JPH0710764A - Antiallergic agent - Google Patents

Abstract

PURPOSE:To obtain an antiallergic and an antiinflammatory agents, containing an extract of Corydalis bulbosa C. ternata as an active ingredient and orally administrable with hardly any side effects. CONSTITUTION:This antiallergic agent contains an extract prepared by extracting a tuber, etc., of Corydalis bulbosa C. ternata which is a perennial herb naturally growing in the eastern part of China with one or two or more polar solvents, e.g. an alcohol such as methanol or ethanol, water, ether or chloroform in combination as an active ingredient. The extract of the Corydalis bulbosa C. ternata is directly administered or prepared into a pharmaceutical preparation such as an oral agent, e.g. a tablet, a capsule, a granule, a powder or a solution or a parenteral agent, e.g. an parenteral injection, an external preparation, a suppository, an inhalant, a nasal drop or an eye drop and administered. The agent is useful for treating allergic rhinitis, allergic conjunctivitis, eczematous dermatitis, urticaria, conjunctivitis, bronchial asthma, chronic articular rheumatism, etc.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to an antiallergic agent and an antiinflammatory agent which have excellent antiallergic and antiinflammatory effects.

[0002]

2. Description of the Related Art Conventionally, the problems to be solved by the invention
As the anti-allergic agent and anti-inflammatory agent, glucocorticoid-type adrenocortical hormone agents, non-steroidal anti-inflammatory agents such as indomethacin, ibuprofen and the like are used. However, steroid hormone agents have an antiallergic action and an anti-inflammatory action, but have a problem that side effects are strong, and indomethacin, ibuprofen and the like are not useful as antiallergic agents because they have only an antiinflammatory action.

Therefore, an orally administrable drug having few side effects and excellent antiallergic and antiinflammatory effects has been desired.

On the other hand, it has been known that cholangae has analgesic and antispasmodic effects, and has been used for a long time in combination with Chinese medicine gastrointestinal medicine and the like.

[0005]

Under such circumstances, the inventors of the present invention have conducted diligent research and, as a result, have found that the use of a polar solvent extract of Chorella chinensis has few side effects and excellent anti-allergic and anti-inflammatory effects. The present invention has been completed by finding that an orally administrable anti-allergic agent and anti-inflammatory agent can be obtained.

[0006] That is, the present invention provides an anti-allergic agent and an anti-inflammatory agent containing a polar solvent extract of chordae cord as an active ingredient.

[0007] Coryda used in the present invention
lis turtschaninovii forma
yanhusuo) is a perennial that grows naturally in eastern China, and its tubers and the like can be used. As a polar solvent for extracting the active ingredient from these cords,
Examples thereof include alcohols such as methanol and ethanol, water, ether, chloroform and the like, and one or more of these are used in combination, and an extract is obtained by single or multiple extractions by a conventional method. be able to.

Examples of the polar solvent extract of chordae cord obtained in this manner include the following. (1) Alcohol extract of Encho (2) Water extract of alcohol extract (3) Ether extract of (2) (This extract contains a tertiary alkaloid fraction) (4) (3 ) Water extract (this extract contains a tertiary phenolic alkaloid fraction) (5) An extract after extracting (4) from (3) Phenolic alkaloid fraction is included) (6) Chloroform extract of (2)
Grade alkaloid fraction is included)

Further, alkaloid components can be further isolated from the alkaloid fractions (4) to (6), and these components can be used alone or in combination. For example, by recrystallizing the extract of (4) using chloroform, ethanol and ether, l-
Tetrahydrocorumbamine can be isolated. In addition, from the extract of (5), by performing column chromatography and a recrystallization method in combination, d-corridalin, d-glaucine, protopine, 1-tetrahydrocoptisine and dl-tetrahydropalmatine were respectively obtained. It can be isolated. Furthermore, dehydrocoridaline can be isolated from the extract of (6) by column chromatography and methanol recrystallization.

[0010] The extract of chordae cord obtained in this manner has excellent antiallergic and antiinflammatory effects as shown in Examples below, and since it is highly safe, allergic rhinitis, allergic conjunctivitis, It is useful as a therapeutic agent for eczema dermatitis, urticaria, acute or chronic conjunctivitis, bronchial asthma, rheumatoid arthritis and the like. Regarding the safety, the results of examination of the presence or absence of antigenicity in the extract of Chordae chinensis by active systemic anaphylaxis (ASA) using guinea pigs and passive cutaneous anaphylaxis (PCA) using guinea pigs with sensitized guinea pig isolated serum In addition, it was confirmed that the Enchocord extract was not antigenic.

[0011] Such an extract of chordae cord can be administered as it is or in various dosage forms. The administration form of the antiallergic agent and the antiinflammatory agent of the present invention is not particularly limited, and oral agents such as tablets, capsules, granules, fine granules, powders and liquids, injections, external preparations, suppositories, It can be administered by any of parenteral agents such as inhalants, nasal drops, and eye drops.

Examples of solid oral carriers include starch,
Lactose, sucrose, mannitol, carboxymethyl cellulose, corn starch, inorganic salts and the like can be mentioned, and if necessary, further, a binder, a disintegrating agent, a surfactant, a lubricant, a fluidity promoter, a flavoring agent, a coloring agent, a flavoring agent. Etc. can be used. More specifically, starch as a binder, dextrin, gum arabic powder, gelatin, hydroxypropyl starch, methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose,
Crystalline cellulose, ethyl cellulose, polyvinylpyrrolidone, macrogol, etc .; starch as a disintegrant, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose,
Carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, etc .; sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80, etc. as surfactants; talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, as lubricants, Aluminum stearate, polyethylene glycol, etc .; Light anhydrous silicic acid as a fluidity promoter, dried aluminum hydroxide gel,
Synthetic aluminum silicate, magnesium silicate, etc. can be used.

Examples of oral liquid preparations include suspensions, emulsions, syrups, elixirs, and other dosage forms. These various dosage forms include flavoring agents, flavoring agents, and coloring agents. Can also be blended.

Further, as a parenteral liquid carrier, distilled water for injection, physiological saline, glucose aqueous solution, vegetable oil for injection,
Sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. are used,
Further, if necessary, a bactericidal agent, a preservative, a stabilizer, an isotonic agent, a soothing agent and the like can be added.

In the present invention, the dose of the active ingredient depends on the properties of the extract and the like, the route of administration, and other factors, but in the case of the alcohol extract of Chrysanthemum japonicum, about 1 to 10 mg / kg per day; alkaloids In the case of a fraction, it is preferably about 0.2 to 2 mg / kg per day; in the case of an alkaloid component, it is preferably about 0.1 to 1 mg / kg per day. In addition, these daily doses can be divided into 2-3 times for administration if desired.

[0016]

INDUSTRIAL APPLICABILITY The antiallergic and antiinflammatory agents of the present invention have few side effects and are excellent in antiallergic action and antiinflammatory action.

[0017]

EXAMPLES Next, the present invention will be further described with reference to examples.

Example 1 Anti-inflammatory Action of Cordyceps sinensis: (1) Preparation of Methanol Extract of Cordyceps sinensis L. Corydalis turtscha from China
The tuber of ninovii forma yanhusuo (manufactured by Nippon Powder Chemicals Co., Ltd.) was cut into small pieces, extracted twice with 10 times the amount of methanol for 2 hours, and filtered while hot. After distilling off methanol under reduced pressure, a methanol-extracted extract was obtained.
The obtained methanol extract of Enchoronus sinensis (hereinafter referred to as CT-e
The yield of (abbreviated as xt) was 2.95%. In the following examples, the obtained CT-ext was dissolved in an appropriate concentration as needed.

(2) Effect on Carrageenin Edema Male Wistar rats were injected with 0.1 ml of 1% λ-carrageenan physiological saline solution / rat subcutaneously in the footpad of the right hind leg,
The edema generated was measured after 15, 30, 45 minutes, and 2 and 3 hours, and the edema rate was calculated. A subject (suspended in 0.5% sodium carboxymethylcellulose (hereinafter, abbreviated as CMC.Na)) was orally administered once 1 hour before injection of a carrageenin physiological saline solution (Table 1).

[0020]

[Table 1]

As is clear from the results shown in Table 1, CT-e
In the xt500 mg / kg administration group, there was a tendency to suppress early edema formation, and an effect of significantly suppressing edema a few hours after carrageenin administration was observed.

(3) Effect on compound 48/80 edema 0.01% compound 4 in Wistar male rats
An edema rate was calculated by injecting 0.1 ml of an 8/80 physiological saline solution / rat subcutaneously into the footpad of the right hind leg and measuring the edema generated 15 and 30 minutes later. Subject (0.5% CMC / Na
Suspension) was orally administered once 1 hour before administration of compound 48/80 (Table 2).

[0023]

[Table 2]

As is clear from the results of Table 2, when compound 48/80 was subcutaneously injected into the right hind footpad of rats, marked edema was observed 15 and 30 minutes later, but CT
In the -ext administration group, an effect of significantly suppressing compound 48 / 80-induced acute edema was observed.

(4) Effect on Capillary Permeability 1 hour after oral administration of the subject (suspended in 0.5% CMC / Na) to male ddY mice, 10 ml / kg of 4% Pontamine Sky Blue physiological saline solution Was intravenously injected, and 15 minutes later, an intraperitoneal injection of 10 ml / kg of a 1% acetic acid physiological saline solution was performed. Furthermore, after 20 minutes, the mice were killed by decapitation, and the dye exuded into the abdominal cavity was washed and collected with physiological saline to make a total volume of 10 ml, and then 0.1 ml of 1N sodium hydroxide solution was added.
Absorbance was measured at 90 nm (Table 3).

[0026]

[Table 3]

As is clear from the results shown in Table 3, CT-e
In the xt-administered group, an action of significantly suppressing acetic acid-induced increase in capillary permeability was observed.

(5) Effect of histamine on guinea pig isolated ileal contraction Hartley fasted for 24 hours according to the Magnus method
Ileum about 1.5 after bleeding from a male guinea pig
The cm was suspended in the organ bath of the Magnus apparatus. Tyrode solution (Na
Cl 8 g, KCl 0.2 g, CaCl ₂ 0.2
g, MgCl ₂ 0.1 g was dissolved in water 1 L, and then NaH ₂ P was added.
O ₄ 0.05g, NaHCO ₃ 1g, glucose 1g
Was added and dissolved), and the temperature was kept at 37 ± 1 ° C., and air was constantly passed through. Histamine 10 ^-9 -10 ^-4 mm
The sample showed a concentration-dependent height of contraction at ol / ml. Then, histamine was injected 2 minutes after the CT-ext treatment as a subject, and their concentration-effect curves were determined. In addition,
The sample rest time between tests was set to 10 minutes (Fig. 1).

As is clear from the results shown in FIG. 1, histamine 10 ^-9 -10 ^-4 mmol / ml was found to have a concentration-dependent action to increase the contraction rate, and its ileal contractile action was CT-e.
Pretreatment with xt antagonized the contractile effects of histamine.

(6) Effect on Adjuvant Arthritis (i) Induction of Adjuvant Arthritis and Administration Method of Subject:
Dry tuberculosis killed cells (Mycobacterium but)
Yricum, manufactured by Difco) was ground in an agate mortar, and mineral oil (Bayol F) was added to prepare a 1% suspension. This was sterilized in an autoclave and used as an adjuvant. 0.05 ml of this adjuvant was intradermally injected into the right hind footpad and tail of a female SD rat about 3 cm from the ridge to induce adjuvant arthritis.
The subject (suspended in 0.5% CMC · Na) was orally administered once a day for 21 consecutive days immediately after the adjuvant injection. The body weight was measured daily. (Ii) Observation of arthritis symptoms: The arthritis symptoms were calculated from the edema rate calculated by measuring the volume of the right hind paw daily for 21 days (Table 4).

[0031]

[Table 4]

As shown in Table 4, when the adjuvant was injected, significant edema was observed in the right hind limb from the 1st day, and the edema was maximized after 5 days and slightly disappeared until 9 days, but after 11 days. The edema increased again. When the subject was orally administered once daily once daily immediately after the adjuvant injection, CT
-Ext 500 mg / kg showed a tendency to suppress edema of the right hind limb, and significantly suppressed edema of the right hind limb 3, 5, 9, 13, 15 days after the adjuvant injection. In addition, administration of 500 mg / kg of CT-ext for 21 days did not affect the body weight change of rats in which adjuvant arthritis was induced.

Example 2 Antiallergic effect of Chorella chinensis extract: (1) Effect on histamine release (i) Preparation of sensitized rat peritoneal mast cell suspension: Wist
Sensitization was carried out by intraperitoneally administering 0.5 ml of 2-fold diluted anti-ovalbumin (hereinafter abbreviated as EWA) rat serum to male ar rats. Uvnas & T after decapitation after 24 hours
hon's method (B. Uvnas and IL Th.
on, Exp. Cell. Res. , 18 , 512 (1
959)), and a mast cell suspension was prepared. That is, immediately after decapitation, 10 ml of Hank's solution (containing 10 μg / ml heparin) was intraperitoneally injected, the abdomen was gently massaged for about 90 seconds, and the intraperitoneal fluid was collected and gently overlaid with 2 ml of 40% Ficoll solution. Then, after leaving it at room temperature for 30 minutes, centrifuge at 5 ° C, 1,200 rpm for 10 minutes,
Mast cells on the Ficoll layer were collected. The mast cells were suspended in a phosphate buffer (hereinafter abbreviated as PBS) (pH 7.0), washed by centrifugation three times, and then again PBS.
The cells were suspended (the number of mast cells was 2.9 × 10 ⁶ cells / ml). It was confirmed that the content of mast cells in this suspension was 85-90%, and the survival rate was 90% or more by the toluidine blue (0.1%, 50% ethanol solution) staining method.

(Ii) Histamine release reaction test: 1.8 ml of the mast cell suspension obtained in (i) was preincubated at 37 ° C. for 10 minutes, and then the test solution (10% dimethyl sulfoxide (hereinafter abbreviated as DMSO) was used. ) / Dissolved in PBS], and incubated for 10 minutes.
A mixture of A and phosphatidyl-L-serine (final concentration: EWA 2 mg / ml, phosphatidyl-L-serine 1
0.1 μl (00 μg / ml) was added and incubated for 15 minutes. Stop the reaction by cooling with ice, 5 ℃, 1,200rp
The mixture was centrifuged for 5 minutes, and the amount of histamine in the supernatant was measured. That is, 0.7 ml of supernatant was added to 1.4 ml of water and 1N-
0.4 ml of NaOH, 1% o-phthaldialdehyde /
After adding 0.1 ml of methanol solution and leaving it for 4 minutes, 3N-
The reaction was stopped with 0.2 ml of HCl solution. 10 minutes after completion of the reaction, centrifugation was performed at 5 ° C., 3,000 rpm for 5 minutes,
A supernatant and a precipitate were obtained. The fluorescence of the supernatant has an excitation wavelength of 360 nm,
Measured at a fluorescence wavelength of 450 nm (Hitachi Fluor
esence Spectrophotometer
650-10S), and the amount of histamine in the supernatant was determined from the calibration curve of histamine of known concentration. Further, the amount of histamine remaining in the mast cells was determined by adding 2 ml of PBS to the precipitate, sonicating, and further releasing histamine from the mast cells by a freeze-thaw method, and measuring by the same method as above to determine the histamine release rate (Table. 5).

[0035]

[Table 5]

As is clear from the results in Table 5, CT-e
xt significantly suppressed histamine release from mast cells due to antigen-antibody reaction.

(2) Effect on picryl chloride-induced contact dermatitis (hereinafter abbreviated as PC-CD) (a) Effect on PC-CD (inflammatory process) ICR female mice (body weight: 30-32 g, shaved the day before) )
Sensitization was carried out by applying 0.1 ml of a 7% picryl chloride ethanol solution to the abdomen of the. Six days later, 0.02 ml of a 1% picryl chloride-olive oil solution was applied to one ear to induce contact dermatitis. The subject was orally administered from 2 days before the sensitization day for 7 days. The thickness of the ear 24 hours after the induction
nest gauge (Ozaki MFG. Co. L.
td. ), And the swelling rate was calculated (Table 6).

[0038]

[Table 6]

As is clear from Table 6, significant inhibition of swelling induced by PC-CD was observed in the CT-ext administration group.

(B) PC-CD (establishment process of induced swelling)
Effects on (a) were sensitized once, and after 24 hours after induction, mice showing sufficient swelling were selected, and sensitized again 3 days later.
Induction was repeated, and ear thicknesses before and 24 hours after induction were measured by dial thickness gauge (Oza).
ki MFG. Co. Ltd. ), And the swelling rate was calculated. Subjects received two oral doses immediately before and 16 hours after induction (Table 7).

[0041]

[Table 7]

As is clear from Table 7, significant suppression of swelling induced by PC-CD was observed in the CT-ext administration group.

(C) Therapeutic effect on PC-CD Twenty-four hours after the induction, the subject was orally administered once, and the swelling rate was measured five times at two-hour intervals from two hours after the administration. PC-CD
After 24 hours when the swelling became maximum, the subject was orally administered once (Table 8).

[0044]

[Table 8]

As is clear from Table 8, PC-CD was administered 6 to 10 hours after oral administration to the CT-ext administration group.
The effect of significantly abolishing the swelling induced by was observed.

(3) Effect on passive cutaneous anaphylaxis (hereinafter abbreviated as PCA) reaction in rats (i) Preparation of anti-EWA rat serum: 1 mg EWA in 20 mg aluminum hydroxide gel and pertussis diphtheria tetanus mixed vaccine 0.5 ml The mixture was mixed with and mixed into the foot pad of Wistar male rat in four divided doses. After 14 days, blood was collected from the carotid artery to obtain anti-EWA serum. (Ii) PCA reaction test: Wistar male rats were shaved on the back and diluted intradermally with anti-EWA serum 8-fold or 16-fold at a rate of 0.05 ml / mouse, 2 each.
A total of 4 points were injected. After 48 hours, 0.5 ml of a 1% Evans blue physiological saline solution containing 2 mg of EWA was injected from the tail vein, and 30 minutes later, the animals were killed by exsanguination, and the area of locus ceruleus produced by 8-fold diluted serum was measured. Further, this blue spot was cut out, dissolved in a 1N-KOH solution, extracted with a phosphoric acid-acetone mixed solution, and the pigment amount was determined from a calibration curve of Evans blue prepared in advance. The subject was orally administered 1 hour before PCA induction (Table 9).

[0047]

[Table 9]

As is clear from the results in Table 9, CT-e
In the xt administration group, significant suppression of PCA reaction was observed.

(4) Effect on IgE antibody production (i) IgE antibody production: 0.2 ml of a mixed solution of 1 mg EWA and 1.5 mg aluminum hydroxide gel was intraperitoneally administered to BALB / c female mice. Blood was collected from the fundus sinus 7 and 14 days after immunization, and the serum obtained by a conventional method was used for the measurement of IgE antibody titer. The subject was orally administered once daily every day from the day of immunization (suspended in 0.5% CMC / Na). (Ii) Measurement of IgE antibody titer: It was measured by a rat homologous PCA reaction for 48 hours. That is, the back of Wistar male rats was shaved, and 0.1 ml of each serum intradermally diluted with a doubling dilution system was injected.
Forty-eight hours later, 0.5 ml of a 1% Evans blue saline solution containing 2 mg of EWA was injected through the tail vein, and 30 minutes later, the animals were killed by exsanguination, the back was peeled off, and the diameter produced by the serum was 5
Blue spots having a size of mm or more were determined as positive and determined as IgE antibody titers (Table 10).

[0050]

[Table 10]

As shown in the results of Table 10, CT-ext50
In the 0 mg / kg administration group, significant suppression of IgE antibody production was observed 1 and 2 weeks after the antigen sensitization. Also,
In each of the CT-ext50 and 200 mg / kg administration groups, significant suppression was observed only after 1 week.

Example 3 (1) Preparation of Alkaloid Fraction of Cordyceps sinensis Corydalis turtscha from China
After cutting 5 kg of tuber of ninovii forma yanhusu) into small pieces, 2 times with 10 times amount of 100% methanol.
The mixture was extracted twice under reflux for 2 hours and filtered while hot. After distilling off methanol under reduced pressure, a methanol extract (yield: 2.95
%) Was obtained. The resulting extract was added with 10% acetic acid solution 4,000
After dissolving in 0 ml and 600 ml of ether, transfer to a separating funnel,
Degreased by shaking (ether layer). The 10% acetic acid layer was filtered (insoluble matter was removed), and the filtrate was saturated with ammonia gas under ice cooling to adjust the liquidity to pH 8.6 to 9.0 (using pH test paper). Irrespective of the resulting precipitate, the mixture was extracted with 1,000 ml of ether by shaking 20 times. This ether extract (tertiary alkaloid fraction) was concentrated to 1/5 and
The mixture was extracted with 1,000 ml of a 10% sodium hydroxide solution by shaking 10 times. The ether layer was dried over calcium carbonate, filtered, and the solvent was distilled off under reduced pressure to obtain a tertiary phenolic alkaloid fraction (yield: 0.162%). The 10% sodium hydroxide layer was saturated with ammonium chloride, filtered, and extracted with ether. The extract was dried over calcium carbonate, filtered, and the solvent was distilled off under reduced pressure to obtain a tertiary non-phenolic alkaloid fraction (yield: 0.0196%). The aqueous layer from which the tertiary alkaloids have been removed is adjusted to pH 6.6 to 7.0 (with pH test paper) with hydrochloric acid, and then
The mixture was extracted with 1,000 ml of chloroform by shaking 20 times, dehydrated with sodium sulfate, and then the chloroform was distilled off under reduced pressure to obtain a quaternary alkaloid fraction (yield: 0.154%).

(2) Isolation of Alkaide Component from Tertiary Non-Phenolic Alkaloid Fraction Tertiary non-phenolic alkaloid fraction (hereinafter CT-1
Abbreviated to basic alumina (230-400 mesh)
Column chromatography using ASTM, MERCK), developed and eluted with ether, ether / chloroform (5: 1) and chloroform, and thin layer chromatography [thin layer plate; silica gel 60F].
₂₅₄ (manufactured by MERCK), developing solvent; cyclohexane:
Ethyl acetate: diethylamine = 80: 15: 5 or chloroform, color reagent solution; Dragendorff reagent solution] as a monitor, and 1-tetrahydrocoptisine-containing fraction, d-
A corydarin-containing fraction, d-corydarin, dl-tetrahydropalmatine and protopine-containing fraction, a dl-tetrahydropalmatine-containing fraction, and a 1-glaucin-containing fraction were obtained. l-Tetrahydrocoptisine and d-coridarine were obtained by a recrystallization method using chloroform, ethanol and ether from the fractions mainly containing them. The fractions containing d-corridalin, dl-tetrahydropalmatine and protopine were silica gel (230-400 mesh AST
M, MERCK) column chromatography using n-hexane / ethyl acetate (1: 1),
Elution was carried out sequentially with (1: 3) and (1: 5), and using the above-mentioned thin layer chromatography as a monitor, d-corridalin and dl-tetrahydro were extracted from each fraction by recrystallization using chloroform, ethanol and ether. Palmatin,
I got protopine. Further, the d-glaucine-containing fraction was silica gel (230-400 mesh ASTM, MER
CK) and column chromatography,
Chloroform, 5% chloroform / methanol, 10%
Elute sequentially with chloroform / methanol, ethanol,
d-Glaucin was obtained by a recrystallization method using n-hexane. The mother liquor was subjected to silica gel chromatography and recrystallization to isolate the above alkaloid component. The yield of each component was 0.0511% for d-coridaline and 0.02 for d-glaucine.
43%, protopine was 0.0104%, 1-tetrahydrocoptisine was 0.00648%, and dl-tetrahydropalmatine was 0.0162%.

(3) Isolation of Alkaide Component from Tertiary Phenolic Alkaloid Fraction The tertiary phenolic alkaloid fraction (hereinafter abbreviated as CT-2) was recrystallized with chloroform, ethanol and ether to give l- Tetrahydrocorumbamine was isolated. The yield was 0.0100%.

(4) Isolation of alkaloid components from quaternary alkaloid fraction Quaternary alkaloid fraction (hereinafter abbreviated as CT-3) was treated with silica gel (230-400 mesh ASTM, MERCK).
(Manufactured by K.K.) and developed and eluted with chloroform. Using the above thin layer chromatography as a monitor, a dehydrocoridarin-containing fraction was obtained. Dehydrocorridalin was obtained from this fraction by a recrystallization method using methanol. The yield was 0.0531%.

(5) Effect of each fraction on histamine release CT-1, CT-2, CT-obtained in (1) to (4)
Example 2 for 3 fractions and each alkaloid component
A rat peritoneal mast cell suspension was prepared in the same manner as in (1) and used as a histamine-releasing compound 48/80.
A histamine release reaction test was conducted in the same manner as in Example 2 (1) using (final concentration 10 μg / ml). (Table 11,
Table 12).

[0057]

[Table 11]

[0058]

[Table 12]

From the results of Table 11, CT-1 and CT-3
The histamine release inhibitory effect was observed in the fractions. Table 1
From the results of 2, the strongest inhibition of histamine release was observed in dehydrocoridaline, and then the inhibition was also observed in d-coridarine, d-glaucine, 1-tetrahydrocoptisine, and protopine.

Example 4 Eye Drops: A 0.5% aqueous solution of CT-ext was prepared, and 1 was given to 50 healthy adult males with hyperemic eyes (red eyes) and Kayumi's symptoms.
The eye drops were applied several times a day (2 to 3 drops / time). When judged 7 days after the start of instillation, 42 out of 50 (84
%), And for Kayumi, 45 out of 50 (90%) were effective. No side effects were observed.

Example 5 Ophthalmic Solution: 30 ml of the following (1) stock solution and 0.3 ml of (2) stock solution were mixed, and CT-ex was added to this so that the final concentration was 0.5%.
t was added. Adjust the pH to 6.8 (1) Stock solution,
(2) Adjust the stock solution and centrifuge the resulting solution (10,000
The supernatant was collected at 0 rpm, 4 ° C, 10 minutes) and filtered through a 0.22 µm filter. The filtrate was put in a sterilized container to obtain an eye drop.

[Table 13] Buffer (1) Acidic stock solution Boric acid 12.4 g Potassium chloride 7.4 g Sterilized purified water 1000.0 ml (2) Alkaline stock sodium carbonate 21.2 g Sterilized purified water 1000.0 ml

Example 6 Eye drops:

Table 14 CT-ext 0.5 g Concentrated glycerin 1.5 g Polyoxyethylene hydrogenated castor oil 1.0 g Benzalkonium chloride 0.005 g Sodium edetate 0.01 g Dilute hydrochloric acid qs Sodium hydroxide qs Sterile purified water Total 100 ml

Example 7 Eye drops:

[Table 15] CT-ext 1.0 g Concentrated glycerin 1.25 g Polysorbate 80 2.0 g Methyl paraoxybenzoate 0.026 g Propyl paraoxybenzoate 0.014 g Dilute hydrochloric acid Adequate amount Sodium hydroxide Adequate amount Sterilized purified water Total amount 100 ml

Example 8 Eye ointment:

Table 16 CT-ext 1.0 g Liquid paraffin 10 g White petrolatum 89 g

Example 9 Eye ointment:

Table 17 CT-ext 2.0 g Liquid paraffin 10 g White petrolatum 88 g

[Brief description of drawings]

FIG. 1 is a graph showing the effect of histamine on guinea pig isolated ileal contraction in Example 1 (5).

Claims (8)

[Claims] 1. An anti-allergic agent which comprises a polar solvent extract of chordae cord as an active ingredient. 2. An anti-inflammatory agent containing a polar solvent extract of chordae cord as an active ingredient. 3. An anti-allergic agent which comprises an alcohol extract of chompae cord as an active ingredient. 4. An anti-inflammatory agent comprising an alcoholic extract of chordae cord as an active ingredient. 5. An anti-allergic agent comprising an alkaloid fraction of Enchoronae as an active ingredient. 6. An anti-inflammatory agent comprising an alkaloid fraction of chordae cord as an active ingredient. 7. An anti-allergic agent comprising as an active ingredient one or more selected from d-coridaline, d-glaucine, protopine, l-tetrahydrocoptisine and dehydrocoridaline. 8. An anti-inflammatory agent comprising as an active ingredient one or more selected from d-coridaline, d-glaucine, protopine, 1-tetrahydrocoptisine and dehydrocoridaline.