JP4385243B2 - Composition for preventing or treating inflammation involving type IV allergic reaction - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a composition for preventing or treating inflammation involving type IV allergic reaction, and an external preparation for skin comprising the composition.
[0002]
[Prior art]
Atopic dermatitis is considered to be caused by both type I and type IV allergic reactions. Acute lesions are caused by type I allergic reactions and chronic lesions are caused by type IV allergic reactions. Yes.
[0003]
Type I allergic reaction is an immediate type of reaction in which antigen recombines with IgE antibody bound to mast cells, a chain reaction occurs in mast cells, and a chemical transmitter is released to cause tissue damage. It is an allergic reaction. Most common allergic diseases such as bronchial asthma, atopic dermatitis, and allergic rhinitis occur with the involvement of this type I allergic reaction.
[0004]
In contrast, type IV allergic reactions do not involve antibodies, and T cells and macrophages are the main working cells. When a living body is sensitized with an antigen, sensitized cells produce various cytokines, resulting in local wetting of macrophages and T cells. Since this reaction takes 1-2 days to complete, it is also called a delayed reaction. Tuberculin reaction, tuberculosis lesion, rejection after organ transplantation, contact dermatitis such as cosmetic rash, etc. are said to be caused by type IV allergic reaction.
[0005]
Many components having an action of suppressing type I allergic reaction have been found so far, but there have been few known ingredients having an action of suppressing type IV allergic reaction.
[0006]
On the other hand, it has recently been clarified that the koji leaf extract has an antiallergic action, particularly an inhibitory action against type I allergy, and astragalin or the like has been reported as an active ingredient thereof.
[0007]
[Problems to be solved by the invention]
The main object of the present invention is to provide a composition for preventing or treating inflammation associated with type IV allergic reaction, obtained from an extract of persimmon leaf ethanol, and an external preparation for skin comprising the composition. And
[0008]
[Means for Solving the Problems]
As a result of diligent research on components having an action of suppressing type IV allergic reaction, the present inventor has determined that pentacyclic triterpenic acid having a specific structure, particularly barbinervic acid (Barbinervic acid) ) Has the effect of significantly suppressing type IV allergic reaction, and the present invention has been completed.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be specifically described.
[0010]
Composition for preventing or treating inflammation involving type IV allergic reaction The composition for preventing or treating inflammation involving type IV allergic reaction according to the present invention has the following formula 1:
[0011]
[Chemical 3]
[0012]
The pentacyclic triterpenic acid represented by these is contained.
[0013]
The pentacyclic triterpenic acid may be contained in the form of a salt or a derivative thereof.
[0014]
Examples of the salt include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, aluminum salt, and organic amine salt.
[0015]
Examples of the derivative include an acetyl derivative in which a hydroxyl group is acetylated.
[0016]
The pentacyclic triterpenic acid represented by Formula 1 is contained in, for example, an extract obtained by extracting bamboo leaves, leopard, kuroganemochi, etc. with ethanol or a water / ethanol solvent.
[0017]
Examples of the pentacyclic triterpenic acid represented by Formula 1 include barbinervic acid, rotundic acid, and rotungenic acid.
[0018]
In particular,
Formula 2:
[0019]
[Formula 4]
[0020]
Barbinerbic acid, which is a compound represented by the formula (2), is preferable in that it exhibits a more excellent type IV allergic reaction inhibitory action.
[0021]
The pentacyclic triterpenic acid represented by the formula 1 or the formula 2 can be obtained, for example, by extracting cocoon leaves with ethanol or a water / ethanol solvent as follows.
[0022]
The koji leaf may be used for extraction as it is after being collected, but it is usually subjected to extraction after pretreatment according to a commonly used method. For example, it can be collected, washed with water, cut into a suitable size (usually about 3 mm width), steamed with steam and dried. You may cut | disconnect immediately after extract | collecting and dry it according to a conventional method. Or like normal green tea, what passed through each process, such as steamed leaves, twisting, and roasting, may be used. Moreover, you may use the residue after extracting a bamboo leaf with water. As the extraction solvent, ethanol or water / ethanol solvent is used. In particular, the use of a solvent in which the mixing ratio of ethanol in a water / ethanol solvent is 50% by weight or more and less than 100% by weight, and more preferably 70% by weight or more and 90% by weight or less is a more excellent IV allergic reaction inhibitory action It is preferable at the point from which the extract which has this is obtained.
[0023]
Although extraction temperature is not specifically limited, About 30 degreeC-a boiling point are preferable, and about 60-80 degreeC is more preferable.
[0024]
The extraction time can be appropriately set according to the ethanol content and the extraction temperature, but is usually about 20 minutes to 24 hours, preferably about 25 minutes to 4 hours, and preferably about 30 minutes to 2 hours. It is more preferable that
[0025]
The ratio of the koji leaf to the extraction solvent is not particularly limited and can be set as appropriate. However, the solvent is preferably about 3 to 30 parts by weight, more preferably about 5 to 20 parts by weight with respect to 1 part by weight of the koji leaf. About 5 to 10 parts by weight is more preferable.
[0026]
The extract (extract) obtained by the extraction method as described above can be separated from the koji leaf according to conventional methods such as filtration and centrifugation. The extract may be used as it is or after being concentrated. Concentration can be performed according to a conventional method, for example, at about 30 to 50 ° C. under reduced pressure of about 30 to 40 mmHg. Further, after concentration, the extract powder can be obtained by commonly used means such as freeze drying, spray drying and granulation drying.
[0027]
Further, the extract and extract powder are represented by Formula 1 or Formula 2 by conventional separation and fractionation techniques such as liquid-liquid distribution using various solvents, recrystallization, gel filtration, chromatography, and the like. Moreover, it can be set as the refined product extracted from the bamboo leaf ethanol which increased content of pentacyclic triterpenic acid. Furthermore, barbinerbic acid can also be isolated by repeating the above operation.
[0028]
Although the method of obtaining the composition of this invention is not specifically limited as long as the pentacyclic triterpenic acid represented by Formula 1 or Formula 2 is contained, (i) A bamboo leaf is ethanol or water / ethanol solvent. What is obtained through the step of extracting, and (ii) the step of purifying the extract and increasing the content of the pentacyclic triterpenic acid represented by Formula 1 or Formula 2 is superior in suppressing type IV allergic reaction It is preferable at the point from which the composition which has an effect | action is obtained.
[0029]
In the step (ii), the degree of increasing the content of the pentacyclic triterpenic acid is about 10% by weight or more, preferably 20% by weight or more, more preferably 40% by weight or more with respect to the total amount of the extract. is there. Moreover, it is carried out until it becomes 100% by weight, and the pentacyclic triterpenic acid may be isolated and used.
[0030]
External preparation for skin The external preparation for skin of the present invention is obtained by blending the above-described composition, for example, by mixing the composition with an appropriate carrier.
[0031]
The content of the pentacyclic triterpenic acid represented by Formula 1 or Formula 2 in the external preparation for skin is not particularly limited as long as the desired effect is obtained, but is preferably about 0.001 to 5% by weight, particularly Preferably, it is about 0.01 to 3 weight%.
[0032]
Examples of the carrier include binders, disintegrants, surfactants, absorption accelerators, humectants, adsorbents, lubricants, fillers, extenders, diluents and excipients such as wetting agents, and the like. It is done.
[0033]
The form of the external preparation for skin can be appropriately selected according to the purpose, and representative examples thereof include powder preparations such as powders, solutions, suspensions, emulsions, lotions, aerosols, ointments and the like. Examples include creamy preparations, cataplasms and the like.
[0034]
Since the external preparation for skin of the present invention comprises the above-mentioned composition, it has an inhibitory action on inflammation associated with type IV allergic reaction, specifically, an anti-inflammatory or rough skin improving action. Examples of inflammation involving type IV allergic reaction include atopic dermatitis, allergic contact dermatitis, urticaria and the like.
[0035]
Because of these effects, the external preparation for skin of the present invention can be suitably used for the prevention or treatment of inflammation involving type IV allergic reaction, specifically for anti-inflammation or rough skin improvement.
[0036]
【Example】
The present invention will be described with reference to examples and formulation examples, but the present invention is not limited thereto. Unless otherwise specified, “%” represents “% by weight”.
[0037]
Measurement Method for Examining Inhibitory Action on Type IV Allergic Reaction A 7-week-old Balb / c female mouse was purchased from Japan SLC. Breeding conditions are room temperature 23 ° C. ± 3 ° C., humidity 55 ± 15%, light / dark cycle 12 hours (light period 7:00 to 19:00), and solid feed (Lab MR Stock, Nippon Agricultural Industrial Co., Ltd.) The drinking water was taken freely from the tap water. After preliminary breeding, the animals were divided into 3 animals / group and sensitized by applying 25 μl of a 1.0% acetone / olive oil (4: 1) solution of oxazolone to the abdomen of the shaved mice. After 5 days, an acetone / olive oil (4: 1) solution of 0.25% oxazolone was applied to both ears of 25 μl mice to induce an allergic reaction. An evaluation sample was applied 30 μl only once 1 hour before induction. In order to determine the intensity of the allergic reaction, the thickness of the auricle 24 hours after the onset and the thickness of the auricle before the onset were measured with a digital dial gauge DG-205 (Ozaki Mfg. Co., Ltd.). The inhibition rate of the ear edema of the test substance was determined. The evaluation sample was adjusted to 5% with ethanol and used, but in the experiment of FIG. 2, it was adjusted to 0.5% with ethanol and in the experiment of FIG. The measured value was expressed as an average value and standard error. Statistical analysis between each group used a t-test, and a risk rate of 5% or less was considered significant.
[0038]
Auricular edema rate (%) =
[Thickness of the auricle after evoking−thickness of the auricle before evoking / thickness of the auricle before evoking] × 100
Auricular edema suppression rate of test substance (%) =
[Average edema rate of test substance applied group−average edema rate of solvent applied group / average edema rate of test substance applied group] × 100
Example 1
6 L of ethanol was added to 300 g of dried bamboo leaves, and extraction by heating reflux was performed twice to obtain an extract, and then this extract was concentrated to dryness to obtain 29.2 g of bamboo leaf ethanol extract. Using a silica gel column (Chromatolex-ODS) for 1 g of kashiwa ethanol extract, changing the ratio of methanol and distilled water from 20% to 100%, fractionating, concentrating to dryness, fraction 1 (20% ethanol) 248 mg, fraction 2 (40% ethanol elution part) 184 mg, fraction 3 (60% ethanol elution part) 87 mg, fraction 4 (80% ethanol elution part) 153 mg, and fraction 5 (elution part) 263 mg of 100% ethanol elution part) was obtained. Each fraction was evaluated for its type IV allergic reaction inhibitory effect by the method described above. The increase in the thickness of the auricle in the untreated group (solvent application group) was 0.168 ± 0.0302 mm, whereas the increase in the thickness of the auricle in the kashiwa ethanol extract application group was 0.0938 ± 0.0251 mm. Yes, the suppression rate of auricular edema in the ethanol extract application group was 43.1%. Amber ethanol extract significantly suppressed ear edema when applied before induction of allergic reaction. Of each fraction, the increase in auricle thickness in the fraction fraction 4 application group was 0.0383 ± 0.0137 mm, and the suppression rate of auricular edema was 76.6%. Fraction 4 showed the highest inhibitory effect (see FIG. 1).
[0039]
Example 2
Fraction 4 in Example 1 was isolated and purified by silica gel column treatment and recrystallization to obtain a compound considered as an active ingredient. This compound was derived into a methyl ester form according to a known method, and the structure of the compound was determined from the physical property values of ¹ H-NMR. As shown in Table 1, the value of the pentacyclic triterpenic acid barbinerbic acid contained in Ryubu (Clethra Barbinervis Sieb.et Zucc.) (Chem. Pharm. Bull., 25 (5), 1977, p. 981-p .985), the active ingredient having an inhibitory effect on type IV allergic reaction was identified as barbinerbic acid.
[0040]
[Table 1]
[0041]
Example 3
Evaluation on the inhibitory action of type IV allergic reaction of barbinerbic acid and structurally similar compounds was performed by the above-described measurement method. The increase in pinna thickness in the barbinerbic acid application group was 0.0641 ± 0.017 mm, which significantly suppressed pinna edema compared to the control. On the other hand, a representative compound having a similar skeleton suppressed auricular edema, but the effect was low (see FIG. 2). Moreover, the inhibitory effect of auricular edema was also low in the glycyrrhizic acid application group (see FIG. 3).
[0042]
Reference example 1
Evaluation on the IV type allergic reaction inhibitory effect of the kashiwa hot water extract (the koji leaf extracted with hot water) and the kashiwa ethanol extract was performed by the above-described measurement method. The acupuncture ethanol extract application group significantly suppressed auricular edema. On the other hand, the Kashiwa hot water extract did not suppress auricular edema very much (refer FIG. 4).
[0043]
Formulation examples 1 to 5 of the present invention are shown below. These can be produced according to conventional methods.
[0044]
[0045]
【The invention's effect】
The composition of the present invention containing a pentacyclic triterpenic acid represented by Formula 1 or Formula 2 has an action of significantly suppressing type IV allergic reaction, and prevention or treatment of inflammation involving type IV allergic reaction It can be effectively used as a composition for use.
[0046]
Further, the external preparation for skin of the present invention comprising the above composition has an inhibitory action on type IV allergic reaction, and further has an anti-inflammatory or rough skin improving action, and prevents or promotes inflammation involving type IV allergic reaction. It can be usefully used for treatment and further for anti-inflammatory or rough skin improvement.
[Brief description of the drawings]
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a drawing showing the measurement results of the IV type allergic reaction inhibitory effect of bamboo leaf ethanol extract and fractions.
[Fig. 2] Fig. 2 is a drawing showing the results of measurement of the inhibitory action of type IV allergic reaction of barbinervic acid and structurally similar compounds.
FIG. 3 is a graph showing the results of comparing the IV type allergic reaction inhibitory action of barbinerbic acid and glycyrrhizic acid with control.
FIG. 4 is a drawing showing the results of comparison of the IV type allergic reaction inhibitory action of bamboo leaf hot water extract and bamboo leaf ethanol extract.

Claims (6)

Formula 1:
It is a pentacyclic triterpenic acid represented by the following, characterized in that it contains pentacyclic triterpenic acid selected from the group consisting of barbinerbic acid, rotundic acid and rotungenic acid, a salt or a derivative thereof as an active ingredient , A composition for preventing or treating inflammation involving type IV allergic reaction ,
i) Extracting bamboo leaves with ethanol or water / ethanol solvent, and (ii) Steps of setting the content of pentacyclic triterpenic acid, its salt or derivative to 10% by weight or more based on the total amount of the extract The resulting composition . Pentacyclic triterpenic acid, salt or derivative thereof,
Formula 2:
The composition of Claim 1 which is the barbinervic acid represented by these, its salt, or a derivative . Mixing ratio of ethanol in water / ethanol solvent is less than 50 wt% 100 wt%, the composition according to claim 1 or 2. The skin external preparation formed by mix | blending the composition in any one of Claims 1 thru | or 3 . The skin external preparation of Claim 4 which has an anti-inflammatory or rough skin improvement effect. The skin external preparation according to claim 4, wherein the content of pentacyclic triterpenic acid, a salt or a derivative thereof in the external skin preparation is 0.001 to 5% by weight.