JP2000239161A - Allergic disease therapeutic agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a therapeutic agent having excellent antiallergic action with slight side effects, and useful as e.g. a medicament for preventing or treating diseases caused by various kinds of allergy by including a specific naphthoquinone derivative. SOLUTION: This allergic disease therapeutic agent contains a naphthoquinone derivative of formula I, II or III [2,2-"-methylenebis(3- hydroxy-1,4-naphthoquinone), 2,2'-ethylidenebis(3-hydroxy-1,4-naphthoquinone), 2-hydroxyethyl)-3-methoxy-1,4-naphthoquinone, respectively]. Each of these naphthoquinone derivatives can be extracted and isolated from lmpatience balsamina L.; it is obtained, for example, by cutting the whole grass, leaves, stems, rinds, or petals of the above plant with or without drying and subjecting them to extraction at normal or higher temperatures with a solvent (e.g. water, an organic solvent, a mixture thereof), or by synthesis. The daily dose of this agent is normally 0.1-500 or 0.01-100 mg/adult for external use or internal use, respectively.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a remedy for allergic diseases characterized by containing a naphthoquinone derivative effective for allergic diseases such as atopic dermatitis.

[0002]

2. Description of the Related Art In recent years, with changes in living environments and lifestyles, humans have been exposed to various external substances.
For example, factory smoke, car exhaust, house dust,
House dust mites, various food additives and detergents containing chemical substances can be said to be products of modern society that have been generated with the improvement of living standards. On the other hand, in an attempt to protect the body from these external substances, an antigen-antibody reaction frequently occurs in a human body, which is a major factor causing various allergic diseases. Allergies are caused by the inability of humans to cope with too many external substances in an antigen-antibody reaction to protect the body from external substances, resulting in pathological symptoms. It is a modern disease.

[0003] Diseases caused by allergies are diverse,
For example, allergic rhinitis / bronchial asthma / dermatitis, hay fever, eczema, contact dermatitis, atopic dermatitis and the like can be mentioned. Above all, atopic dermatitis is a disease with severe itching from redness and eczema, and these symptoms are chronic for many years. It is. Furthermore, atopic dermatitis has been increasing in recent years, and in Japan, one out of every twenty adults and one in five children are said to be atopic patients, and it is a large social disease that is widely distributed from infants to adults. It is a problem.

[0004] The itch due to atopic dermatitis is persistent, spreads throughout the body for some people, and its symptoms are so severe that it also interferes with daily life such as sleep. Therefore, it is necessary for patients with atopic dermatitis to reduce the itch as soon as possible in order to prevent the deterioration of the symptoms due to the scratching behavior (scratching, scratching) accompanying itching.

At present, as a therapeutic agent for allergic diseases such as atopic dermatitis, one of chemical mediators (chemical messengers) released (degranulated) from mast cells in a living body by an allergic reaction, An antihistamine which is an antagonist of histamine which is a substance causing allergic symptoms, an antiallergic agent which is a release inhibitor of a chemical mediator containing histamine, and the like are generally used.

As the antihistamine, diphenhydramine and chlorpheniramine maleate are used as the first generation, and mequitazine and terfenadine are used as the second generation. In addition, as an antiallergic agent,
Ketotifen fumarate, oxatomide, sodium cromoglycate and the like are used.

[0007]

However, the above-mentioned antihistamines have many problems because of their strong side effects such as drowsiness due to central sedation, dryness due to dizziness and anticholinergic effects, and gastric disorders. Also, even in the above antiallergic agent,
As with the antihistamines, attention must be paid to side effects such as drowsiness and gastric disorders. Furthermore, at present, these antihistamines or antiallergic agents are not always satisfactory.

[0008]

Means for Solving the Problems In view of the above problems, the present inventors have searched various natural materials having few side effects, and have found out that impatiens Balsamina which has been used as a folk medicine for anti-itch in the Shikoku region since ancient times. L.). Various pharmacological effects have been confirmed by the present inventors' research on the Balsam extract and its components.
That is, the extract of the white petals of the Balsam monkey and its isolated components may have an anti-anaphylactic effect [Phytot
herapy Res., 11 , 48-50 (1997)]. Also, whole plant extracts and their isolated components may have an inhibitory effect on testosterone 5α-reductase [JP-A-11-21245].
No.] has been confirmed.

Further, the present inventors have conducted various studies on the pharmacological effect of the ingredient using the antiallergic action of the component of the balsam as an indicator. As a result, three new naphthoquinone derivatives were newly isolated from the balsam extract, The present invention was completed after confirming allergic effects.

That is, the present invention provides the following formulas (I) and (I)
It is intended to provide a therapeutic agent for allergic diseases, comprising a naphthoquinone derivative represented by I) or (III) as an active ingredient.

[0011]

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[0012]

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[0013]

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Further, the present invention provides a therapeutic agent for atopic dermatitis, comprising a naphthoquinone derivative represented by the above formula (I), (II) or (III) as an active ingredient. is there.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION The naphthoquinone derivative of the present invention (hereinafter, referred to as "the substance of the present invention") can be extracted and isolated from balsam. The origin of Balsam is an annual plant that grows naturally in Asia, including Japan, and is a plant that has the scientific name of Impatiens Balsamina L .. The substance of the present invention is a commonly used separation from an extract obtained by extracting the balsam with a solvent,
It can be isolated by purification means.

The extract of the Balsam may be, for example, the whole plant of the Balsam or any one of leaves, stems, pericarp, and petals
It can be obtained by drying or cutting without drying at more than two places (hereinafter referred to as "the drug substance") and extracting with a solvent at room temperature or under heating.

The solvents used here include water, organic solvents and mixtures thereof. Specific examples of these organic solvents include lower alcohols such as methanol, ethanol, and butanol, or a mixture of these lower alcohols and water (lower alcohol concentration 10 to 90 V / V
%, Preferably 20 to 70 V / V%), ketones such as acetone and methyl ethyl ketone, or a mixture of these ketones and water (ketones concentration 10 to 90 V / V%, preferably 20 to 7%)
0V / V%), hydrocarbons such as hexane, benzene, toluene, petroleum ether, chloroform, dichloromethane, 1,
Examples include halogenated hydrocarbons such as 2-dichloroethane, ethers such as diethyl ether and tetrahydrofuran, esters such as methyl acetate and ethyl acetate, and mixtures thereof.

As a specific example of a preferred extraction method from the drug substance, after cutting the drug substance, an appropriate organic solvent (preferably lower alcohols, hydrated lower alcohols, ketones, hydrated ketones, hydrocarbons) Or esters)
And extracting the solvent, or after cutting the drug substance, extracting with a solvent such as anhydrous or hydrated lower alcohol, and then extracting the extract with a water-immiscible solvent such as ethyl acetate and butanol. The method includes subjecting the mixture to liquid-liquid extraction to be used, and further distilling off the solvent from the organic layer or the aqueous layer, but is not particularly limited thereto. In addition, for the efficient extraction of the substance of the present invention, whole plant or pericarp of balsam was used as the drug substance, and the extraction ratio of the drug substance and the solvent was 1 to 80 W / V%, preferably 10 to 60 W / V%. Is preferred.

The substance of the present invention can be isolated from the balsam extract obtained by the above-mentioned extraction method by repeatedly carrying out separation and purification means usually used, for example, column chromatography, thin-layer chromatography or recrystallization. it can.

The substance of the present invention can also be obtained by organic synthesis. That is, 2,
2'-methylenebis (3-hydroxy-1,4-naphthoquinone)
2-hydroxy-1,4-naphthoquinone (sometimes called Lawson) and formaldehyde with a suitable organic solvent,
It is preferably produced by condensation in a solvent such as lower alcohol, dimethylformamide or acetonitrile at ambient temperature or under heating, preferably at 50 to 80 ° C. Similarly, 2,2′-ethylidenebis (3-hydroxy-1,4-naphthoquinone) represented by the formula (II) is produced by condensing Lawesson with acetaldehyde or acetal. Further, the 2- (1-) represented by the formula (III)
(Hydroxyethyl) -3-methoxy-1,4-naphthoquinone is 2
Produced from 3-chloro-1,4-naphthoquinone in three steps. That is, 2-chloro-1,4-naphthoquinone and malonic acid monoethyl ester potassium salt are condensed in a suitable solvent, preferably in aqueous acetonitrile, in the presence of silver nitrate and alkali persulfate, preferably ammonium persulfate to form 2-
Chloro-3-ethoxycarbonylmethyl-1,4-naphthoquinone and then 2-chloro-3- obtained by reduction with a metal hydride, preferably lithium aluminum hydride, in an ether organic solvent, preferably in tetrahydrofuran (1-
Hydroxyethyl) -1,4-naphthoquinone is synthesized by methoxylating a chloro group with a methanol solution of an alkali hydroxide, preferably a methanol solution of potassium hydroxide. The crude product of each substance of the present invention obtained by the above synthesis method is isolated and purified by commonly used separation and purification means, for example, extraction, concentration, column chromatography, thin layer chromatography, recrystallization and the like.

The naphthoquinone derivative represented by the formula (I) of the present invention has been reported as a synthetic product for its antitumor effect [Cancer Letters, 113 , 4].
7 (1997).]. In addition, the naphthoquinone derivative represented by the formula (II) has also been isolated as a balsam component by the present inventors [JP-A-11-21245].
issue]. Further, as for the naphthoquinone derivative represented by the formula (III), although it is a known substance as a synthetic product, [Synlet
t, (6), 355 (1990).] This is the first time that it has been isolated from a natural product, and there is no report on its pharmacological action.

As described above, no antiallergic effect of the substance of the present invention has been reported so far, and this is a useful pharmacological action first discovered by the present inventors.

The naphthoquinone derivatives represented by the formulas (I), (II) or (III) thus obtained each have an excellent antiallergic effect with few side effects,
Its applications include medicines, quasi-drugs, cosmetics and health care for various allergic diseases such as allergic rhinitis, bronchial asthma, dermatitis, hay fever, eczema, contact dermatitis and atopic dermatitis. When administered as an oral preparation, injection or external preparation of food, it can be used for its prevention or treatment, but is not particularly limited thereto.

The naphthoquinone derivative represented by the formula (I), (II) or (III) of the present invention is usually used in an amount of 1 per adult.
As a daily dose, in the case of an external preparation, 0.1 mg to 500 mg, preferably 1 mg to 50 mg, and in the case of an internal preparation, 0.01 to 100 m
g, preferably 0.1 to 50 mg. However, the dosage is age, body weight, symptoms, therapeutic effect, administration method,
Since the amount varies depending on the treatment time and the like, an amount smaller than the above-mentioned administration range may be sufficient in some cases, and it may be necessary to administer beyond the range.

The dosage form of the therapeutic agent for allergic diseases of the present invention is
It is not particularly limited, and examples thereof include those commonly used for skin, such as ointments, lotions, creams, gels, and emulsions for external use. For internal use, there may be mentioned those usually used as oral preparations or health foods of pharmaceuticals such as tablets, capsules, granules, fine granules, powders, and liquids.

The therapeutic agent for allergic diseases of the present invention may, if necessary, impair the effects of the present invention, in addition to the above-mentioned naphthoquinone derivative represented by the formula (I), (II) or (III). To the extent that is not in the case of external preparations, usually applied hydrocarbons, waxes, fats and oils, higher fatty acids, lower or higher alcohols, surfactants, fragrances, pigments,
Preservatives, antioxidants, UV absorbers, pH regulators, and, for oral preparations, appropriate excipients such as disintegrants, binders, lubricants, coating agents, coloring agents, etc. can do.

[0027]

The present invention will be described below by way of examples, but the present invention is not limited to these examples.

Extraction Example 1 Whole grass (aboveground part) (15 kg) of balsam pears cut and shaded all day and night is soaked in 35 V / V% ethanol (30 liters),
Leached for one month. Next, the whole plant was filtered off and aged for another 2 weeks. The extract (2
(6 liters) was concentrated under reduced pressure to about 2 liters, and then liquid-liquid extracted twice with ethyl acetate (1 liter). Next, the ethyl acetate extracts were combined, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure to obtain an ethyl acetate extract (8.6 g).

<2,2'-methylenebis (3-hydroxy-1,4-
Production method of naphthoquinone) (I)> Production example 1 The ethyl acetate extract (1.53 g) of balsam obtained by the method of extraction example 1 was separated and purified by silica gel column chromatography (developing solvent: chloroform). Chloroform was added to the residue, and the insolubles were collected by filtration. Next, the insoluble crystals collected by filtration were recrystallized with a dimethylformamide / ethanol mixture to give 2,2′-methylenebis (3-
Hydroxy-1,4-naphthoquinone) (18 mg) was obtained. IR (KBr) cm ^-1 : 3360, 1680, 1611, 1574, 1352, 1213, 7
^{72, 735. 1 H-NMR (} DMSO-d 6) δ: 3.75 (2H, s, -CH 2 -), 7.75-8.04 (8
. H, m, 5,6,7,8-H of naphthoquinone) 13 C-NMR (DMSO-d 6) δ: 18.0 (-CH 2 -), 122.0 (2,2 '), 125.
6 (5,5 '), 125.9 (8,8'), 129.9 (4a, 4'a), 132.0 (8a, 8'a),
133.1 (6,6 '), 134.5 (7,7'), 155.0 (3,3 '), 180.7 (4,
4 '), 183.6 (1, 1'). EI-MS: m / z 360 [M ⁺ ] mp 245 ° C (decomp.).

Production Example 2 A solution of Lawson (30.0 g), 37% formalin (30.0 mL) and triethylamine (34.8 g) in ethanol (500 mL) was added at 70 ° C. for 16 hours.
Stirred for hours. After cooling, the reaction solution was acidified by adding 6N-hydrochloric acid. The precipitated crystals were collected by filtration, washed with hot ethanol, and recrystallized with a dimethylformamide / ethanol mixture to give 2,2′-methylenebis (3-hydroxy-1,4-naphthoquinone) (8.87) as yellow prism crystals. g) was obtained. Physical property data (IR, ¹ H-NMR, ¹³ C-NMR, mass spectrum, mp) of the compound obtained in this Production Example were identical to those obtained in Production Example 1 extracted and isolated from balsam. <Production method of 2,2′-ethylidenebis (3-hydroxy-1,4-naphthoquinone) (II)> Production Example 3 Chloroform was added to ethyl acetate extract (10.0 g) of balsam obtained by the method of Extraction Example 1. And insolubles were filtered off.
Then, the filtrate is concentrated under reduced pressure, and the residue obtained is separated and purified twice by silica gel column chromatography (developing solvent: chloroform), recrystallized with ethyl acetate, and 2,2′-ethylidenebis (yellow prism crystal) 3-hydroxy-1,4-naphthoquinone) (1.03 g) was obtained. IR (KBr) cm ^-1 : 3296, 1664, 1637, 1359, 1344, 1278, 7
_{^{29. 1 H-NMR (CDCl 3}} ): 1.73 (3H, d, J = 7.5Hz, = CHC H 3), 4.83
(1H, q, J = 7.5Hz , = C H CH 3), 7.54-7.83 (4H, m, 6,7-H
of naphthoquinone), 7.96-8.14 (4H, m, 5,8-H of naph
thoquinone). Here, "= C" means the carbon atom at the 1-position of alkylidene. _{^{13 C-NMR (CDCl 3)}} δ: 16.6 (= CH C H 3), 28.5 (= C HCH 3), 12
5.0 (2,2 '), 126.1 (5,5'), 127.0 (8,8 '), 129.4 (4a, 4'
a), 132.9 (6,6 ', 8a, 8'a), 134.9 (7,7'), 153.8 (3,3 '),
181.7 (4,4 '), 184.7 (1,1'). Here, "= C" means the carbon atom at the 1-position of alkylidene. EI-MS: m / z 374 [M ⁺ ] mp 196-198 ° C.

Production Example 4 Lawson (10.0 g) and acetal (14.2 g) were replaced with argon.
A solution of 8 g) in dimethylformamide (50 mL) was stirred at 80 ° C. for 4.5 hours. After cooling, ethyl acetate was added to the reaction solution, washed with diluted hydrochloric acid and saturated saline in this order, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was crystallized from ethyl acetate and further recrystallized from ethyl acetate to obtain 2,2′-ethylidenebis (3-hydroxy-1,4-naphthoquinone) (9.10 g) as yellow prism crystals. . Physical property data of the compound obtained in this Production Example (IR, ¹ H-NMR, ¹³ C-NMR, mass spectrum, m.
p.) was consistent with that obtained in Production Example 3 which was extracted and isolated from Balsam. Extraction Example 2 Undried pericarp (excluding seeds in pericarp) (75
g) was immersed in 50% ethanol (400 mL) and leached for 3 days, and the 50% ethanol extract obtained by filtering the pericarp was concentrated under reduced pressure to about 30 mL. Next, the filtrate obtained by filtering off the precipitated insolubles was evaporated under reduced pressure to obtain a 50% ethanol extract (2.9 g).

<2- (1-hydroxyethyl) -3-methoxy-1,
Production Method of 4-Naphthoquinone (III)> Production Example 5 The 50% ethanol extract (2.9 g) obtained in Extraction Example 2 was subjected to silica gel column chromatography (developing solvent: chloroform: methanol = 30: 1), followed by thinning. Layer chromatography (developing solvent: chloroform: methanol = 10: 1)
And then recrystallized with a mixed solution of n-hexane / ethyl acetate to give 2- (1-hydroxyethyl) as pale orange needles.
3-Methoxy-1,4-naphthoquinone (6.0 mg) was obtained. IR (KBr) cm ^-1 : 3329, 2949, 1674, 1655, 1615, 1341, 1
. 217, 1080, 1042, 990,727 1 H-NMR (CDCl 3): 2.91 (2H, t, J = 6.2Hz, C H 2 CH 2 OH), 3.8
2 (2H, t, J = 6.2Hz, CH ₂ C H ₂ OH), 4.17 (3H, s, OC H ₃ ), 7.
65-7.75 (2H, m, 6,7-H of naphthoquinone), 8.01-8.13
. (2H, m, 5,8- H of naphthoquinone) 13 C-NMR (CDCl 3) δ: 27.3 (C H 2 CH 2 OH), 61.4 (OCH 3), 61.6
(CH ₂ C H ₂ OH), 126.1 (5,8), 131.5 (2), 131.8 (4a, 8a), 13
3.3 (6), 133.8 (7), 158.6 (3), 181.3 (4), 186.0 (1). EI-MS: m / z 274 [M ⁺ ] mp 107-108 ° C.

Production Example 6 <2-Chloro-3-ethoxycarbonylmethyl-1,4-naphthoquinone> 2-Chloro-1,4-naphthoquinone (27.17 g), potassium malonate monoethyl ester (36.02 g) and silver nitrate (7.19 g) in acetonitrile (270 mL) was heated to 70 ° C., and then an aqueous solution (270 mL) of ammonium persulfate (48.29 g) was added to 30 ° C.
After dropwise addition over a period of minutes, the mixture was stirred at the same temperature for 1.5 hours. After cooling, chloroform was added to the reaction solution, and insolubles were filtered off. The organic layer of the filtrate was separated, washed with saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was separated and purified by silica gel column chromatography (developing solvent: benzene) to give 2-chloro-3-ethoxycarbonylmethyl-1,4-naphthoquinone (8.5%) as pale orange crystals.
2g) was obtained. IR (KBr) cm ^-1 : 2986, 1723, 1682, 1665, 1613, 1589, 1
406, 1373, 1339, 1281,1202, 1157. 1 H-NMR (CDCl 3): 1.27 (3H, t, J = 7.2Hz, CO 2 CH 2 C H 3), 3.
88 (2H, s, C H ₂ CO ₂ Et), 4.20 (2H, q, J = 7.2Hz, CO ₂ C H ₂ CH
₃ ), 7.72-7.88 (2H, m, 6,7-H of naphthoquinone), 8.0
9-8.24 (2H, m, 5,8-H of naphthoquinone).

<2-chloro-3- (1-hydroxyethyl) -1,4-
Naphthoquinone> A suspension of lithium aluminum hydride (222 mg) in tetrahydrofuran (20 mL) was added under ice-cooling to 2-chloro-3.
-Ethoxycarbonylmethyl-1,4-naphthoquinone (1.08g)
Of tetrahydrofuran (10 mL) was added thereto, and the mixture was stirred at room temperature for 3 hours while replacing with argon. Then, lithium aluminum hydride (50 mg) was added, and the mixture was further stirred at room temperature for 1.5 hours. Ethyl acetate was added to the reaction solution, washed with diluted hydrochloric acid and saturated saline in this order, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue is subjected to silica gel column chromatography (developing solvent: chloroform: methanol = 4).
0: 1) to separate and purify 2-chloro-3- (1
-Hydroxyethyl) -1,4-naphthoquinone (511 mg) was obtained. ^{IR (KBr) cm -1: 3355} , 1665, 1593, 1281, 1047, 716. 1 H-NMR (CDCl 3): 3.14 (2H, t, J = 6.2Hz, C H 2 CH 2 OH), 3.8
6 (2H, t, J = 6.2Hz, CH ₂ C H ₂ OH), 7.71-7.86 (2H, m, 6,7-
H of naphthoquinone), 8.08-8.22 (2H, m, 5,8-Hof nap
hthoquinone) ..

<2- (1-hydroxyethyl) -3-methoxy-1,
4-naphthoquinone> 2-chloro-3- (1-hydroxyethyl) -1,
To a solution of 4-naphthoquinone (725 mg) in tetrahydrofuran (15 mL) was added a methanol solution of 2.5% potassium hydroxide (3.6 mL), and the mixture was stirred at room temperature for 30 minutes. Next, a methanol solution (3.6 mL) of 2.5% potassium hydroxide was added, and the mixture was further stirred at room temperature for 30 minutes. Ethyl acetate was added to the reaction solution, washed with brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was separated and purified by silica gel column chromatography (developing solvent: benzene: ethyl acetate = 20: 1), and then recrystallized from a mixed solution of n-hexane / ethyl acetate to give light orange prism crystals of 2- (1 -Hydroxyethyl) -3-methoxy-1,4-naphthoquinone (320 mg) was obtained. The physical property data (IR, ¹ H-NMR, ¹³ C-NMR, mass spectrum, mp) of the compound obtained in this Production Example were consistent with those obtained in Production Example 5 which was extracted and isolated from balsam.

Experimental Example 1 (Acute toxicity test) Two groups of five ICR mice (male, 5 weeks old, body weight 20 to 25 g)
In addition, 2,2'-methylenebis (3-hydroxy-1,4-
Naphthoquinone) was orally administered at doses of 1000 mg / kg and 2000 mg / kg. As a result, at 14 days after administration, 1000
No deaths were observed at mg / kg, and 1 out of 5 animals at 2000 mg / kg
One of the animals died. Therefore, the LD ₅₀ value of the substance of the present invention in Production Example 1 was 2000 mg / kg or more.

Each group of 5 ICR mice (male, 5 weeks old,
Weight 20-25 g), 2,2'-ethylidenebis (3-hydroxy-1,4-naphthoquinone) of Production Example 3 from 250 mg / kg to 1000 mg / kg
Oral administration at doses between As a result, the LD ₅₀ value of the substance of the present invention in Production Example 3 was 514 mg / kg.

[0038] Five ICR mice (male, 5 weeks old,
2- (1-hydroxyethyl) -3 of Production Example 5
-Methoxy-1,4-naphthoquinone was administered orally at a dose between 280 mg / kg and 480 mg / kg. As a result, the LD ₅₀ value of the substance of the present invention in Production Example 5 was 374 mg / kg.

Experimental Example 2 (Anti-itch test) Using an SPF (Specific pathogen free = no specific pathogen) ddy male mouse (7 weeks old) as an experimental animal,
According to the following method, the anti-itch test of the substance of the present invention obtained in Production Examples 1, 3 and 5 was performed.

Each of the substances of the present invention suspended in water was orally administered at 10 mg / kg to three groups of 7 mice each, and 24 mg after administration.
After a lapse of time, compound 48/80, a degranulation-inducing substance, was injected subcutaneously at 3 mg / kg. Similarly, do not administer each substance 1
A group of 7 mice was set as a control group, and compound 48/80
The number of times of scratching the whole body of the mouse for 20 minutes after the administration was measured. Further, as a normal (normal) group, the number of times of scratching of 7 mice per group to which none of each substance and compound 48/80 was administered was measured in the same manner.

FIG. 1 shows the results. In FIG. 1, each value is represented by the average value ± standard error of 7 mice in each group.
The asterisk indicates a significant difference from the control group by the Student's t-test ( ^* P <0.05, ^** P <0.02, ^*** P <
0.0001).

FIG. 1 shows that the substances of the present invention exhibited an inhibitory effect on the increase in the number of times of mouse scratching due to degranulation caused by an allergic reaction. Movement was significantly suppressed. Therefore, it was suggested that it is useful for alleviating various itches caused by allergic reactions including atopic dermatitis.

Experimental Example 3 (Platelet activating factor PAF (= Platele
Inhibition test for blood flow decrease by t-activating factor) Using SPF (Specific pathogen free = absence of specific pathogen) ddy male mouse (5 weeks old) as an experimental animal,
According to the following method, an inhibitory test of the substance of the present invention obtained in Production Examples 1, 3 and 5 against reduction in blood flow due to PAF was performed.

To three groups of 7 mice each, 10 mg / kg of each of the substances of the present invention of Production Examples 1, 3 and 5 suspended in water was orally administered, and 24 hours after the administration, PAF was administered at 10 μg / kg iv. Injection. Similarly, a group of 7 mice to which the substance was not administered was used as a control group, and the blood flow of microvessels in the tail of the mouse for 30 minutes after PAF administration was measured at intervals of 2 minutes using a laser blood flow meter (FLO-N1, type ) Neuroscience). The results are shown in FIGS.
As a positive control, a similar test was conducted using CV-3988, a commercially available anti-PAF agent. Since CV-3988 is not absorbed by oral administration, it was administered by intravenous injection at a dose of 10 mg / kg in this test, and PAF was intravenously injected 45 minutes after administration, and blood flow was measured. The results are shown in FIG.

In FIGS. 2 to 5, each value is represented by the average value ± standard error of 7 mice in each group. In addition, * mark is Student
Represents a significant difference from the control group by t-test of t ( ^* P <0.05, ^** P <0.01, ^*** P <0.001).

The platelet activating factor PAF is an in vivo autakoid having various actions such as a platelet activating action, a hypotensive action, a vascular permeability enhancing action, and an intestinal motility enhancing action, and was sensitized with immunoglobulin IgE. It is a substance produced by basophils and neutrophils in blood, and is known as a very strong shock inducer. PAF is 1/1000 of histamine
The following concentrations cause hyperpermeability of microvessels such as skin and respiratory tract, and studies on the relationship with allergies are in progress. In recent reports, this PAF is involved in so-called chemical mediators such as histamine and prostaglandin, which are commonly known, as a causative agent of anaphylactic shock, which is a type of immediate (type I) allergic reaction. [Platelet activating factor-biochemistry / physiology / pathology-, published by Tokyo Kagaku Dojin, page 190 (1989)]. In addition, the present inventors have confirmed that CV-3988, a PAF antagonist, suppresses a decrease in blood pressure in mice due to anaphylactic shock [Phytotherapy Research, 8 , 301.
(1994).].

As shown in FIGS. 2 to 5, the substance of the present invention significantly suppressed the decrease in blood flow caused by anaphylaxis due to PAF in mice by oral administration, and its effect was more than that of CV-3988 by intravenous administration. Was. Therefore, it is considered that all the substances of the present invention strongly suppress anaphylaxis, which is one of allergic reactions, and it was suggested that they are useful as therapeutic agents for various diseases caused by allergic reactions.

Experimental Example 4 (Inhibition test on decrease in blood flow due to degranulation-inducing substance) As a test animal, SPF (Specific pathogen free = no specific pathogen-free) ddy male mouse (5 weeks old) was used.
According to the following method, an inhibitory test was performed on the reduction in blood flow of the substance of the present invention obtained in Production Examples 1 and 5 due to a degranulation-inducing substance.

Two groups of 7 mice were orally administered 10 mg / kg of each of the substances of the present invention of Suspension Examples 1 and 5 suspended in water, and 24 hours after administration, 1 mg of compound 48/80 was administered.
/ kg injected intravenously. Similarly, a group of 7 mice to which the substance of the present invention was not administered was used as a control group, and compound 48 /
The blood flow in the microvessels of the mouse tail for 30 minutes after the 80 administration was measured at intervals of 2 minutes using a laser blood flow meter (FLO-N1 type, manufactured by Neuroscience Co., Ltd.). The result is shown in FIG.
And 7.

In FIGS. 6 and 7, each value is represented by the average value ± standard error of 7 mice in each group. * Mark is Student
Represents a significant difference from the control group by the t-test ( ^* P <0.05).

6 and 7 that the substances of the present invention of Production Examples 1 and 5 all suppressed the decrease in blood flow due to degranulation due to allergic reaction in mice by oral administration. That is, it was suggested that suppressing release of chemical mediators such as histamine exhibited an antiallergic effect.

Experimental Example 5 (Atopic dermatitis model NC / Jic
Pharmacological evaluation test using Jcl mice) <Induction of dermatitis> 4 groups of 7 SPF experimental animals
(Specific pathogen free = absence of specific pathogen) NC / JicJc
l Using male mice (5 weeks old), Matsuda et al.'s method [96th Annual Meeting of the Japanese Dermatological Association / Academic Conference, Luncheon Seminar Records, pp. 5-15 (1997). Reference] was partially modified to cause chronic dermatitis. That is, a 1: 1 solution of 5% picryl chloride in acetone: olive oil (100%) was applied to the shaved mouse abdomen.
μL), acetone was evaporated with a drier to sensitize. Five days later, under anesthesia with Nembutal injection, 1% picryl chloride in acetone: olive oil =
A 1:10 solution (20 μL) was applied, and this operation was repeated seven times a week until atopic dermatitis-like dermatitis became chronic.

<Heterologous P using rat back skin
Measurement of IgE Antibody Amount by CA Reaction> Using the serum of these dermatitis-induced mice, the amount of IgE antibody associated with the development of atopic dermatitis of the substance of the present invention obtained in Production Examples 1 and 3 by the following method was determined. An inhibitory effect test on increase was performed.
Each substance of the present invention was orally administered at a dose of 10 mg / kg to each group four times a week from four days before sensitization until blood collection. In addition, a commercially available antiallergic agent, epinastine hydrochloride, was orally administered in the same manner at a dose of 5 mg / kg, and a group to which these drugs were not administered was set as a control group.

At the end of the atopic dermatitis induction operation seven times, blood was collected from the subocular vein sac of each group of mice and centrifuged (3000 rpm).
, 10 minutes), and the supernatant (serum) was diluted 5-fold with physiological saline to obtain a sample. These diluted sera (50 μL) were injected intradermally into the back of shaved Wistar rats (10 weeks old),
After an hour, 0.5% Evans blue saline solution (0.5 mL) containing anti-mouse IgE antibody was injected into the tail vein of the rat. Thirty minutes later, the rats were sacrificed, and the diameter of the blue spot due to pigment leakage behind the back skin was measured using a caliper.
The antibody amount was used. The result is shown in FIG.

In FIG. 8, each value is represented by the average value ± standard error of the diameter of each locus on the back side of the skin on the back of the rat obtained using the serum of 7 mice in each group. The asterisk indicates a significant difference from the control group by the Student's t-test ( ^* P <
0.05).

FIG. 8 shows that the substances of the present invention of Production Examples 1 and 3 significantly suppressed the increase in IgE antibody production in mice that developed atopic dermatitis by oral administration. In particular, it was suggested to be useful as a therapeutic agent for atopic dermatitis.

Example 1 (skin lotion 1)% by weight A Compound of Production Example 1 0.1% Ethanol 10.0% Methyl parahydroxybenzoate 0.1% B Glycerin 3.0% 1,3-butylene glycol 2.0% Remaining purified water Total 100.0% After uniformly dissolving the component A, the solution was gradually added to the component solution B, and uniformly mixed and dissolved to obtain a skin lotion.

Example 2 (skin lotion 2)% by weight A Compound of Preparation Example 3 0.1% Ethanol 10.0% Methyl parahydroxybenzoate 0.1% B Glycerin 3.0% 1,3-butylene glycol 2.0% Remaining purified water Total 100.0% After uniformly dissolving the component A, the solution was gradually added to the component solution B, and uniformly mixed and dissolved to obtain a skin lotion.

Example 3 (skin lotion 3)% by weight A Compound of Production Example 5 0.1% Ethanol 10.0% Methyl parahydroxybenzoate 0.1% B Glycerin 3.0% 1,3-butylene glycol 2.0% Remaining purified water Total 100.0% After uniformly dissolving the component A, the solution was gradually added to the component solution B, and uniformly mixed and dissolved to obtain a skin lotion.

Example 4 (Skin cream 1)% by weight A Compound of Preparation Example 1 0.1% Salicone beeswax 5.0% Glycerin monostearate 3.5% Behenyl alcohol 3.0% Squalene 6.0% Methyl paraoxybenzoate 0.1% Propyl paraoxybenzoate 0.1% B Glycerin 10.0% Purified water Remainder Total 100.0% According to the above formula, each of A and B components was heated to about 80 ° C, A was gradually added to B with stirring, mixed uniformly, and then cooled to form a skin cream. .

Example 5 (skin cream 2)% by weight A Compound of Production Example 3 0.1% Salami beeswax 5.0% Glycerin monostearate 3.5% Behenyl alcohol 3.0% Squalene 6.0% Methyl parahydroxybenzoate 0.1% Propyl paraoxybenzoate 0.1% B Glycerin 10.0% Purified water Remainder Total 100.0% According to the above formula, each of A and B components was heated to about 80 ° C, A was gradually added to B with stirring, mixed uniformly, and then cooled to form a skin cream. .

Example 6 (skin cream 3)% by weight A Compound of Preparation Example 5 0.1% Salicone beeswax 5.0% Glycerin monostearate 3.5% Behenyl alcohol 3.0% Squalene 6.0% Methyl paraoxybenzoate 0.1% Propyl paraoxybenzoate 0.1% B Glycerin 10.0% Purified water Remainder Total 100.0% According to the above formula, each of A and B components was heated to about 80 ° C, A was gradually added to B with stirring, mixed uniformly, and then cooled to form a skin cream. .

Example 7 (Emulsion 1)% by weight A Compound of Production Example 1 0.1% Glycerin monostearate 2.0% Behenyl alcohol 65 2.0% Adsorbed and purified lanolin 1.0% Squalene 5.0% Methyl parahydroxybenzoate 0.1% Propyl paraoxybenzoate 0.1% B Polyethylene glycol 2.0% Glycerin 10.0% Purified water Remainder Total 100.0% According to the above formula, each of A and B components is heated to about 80 ° C, A is gradually added to B with stirring, mixed uniformly, and then cooled. To make an emulsion.

Example 8 (Emulsion 2)% by weight A Compound of Preparation Example 3 0.1% Glycerin monostearate 2.0% Behenyl alcohol 65 2.0% Adsorption purified lanolin 1.0% Squalene 5.0% Methyl parahydroxybenzoate 0.1% Propyl paraoxybenzoate 0.1% B Polyethylene glycol 2.0% Glycerin 10.0% Purified water Remainder Total 100.0% According to the above formula, each of A and B components is heated to about 80 ° C, A is gradually added to B with stirring, mixed uniformly, and then cooled. To make an emulsion.

Example 9 (Emulsion 3)% by weight A Compound of Preparation Example 5 0.1% Glycerin monostearate 2.0% Behenyl alcohol 65 2.0% Adsorption purified lanolin 1.0% Squalene 5.0% Methyl parahydroxybenzoate 0.1% Propyl paraoxybenzoate 0.1% B Polyethylene glycol 2.0% Glycerin 10.0% Purified water Remainder Total 100.0% According to the above formula, each of A and B components is heated to about 80 ° C, A is gradually added to B with stirring, mixed uniformly, and then cooled. To make an emulsion.

Example 10 (Ointment 1)% by weight A Compound of Production Example 1 0.1% B Polyethylene glycol 400 15.0% (Average molecular weight 400) C Macrogol ointment 84.9% Total 100.0% According to the above-mentioned formula, the A component is uniformly dispersed in B. After dispersing in
Was added and mixed to obtain an ointment.

Example 11 (Ointment 2)% by weight A Compound of Preparation Example 3 0.1% B Polyethylene glycol 400 15.0% (Average molecular weight 400) C Macrogol ointment 84.9% Total 100.0% According to the above-mentioned formula, the A component is uniformly dispersed in B. After dispersing in
Was added and mixed to obtain an ointment.

Example 12 (Ointment 3)% by weight A Compound of Production Example 5 0.1% B Polyethylene glycol 400 15.0% (Average molecular weight 400) C Macrogol ointment 84.9% Total 100.0% According to the above-mentioned formula, the A component was uniformly dispersed in B. After dispersing in
Was added and mixed to obtain an ointment.

Example 13 (Tablet 1) Compound of Production Example 1 5 mg Lactose 73 mg Corn starch 20 mg Hydroxypropylcellulose 1 mg Magnesium stearate 1 mg Total 100 mg Each component is uniformly mixed so that 1 tablet has the above ratio. And then tableted into tablets.

Example 14 (Tablet 2) Compound of Production Example 3 5 mg Lactose 73 mg Maize starch 20 mg Hydroxypropylcellulose 1 mg Magnesium stearate 1 mg A total of 100 mg Each component was homogenized so that 1 tablet would have the above ratio. And then tableted into tablets.

Example 15 (Tablet 3) Compound of Production Example 5 5 mg Lactose 73 mg Maize starch 20 mg Hydroxypropylcellulose 1 mg Magnesium stearate 1 mg Total 100 mg Each component is uniformly mixed so that 1 tablet has the above ratio. And then tableted into tablets.

Example 16 (Capsule 1) Compound of Production Example 1 10 mg Lactose 188 mg Magnesium stearate 2 mg Total 200 mg The components were uniformly mixed so that 1 capsule had the above ratio, and then filled into capsules. .

Example 17 (Capsule 2) Compound of Production Example 3 10 mg Lactose 188 mg Magnesium stearate 2 mg Total 200 mg The components were uniformly mixed so that 1 capsule had the above ratio, and then filled into capsules. .

Example 18 (Capsule 3) Compound of Production Example 5 10 mg Lactose 188 mg Magnesium stearate 2 mg Total 200 mg The components were uniformly mixed so that 1 capsule had the above ratio, and then filled into capsules. .

[0075]

The substances of the present invention represented by the general formulas (I) and (II)
And the naphthoquinone derivative represented by (III) has high safety and excellent antiallergic activity,
By blending the substance as an active ingredient, it is effective in preventing and treating various diseases caused by allergy such as atopic dermatitis.

[Brief description of the drawings]

FIG. 1 is a graph showing the results of an anti-itch test of Experimental Example 2.

FIG. 2 is a graph showing test results of the compound of Production Example 1 in Experimental Example 3.

FIG. 3 is a graph showing test results of the compound of Production Example 3 in Experimental Example 3.

FIG. 4 is a graph showing the test results of the compound of Production Example 5 in Experimental Example 3.

FIG. 5 shows a positive control compound (CV-3988) in Experimental Example 3.
Graph showing test results of

FIG. 6 is a graph showing the test results of the compound of Production Example 1 in Experimental Example 4.

FIG. 7 is a graph showing the test results of the compound of Production Example 5 in Experimental Example 4.

FIG. 8 is a graph showing test results of Experimental Example 5;

 ──────────────────────────────────────────────────の Continuing on the front page (72) Inventor Toyoya Kato 270-11 Atsuhara, Fuji-shi, Shizuoka Japan F-term in Limev Co., Ltd. 4C088 AB12 BA32 NA14 ZA89 ZB13 4C206 AA01 AA02 CB28 MA01 MA04 NA14 ZA89 ZB13 4H006 AA03 AB20

Claims (3)

[Claims] 1. A therapeutic agent for an allergic disease, comprising a naphthoquinone derivative represented by the following formula (I), (II) or (III) as an active ingredient. Embedded image Embedded image Embedded image 2. The therapeutic agent for allergic diseases according to claim 1, wherein the naphthoquinone derivative represented by the formula (I), (II) or (III) is extracted and isolated from balsam. 3. The therapeutic agent for an allergic disease according to claim 1, wherein the allergic disease is atopic dermatitis.