JP4863870B2 - Antiallergic agent - Google Patents

Description

  The present invention relates to an antiallergic agent. The antiallergic agent of the present invention can be administered not only as a pharmaceutical product, but also in various forms such as health food (preferably functional food) or feed as food and drink. The food includes a beverage. Furthermore, an oral hygiene composition, for example, temporarily contained in the mouth, but most of it is given in the form of exhaling from the mouth, for example, in the form of a dentifrice, mouthwash, chewing gum, or mouthwash, or It is also possible to give in the form of an inhalant to be aspirated from the nose.

It is known that basidiomycetes belonging to the genus Kawaratake of the Porous Mycology family contain various physiologically active substances. For example, Patent Document 1 discloses that a glycoprotein complex contained in Kawaratake belonging to the genus Kawaratake has an antitumor action, an immunoregulatory action, and a growth factor inhibitory action. Patent Document 2 discloses that a bacterium belonging to the porous family, for example, Kawaratake, contains an antitumor component.
JP-A-8-208704 JP 52-61214 A

The present inventor has intensively studied for the purpose of finding a physiologically active substance contained in basidiomycetes belonging to the genus Kawaratake, and found that the fraction derived from Kawaratake exhibits anti-allergic activity. 3,8,9-tetrahydroxybenzobis [1,2-b, 4,5-b ′] benzofuran-6,12-quinone (2,3,8,9-Tetrahydroxybenzobis [1,2-b, 4, 5-b '] benzofuran-6,12-quinone; hereinafter referred to as teleholic acid). The present invention is based on such knowledge.
An object of the present invention is to provide a novel antiallergic agent.

［式中、Ｒ^１、Ｒ^２、Ｒ^３、及びＲ^４は、それぞれ独立して、水素原子、炭素数１〜４の低級アルキル基、アセチル基、又はベンゾイル基である］
で表されるテレホール酸化合物若しくはその塩、あるいは、前記テレホール酸化合物若しくはその塩とタンパク質との結合物、
（２）（ａ）カワラタケ属に属する担子菌のアルカリ水溶液抽出液を酸性に調整することにより得られる、アルカリ水溶液抽出液の酸性条件下沈殿画分（以下、画分Ｆ１と称することがある）、
（ｂ）前記画分（ａ）を極性有機溶媒存在下で還流して得られる、極性有機溶媒抽出画分（以下、画分Ｆ２と称することがある）、
（ｃ）前記画分（ｂ）から熱水可溶部及び非極性有機溶媒可溶部を除去して得られる、熱水可溶部及び非極性有機溶媒可溶部除去画分（以下、画分Ｆ３と称することがある）、
（ｄ）前記画分（ｃ）をアセチル化して得られる、アセチル化画分（又はアセチル化物）（以下、画分Ｆ４と称することがある）、
（ｅ）前記画分（ｄ）から酸性水溶液可溶部、アルカリ水溶液可溶部、及び非極性有機溶媒可溶部を除去して得られる、酸性水溶液可溶部、アルカリ水溶液可溶部、及び非極性有機溶媒可溶部除去画分（以下、画分Ｆ５と称することがある）、
（ｆ）前記画分（ｅ）を脱アセチル化して得られる、脱アセチル化画分（又は脱アセチル化物）（以下、画分Ｆ６と称することがある）、若しくは
（ｇ）前記画分（ｆ）の有機溶媒抽出画分（以下、画分Ｆ７と称することがある）、又は（３）（ｈ）カワラタケ属に属する担子菌を極性有機溶媒により還流抽出した後、得られた極性有機溶媒抽出画分から非極性有機溶媒可溶部及び熱水可溶部を除去して得られる、極性有機溶媒抽出画分の熱水可溶部及び非極性有機溶媒可溶部除去画分（以下、画分Ｆ８と称することがある）、
（ｉ）前記画分（ｈ）をアセチル化して得られる、アセチル化画分（又はアセチル化物）（以下、画分Ｆ９と称することがある）、
（ｊ）前記画分（ｈ）から酸性水溶液可溶部、アルカリ水溶液可溶部、及び非極性有機溶媒可溶部を除去して得られる、酸性水溶液可溶部、アルカリ水溶液可溶部、及び非極性有機溶媒可溶部除去画分（以下、画分Ｆ１０と称することがある）、
（ｋ）前記画分（ｊ）を脱アセチル化して得られる、脱アセチル化画分（又は脱アセチル化物）（以下、画分Ｆ１１と称することがある）、若しくは
（ｌ）前記画分（ｋ）の有機溶媒抽出画分（以下、画分Ｆ１２と称することがある）、又は
（４）（ｍ）前記画分（ａ）〜（ｌ）の少なくとも１つの画分と、タンパク質との結合物を有効成分として含有することを特徴とする、抗アレルギー剤により解決することができる。The object is achieved by the present invention in accordance with (1) the general formula (I):

[Wherein, R ¹ , R ² , R ³ , and R ⁴ are each independently a hydrogen atom, a lower alkyl group having 1 to 4 carbon atoms, an acetyl group, or a benzoyl group]
A teleholic acid compound or a salt thereof represented by the above, or a conjugate of the teleholic acid compound or a salt thereof and a protein,
(2) (a) Precipitation fraction under acidic conditions of an aqueous alkaline solution obtained by adjusting the aqueous alkaline solution of a basidiomycete belonging to the genus Kawaratake (hereinafter sometimes referred to as fraction F1). ,
(B) a polar organic solvent extraction fraction obtained by refluxing the fraction (a) in the presence of a polar organic solvent (hereinafter sometimes referred to as fraction F2),
(C) Hot water soluble part and nonpolar organic solvent soluble part removal fraction (hereinafter referred to as fraction) obtained by removing the hot water soluble part and nonpolar organic solvent soluble part from the fraction (b). Sometimes referred to as minute F3),
(D) an acetylated fraction (or acetylated product) obtained by acetylating the fraction (c) (hereinafter sometimes referred to as fraction F4),
(E) an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part obtained by removing an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part, and a nonpolar organic solvent soluble part from the fraction (d), and Nonpolar organic solvent soluble part removal fraction (hereinafter sometimes referred to as fraction F5),
(F) a deacetylated fraction (or deacetylated product) (hereinafter sometimes referred to as fraction F6) obtained by deacetylating the fraction (e), or (g) the fraction (f ) Organic solvent extraction fraction (hereinafter sometimes referred to as Fraction F7), or (3) (h) basidiomycete belonging to the genus Kawaratake is extracted by refluxing with a polar organic solvent, and the obtained polar organic solvent extraction The fraction obtained by removing the non-polar organic solvent soluble part and the hot water soluble part from the fraction, and the fraction obtained by removing the hot water soluble part and the non-polar organic solvent soluble part of the polar organic solvent extraction fraction (hereinafter referred to as fraction). Sometimes called F8),
(I) an acetylated fraction (or an acetylated product) obtained by acetylating the fraction (h) (hereinafter sometimes referred to as fraction F9),
(J) An acidic aqueous solution soluble part, an alkaline aqueous solution soluble part obtained by removing an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part, and a nonpolar organic solvent soluble part from the fraction (h), and Nonpolar organic solvent soluble part removal fraction (hereinafter sometimes referred to as fraction F10),
(K) a deacetylated fraction (or deacetylated product) (hereinafter sometimes referred to as fraction F11) obtained by deacetylating the fraction (j), or (l) the fraction (k ) Organic solvent extract fraction (hereinafter also referred to as fraction F12), or (4) (m) at least one fraction of the fractions (a) to (l) and a conjugate of protein Can be solved by an antiallergic agent characterized in that it contains as an active ingredient.

The present invention also provides:
(A) a precipitate fraction under acidic conditions of an aqueous alkaline solution obtained by adjusting the aqueous alkaline solution of a basidiomycete belonging to the genus Kawaratake
(B) a polar organic solvent extraction fraction obtained by refluxing the fraction (a) in the presence of a polar organic solvent,
(C) A hot water soluble part and a nonpolar organic solvent soluble part removal fraction obtained by removing a hot water soluble part and a nonpolar organic solvent soluble part from the fraction (b),
(D) an acetylated fraction (or acetylated product) obtained by acetylating the fraction (c),
(E) an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part obtained by removing an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part, and a nonpolar organic solvent soluble part from the fraction (d), and Non-polar organic solvent soluble part removal fraction,
(F) a deacetylated fraction (or deacetylated product) obtained by deacetylating the fraction (e),
(G) an organic solvent extraction fraction of the fraction (f),
(H) A polar organic compound obtained by reflux-extracting basidiomycetes belonging to the genus Kawaratake with a polar organic solvent and then removing the nonpolar organic solvent soluble part and the hot water soluble part from the obtained polar organic solvent extract fraction Hot water soluble part and nonpolar organic solvent soluble part removal fraction of solvent extraction fraction,
(I) an acetylated fraction (or acetylated product) obtained by acetylating the fraction (h),
(J) An acidic aqueous solution soluble part, an alkaline aqueous solution soluble part obtained by removing an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part, and a nonpolar organic solvent soluble part from the fraction (h), and Non-polar organic solvent soluble part removal fraction,
(K) a deacetylated fraction (or deacetylated product) obtained by deacetylating the fraction (j), or (l) an organic solvent extract fraction of the fraction (k), or (m) The present invention relates to a bound product of at least one of the fractions (a) to (l) and a protein.

The present invention also provides a pharmaceutical comprising at least one of the fractions (a) to (l) or the conjugate (m) and a pharmaceutically or veterinarily acceptable carrier or diluent. Relates to the composition.
Further, the present invention includes administering at least one of the substance (1), fractions (a) to (l), or the conjugate (m) in an effective amount to a subject in need of allergy suppression. , Allergy control method.
In addition, the present invention is effective for at least one of the substances (1), fractions (a) to (l), or the conjugate (m) for a subject in need of treatment or prevention of a disease associated with allergy. The present invention relates to a method for treating or preventing the above-mentioned diseases, which comprises administering in an amount.
The present invention also relates to the use of at least one of the substances (1), fractions (a) to (l) or the conjugate (m) for producing an antiallergic agent.

  According to the antiallergic agent of the present invention, allergy can be suppressed. Therefore, the antiallergic agent of the present invention is useful for the treatment and / or prevention of diseases related to allergy.

It is the spectrum acquired by ¹ H-NMR measurement of the fraction derived from Kawaratake. It is the spectrum obtained by the ¹³ C-NMR measurement of a Kawaratake-derived fraction. It is IR spectrum of a fraction derived from Kawaratake. It is the spectrum acquired by MS analysis of the fraction derived from Kawaratake.

［式中、Ｒ^１、Ｒ^２、Ｒ^３、及びＲ^４は、それぞれ独立して、水素原子、炭素数１〜４の低級アルキル基、アセチル基、又はベンゾイル基である］
で表されるテレホール酸化合物若しくはその塩、あるいは、前記テレホール酸化合物若しくはその塩とタンパク質との結合物、又は（２）カワラタケ属に属する担子菌に由来する本発明の各種画分（Ｆ１〜Ｆ１２）の少なくとも１つ、あるいは、前記画分Ｆ１〜Ｆ１２の少なくとも１つの画分とタンパク質との結合物を含有する。The antiallergic agent of the present invention comprises (1) general formula (I):

[Wherein, R ¹ , R ² , R ³ , and R ⁴ are each independently a hydrogen atom, a lower alkyl group having 1 to 4 carbon atoms, an acetyl group, or a benzoyl group]
Or a combination of the teleholic acid compound or a salt thereof and a protein, or (2) various fractions of the present invention derived from a basidiomycete belonging to the genus Kawaratake (F1 to F12) Or at least one fraction of the fractions F1 to F12 and a bound product of protein.

あるいは、イボタケ科に属するキノコ［例えば、コウタケ（Sarcodon aspratus (Berk), S.Ito）、ケロウジ（Sarcodon scabrosus
(Fr.) Karst.）、又はシシタケ（Sarcodon imbricatum (L. ex Fr.) Karst.）］を挙げることができる。Teleholic acid, which is one of the active ingredients in the antiallergic agent of the present invention, is a compound in which R ¹ , R ² , R ³ , and R ⁴ are all hydrogen atoms in general formula (I). Teleholic acid can be chemically synthesized by, for example, the method described in J. Gripenberg, Tetrahedron, vol. 10, pp. 135-143 (1960), or separated from natural mushrooms or mycelium of artificial culture. You can also Examples of mushrooms containing teleholic acid include Kawaratake

Alternatively, mushrooms belonging to the family Amanita [for example, Sarcodon aspratus (Berk), S. Ito ), Keroji ( Sarcodon scabrosus
(Fr.) Karst.), Or Shishitake (Sarcodon imbricatum (L. ex Fr.) Karst.)] Can be exemplified.

The solubility of teleholic acid is
Pyridine, dimethyl sulfoxide (DMSO), or dioxane>acetone> methanol, ethanol, ethyl acetate> ether (for example, diethyl ether), esters>
Water, benzene, hexane, and carbon tetrachloride. The solubility of the acetylated form of teleholic acid is soluble in pyridine, DMSO, or dioxane, but insoluble in other major solvents.

  The lower alkyl group having 1 to 4 carbon atoms in the general formula (I) includes a linear alkyl group and a branched alkyl group. For example, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, n -A butyl group, an isobutyl group, a sec-butyl group, or a tert- butyl group can be mentioned, A methyl group or an ethyl group is preferable.

  The salt of teleholic acid, which is one of the active ingredients in the present invention, is not particularly limited as long as it is a pharmaceutically acceptable salt. Examples thereof include salts with inorganic bases or organic bases. Can do. Inorganic bases suitable for salt formation are, for example, hydroxides such as ammonia, potassium, sodium, lithium, calcium, magnesium, or aluminum, carbonates, bicarbonates, and the like. Examples of salts with organic bases include mono-, di-, or tri-alkylamine salts such as methylamine, dimethylamine, and triethylamine, mono-, di-, or tri-hydroxyalkylamine salts, guanidine salts, N-methylglucosamine salt, amino acid salt, etc. can be mentioned.

又はハカワラタケ［Coriolus pargamenus (Fr.) Pat.］を挙げることができるが、とりわけカワラタケが好ましい。Fractions F1 to F7 and fractions F8 to F12 derived from basidiomycetes belonging to the genus Kawaratake, which can be used as active ingredients in antiallergic agents in the present invention, are novel substances themselves.
Examples of basidiomycetes belonging to the genus Kawaratake include, for example, Kawaratake

Alternatively, there may be mentioned [ Tatsunotake ] [ Coriolus pargamenus (Fr.) Pat.].

  Examples of basidiomycetes (preferably Kawaratake) belonging to the genus Kawaratake used in the present invention include the fruit bodies or mycelia of natural basidiomycetes, or mycelia or cultures (Broth) of basidiomycetes obtained by culturing. Can do. When a fruit body or mycelium is used as the basidiomycete, it is preferably processed into a crushed material or powder so as to improve extraction efficiency.

  The precipitate fraction under acidic conditions of the present invention (that is, the precipitate fraction under acidic conditions obtained by the step of adjusting the alkaline aqueous solution extract of basidiomyceae belonging to the genus Kawaratake; fraction F1) belongs to the genus Kawaratake. The basidiomycete can be extracted with an alkaline aqueous solution, and the resulting alkaline aqueous solution extract is adjusted to be acidic and left under acidic conditions. As the alkaline aqueous solution, for example, a saturated sodium borate aqueous solution can be used. Extraction with an aqueous alkali solution can be performed, for example, at 0 to 100 ° C. for 0.5 to 72 hours under alkaline conditions of pH 8 to 14. The acidic condition can be adjusted to, for example, pH 1 to 6, and for example, the precipitate can be recovered by centrifuging after standing at 0 to 100 ° C. for 0.5 to 72 hours. The centrifugation can be performed, for example, at 10,000 to 50,000 × g for 10 to 30 minutes. The obtained precipitate is preferably washed repeatedly using a suitable washing liquid (for example, water, preferably distilled water) until the washing liquid becomes neutral (pH 7).

  Extraction fraction of polar organic solvent of the present invention (that is, the step of adjusting the alkaline aqueous solution extract of basidiomycete belonging to the genus Kawaratake to acidity, and refluxing the precipitate fraction obtained by the above step in the presence of the polar organic solvent) The polar organic solvent extraction fraction; fraction F2) obtained by the above step can be obtained by adding a polar organic solvent to the precipitate fraction under the acidic condition and refluxing. The polar organic solvent is not particularly limited as long as it exhibits water-soluble properties. For example, acetone, methanol, ethanol, ethyl acetate, or dimethyl ether can be used, and acetone is preferable. The refluxing is preferably carried out until the color of the organic solvent extract disappears, and can be carried out, for example, at 50 to 150 ° C. for 1 to 24 hours. The polar organic solvent extraction fraction of the present invention includes both the extract after reflux and its dried product.

  Fraction removing hot water soluble part and non-polar organic solvent soluble part of the present invention (that is, a step of adjusting the aqueous alkaline solution extract of basidiomyceae belonging to the genus Kawaratake to acidic, a precipitate under acidic conditions obtained by the step A hydrothermally soluble part obtained by a step of refluxing the fraction in the presence of a polar organic solvent, and a process of removing the hot water soluble part and the nonpolar organic solvent soluble part from the polar organic solvent extraction fraction obtained by the process And non-polar organic solvent soluble part removal fraction; fraction F3), for example, after contacting the polar organic solvent extraction fraction and hot water, the hot water soluble part is removed, for example, by centrifugation Subsequently, after contacting the obtained hot water soluble part removal fraction and the nonpolar organic solvent, the nonpolar organic solvent soluble part can be obtained, for example, by removing by centrifugation. . The contact with hot water and the contact with the nonpolar organic solvent can be carried out in this order, or can be carried out in the reverse order.

Contact with hot water can be carried out, for example, at 50 to 100 ° C. for 10 to 60 minutes. The contact with hot water is preferably repeated 5 to 10 times.
The contact with the nonpolar organic solvent can be carried out, for example, for 10 to 60 minutes. As the nonpolar organic solvent, for example, chloroform, hexane, benzene, carbon tetrachloride, or methylene chloride can be used. The contact with the nonpolar organic solvent is preferably repeated 5 to 10 times for one nonpolar organic solvent. Moreover, only one type of nonpolar organic solvent can be used, or different nonpolar organic solvents can be used sequentially.

  The acetylated fraction (or acetylated product) of the present invention [that is, the step of adjusting the alkaline aqueous solution extract of basidiomycetes belonging to the genus Kawaratake to acidity, the precipitate fraction obtained under the acidic conditions obtained by the above step in the presence of a polar organic solvent In which the hot water soluble part and the nonpolar organic solvent soluble part are removed from the polar organic solvent extraction fraction obtained by the process, and the hot water soluble part and the nonpolar organic solvent obtained by the process The acetylated fraction (or acetylated product); fraction F4] obtained by the step of acetylating the soluble portion-removed fraction is obtained by acetylating the hot water soluble portion and the nonpolar organic solvent soluble portion removed fraction. Can be obtained. In the acetylation, for example, acetic anhydride and a suitable organic solvent [for example, pyridine, dimethyl sulfoxide (DMSO), or dioxane] are added to the hot water soluble part and the nonpolar organic solvent soluble part removal fraction. It can be carried out by heating at 0 to 100 ° C. for 1 to 24 hours.

  The acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction of the present invention (that is, the step of adjusting the alkaline aqueous solution extract of basidiomycetes belonging to the genus Kawaratake to the acid, A step of refluxing a precipitate fraction in the presence of a polar organic solvent under acidic conditions, a step of removing a hot water-soluble portion and a nonpolar organic solvent-soluble portion from the polar organic solvent extraction fraction obtained by the step, and the step From the step of acetylating the hot water soluble part and the nonpolar organic solvent soluble part removal fraction obtained by the above, and from the acetylated fraction obtained by the above step, an acidic aqueous solution soluble part, an alkaline aqueous solution soluble part, The acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction obtained from the step of removing the polar organic solvent soluble part; fraction F5) is the acetylated fraction. After contacting with the acidic aqueous solution, the acidic aqueous solution soluble part is removed, and then the obtained acidic aqueous solution soluble part removing fraction and the alkaline aqueous solution are contacted, and then the alkaline aqueous solution soluble part is removed. Furthermore, after making the acidic aqueous solution soluble part and alkaline aqueous solution soluble part removal fraction and nonpolar organic solvent which were obtained contact, the nonpolar organic solvent soluble part can be removed. Contact with the acidic aqueous solution, the alkaline aqueous solution, and the nonpolar organic solvent can be performed in this order, or can be performed in any other order.

  Contact with acidic aqueous solution can be implemented at 0-100 degreeC for 10 to 60 minutes, for example. As said acidic aqueous solution, hydrochloric acid, a sulfuric acid, phosphoric acid, or nitric acid can be used, for example, The density | concentration can be 0.5-2N. The contact with the acidic aqueous solution is preferably repeated 5 to 10 times.

  The contact with the alkaline aqueous solution can be carried out, for example, at 0 to 100 ° C. for 10 to 60 minutes. As the alkaline aqueous solution, for example, a saturated sodium hydrogen carbonate aqueous solution, a 10-20% sodium carbonate aqueous solution, or a saturated potassium hydrogen carbonate aqueous solution can be used, and its pH can be 7.5-10. The contact with the acidic aqueous solution is preferably repeated 5 to 10 times.

  The contact with the nonpolar organic solvent can be carried out, for example, at 0 to 100 ° C. for 10 to 60 minutes. As the nonpolar organic solvent, for example, chloroform, ethyl acetate, hexane, benzene, carbon tetrachloride, or methylene chloride can be used. The contact with the nonpolar organic solvent is preferably repeated 5 to 10 times for one nonpolar organic solvent. Moreover, only one type of nonpolar organic solvent can be used, or different nonpolar organic solvents can be used sequentially.

  Deacetylated fraction (or deacetylated product) of the present invention (that is, a step of adjusting the alkaline aqueous solution extract of basidiomycetes belonging to the genus Kawaratake to acid, the precipitate fraction obtained under the acidic conditions obtained by the step described above as a polar organic solvent The step of refluxing in the presence, the step of removing the hot water soluble part and the nonpolar organic solvent soluble part from the polar organic solvent extraction fraction obtained by the step, the hot water soluble part and the nonpolar organic obtained by the step The step of acetylating the solvent-soluble portion removal fraction, the step of removing the acidic aqueous solution-soluble portion, the alkaline aqueous solution-soluble portion, and the nonpolar organic solvent-soluble portion from the acetylated fraction obtained by the above-mentioned step, and Deacetylation fraction (or deaeration) obtained from the step of deacetylating the acidic aqueous solution soluble portion, alkaline aqueous solution soluble portion, and nonpolar organic solvent soluble portion removal fraction obtained from the step Chill products); Fraction F6), the acidic aqueous solution-soluble portion, alkaline aqueous solution-soluble portion, and a non-polar organic solvent-soluble portion removed fraction may be obtained by deacetylation. In the deacetylation, for example, an alkoxide (for example, sodium methoxide) is added to an appropriate organic solvent (for example, sodium methoxide) in the acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction. It can carry out by adding the solution melt | dissolved in absolute ethanol, pyridine, DMSO, or a dioxane and stirring at 70-100 degreeC for 1 to 24 hours.

  The organic solvent extraction fraction of the present invention (that is, the step of adjusting the alkaline aqueous solution extract of basidiomyceae belonging to the genus Kawaratake to acidity, the step of refluxing the precipitate fraction obtained under the acidic conditions in the presence of a polar organic solvent) , The step of removing the hot water soluble part and the nonpolar organic solvent soluble part from the polar organic solvent extraction fraction obtained by the step, the hot water soluble part and the nonpolar organic solvent soluble part removal fraction obtained by the step The step of acetylating the fraction, the step of removing the acidic aqueous solution soluble part, the alkaline aqueous solution soluble part and the non-polar organic solvent soluble part from the acetylated fraction obtained by the above process, the acidic aqueous solution obtained from the above process A step of deacetylating a soluble portion, an aqueous alkali solution soluble portion, and a non-polar organic solvent soluble portion removed fraction, and a step of extracting a deacetylated fraction obtained from the step with an organic solvent More obtained organic solvent extract fraction; fractions F7), the contacting the deacetylation fraction and an organic solvent, can be obtained by recovering an organic solvent phase. The organic solvent is not particularly limited as long as it exhibits hydrophobic properties. For example, viridine, DMSO, or ethyl acetate can be used. The extraction by contact with an organic solvent is preferably carried out until no coloration is observed in the organic solvent phase.

  The fraction obtained by removing the hot water soluble part and the nonpolar organic solvent soluble part of the fraction extracted from the polar organic solvent of the present invention (that is, the basidiomycete belonging to the genus Kawaratake was refluxed and extracted with a polar organic solvent, A hot water soluble part and a nonpolar organic solvent soluble part removal fraction obtained by a process of removing a nonpolar organic solvent soluble part and a hot water soluble part from a solvent extraction fraction; The fraction F8) is obtained by reflux-extracting basidiomycetes belonging to the genus Kawaratake with a polar organic solvent, for example, bringing the obtained polar organic solvent-extracted fraction into contact with a nonpolar organic solvent, and then a nonpolar organic solvent soluble part Subsequently, after the obtained nonpolar organic solvent removal fraction and hot water are brought into contact with each other, the hot water-soluble part can be removed. The contact with the non-polar organic solvent and the contact with hot water can be performed in this order, or can be performed in the reverse order.

  The polar organic solvent is not particularly limited as long as it exhibits water-soluble properties. For example, acetone, methanol, ethanol, ethyl acetate, or dimethyl ether can be used, and acetone is preferable. The refluxing is preferably carried out until the color of the organic solvent extract disappears, and can be carried out, for example, at 50 to 150 ° C. for 1 to 24 hours. The polar organic solvent extraction fraction of the present invention includes both the extract after reflux and its dried product.

The contact with the nonpolar organic solvent can be carried out, for example, for 10 to 60 minutes. As the nonpolar organic solvent, for example, chloroform, hexane, benzene, carbon tetrachloride, or methylene chloride can be used. The contact with the nonpolar organic solvent is preferably repeated 5 to 10 times for one nonpolar organic solvent. Moreover, only one type of nonpolar organic solvent can be used, or different nonpolar organic solvents can be used sequentially.
Moreover, a contact with hot water can be implemented at 50-100 degreeC for 10 to 60 minutes, for example. The contact with hot water is preferably repeated 5 to 10 times.

  Acetylated fraction (or acetylated product) of the present invention [that is, basidiomycetes belonging to the genus Kawaratake are extracted by refluxing with a polar organic solvent, and then a non-polar organic solvent soluble part and hot water are obtained from the obtained polar organic solvent extract fraction. The acetylated fraction (or acetylated product) obtained by the step of removing the soluble part and the step of acetylating the fraction obtained by the above step; the fraction F9] is the heat of the polar organic solvent extraction fraction. It can be obtained by acetylating the water-soluble part and the nonpolar organic solvent-soluble part removal fraction. The acetylation can be carried out in the same manner as the acetylation carried out to prepare the fraction F4 of the present invention except that the starting materials are different.

The acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction of the present invention (ie, the polar organic obtained after reflux extraction of basidiomycetes belonging to the genus Kawaratake with a polar organic solvent) The step of removing the non-polar organic solvent soluble part and the hot water soluble part from the solvent extraction fraction, the step of acetylating the fraction obtained by the above step, and the acidic aqueous solution soluble from the acetylated fraction obtained by the above step Fraction, acidic aqueous solution soluble part, alkaline aqueous solution soluble part, and nonpolar organic solvent soluble part removal fraction obtained from the step of removing the aqueous solution, alkaline aqueous solution soluble part, and nonpolar organic solvent soluble part; F10), after contacting the acetylated fraction and the acidic aqueous solution, the acidic aqueous solution soluble part was removed, and then the obtained acidic aqueous solution soluble part removing fraction was contacted with the alkaline aqueous solution. After, al The aqueous solution soluble part is removed, and after the acidic aqueous solution soluble part and alkaline aqueous solution soluble part removal fraction are brought into contact with the nonpolar organic solvent, the nonpolar organic solvent soluble part is removed. Can be obtained. Contact with the acidic aqueous solution, the alkaline aqueous solution, and the nonpolar organic solvent can be performed in this order, or can be performed in any other order.
Each contact with the acidic aqueous solution, the alkaline aqueous solution, and the nonpolar organic solvent can be carried out in the same manner as each contact carried out for preparing the fraction F5 of the present invention except that the starting materials are different. .

  The deacetylated fraction (or deacetylated product) of the present invention (that is, basidiomyceae belonging to the genus Kawaratake) after reflux extraction with a polar organic solvent, from the obtained polar organic solvent extract fraction, From the step of removing the hot water soluble portion, the step of acetylating the fraction obtained by the above step, and the acetylated fraction obtained by the above step, the acidic aqueous solution soluble portion, the alkaline aqueous solution soluble portion, and the nonpolar organic Deacetylation obtained from the step of removing the solvent soluble part, and the step of deacetylating the acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction obtained from the above step The fraction (or deacetylated product); the fraction F11) is to deacetylate the acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction. Ri may be obtained. The deacetylation can be carried out in the same manner as the deacetylation carried out to prepare the fraction F6 of the present invention, except that the starting materials are different.

  The organic solvent extract fraction of the present invention (that is, basidiomycetes belonging to the genus Kawaratake are extracted by refluxing with a polar organic solvent, and then the nonpolar organic solvent soluble part and the hot water soluble part are extracted from the obtained polar organic solvent extract fraction. The step of removing, the step of acetylating the fraction obtained by the above step, and the acetylated fraction obtained by the above step remove the acidic aqueous solution soluble portion, the alkaline aqueous solution soluble portion, and the nonpolar organic solvent soluble portion. A step of deacetylating the acidic aqueous solution soluble part, the alkaline aqueous solution soluble part, and the nonpolar organic solvent soluble part removal fraction obtained from the step, and the deacetylation fraction obtained from the step The organic solvent extraction fraction obtained by the step of extracting with an organic solvent; fraction F12) is obtained by contacting the deacetylated fraction with an organic solvent and recovering the organic solvent phase. Rukoto can. The contact with the organic solvent can be carried out in the same manner as the contact with the organic solvent carried out for preparing the fraction F7 of the present invention, except that the starting materials are different.

As long as the conjugate of the present invention is a conjugate of a protein with at least one fraction of the fractions F1 to F12 of the present invention, or a telephoric acid compound represented by the general formula (I) or a salt thereof, It is not particularly limited.
The protein is not particularly limited as long as it can form a complex with the teleholic acid and the complex exhibits antiallergic activity, and examples thereof include albumin and globulin.

The fractions F1 to F12 of the present invention derived from basidiomycetes belonging to the genus Kawaratake are, for example, as they are in a state where insoluble matters are mixed, or in a state where insoluble matters are removed, and further suitable treatment (for example, concentration) Alternatively, it can be used as an active ingredient of the antiallergic agent of the present invention after being diluted, dried, or redissolved). Of the fraction F7 of the present invention, the fraction prepared according to the procedure of Example 1 substantially contains only teleholic acid.
In the present invention, when the fractions F1 to F12 of the present invention are used as active ingredients, the present inventor believes that the teleholic acid contained in the fraction exhibits antiallergic activity. It is considered that components other than teleholic acid in the fraction also exhibit antiallergic activity, and the scope of the present invention is not limited to the above mechanism.

  The antiallergic agent of the present invention comprises, as an active ingredient, (1) a teleholic acid compound represented by the general formula (I) or a salt thereof, a conjugate of the teleholic acid compound or a salt thereof and a protein, or (2 ) It contains at least one of the various fractions (F1 to F12) of the present invention derived from basidiomycetes belonging to the genus Kawaratake, or a conjugate of at least one fraction of the fractions F1 to F12 and a protein. The antiallergic agent of the present invention can contain any one of the above active ingredients, and can contain two or more in any combination. In the present invention, the above-mentioned active ingredient alone, or preferably together with a usual carrier or diluent that can be pharmaceutically or veterinarily acceptable, suppresses allergy or treats an allergy-related disease. Alternatively, it can be administered to a subject in need of prevention [eg, an animal, preferably a mammal (particularly a human)] in an effective amount. Moreover, the said active ingredient can be used in order to manufacture the antiallergic agent or the treatment or prevention agent of the disease relevant to allergy.

Diseases related to allergy include, for example, asthma, atopic dermatitis, chemical sensitivity, hay fever, food allergy, latex allergy, metal allergy, tick allergy, drug rash, bronchial asthma, allergic rhinitis, hives, allergies Bronchial asthma, drug anaphylactic shock, gastrointestinal allergy, allergic subsepsis, allergic gastroenteropathy, allergic bronchopulmonary aspergillosis, allergic stomatitis, allergic purpura, allergic granulomas, allergic pneumonia, allergic Examples thereof include pulmonary eosinophilic disease, allergic alveolitis, allergic food poisoning, or allergic granulomatous vasculitis.

  The dosage form of the antiallergic agent of the present invention is not particularly limited, and examples thereof include powders, fine granules, granules, tablets, capsules, suspensions, emulsions, syrups, extracts, or pills. Or parenteral preparations such as injections, external solutions, ointments, suppositories, topical creams, or eye drops.

  These oral preparations include, for example, sodium alginate, starch, corn starch, sucrose, lactose, glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid ester, talc, magnesium stearate, Excipients such as polyethylene glycol, magnesium silicate, anhydrous silicic acid, or synthetic aluminum silicate, binders, disintegrants, surfactants, lubricants, fluidity promoters, diluents, preservatives, colorants, It can be produced according to a conventional method using a fragrance, a flavoring agent, a stabilizer, a moisturizing agent, an antiseptic, or an antioxidant.

Examples of parenteral administration methods include injection (subcutaneous, intravenous, etc.) or rectal administration. Of these, the injection is most preferably used.
For example, in the preparation of an injection, in addition to the active ingredient, for example, a water-soluble solvent such as physiological saline or Ringer's solution, a water-insoluble solvent such as vegetable oil or fatty acid ester, an isotonic agent such as glucose or sodium chloride, Solubilizers, stabilizers, preservatives, suspending agents, or emulsifiers can be optionally used.
Further, the antiallergic agent of the present invention may be administered by using a method of sustained release preparation using a sustained release polymer or the like. For example, the active ingredient in the present invention can be incorporated into a pellet of ethylene vinyl acetate polymer and the pellet can be surgically implanted into the tissue to be treated or prevented.

The antiallergic agent of the present invention is not limited to this, but the active ingredient in the present invention can be contained in an amount of 0.01 to 99% by weight, preferably 0.1 to 80% by weight. .
The dosage in the case of using the antiallergic agent of the present invention can be appropriately determined depending on the type of disease, the age, sex, weight, symptom of the patient, or the administration method, orally or parenterally. Can be administered as a whole.

Further, the administration form is not limited to pharmaceuticals, and various forms such as health food (preferably functional food) or feed can be given in the form of food and drink. The food includes a beverage.
In addition to (1) function as a nutrient (primary function), (2) function to appeal to the human senses (secondary function), (3) human health, physical ability, or psychological state For the maintenance of health and recovery of health by regulating physiological systems such as the digestive system, circulatory system, endocrine system, immune system, or nervous system. It is known to have an effect. In the present specification, “health food” means a food that has some effect on health or can be expected to have an effect, and “functional food” refers to the above-mentioned “health food”. Foods designed and processed to fully express various bioregulatory functions (i.e., regulatory functions of digestive, circulatory, endocrine, immune, or nervous systems) Means.

  Furthermore, an oral hygiene composition, for example, temporarily contained in the mouth, but most of it is given in the form of exhaling from the mouth, for example, in the form of a dentifrice, mouthwash, chewing gum, or mouthwash, or It is also possible to give in the form of an inhalant to be aspirated from the nose. For example, the active ingredient in the present invention may be added as an additive (for example, a food additive) to a desired food (including beverage), feed, dentifrice, mouthwash, chewing gum, gargle, or the like. it can.

  EXAMPLES Hereinafter, the present invention will be specifically described by way of examples, but these do not limit the scope of the present invention.

<< Example 1: Separation and identification of teleholic acid fraction from dried fruit body of Kawaratake >>
Add 2 L of saturated sodium borate aqueous solution to 15 g of dried fruit body powder of Kawaratake and stir and extract in a 5 L beaker at 25 ° C. for 24 hours, and then centrifuge (12,000 × g, 30 minutes) to recover the supernatant did. The pH of the supernatant was adjusted to 2.0 with hydrochloric acid, allowed to stand at room temperature overnight, and then centrifuged (40,000 × g, 30 minutes) to recover the precipitate. Distilled water (500 mL) was added to the precipitation part, suspended sufficiently, and after repeating the washing operation until the pH of the washing solution reached 7.0, centrifugation (40,000 × g, 30 minutes) was performed to recover the precipitation part, Lyophilization gave 1.2 g of powder.

  1.2 g of lyophilized product was added to Dimroth (manufactured by PYREX), 10 mL of acetone was added, and the mixture was heated to reflux at 80 ° C for 3 hours. Extraction was repeated until the color of the extract disappeared, and then the extract was collected and concentrated to dryness using an evaporator to obtain a dried product. Distilled water (10 mL) was added to the dried product, heated at 80 ° C. for 5 minutes, and the hot water soluble part was removed by centrifugation. After this operation was repeated 10 times, the same operation was performed 10 times each by replacing the solvent with hexane and then chloroform to obtain 8 mg of a blue-purple precipitate fraction.

  1 mL of acetic anhydride was added to this fraction, heated in a 50 mL eggplant flask containing 1 mL of pyridine at 70 ° C. for 24 hours, and then concentrated to dryness using an evaporator. Next, 10 mL of 1N hydrochloric acid was added to remove the hydrochloric acid soluble part, and this operation was repeated three times. Then, instead of hydrochloric acid, using a saturated aqueous sodium bicarbonate solution, hexane, chloroform, and ethyl acetate in this order, the same operation was repeated to remove each soluble part, and then the insoluble part was dissolved in pyridine. Then, it was concentrated to dryness using an evaporator to obtain 5.9 mg of an acetylated fraction.

To the acetylated fraction, 1 mL of a solution in which CH ₃ ONa was dissolved in absolute ethanol to 1 N was added, and the mixture was stirred in a 50 mL Erlenmeyer flask at room temperature for 1 hour. Next, after concentrating to dryness using an evaporator, 20 mL of 1N hydrochloric acid solution was added and put into a separatory funnel. Next, an equal amount of ethyl acetate was added, and extraction operation was performed. This operation was repeated until no coloration was observed in the ethyl acetate phase. Then, the ethyl acetate phase was recovered and concentrated to dryness using an evaporator to obtain 4 mg of a blue deacetylated product.

The obtained fraction was subjected to structural analysis by ¹ H-NMR and ¹³ C-NMR (UNITY INOVA 500; Varian). As a result, in ¹ H-NMR (FIG. 1), δ7.14 and 7.35 ppm were derived from a benzene ring. A signal derived from a hydroxyl group was observed at the root signal, δ9.81, 10.08 ppm. Further, in ¹³ C-NMR (FIG. 2), a signal derived from a benzene ring in the vicinity of δ 98.49, 104.82, 146.43, 149.24, 150.79, 151.326 ppm, and quinone derived from δ 172.38 ppm. A signal was confirmed. From the above, the presence of aromatics having a quinone skeleton was confirmed.

Next, in the IR spectrum (FT / IR VALORIII; JASCO), 3204 cm ⁻¹ absorbs hydroxyl group, 1634 cm ⁻¹ absorbs quinone derived carbonyl, 867 cm ⁻¹ and 801 cm ⁻¹ absorbs furan ring. (FIG. 3). MS analysis (Hybrid Mstation JMS-700QQ; JEOL) showed m / zC ₁₈ H ₈ O ₈ (M ⁺ ) 352.0240, which was consistent with teleholic acid (FIG. 4).

<< Example 2: Preparation of a conjugate of a teleholic acid fraction and a protein >>
The teleholic acid fraction prepared in Example 1 was dissolved in dimethyl sulfoxide (DMSO) and adjusted to 1 mmol / L to prepare a teleholic acid solution. This solution was added to 10 mg / mL of human albumin or human globulin in PBS (pH 7.2) and reacted at room temperature for 3 hours. The resulting reaction solution was dialyzed against distilled water, and the content solution was collected and lyophilized to obtain a telephoric acid / human albumin conjugate and a teleholic acid / human globulin conjugate.

<< Example 3: Evaluation of antiallergic activity using PCA reaction as an index >>
(1) Preparation of sensitizing sample solution and each administration solution As a sample solution, 1 mg of a sample (that is, the teleholic acid fraction prepared in Example 1) was dissolved in 250 μL of a 10 mmol / L sodium borate aqueous solution and sonicated. Thereafter, 250 μL of distilled water was added to obtain a sample solution (1).

The administration liquid I to the administration liquid V were prepared by the following procedure. In addition, preparation of each administration liquid was performed at the time of use.
The administration liquid I comprises the sample liquid (1), an anti-ovalbumin antibody (rabbit antiserum) diluted 1000 times with 10 mmol / L phosphate buffered saline (hereinafter referred to as PBS) (pH 7.2), and Were mixed to prepare an equal amount.
The dosing solution II was prepared by mixing equal amounts of the sample solution (1) diluted 2-fold with distilled water and the anti-ovalbumin antibody diluted 1000-fold with 10 mmol / L PBS (pH 7.2).
The administration liquid III and the administration liquid VI were prepared by mixing equal amounts of a 10 mmol / L sodium borate aqueous solution and an anti-ovalbumin antibody diluted 1000 times with 10 mmol / L PBS (pH 7.2).

The administration liquid IV was prepared by dissolving 1 mg of diphenhydramine hydrochloride (positive control) in 250 μL of 10 mmol / L sodium borate aqueous solution, sonicating, and then adding 250 μL of distilled water, and 10 mmol / L An equal amount of an anti-ovalbumin antibody diluted 1000 times with PBS (pH 7.2) was prepared.
The dosing solution V was prepared by mixing equal amounts of the sample solution (2) diluted 2-fold with distilled water and the anti-ovalbumin antibody diluted 1000-fold with 10 mmol / L PBS (pH 7.2).

(2) Sensitization method The back of the guinea pig (about 15 × 8 cm) was shaved with a clipper. Each administration solution (0.1 mL) was injected into the back skin of three guinea pigs. The administration sites were administration liquids I to III from the head on the right side, and administration liquids IV to VI on the left side of the back-middle line.

(3) Induction method 48 hours after the intradermal injection, the inducing antigen solution was mixed with 1% Evans blue (Wako Pure Chemicals) / physiological saline solution in equal amounts, and 1 mL was injected into the hindlimb hind vein of the guinea pig. .

(4) Determination method Guinea pigs were sacrificed 30 minutes after the dye solution injection, and the epidermis around the injection site was peeled off. The site of intradermal injection was observed, the major axis and minor axis of pigment spots were measured, and A [= major axis x minor axis (mm ² )] was calculated. Next, the ratio B [= A / A ′] of the sample treatment site A to the control sensitization site (site III or VI) A ′ on the same side with the back line as a boundary was determined.
PCA (passive cutaneous anaphylaxis) reaction inhibition rate was calculated from the following equation.
C = 100−B × 100 (%)

(5) Results As shown in Table 1, the sample prepared in Example 1 (Agaricus-derived teleholic acid) suppressed the PCA reaction.

<< Example 4: Evaluation of anti-allergic activity using IgE production as an index >>
(1) Experimental Method Female BALB / c mice purchased from Charles River Japan (Kanagawa) were preliminarily raised and subjected to experiments at 8 weeks of age (5 per group).
The telephoric acid prepared in Example 1 was dissolved in dimethyl sulfoxide (DMSO), adjusted to a predetermined concentration with polyethylene glycol (molecular weight 400; Sigma-Aldrich), and sensitized for 1 week in an amount of 5 mg / kg. From before until the end of the experiment, subcutaneous injection was performed 3 times a week. As a control, a polyethylene glycol subcutaneous injection group was provided.
For antigen sensitization, dinitrophenyl-linked keyhole limpet hemocyanin (hereinafter abbreviated as DNP-KLH; Cosmo Bio, Tokyo) was dissolved in a phosphate buffer and adjusted to a concentration of 10 mg / mL. 2 μL was added to 1 mL of a 20 mg / mL ALUM (Cosmo Bio) solution and mixed. Antigen sensitization was performed by injecting 0.2 mL of this mixed solution into a mouse abdominal cavity.
After 3, 7, 14, and 21 days of antigen sensitization, the mouse orbital vascular plexus was lightly damaged with a capillary, a partial blood sample was collected, and the serum was separated. Serum IgE levels in the serum were measured using a commercially available ELISA kit.

The measurement of free histamine was carried out by the following procedure. That is, mice 14 days after sensitization were killed, and the peritoneal cavity was washed and collected with RPMI 1640 medium, and centrifuged (700 rpm, 10 minutes) to obtain peritoneal cells. The peritoneal cells were suspended in RPMI 1640 medium and subjected to specific gravity centrifugation with bovine serum albumin (BSA) / saline gradient solution (d = 1.07) to obtain mast cells. The cells were washed with RPMI1640 medium, adjusted to a concentration of 1 × 10 ⁵ mL, added with 250 ng / mL of DNP-KLH, and reacted in a 5% carbon dioxide culture vessel at 37 ° C. for 30 minutes. The histamine level in the culture supernatant was quantified using a commercially available ELISA kit (Immunotech). The histamine release rate is given by the following formula:
(Total amount of histamine in DNP-KLH-stimulated group) × 100 / (Total amount of histamine in non-stimulated DNP-KLH group)
Calculated by

(2) Results In control mice sensitized with DNP-KLH, IgE production was observed from 3 days after sensitization, and persisted until 21 days after sensitization. On the other hand, in the group in which the sample (teleholic acid fraction) obtained in Example 1 was administered subcutaneously (5 mg / kg), IgE production was significantly suppressed throughout the observation period compared to the control group.
Further, when intraperitoneal mast cells were isolated from mice 14 days after sensitization with DNP-KLH, and the amount of histamine released when restimulated with DNP-KLH was measured, the amount of histamine released in the example sample administration group was It was significantly suppressed compared to.
As a result of the above, it is considered that teleholic acid suppresses blood IgE production in DNP-KLH-sensitized mice, and thus suppresses degranulation induction of mast cells.

Example 5 Effect of teleholic acid on histamine release from basophils
In this example, the rat basophilic leukemia cell line RBL-2H3 [Rintaro
Yamanishi et al., Bioscience, Biotechnology and Biochemistry, 61, 19-23 (2002)] were used as histamine-secreting cells. The cell line RBL-2H3 was obtained from Tohoku University, Aging Research Institute, Medical Cell Resource Center (Sendai, Miyagi Prefecture), and in Kureha Chemical Industry Co., Ltd. Biomedical Research Institute in 10% FBS-added RPMI1640 medium. It is the one that has been passaged and maintained. The cell line RBL-2H3 was seeded on a 48-well culture plate at 2.5 × 10 ⁵ cells per well (capacity 500 μL), cultured in a 5% carbon dioxide incubator overnight, and then 11 μg / 50 mL of mL anti-DNP-DNP-Ascaris IgE (Seikagaku Corporation, Tokyo) solution was added (final concentration: 1 μg / mL) and cultured for 3 hours.

  After completion of the culture, each well was washed with PBS, and then 500 μL of a medium containing 0, 0.1, 1 or 10 μmol / L of teleholic acid was dispensed into each well and incubated for 30 minutes. Subsequently, 10 μL of a 40 μg / mL DNP-BSA (Cosmo Bio, Tokyo) solution was added (final concentration 0.8 μg / mL), followed by further culturing for 1 hour. Then, the culture supernatant was collected by centrifugation and stored frozen at -20 ° C.

The histamine content in the culture supernatant was measured using a commercially available ELISA kit (Histamine EIA
KIT; SPI-BIO, Montigny Le Bretonneux, France). The measurement results are shown in Table 2. Compared with the control group, the histamine level in the supernatant was significantly lower in the teleholic acid addition group. That is, it was suggested that teleholic acid may suppress allergy by suppressing histamine secretion.

Example 6 Inhibition of Passive PCA Reaction by Teleholic Acid
1 mg of the test substance teleholic acid was dissolved in 250 μL of 10 mmol / L sodium borate aqueous solution, and then 250 μL of distilled water was added to make 0.2 mg / mL. The positive control sodium cromoglycate (Fujisawa Pharmaceutical, Osaka) was dissolved in physiological saline to a concentration of 0.2 mg / mL.

  Into the back skin of 8-week-old male SD rat (Claire Japan), 0.1 mL of anti-DNP-DNP-Ascaris IgE (Seikagaku Corporation, Tokyo) diluted 800-fold was injected. 48 hours later, the telephoric acid or cromoglycate sodium was intravenously injected (1 mg / kg amount), and after 1 minute, a DNP-Ascaris (Cosmo Bio) solution having a final concentration of 0.3 mg / mL And a final concentration of 5 mg / mL of Evans blue solution (Sigma Aldrich, Tokyo) was intravenously injected (1 mL / animal). After 30 minutes, the rats were exsanguinated under ether anesthesia, the skin around the injection site was peeled off, and the pigment leakage part was cut out with scissors. This was treated in 2 mL of 2 mol / L aqueous potassium hydroxide solution at 37 ° C. for 21 hours, and 6 mL of a 7.4% phosphoric acid / acetone mixture (1: 3) was added and mixed vigorously. After centrifuging at 3,000 rpm for 15 minutes, the absorbance at 630 nm of 100 μL of the collected supernatant was measured with a spectrophotometer, and the dye concentration was calculated from a calibration curve. The calibration curve was obtained by adding 2 mL of 2 mol / L potassium hydroxide aqueous solution so that Evans blue had a concentration of 0, 3.125, 6.25, 12.5, 25, 50, 100, and 200 μg / mL. This was prepared by measuring the absorbance at 630 nm of a dye standard solution prepared by adding and dissolving rat skin of an untreated control.

  The results are shown in Table 3. The passive PCA reaction was suppressed by administration of teleholic acid. That is, it was suggested that teleholic acid has antiallergic activity.

The antiallergic agent of the present invention can be applied to the use for treatment or prevention of allergy.
As mentioned above, although this invention was demonstrated along the specific aspect, the deformation | transformation and improvement obvious to those skilled in the art are included in the scope of the present invention.

Claims (2)

The following general formula (I):

[Wherein, R ¹ , R ² , R ³ , and R ⁴ are each independently a hydrogen atom, a lower alkyl group having 1 to 4 carbon atoms, or an acetyl group . ]
Antiallergic agents you characterized by containing Terehoru acid compound or a salt thereof as an active ingredient represented in. 2. The teleholic acid compound or a salt thereof is obtained by extraction from at least one of basidiomycetes belonging to the genus Kawaratake, mushrooms belonging to the family Amanita, Koshiji, or shiitake mushroom. Antiallergic agent.

Images