JP2001114686A - Th2 CYTOKININ EXPRESSION SUPPRESSANT - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a medicinal composition having excellent Th2 cytokinin expression-suppressing effects. SOLUTION: This Th2 cytokinin expression suppressant contains a flavonoid compound expressed by general formula (1) [R1, R2, R3, R4, R5, R6 and R7 are each the same or different and express H or OH; R is H, OH or a glycosyl;... is a single bond or a double bond.] as an effective ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a Th2 cytokine expression inhibitor containing a flavonoid compound as an active ingredient.

[0002]

2. Description of the Related Art Th2 cytokines including interleukin-4, interleukin-5, and interleukin-13 have various functions, and various effects can be expected by suppressing the expression of Th2 cytokines ( For example, since interleukin 4 has an effect of promoting the production of IgE, suppression of the expression of interleukin 4 can be expected to prevent allergic symptoms associated with IgE). Also, the Th2 cytokine mR
It is considered that production of Th2 cytokine is suppressed by suppressing the expression of NA.

[0003]

SUMMARY OF THE INVENTION The present invention provides an excellent Th
2. An object of the present invention is to provide a pharmaceutical composition having an effect of suppressing cytokine expression.

[0004]

Means for Solving the Problems The present inventors have found that a flavonoid compound represented by the general formula (1) can be used for screening Th2 cytokines, specifically, interleukin-4 (IL-4) in screening for Th2 cytokine expression. ,
It shows an excellent inhibitory effect on the expression of mRNA of interleukin-5 (IL-5) and interleukin-13 (IL-13), and inhibits the production of these interleukins by suppressing the expression of mRNA of these interleukins. The present invention was found to have an effect, and the present invention was completed.

That is, the present invention provides the inventions described in the following items.

Item 1 General Formula (1)

[0007]

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[Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and each represent a hydrogen atom or a hydroxyl group. R ⁸ represents a hydrogen atom, a hydroxyl group or a glycosyl group. --- indicates a single bond or a double bond. And a flavonoid compound represented by the formula:

Item 2 The flavonoid compound represented by the general formula (1) is represented by the following general formula (2)

[0010]

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[Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and each represent a hydrogen atom or a hydroxyl group. R ⁸ ′ represents a hydroxyl group or a glycosyl group. ], The following general formula (3)

[0012]

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[Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and represent a hydrogen atom or a hydroxyl group. Or the following general formula (4)

[0014]

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[Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and represent a hydrogen atom or a hydroxyl group. R ⁸ represents a hydrogen atom, a hydroxyl group or a glycosyl group. The expression inhibitor according to item 1, which is a flavonoid compound represented by the formula:

Item 3. The expression inhibitor according to item 1, wherein the flavonoid compound represented by the general formula (1) is fisetin, quercetin, kaempferol, apigenin or luteolin.

Item 4. The expression inhibitor according to any one of Items 1 to 3, which is an interleukin-4 expression inhibitor.

Item 5. The expression inhibitor according to any one of Items 1 to 3, which is an interleukin-5 expression inhibitor.

Item 6. The expression inhibitor according to any one of Items 1 to 3, which is an interleukin-13 expression inhibitor.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION The Th2 cytokine expression inhibitor of the present invention has the general formula (1)

[0021]

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[Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ ,
R ⁷ , R ⁸ and --- are the same as above. ] As an active ingredient.

The glycosyl group represents a sugar residue such as a monosaccharide such as glucose, galactose, fructose, and rhamnose, and a disaccharide such as rutinose, bisianose, lactose, maltose, and sucrose. Therefore, the glycosyl group indicates, for example, a glucosyl group, a galactosyl group, a fructosyl group, a rhamnosyl group and the like. Glycosyl groups include any combination of these groups, 1 → 2
Also included are groups linked by a bond, 1 → 3 bond, 1 → 4 bond or 1 → 6 bond to form a disaccharide.

The compound represented by the general formula (1) includes
Preferred examples include compounds represented by the following general formula.

General formula (2)

[0026]

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Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above. R ⁸ ′ represents a hydroxyl group or a glycosyl group. (In the general formula (1), R ⁸ is a hydroxyl group or a glycosyl group, and corresponds to a compound in which --- is a double bond).

The flavonoid compound represented by the general formula (2) is a flavonol compound having a hydroxyl group at the 3-position of flavone or a glycoside thereof.
Examples include galangin, morin, myricetin, quercetin, fisetin, kaempferol, astragalin, rutin, and isoquercitrin.

General formula (3)

[0030]

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Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same as above. Flavonoid compound represented by In (formula (1), R ⁸ is a hydrogen atom, ---
Corresponds to a compound having a double bond. ).

As the compound represented by the general formula (3),
Specific examples include chrysin, apigenin, luteolin and the like.

General formula (4)

[0034]

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[Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ ,
R ⁷ and R ⁸ are the same as above. (In the general formula (1), the flavonoid compound corresponds to a compound in which --- is a single bond).

The compound represented by the general formula (4) includes
Specific examples include fustin, eriodictyol, and the like.

Among these specifically listed compounds, the active ingredient of the present invention is preferably fisetin, apigenin, luteolin, kaempferol, myricetin, quercetin and rutin, and fisetin, kaempferol, quercetin, apigenin and luteolin. More preferred are fisetin, apigenin and luteolin.

It should be noted that a compound in which a cinnamoyl group or a caffeoyl group is substituted on the flavonoid skeleton can be expected to have the same effect as the flavonoid compound represented by the general formula (1).

The active ingredient of the present invention represented by the general formula (1) may be a synthesized one or a plant-derived one.

Plants containing a large amount of the active ingredient of the present invention include:
Persimmon leaves, mugwort, cedar leaves, mint, loofah, birch,
Grape seeds, amachaworu, gymnema, guava, wolfberry,
Kumazasa, jasmine, horsetail, dogwood, adlay,
Loquat leaves, moroheiya, udo, saffron, mulberry, rafuma, safflower, eucommia, lily of the valley, spring and the like. Of these, persimmon leaves, mugwort, cedar leaves, mint, loofah, birch and grape seeds are preferred, and persimmon leaves are particularly preferred.

In the present invention, these plant extracts obtained according to a conventional method may be used as they are or may be used after being purified.

The Th2 cytokine expression inhibitor of the present invention comprises a flavonoid compound of the general formula (1) as an essential component and a suitable pharmaceutically acceptable carrier, and is used in the form of a general pharmaceutical preparation. Practical as.

As the dosage unit form of the above pharmaceutical preparation, various forms can be selected according to the purpose of treatment, and typical examples thereof include solid preparations such as tablets, pills, granules, capsules, and troches. Powder preparations such as internal powders, external powders and powders; liquid preparations such as solutions, suspensions, emulsions, injections (solutions, suspensions, etc.), syrups, lotions, aerosols, and eye drops; Creamy preparations such as ointments; cataplasms and the like.

The pharmaceutically acceptable carrier utilized in the Th2 cytokine expression inhibitor of the present invention includes binders, disintegrants, surfactants, absorption enhancers which are usually used depending on the form of the preparation. Agents, humectants, adsorbents, lubricants, fillers, extenders, diluents or excipients such as humectants,
These are appropriately selected and used according to the dosage unit form of the preparation.

The Th2 cytokine expression inhibitor of the present invention may further contain, if necessary, a preservative, a sweetening agent, a coloring agent, an antioxidant, a preservative, a fragrance, a flavoring agent, and other agents. Can also be prepared.

The above pharmaceutical preparation can be prepared according to a conventional method in the above-mentioned form.

The administration method of the above pharmaceutical preparation is not particularly limited, and it is administered according to the form of the preparation. For example, tablets, pills, granules, capsules, troches, powders for internal use, solutions, suspensions, Emulsions, syrups and the like are orally administered, and external powders, powders, lotions, ointments and cataplasms are transdermally administered. Injections are given intravenously or intramuscularly
It is administered intradermally, subcutaneously or intraperitoneally. Aerosols can be administered as nasal drops.

The dose of the above pharmaceutical preparation is appropriately selected depending on the form of the preparation, the age of the patient, gender and other conditions, the degree of the disease, and the like.
The dose should be about 0.025 to 3 mg, preferably about 0.05 to 1.5 mg per 1 kg of body weight per day.
It can be administered in two or four divided doses.

The flavonoid compound represented by the general formula (1) is a Th2 cytokine (specifically, IL-4, I-4
L5 and IL-13) have the effect of suppressing the expression of mRNA, and therefore, Th2 cytokines (specifically, I-5
L2, IL-5 and IL-13) are suppressed (inhibited) to suppress the expression of Th2 cytokines (specifically, IL-4, IL-5 and IL-13). can do. Therefore, a pharmaceutical preparation containing the flavonoid compound represented by the general formula (1) as an active ingredient has a T
h2 cytokine expression inhibitor, specifically IL-4, I
It can be used as an L-5 and IL-13 expression inhibitor.

Since IL-4, IL-5, and IL-13 are involved in IgE production and induction of chronic inflammation, the Th2 cytokine expression inhibitor of the present invention may be used in various conditions associated with them, specifically, allergic conditions. It can be expected to act as a controller (regulator) for immediate allergic reactions and chronic allergic inflammation (eg, allergic rhinitis, hay fever, atopic dermatitis, etc.) in sexually transmitted diseases.

The plant extract (or plant extract) exemplified above as a plant containing the flavonoid compound represented by the general formula (1)
If the composition containing the extract is taken for a long time, Th
2 cytokines (specifically IL-4, IL-5 and I
L-13) suppresses the production of IgE and suppresses inflammation based on the suppression of expression of L-13), thereby preventing or ameliorating various symptoms related to production and inflammation of IgE, such as allergy (allergic rhinitis, pollinosis, atopy Dermatitis, etc.), the prevention or improvement of rough skin, and the improvement of itching can be expected.

[0052]

EXAMPLES The present invention will be described with reference to experimental examples and prescription examples, but the scope of the present invention is not limited only to these examples.

Experimental Example 1: IL-4, IL-5 and IL by RT-PCR
Expression of -13 mRNA The expression levels of IL-4, IL-5 and IL-13 mRNA were measured according to a conventional method.

The human basophil cell line (KU812) was cultured in RPMI1640 medium (Gibco) containing 10% fetal bovine serum (treated at 56 ° C. for 30 minutes) at 37 ° C. under 5% CO ₂ . KU812 cells (5
× 10 ⁵ cells / ml) were stimulated with the flavonoid compounds at the concentrations shown in Tables 1 and 2 and 1 μMA23187 ionophore for 3 hours. After washing, total RNA was collected by RNAzol (Biotex, USA). Of the total RNA collected, 800 ng
The mixture was mixed with xture (Rerkin ElmerCetus, USA) and incubated at 99 ° C. for 5 minutes and at 37 ° C. for 60 minutes. After each RT product was obtained, PCR amplification was performed using IL-4, IL-5 and IL-13 specific primers and β-actin specific primers. The product was electrophoresed on a 2% agarose gel, stained with ethidium bromide, photographed, measured with a densitometer, and the expression level was evaluated as-, ±, +, ++, or +++. The results are shown in Tables 1 and 2 below.

[0055]

[Table 1]

[0056]

[Table 2]

The expression levels in Tables 1 and 2 are shown by the following criteria. +++; very large, ++; large, +: small, ±; very small, −; none By administering each flavonoid compound shown in Table 1 to basophil cells, IL-4 of basophil cells was reduced. The effect of suppressing the expression of mRNA was observed. In particular, remarkable inhibitory effects were observed with fisetin, kaempferol, quercetin, apigenin and luteolin. Fisetin was found to suppress the expression of IL-4, IL-5 and IL-13.

The following is a formulation example of the interleukin expression inhibitor of the present invention. These can be manufactured according to a usual method in the form.

Formulation Example 1: Chewable tablets (mg) Fisetin 5 Xylitol 300 Aspartame 4 Magnesium stearate 10 Fragrance 1 Formulation Example 2: Ointment (g) Apigenin 0.5 Polyethylene glycol 400 49.5 Polyethylene glycol 4000 49.8 Acrynol powder 0.2

[0060]

The flavonoid compound represented by the general formula (1), specifically, the general formula (2), the general formula (3) or the general formula (4) can be used for basophil or mast cell IL-
4. It has an effect of suppressing the expression of mRNA of IL-5 and IL-13, and has an effect of suppressing the expression of IL-4, IL-5 and IL-13. Therefore, a pharmaceutical composition containing the flavonoid compound of the general formula (1) is a Th2 cytokine, specifically, IL-4, IL-5 and IL-13.
It can be used as an expression inhibitor. IL-4,
Th2 cytokines such as IL-5 and IL-13 are Ig
It is involved in E production and induction of chronic inflammation. Therefore, the Th2 cytokine expression inhibitor of the present invention can be used as a controller (regulator) of an immediate allergic reaction or chronic allergic inflammation (eg, allergic rhinitis, hay fever, atopic dermatitis, etc.) in allergic diseases. Action can be expected. Alternatively, a plant extract containing a flavonoid compound represented by the general formula (1) (or a composition containing the extract) is converted into IgE production based on suppression of expression of IL-4, IL-5 and IL-13. Allergies due to suppression (eg, allergic rhinitis, hay fever, atopic dermatitis)
Can be taken over a long period of time for the purpose of prevention of skin irritation, prevention or improvement of rough skin, improvement of itching, and the like.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 43/00 111 A61P 43/00 111 // A61K 35/78 A61K 35/78 CF term (Reference) 4C086 AA01 AA02 BA08 EA11 GA17 MA01 MA04 NA14 ZA33 ZA34 ZA89 ZB13 ZC41 4C088 AB03 AB12 AB17 AB18 AB19 AB20 AB24 AB25 AB26 AB29 AB34 AB38 AB47 AB48 AB51 AB52 AB56 AB64 AB72 AB76 AB77 AB85 AC04 AC05 BA14 NA14 ZA89 ZA33

Claims (6)

[Claims] 1. A compound of the general formula (1) [Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and represent a hydrogen atom or a hydroxyl group. R ⁸ is
It represents a hydrogen atom, a hydroxyl group or a glycosyl group. --- indicates a single bond or a double bond. And a flavonoid compound represented by the formula: 2. The flavonoid compound represented by the general formula (1) is represented by the following general formula (2): [Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and represent a hydrogen atom or a hydroxyl group. R ⁸ '
Represents a hydroxyl group or a glycosyl group. ], The following general formula (3): [Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and represent a hydrogen atom or a hydroxyl group. Or the following general formula (4): [Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are the same or different and represent a hydrogen atom or a hydroxyl group. R ⁸ is
It represents a hydrogen atom, a hydroxyl group or a glycosyl group. The expression inhibitor according to claim 1, which is a flavonoid compound represented by the formula: 3. The expression inhibitor according to claim 1, wherein the flavonoid compound represented by the general formula (1) is fisetin, quercetin, kaempferol, apigenin or luteolin. 4. The expression inhibitor according to claim 1, which is an interleukin-4 expression inhibitor. 5. The expression inhibitor according to claim 1, which is an interleukin-5 expression inhibitor. 6. The expression inhibitor according to claim 1, which is an interleukin-13 expression inhibitor.