JPH08113536A - Inhibitor of arachidonic acid metabolism - Google Patents

Abstract

PURPOSE: To obtain an inhibitor of the arachidonic acid metabolism, having excellent inhibiting actions on lipoxygenases and cyclooxygenases and useful as an antiinflammatory and an antiallergic agents, etc. CONSTITUTION: This inhibitor of the arachidonic acid metabolism contains mollugin or its derivative of the formula (R<1> is OH or an alkoxy; R<2> is OH, an alkoxy or an alkoxycarbonyl) as an active ingredient. The mollugin or its derivative is obtained by extracting Rubia cordifolia with diethyl ether, ethyl acetate, acetone, methanol, hexane, water, etc., distilling off the solvent from the resultant extract solution, suitably dissolving the prepared residue in a solvent such as methanol, ethanol or ethyl acetate, treating and fractionating the resultant solution by a column chromatography. The daily dose of the compound is preferably within the range of 0.01-500mg, especially preferably 0.1-300mg based on 1kg body weight.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an arachidonic acid metabolism inhibitor useful as an anti-allergic agent, an anti-inflammatory agent and the like.

[0002]

BACKGROUND OF THE INVENTION Arachidonic acid is an unsaturated fatty acid having 20 carbon atoms that is released from cell membranes in response to various stimuli. It is converted into prostaglandins by a cyclooxygenase system and leukotrienes by a lipoxygenase system. And HETE (hydroxyeicosatetraenoic acid) are metabolized, and each metabolite is known to be deeply involved in various allergic and inflammatory diseases (protein, nucleic acid, enzyme, 546- 560,35
(4) 1990; proteins, nucleic acids, enzymes, 136-14
9, 33 (2) 1988; protein, nucleic acid, enzyme, 226.
238, 33 (3) 1988).

In recent years, the number of patients with allergic diseases such as bronchial asthma and hay fever has increased with the occurrence of pollution such as changes in eating habits and living environment and air pollution, which is becoming a big social problem. Asthma is a case of severe airway hypersensitivity in patients with severe respiratory dysfunction, which causes vascular permeability increase, bronchial smooth muscle contraction and secretion increase due to external allergens and nonspecific stimulation of the airway. Is a deadly disease. Steroids and antihistamines have been used as therapeutic agents for asthma, but since these have serious side effects, it is an important issue to overcome them.

With the progress of basic research on asthma in recent years,
It has become clear that, among the arachidonic acid metabolism systems, especially the lipoxygenase metabolites play an important role in the pathological condition of asthma.

Further, psoriasis, which is a kind of skin disease, is a refractory chronic inflammatory keratosis which causes proliferation of epidermis and infiltration of inflammatory cells. Although the cause of psoriasis has not been clarified yet, the amount of lipoxygenase metabolites leukotriene and HETEs increased at the lesion, which strongly suggests the relationship between lesion formation in psoriasis and abnormal arachidonic acid metabolism. (Dermatological Clinic, 1333-1341, 35 (8)
1993).

[0006] When the skin is exposed to ultraviolet rays, erythema (sunburn) occurs, which activates the arachidonic acid metabolic system and causes prostaglandin E2, F2α, 12-H.
It is known that substances such as ETE are produced (Seminarsin Dermatolog
y, 11 (2) 114-120 (1992); Nichikaikai, 91 (6) 645-652 (1981); Photo.
dermatology, 2,359-366 (19
85)).

As described above, arachidonic acid metabolites are considered to play an extremely important role in various allergic and inflammatory diseases. Inhibitors of these arachidonic acid metabolizing enzymes are arachidonic acid such as those mentioned above. It is considered to be useful as a preventive or therapeutic agent for various diseases associated with metabolic disorders.

Therefore, an object of the present invention is to provide an arachidonic acid metabolism inhibitor useful for the prevention and treatment of various allergic diseases and inflammatory diseases.

[0009]

In view of such circumstances, the inventors of the present invention have conducted extensive studies to develop a drug effective for treating a disease associated with arachidonic acid metabolism abnormality, and as a result, the maddergrass of Rubiaceae ( We have found that morphine and its derivatives contained in Rubiacordifolia) have an arachidonic acid metabolism inhibitory action, and completed the present invention.

That is, the present invention is represented by the general formula (1);

[0011]

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An arachidonic acid metabolism inhibitor containing morphine represented by the formula (wherein R ¹ represents a hydroxyl group or an alkoxy group and R ² represents a hydroxyl group, an alkoxy group or an alkoxycarbonyl group) or a derivative thereof as an active ingredient. Is provided.

The present invention also provides an antiallergic agent and an antiinflammatory agent containing the above-mentioned morphine or a derivative thereof as an active ingredient.

It is known that morphine is contained in Akane (Chem. Pharm. Bull. 40.
(6) 1504-1509 (1992)), and its effect on the arachidonic acid metabolic system was not known.

The morphine or its derivative used in the present invention is represented by the above-mentioned general formula (1). In the formula, R ¹ and R ² represent an alkoxy group having 1 to 1 carbon atoms. 6 straight chain or branched chain alkoxy groups such as methoxy group, ethoxy group, n-propyloxy group, isopropyloxy group, n-butyloxy group, s
An ec-butyloxy group, an n-pentyloxy group, an n-hexyloxy group and the like can be mentioned. Further, examples of the alkoxycarbonyl group include a methoxycarbonyl group, an ethoxycarbonyl group, a butoxycarbonyl group, a pentyloxycarbonyl group, and the like. Of these, those in which R ¹ is an alkoxy group and R ² is a hydroxyl group are preferred.

Such morphine or its derivative (1)
Can be extracted from, for example, Akane. The extraction is first performed from a solvent selected from diethyl ether, ethyl acetate, acetone, methanol, ethanol, hexane, ethyl acetate and water. Then, the residue obtained by evaporating the solvent from the obtained extract is appropriately dissolved in a solvent such as methanol, ethanol, or ethyl acetate, and further dissolved in water, methanol, ethanol, ethyl acetate, chloroform, dichloromethane, hexane, or acetone. Amberlite XAD-2, Diaion HP-
20, TSK gel HW-40 and other hydrophilic polymers and Sephadex such as LH-20 Sephadex, reverse phase silica gel, silica gel, cellulose, etc., subjected to column chromatography using a carrier, such as thin layer chromatography The desired product can be obtained by fractionation while confirming the desired component. In some cases, purification may be performed by recrystallization using a suitable solvent such as benzene, ether, hexane, acetone, methanol, ethanol, or water.

The morphine or its derivative (1) can be administered to animals and humans as it is or together with a conventional preparation alone. This dose depends on the age and sex of the patient or animal,
It may be appropriately determined depending on the degree of disease, etc., but is usually 0.01 to 500 mg per 1 kg of body weight per day, especially 0.1 to 0.1 mg.
It is preferably in the range of 300 mg.

The dosage form of morphine or its derivative (1) is not particularly limited and may be appropriately selected as needed. The dosage form is an oral preparation such as tablets, capsules, granules and powders; injection. It can be appropriately selected from parenteral agents such as agents, suppositories, sprays and ointments.

In the case of tablets, the carrier is
A wide variety of materials known in the art can be used. This includes excipients such as starch, lactose, sucrose, carboxymethyl cellulose, corn starch, inorganic salts, urea; water,
Binders such as ethanol, propanol, simple syrup, glucose, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone; dry starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, carbonate Disintegrators such as calcium, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch and lactose; disintegration inhibitors such as sucrose, stearin, cocoa butter and hydrogenated oils;
Absorption enhancers such as sodium lauryl sulfate and quaternary ammonium salts; moisturizers such as glycerin and starch; starch, lactose,
Adsorbents such as kaolin, bentonite, colloidal silicic acid; stearates, boric acid powder, purified talc, lubricants such as polyethylene glycol and the like. Further, the tablets are tablets coated with a usual coating as necessary, for example, sugar-coated tablets,
It may be a gelatin-coated tablet, an enteric-coated tablet, a film-coated tablet, a double tablet, or a multi-layer tablet.

When in the form of pills, a wide variety of carriers known in this field can be used, and examples thereof include starch, lactose, glucose, cocoa butter, hydrogenated vegetable oil, kaolin, talc and the like. Shaped agents; gum arabic powder, tragacanth powder, binders such as gelatin and ethanol; disintegrants such as laminaran acanthen.

In the case of a suppository, a wide variety of carriers known in the art can be used, and examples thereof include cocoa butter, gelatin, polyethylene glycol, higher alcohols, higher alcohol esters, and semisynthetic glycerides. Etc. can be mentioned.

When prepared as an injection, the solution and suspension are preferably sterilized and isotonic with blood. In the case of these solutions, suspensions and emulsions,
As the diluent, those commonly used in this field can be used. For example water, ethyl alcohol,
Examples thereof include propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylenated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like. In this case, salt, glucose, glycerin, etc., may be contained in the pharmaceutical preparation in an amount sufficient to prepare an isotonic aqueous solution, and a usual solubilizing agent, buffer, soothing agent, etc. may be added. You may add. Further, if necessary, a colorant, a preservative, a flavor, a flavoring agent, a sweetening agent and other pharmaceuticals may be contained in the pharmaceutical preparation.

In the case of a propellant, the dispersants and propellants known in this field can be widely used.
Examples of the dispersant include soybean lecithin, egg yolk lecithin,
Fatty acids such as oleic acid, linoleic acid and linolenic acid, and sorbitans such as sorbitan trioleate and sorbitan monooleate can be used. Further, as the propellant, for example, a normally incombustible liquefied gas such as Freon 11, Freon 12, Freon 114 can be used.

In the case of the ointment form, those known in the art can be widely used, and examples thereof include water, ethanol, isopropyl alcohol, glycerin, polyethylene glycol, sorbitol, polyvinyl alcohol and other polyhydric alcohols, animal fats and oils, plants. Waxes such as natural fats and oils, mineral oils, hydrogenated oils, beeswax, higher hydrocarbons such as liquid paraffin and paraffin wax, fatty acids such as stearic acid, emulsifiers,
Surface active agents such as anionic surface active agents, cationic surface active agents and amphoteric surface active agents, water-soluble polymer compounds such as xanthan gum, sodium alginate, methyl cellulose, hydroxyethyl cellulose, carboxyvinyl polymer and the like can be used. Also, dyes, preservatives,
Fragrances and the like may be blended as necessary.

The amount of morphine or its derivative (1) to be incorporated in the preparation is not particularly limited and may be appropriately selected within a wide range.
It is preferably 30% by weight.

The administration method of the above-mentioned preparation is not particularly limited, and the preparation is administered according to various preparation forms, age and sex of the patient, other conditions, and degree of the patient. For example, tablets, pills, solutions, suspensions, emulsions, granules and capsules are orally administered. In the case of injection, it is intravenously administered alone or in admixture with a normal replacement fluid such as glucose or amino acid, and further, if necessary, is intramuscularly, intradermally, subcutaneously or intraperitoneally administered alone. In the case of suppositories, they are administered rectally. In the case of a spray, it is sprayed from the mouth or nose and administered to the bronchi. In the case of ointment, it is applied directly to the lesion site.

[0027]

The present invention will be described in more detail with reference to Examples, Production Examples and Test Examples, but the present invention is not limited to these.

Production Example 1 Dry madder (weight 1 kg) is crushed and extracted with 1 l of methanol. The extract was concentrated under reduced pressure and then liquid-liquid partitioned with ethyl acetate-water. The obtained organic layer was concentrated under reduced pressure and further hexane-9.
Partition with 0% methanol. The hexane layer was concentrated to dryness (16 g) and silica gel column chromatography (Wa).
kogel C-200) and 250 mg of morgin (R ¹ ＯOCH ₃ , R ² ＯＨOH in the general formula (1))
I got

Test Example 1 Rat Basophilic Leukemia Cell Line (Rat Basophi)
lic Leukemia Cell: RBL-1) is suspended in D-PBS (+). 2 × 10 ⁵ cells of each tube were placed in a plastic tube, incubated with various concentrations of morphine (Production Example 1) at 37 ° C. for 15 minutes, and then calcium ionophore A23187 (final concentration 2.5 μM) was added. After incubating for another 15 minutes, EDTA (final concentration 4 mM) was added and ice-cooled,
After centrifugation at 2000 rpm for 5 minutes, the supernatant was collected. Next, leukotriene C ₄ (LTC ₄ ) contained in the supernatant was quantified using a radioimmunoassay method, and its production inhibitory effect was evaluated. Table 1 shows the results.

[0030]

[Table 1]

From the results shown in Table 1, it was confirmed that morphine has an excellent lipoxygenase inhibitory action.

Test Example 2 Normal human fibroblasts were suspended in DMEM medium containing 10% FCS, seeded on a 12-well culture plate, and when confluent, the medium was replaced with serum-free DMEM medium. 24 hours later, the medium was replaced with a medium containing morphine (Production Example 1), and after incubation for 30 minutes, interleukin 1α (1
Add 0 unit / ml) and incubate for another 6 hours. After 6 hours, the medium was collected and the amount of prostaglandin E ₂ (PGE ₂ ) released in the medium was quantified by an enzyme immunoassay.
The results are shown in Table 2.

[0033]

[Table 2]

From the results shown in Table 2, it was confirmed that morphine has an excellent cyclooxygenase inhibitory action.

Test Example 3 The back of a Hartley white guinea pig was shaved to 0.2
% Croton oil was applied to induce inflammation. Inflammation induction 2
Before and after 6 hours, 25 μl / (1.5) of an ethanol solution (100 mM) of morphine (Production Example 1) was used as a sample.
(cm × 1.5 cm) was applied, and 24 hours after the induction of inflammation, the degree of erythema was visually determined. The judgment was made according to the following Japanese Dermatological Association standards. Table 3 shows the results.

Japanese Dermatological Association Standard 0 (−) No reaction. 0.5 (±) Mild or partial erythema. 1.0 (+) Clear erythema on the whole surface. 2.0 (++) Erythema and edema. 3.0 (+++) Erythema, edema and small blisters.

[0037]

[Table 3]

From the results shown in Table 3, the excellent anti-inflammatory effect of morphine in the croton oil-induced inflammation model was confirmed.

Test Example 4 The back of a Hartley white guinea pig was shaved and a 100 mM ethanol solution of morphine (Production Example 1) was prepared (sample).
25 μl / (1.5 cm × 1.5 cm) was applied. After 2 hours, wipe the sample with 70% ethanol and UVB
(1.7 mW / cm ² × 9 minutes) was irradiated, and immediately after that, 25 μl / (1.5 cm × 1.5 cm) of the sample was applied. UV
After 6 hours of B irradiation, 25 μl of the sample was applied again. At 24 hours after irradiation, the degree of erythema was evaluated in the same manner as in Test Example 3. The results are shown in Table 4.

[0040]

[Table 4]

From the results shown in Table 4, the excellent anti-inflammatory effect of morphine in the UV inflammation model was confirmed.

Example 1 According to the following formulation, the respective components were uniformly mixed and compression-molded with a tableting machine to obtain 200 mg tablets.

[0043]

[Table 5] Morgin (Production Example 1) 10 g Corn starch 30 g Starch 30 g Carboxymethyl cellulose 5 g Magnesium stearate 5 g Lactose 20 g Total 100 g

Example 2 According to the following formulation, the respective components were uniformly mixed and blended. After granulating with an extrusion granulator, it was dried and sieved to obtain granules.

[0045]

[Table 6] Morgin (Production Example 1) 10 g Crystalline cellulose 50 g 10% Hydroxypropyl cellulose 40 g Ethanol solution total 100 g

Example 3 A propellant was prepared by filling a cylinder with the following composition by a conventional method.

[0047]

[Table 7] Morgin (Production Example 1) 1 g Oleic acid 3 g Freon 11 1.2 g Freon 12 2.5 g Freon 114 1.3 g Total 9 g

Example 4 The components were uniformly mixed according to the following formulation to obtain an ointment.

[0049]

[Table 8] Morgin (Production Example 1) 10 g Squalane 20 g Glycerin 20 g Cetyl alcohol 5 g Magnesium stearate 3 g Propylene glycol 5 g Water 20 g Ethanol 7 g Total 90 g

[0050]

The arachidonic acid metabolism inhibitor of the present invention has an excellent lipoxygenase and cyclooxygenase inhibitory action and is useful as an anti-inflammatory agent, an anti-allergic agent and the like. Therefore, it can be widely used for the prevention and treatment of bronchitis, asthma, allergic rhinitis, gout, arthritis, nephritis, hepatitis, psoriasis, urticaria, contact dermatitis, atopic dermatitis, UV inflammation (sunburn) and the like.

Claims (5)

[Claims] 1. A compound represented by the general formula (1): (In the formula, R ¹ represents a hydroxyl group or an alkoxy group, and R ² represents a hydroxyl group, an alkoxy group, or an alkoxycarbonyl group.) An arachidonic acid metabolism inhibitor containing morphine or a derivative thereof as an active ingredient. 2. An anti-allergic agent comprising the morphine or its derivative according to claim 1 as an active ingredient. 3. An anti-inflammatory agent comprising the morphine or its derivative according to claim 1 as an active ingredient. 4. The arachidonic acid metabolism inhibitor according to claim 1, which is a therapeutic or prophylactic agent for bronchitis, asthma, allergic rhinitis, gout, arthritis, nephritis or hepatitis. 5. The arachidonic acid metabolism inhibitor according to claim 1, which is a preventive or therapeutic agent for psoriasis, urticaria, contact dermatitis, atopic dermatitis or UV inflammation (sunburn).