JP2006096672A - Antiallergic agent and antipruritic agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an antiallergic agent and an antipruritic agent having little adverse actions and obtain an antiallergic agent having both of antiallergic action and antipruritic action. <P>SOLUTION: The antiallergic agent and antipruritic agent contain an extract of Garcinia subelliptica as an active component. The excellent antiallergic agent having both of antiallergic action and antipruritic action contains Garcinia subelliptica or its extract as an active component. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an antiallergic agent and an antipruritic agent containing fukugi as an active ingredient, and an excellent antiallergic agent having both an antiallergic action and an antipruritic action containing fukugi as an active ingredient.

Conventionally, as antiallergic agents and anti-inflammatory agents, glucocorticoid-type corticosteroid agents, non-steroidal anti-inflammatory agents such as indomethacin, and the like have been used. However, steroid hormone agents have anti-allergic and anti-inflammatory effects, but have a problem of strong side effects. Indomethacin and the like have only anti-inflammatory effects, and thus are not useful as anti-allergic agents.
Therefore, an orally administrable drug having few side effects and having an excellent antiallergic action has been desired.

  Furthermore, it often involves itch when suffering from skin allergic diseases such as atopic dermatitis and contact dermatitis, and the scratching action often exacerbates the original disease. Skin pruritus is a troublesome problem for the living body and originates from various causes, but the cause of its onset is not necessarily clarified. As a symptomatic treatment, various antipruritic agents are locally used as external preparations. There are many types of anti-pruritic agents, some of which are effective only against pruritus, but rather affect allergies, reticuloendothelial system, plant nerves or endocrine system, and more often reduce itching as a result of anti-inflammatory effect. .

However, there is a report that an anti-inflammatory agent whose mechanism is inhibition of prostaglandin synthesis, such as indomethacin, rather hates itching. Anti-pruritic agents that are widely used every day include antihistamines, but other anti-plasmin agents, SH preparations (reduced glutathione, sodium thiosulfate, etc.), vitamin agents (group B ₂ ), hormone agents, plant nerve blockers, Examples include liver function enhancers. However, these treatments have not been sufficiently satisfactory as antipruritic agents.

  The onset of atopic dermatitis is an allergic reaction, and steroids are widely used as therapeutic agents. Once it develops, the rash with itching will scratch the area, but the action of scratching causes an infection (mainly a skin infection caused by Staphylococcus aureus), making it increasingly difficult to heal. .

Therefore, an orally administrable drug having few side effects and having antiallergic action and antipruritic action has been desired.
In this situation, the present inventors paid attention to Fukugi. Currently, the following studies have been conducted on Fukugi.

1. Plant taxonomic and ethnopharmacological studies of Fukugi Fukugi is Garcinia subelliptica of the Hypericaceae family. This plant is distributed in the subtropical region and is planted mainly for the purpose of wind protection. It seems that it was not used much as a folk medicine.

2. Fukugi Chemical Research 1,2,5-trihydroxyxanthone, 1,2-dihydroxy-5,6-dimethoxyxanthone, 1,8-dihydroxy-6-methoxyxanthone, garcinoxanthone D, 1,4,5-trihydroxyxanthone Garcinielliptone F and garcinielliptone oxide have been isolated from seeds.

3. Pharmacological studies of Fukugi As pharmacological studies, anti-inflammatory action, antibacterial action, choline acetyltransferase activation action and peroxide scavenging action have been reported so far. Regarding anti-inflammatory activity, garcinielliptone F extracted from seeds is effective in suppressing the production of β-glucuronidase and lysozyme from rat neutrophils, and garcinielliptone I extracted from seeds is also reported to be effective in suppressing NO production. ing. In addition, garsubellin A and garcinielliptin oxide have been reported to inhibit β-glucuronidase and histamine production on mast cells. Garcinielliptin oxide has the effect of inhibiting the formation of neutrophil peroxides. As for the antibacterial action, it has been reported that xanthochymol extracted from Fukugi peel has a strong antibacterial action equivalent to vancomycin against MRSA.
Garsubellin A extracted from wood increases choline acetyltransferase activity in cultured septal nerves of rats, and xanthones such as garciniaxanthone have been confirmed to act as radical scavengers.

  Fukugi is used as an active oxygen scavenger (for example, Patent Document 1), an ultraviolet absorber and a skin external preparation containing the same (for example, Patent Document 2), and an aldose reductase inhibitor (for example, Patent Document 3). Have been.

JP-A-9-255571 JP-A-9-87155 JP-A-6-172340

Under such circumstances, the present inventors have conducted extensive research and found that Fukugi or its extract has excellent anti-allergic action, anti-pruritic action, and both of these two actions, with excellent side effects. It has been found that it has an allergic action, and the present invention has been completed.
Therefore, an object of the present invention is to provide an antiallergic agent, an antipruritic agent having almost no side effects, and an antiallergic agent having both antiallergic and antipruritic effects.

  In order to achieve the above object, the antiallergic agent and antipruritic agent of the present invention comprises fukugi or an extract thereof as an active ingredient.

  The anti-allergic agent of the present invention is an anti-allergic agent having both anti-allergic action and anti-pruritic action comprising fukugi or an extract thereof as an active ingredient.

  Garcinielliptone F isolated from Fukugi suppresses β-glucuronidase and lysozyme production, garcinielliptone I suppresses NO production, garsubellin A and garcinielliptin oxide suppresses β-glucuronidase and histamine production in mast cells, and favors garcinielliptin oxide Radical scavenger action has been reported to inhibit the formation of sphere peroxides, and xanthones including garciniaxanthone, but anti-allergic action and anti-pruritic action are not known.

  Thus, the present invention has excellent antiallergic action and antipruritic action, and also has high safety, so that allergic rhinitis, allergic conjunctivitis, atopic dermatitis, eczema dermatitis, urticaria, acute or chronic It is useful as a therapeutic agent for conjunctivitis, bronchitis, itching and rash.

  Furthermore, the extract of Fukugi can be administered as it is or in various dosage forms. There are no particular restrictions on the dosage form of the antiallergic agent, antibacterial agent and antipruritic agent of the present invention, and oral agents such as tablets, capsules, granules, powders, liquids, injections, external preparations, suppositories, inhalation It can be administered by any of parenteral agents such as an agent, a nasal drop, an eye drop, an ointment, a patch.

Hereinafter, production examples and experimental examples of the present invention will be described.
(Production example)
In the present invention, all parts constituting the plant or a part of leaves, stems, roots, flowers, fruits and the like can be used as they are, and these can be dried and pulverized to be used as a powder.

  In the present invention, the method for obtaining the extract is, for example, a method for obtaining an extract by extracting the fruits, leaves, roots, rhizomes, stems, flowers, etc. of this plant using water and / or a hydrophilic organic solvent. There is. Furthermore, a method of obtaining a powder from such an extract by freeze-drying, spray-drying, distillation under reduced pressure, or the like.

  As a hydrophilic organic solvent, C1-C4 lower alcohols, such as methanol and ethanol, acetone, etc. are mentioned, for example. Ethanol is particularly preferable. These solvents may be used alone or in combination of two or more. Moreover, you may mix and use water and these hydrophilic organic solvents. A preferable extraction solvent includes hydrous alcohol, and hydrous ethanol is particularly preferred. The amount of these extraction solvents to be used is not particularly limited, and for example, a cold immersion method, a digestion method, a percolation method and the like used for producing an extract agent, a tincture agent and the like can be applied.

  Furthermore, a drug can be produced by adding a pharmaceutically acceptable additive (for example, an excipient, a surfactant, etc.) to the powder or the extract as necessary.

(Experimental example 1)
Effect of Fukugi on the triphasic skin reaction Experimental method 1. As experimental animals, ICR female mice (body weight 24-26 g) were used.
2. DNFB-induced atopic dermatitis test
1) Preparation of dinitrophenylated ovalbumin (DNP-OVA): Dissolve 2 g of ovalbumin (EWA) and K ₂ CO ₃ in 100 ml of water, and add 2 g of dinitrobezenesulfonic acid sodium salt to this solution. Stir with a stirrer at 37 ° C. for 24 hours in the dark. The obtained reaction solution was dialyzed against water for 2 days, and the internal solution was lyophilized.

2) DNFB-induced triphasic skin reaction test: ICR female mice were actively sensitized by intraperitoneal administration of 0.2 ml of physiological saline containing 1 mg of aluminum hydroxide gel and 10 μg of DNP-OVA. One week later, 10 μl of 0.1% DNFB was applied to the front and back of both ears. The next day, sensitization was performed again, and a reaction was elicited one week later. The thickness of the auricle was measured using a dial thickness gauge before the reaction was initiated and after 1, 24 hours and 8 days after the reaction, and was calculated as the auricular edema rate. The pruritus behavior was observed for 1 hour after the reaction was triggered. The subject was orally administered every day from 1 hour before the reaction was initiated and from 2 days to 8 days after the initiation.

3. Preparation of specimen Fukugi collected at Amami Oshima was dried, crushed, extracted twice with hot 70% methanol for 2 hours for 2 hours, filtered, evaporated under reduced pressure, and freeze-dried The obtained fukugi extract was used as a subject.
Experimental results When mice were induced to induce DNFB-induced triphasic skin reaction, they peaked at 1 hour (immediate phase; IPR), 24 hours (late phase; LPR), and 8 days (very late phase; vLPR). The shown triphasic ear edema was observed. In addition, scratching (itching) was observed in IPR, and the number of times was observed.

  As shown in Tables 1 and 2, Fukugi extract was found to have the effect of significantly suppressing the scratching behavior in IPR and ear edema of IPR, LPR, and vLPR. The first phase (immediate phase, IPR) ear edema is caused by chemical mediators such as histamine released from mast cells, and the second phase (late phase, LPR) is triggered by various cytokines. It is said to be inflammation. In addition, vLPR is a reaction in which significant infiltration of eosinophils is observed, which is similar to a pathological image seen in individuals with atopic dermatitis.

(Experimental example 2)
Inhibition of Histamine Release from Compound 48 / 80-Induced Mast Cells The antipruritic action of Fukugi was examined using the histamine release inhibition test from mast cells, which is closely related to the induction of itching.

  Preparation of specimen: After cutting dry fukugi leaves or branches, extract twice with hot 70% methanol for 2 hours, filter with filter paper, concentrate under reduced pressure and freeze-dry. The obtained extract was used for the experiment.

Experimental method 1. Preparation of rat peritoneal mast cells; separation of peritoneal mast cells from Wistar male rats was performed according to the method of Uvnas et al. That is, 10 ml of Hank's solution (containing 10 U / ml of heparin) was injected into the abdominal cavity immediately after decapitation of rats. Gently massage the abdomen for about 90 seconds, then collect the intraperitoneal fluid, layer it gently on 40% ficoll solution, leave it at room temperature for 30 minutes, and centrifuge at 5 ° C, 1200 rpm for 10 minutes. The mast cells were collected. The mast cells were suspended in phosphate buffer (PBS, pH 7.0), washed by centrifugation four times, and resuspended in PBS (2.9 × 10 ⁶ cells / ml). The mast cell content in the suspension was 85 to 90%, and the survival rate was confirmed to be 90% or more by the toluidine blue (0.1%, 50% ethanol solution) staining method.

2. Measurement of Histamine release from mast cells: After preincubation of 1.8 ml of mast cell suspension at 37 ° C for 10 minutes, add 0.1 ml of test solution (PBS dissolved), incubate for 5 minutes, and then compound 48/80 (final concentration) 10 μg / ml) 0.1 ml was added and incubated for 10 minutes. The reaction was stopped by cooling with ice, centrifuged at 5 ° C., 1200 rpm for 5 minutes, and the amount of histamine in the supernatant was measured according to the method of Shore.

That is, 1.4 ml of H ₂ O, 0.4 ml of 1N NaOH solution and 0.1 ml of 1% o-phtaldialdehyde-methanol solution are added to 0.7 ml of the supernatant and left for 4 minutes, and then the reaction is stopped with 0.2 ml of 3N HCl solution. Ten minutes after the completion of the reaction, centrifugation was performed at 5 ° C. and 3000 rpm for 5 minutes to obtain a supernatant and a sediment. The supernatant fluorescence was measured at an excitation wavelength of 360 nm and a fluorescence wavelength of 450 nm, and the amount of histamine in the supernatant was determined from a histamine calibration curve with a known concentration. The amount of histamine remaining in the mast cells was measured by adding 2 ml of PBS to the sediment, releasing histamine from the mast cells by sonication and freeze-thawing, and the same method as described above. The histamine release rate (histamine release%) by Compound 48/80 was calculated by the following formula.

Histamine release% ＝ (histamine released with compound 48 / 80−spontaneously released histamine) / total histamine × 100
Inhibition% = (% histamine release without test substance−% histamine release with test substance) /% histamine release without test substance × 100

Experimental Results As shown in Table 3, when compound 48/80 was allowed to act on rat abdominal mast cells, histamine was released from the mast cells by 82.3 ± 0.4%. The positive control drug sodium cromoglycate (SCG) showed an inhibitory effect of 20.9% at a concentration of 500 μg / ml. Fukugi extract showed an inhibitory action of 48.5% at 200 μg / ml and 95.6% at 500 μg / ml.

Effect on compound 48 / 80-induced scratching behavior (anti-pruritic action)
The antipruritic action of Fukugi was examined using a compound 48 / 80-induced scratch model.

Experimental method Preparation of specimen extract; dried fukugi leaves were minced, extracted twice with hot 70% methanol for 2 hours, filtered through filter paper, concentrated under reduced pressure and freeze-dried . The obtained extract was used for the experiment.

Experimental animals: ICR male mice (body weight 26-28 g) were used.

Experimental method: A subject was orally administered to an ICR male mouse fasted for 18 hours, and 1 hour later, 0.1 ml of 48/80 physiological saline was subcutaneously injected to the back of the mouse to induce scratching behavior. The action of scratching the injection site with the hind limb was regarded as an itching action, and the number of scratching actions was determined for 10 minutes immediately after the injection. In addition, diphenhydramine was used as a positive control drug.

Results: As shown in Table 4, when 48/80 physiological saline was subcutaneously administered to the back of the mice, 72.9 ± 7.8 scratching behaviors were observed in the control group. Diphenhydramine, a positive control drug, significantly inhibited scratching by compound 48/80 at a dose of 50 mg / kg. Fukugi extract significantly suppressed this scratching behavior at doses of 200 and 500 mg / kg.

Claims (3)

  An antiallergic agent comprising fukugi or an extract thereof as an active ingredient.   An anti-pruritic agent comprising fukugi leaf or an extract thereof as an active ingredient. An antiallergic agent having both an antiallergic action and an antipruritic action comprising fukugi or an extract thereof as an active ingredient.