition of the present invention is such that 1 to 100 g of the pharmaceutical composition is orally administered to an adult per day.
It may be administered once to several times by rectal or injection, the food composition may be ingested from 1 to 500 g once to several times, and the external preparation for skin or cosmetic may be applied in an appropriate amount once to several times. Good. Since the antibody-producing cell inhibitor of the present invention is excellent in safety as described in Examples below, it is possible to administer such a large amount.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described in detail below with reference to examples, but it goes without saying that the present invention is not limited to these examples.

Example 1 (Production Example) 100 g of silk was finely chopped, dispersed in 500 ml of warm water at 39 ° C., 1 g of pepsin was added thereto, stirred at 39 ° C. for 72 hours, and treated at 100 ° C. for 1 hour to inactivate pepsin. Made into This was freeze-dried to obtain 95 g of antibody-producing cell inhibitor 1.

Example 2 (Production Example) 10 g of the antibody-producing cell inhibitor 1 was placed in 100 ml of this.
50% ethanol aqueous solution was added, the mixture was heated under reflux for 2 hours, filtered, and concentrated under reduced pressure to obtain 1.3 g of the antibody-producing cell inhibitor 2.

Example 3 (Production Example) 1 g of 50% ethanol aqueous solution was added to 200 g of silkworm cocoons, and 2
The mixture was heated under reflux for hours, filtered, and concentrated under reduced pressure to obtain 2.9 g of antibody-producing cell inhibitor 3.

Example 4 (Formulation example: pharmaceutical composition) Granules were prepared according to the following formulation. That is, the prescription ingredients were weighed in a glad granulator and granulated while spraying 20 parts by weight of a 50% ethanol aqueous solution. This was blow-dried at 40 ° C. for 48 hours and sieved to obtain granules. Antibody-producing cell inhibitor 1 30 parts by weight Hydroxypropyl methylcellulose 10 parts by weight Lactose 40 parts by weight Potato starch 20 parts by weight

Example 5 (formulation example: pharmaceutical composition) A granule was prepared according to the following formulation. That is, the prescription ingredients were weighed in a glad granulator and granulated while spraying 20 parts by weight of a 50% ethanol aqueous solution. This was blow-dried at 40 ° C. for 48 hours and sieved to obtain granules. Antibody-producing cell inhibitor 2 30 parts by weight Hydroxypropyl methylcellulose 10 parts by weight Lactose 40 parts by weight Potato starch 20 parts by weight

Example 6 (Formulation example: pharmaceutical composition) Granules were prepared according to the following formulation. That is, the prescription ingredients were weighed in a glad granulator and granulated while spraying 20 parts by weight of a 50% ethanol aqueous solution. This was blow-dried at 40 ° C. for 48 hours and sieved to obtain granules. Antibody-producing cell inhibitor 3 30 parts by weight Hydroxypropyl methylcellulose 10 parts by weight Lactose 40 parts by weight Potato starch 20 parts by weight

Example 7 (blending example: food composition) A candy was prepared according to the following formulation. That is,
The ingredients were heated and mixed at 120 ° C., and the mixture was cast into a mold to form a candy. Antibody-producing cell inhibitor 1 10 parts by weight starch syrup 30 parts by weight Sorbit 60 parts by weight

Example 8 (formulation example: food composition) A candy was prepared according to the following formulation. That is,
The ingredients were heated and mixed at 120 ° C., and the mixture was cast into a mold to form a candy. Antibody-producing cell inhibitor 2 10 parts by weight starch syrup 30 parts by weight Sorbit 60 parts by weight

Example 9 (Formulation example: food composition) A candy was prepared according to the following formulation. That is,
The ingredients were heated and mixed at 120 ° C., and the mixture was cast into a mold to form a candy. Antibody-producing cell inhibitor 3 10 parts by weight starch syrup 30 parts by weight sorbit 60 parts by weight

Example 10 (Formulation example: external preparation for skin) An external preparation for skin was prepared according to the following formulation. That is, the ingredients were heated and kneaded with a kneader and cooled to obtain an ointment. Antibody-producing cell inhibitor 15 parts by weight Vaseline 95 parts by weight

Example 11 (Formulation example: external preparation for skin) An external preparation for skin was prepared according to the following formulation. That is, the ingredients were heated and kneaded with a kneader and cooled to obtain an ointment. Antibody-producing cell inhibitor 25 parts by weight Vaseline 95 parts by weight

Example 12 (Formulation example: external preparation for skin) An external preparation for skin was prepared according to the following formulation. That is, the ingredients were heated and kneaded with a kneader and cooled to obtain an ointment. Antibody-producing cell inhibitor 35 5 parts by weight Vaseline 95 parts by weight

[0028]

【Example】

Example 1 Acute toxicity The acute toxicity of the antibody-producing cell inhibitors 1 to 3 of the present invention was examined using 5 ICR mice per group. That is, the antibody-producing cell suppressors 1 to 3 of the present invention were dispersed or solubilized in physiological saline and orally administered in an amount of 2 g / Kg. 14 after administration
All animals were alive when they were confirmed to be alive or dead on the day. From this, L of the antibody-producing cell inhibitor of the present invention is
It can be seen that the D50 value is larger than 2 g / Kg and the safety is high.

Example 2 Antibody-Producing Cell Inhibitory Action The antibody-producing cell inhibitory action is a hemolytic plaque method (Science, 140, 40) developed by Jern and others.
5, 1963) to identify and count splenocyte antibody-producing cells by plaque formation. That is, 6 ddy male mice per group were injected from the tail vein into SRBC.
(Sheep red blood cells) in phosphate buffered saline 4 × 10 ⁸ cells / m
It was dispersed at a concentration of 1 and 0.25 ml was administered for sensitization.
4 days a day from SRBC administration, 500, 1000, 20
The sample was dispersed or solubilized in physiological saline and administered at a dose of 00 mg / Kg. Twenty-four hours after the final administration, the spleen was removed and antibody-producing cells were counted by the method of Jern. That is, the cells were divided into spleen cells, incubated with SRBC on an agar medium, and further incubated with guinea pig serum to count plaque-forming cells. The results are shown in Table 1. From this, it is understood that the antibody-producing cell inhibitor of the present invention is excellent in the antibody-producing cell inhibitory action.

[0030]

[Table 1]

[0031]

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a composition which suppresses antibody production of antibody-producing cells and is useful for treating and preventing autoimmune diseases.

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Toshiyuki Fukuda 560 Kashio-cho, Totsuka-ku, Yokohama-shi, Kanagawa Pola Chemical Industry Co., Ltd. Totsuka Research Laboratory (72) Inventor Shinobu Abe 560 Kashio-cho, Totsuka-ku, Yokohama, Kanagawa Prefecture Pola Kasei Kogyo Totsuka Institute Inc.

Claims (6)

[Claims] 1. An antibody-producing cell inhibitor comprising a silk-related polymer. 2. The antibody-producing cell inhibitor according to claim 1, wherein the silk-related polymer is silk, a silk extract, or a silk enzymatic degradation product. 3. A composition containing the antibody-producing cell inhibitor according to claim 1 or 2. 4. The composition according to claim 3, which is a food for preventing or treating autoimmune diseases. 5. The composition according to claim 3, which is a pharmaceutical composition for preventing or treating autoimmune diseases. 6. The composition according to claim 4 or 5, wherein the autoimmune disease is atopic dermatitis or systemic lupus erythematosus.