JPH0713010B2 - Anti-allergic drug - Google Patents

Description

TECHNICAL FIELD The present invention relates to an antiallergic drug containing a dihydrochalcone compound as an active ingredient.

(Background of the Invention) In recent years, the involvement of prostaglandins (PG) and related substances in biological functions, particularly in the respiratory system, inflammation, and immunity has been revealed. PG and its related substances are made from arachidonic acid by a specific enzyme and constitute a metabolic system called arachidonic acid cascade. Therefore, it is also known that when arachidonic acid metabolism abnormality is caused, various diseases such as vascular system, respiratory system and immune system are caused.

PG is abundantly contained in the spermatic gland of sheep, and this discovery triggered this.
Six kinds of PGs of PGE ₁ , E ₂ , E ₃ , F ₁ α, F ₂ α and F ₃ α were extracted and isolated, and their chemical structures were clarified. These PG
Is a fat-soluble substance having physiological activities such as effects on contraction and relaxation of smooth muscle, effects on platelet aggregation, effects on adenylate cyclase, effects on gastric acid secretion, effects on reproductive physiology, etc. The highly potent PGE ₁ , E ₂ and F ₂ α have already been formulated, and are used as peripheral vasodilators, antithrombotic agents, and parturition promoters.

In addition, unstable endoperoxides (PGG ₂ and PGH ₂ ) were isolated, their chemical structures were elucidated, and the biosynthetic pathway of PG was elucidated. Furthermore, thromboxane A ₂ , which is known as a physiologically active substance, PG-related substances such as prostacyclin and leukotriene are all biosynthesized from arachidonic acid stored in the phospholipids of cell membranes together with the PG. It plays an extremely important role in regulating functions. Part of arachidonic acid is hydroperoxyeicosatetraenoic acid (Hydroperoxy-eicosatetraenoic acid) produced by lipoxygenase.
(HPETE) and then hydroxyeicosatetraenoic acid (Hyd
It is converted into a physiologically active substance called roxyeicosatetraenoic acid (HETE), and among them, 5-lipoxygenase forms a leukotriene group via 5-HPETE. Leukotriene is produced in the lungs during an asthma attack and has a strong contractile effect on the lungs and bronchial muscles, and also has a physiological role that is closely related to inflammatory reactions such as vascular permeability enhancement and leukocyte migration. It is also known that it is active and that inflamed tissues actually have leukotriene production activity. Thus, the relationship between leukotrienes and asthma and inflammation has been clarified. Therefore, it inhibits the leukotriene synthesis activity without inhibiting the synthesis of PG which also has peripheral circulation promoting action and arteriosclerosis preventing and treating actions.
Finding a specific inhibitor of lipoxygenase is an important factor leading to the causal therapy of allergic diseases such as asthma and inflammation.

From such a background, as a result of searching for compounds having 5-lipoxygenase inhibitory activity and leukotriene production inhibitory activity, the present inventors have found that dihydrochalcone compounds having the structural formulas described below are excellent in inhibiting 5-lipoxygenase activity. The inventors have newly found that they have physiological activity and completed the present invention.

(Object of the Invention) An object of the present invention is to provide a novel anti-allergic drug.

(Structure of Invention) The present invention is an antiallergic drug containing a specific dihydrochalcone compound as an active ingredient.

A dihydrochalcone compound which is an active ingredient of the present invention, 2-
Geranyl-3,4,2 ', 4'-tetrahydroxychalcone has the physical properties described below, and is a plant that grows naturally in various parts of Indonesia, the Artocarpus communis (Artocarpus C
ommunis) (Indonesia name Kulur) is a new compound extracted and isolated. An example of the extraction / isolation method is shown below.

(Extraction / isolation example) About 500 g of the flower part of the Indonesian medicinal plant Altcalps communis is extracted several times with 90% ethanol, and the extract is concentrated under reduced pressure to obtain about 20 g of an oily substance. Then, this oily substance is subjected to column chromatography packed with Diaion HP-20, and eluted successively with 40% water-containing methanol, 60% water-containing methanol, 80% water-containing methanol, methanol and acetone. Next, the methanol elution fraction and the acetone elution fraction were combined and concentrated under reduced pressure to obtain 15.7 g of a brown oily substance. This shows a remarkable growth-inhibitory effect on Yoshida sarcoma cultured cells.

Then, the above oily substance was further subjected to silica gel column chromatography (solvent: hexane / ethyl acetate = 10: 1 to 0:10).
0, methanol only), 11 fractions (AC-1: 2.0 g, A
C-2: 0.6g, AC-3: 0.8g, AC-4: 0.3g, AC-5: 1.0g, AC
−6: 3.0g, AC−7: 0.3g, AC−8: 2.4g, AC−9: 0.2g, AC−
10: 3.0g, AC-11: 1.0g). Each of the obtained fractions was examined for growth inhibitory effect on Yoshida sarcoma cultured cells, and among these fractions, the AC-5 and AC-6 fractions with strong activity (hexane: ethyl acetate = 5: 2) Is further fractionated by high performance liquid chromatography (Nucleosil50-5, solvent; hexane: ethyl acetate = 2: 1, flow rate 9 ml / min), and the peak around retention time 28 minutes is collected and concentrated to 2
-Geranyl-3,4,2 ', 4'-tetrahydroxychalcone (950 mg) was obtained.

The dihydrochalcone compound which is the active ingredient of the present invention has the following structural formula and physical properties.

2-geranyl-3,4,2 ', 4'-tetrahydroxychalcone (2-geranyl) -3,4,2', 4'-tetrahydroxychalco
ne) [Physical properties] MS: m / e410 (M ⁺ ), C ₂₅ H ₃₀ O ₅ IR (KBr): 3450 (OH), 1620 (hydrogen-bonded C = O) cm
^{-1 1} H-NMR (90 MHz, CDCl ₃ , δ): 1.56 (3H, bs), 1.65 (3
H, bs), 1.78 (3H, bs), 2.04 (4H, bs), 2.80-3.20 (4
H, m), 3.40 (2H, d, J = 6.6Hz), 4.88-5,30 (2H, m),
6.32 (1H, dd, J = 2.4,9.5Hz), 6.37 (1H, bs), 6.62,
6,73 (2H, ABq, J = 9.2Hz) ¹³ C-NMR (22.5MHz, CDCl ₃ , ε): 16.1 (q), 17.4
(Q), 25,4 (q), 26.3 (t), 27.7 (t), 39.4
(Two carbon, t), 58.6 (t), 103.4 (d), 108.1
(D), 112.9 (d), 113.3 (s), 121.0 (d), 12
1.8 (d), 123.7 (d), 126.3 (s), 131.3 (s),
131.7 (s), 132.2 (d), 137.7 (s), 142.2
(S), 142.4 (s), 163.2 (s), 164.8 (s), 20
4.3 (s) This compound has no toxicity even after continuous administration to mice at 50 mg / kg.

Next, the dose and administration method of the compound will be described. The drug of the present invention is administered as an oral administration agent such as tablets, capsules and aerosols, and as a parenteral administration agent such as injection (intravenous, subcutaneous), lumber for infusion, suppository preparation, etc. Although it may vary depending on the administration route, dosage form, number of administrations, etc., the effective amount when used for adult treatment is usually about 0.005 to 100 mg / kg of body weight of the above-mentioned compound as an active ingredient per day for oral administration. In the range of, preferably 0.2
〜50 mg / kg of body weight, more preferably 0.5 to 10 mg / kg of body weight is suitable, and in the case of parenteral administration, it is about 0.002 to 200 mg / kg of body weight per day as the compound of the active ingredient.
Preferably 0.02 to 100 mg / kg body weight, more preferably 0.2 to 2
0 mg / kg body weight is appropriate.

When the compound is to be prepared as a preparation for injection or drip, it may be provided in a unit dose ampoule or a dose container together with additives. The formulations may take such forms as suspensions, solutions, emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing or dispersing agents.

As the suspending agent, glucose, gelatin, aluminum stearate gel, etc., and as the stabilizing agent, lecithin,
Almond oil, coconut oil, refined sesame oil as non-aqueous vehicle such as linoleic acid and sorbitan monooleate
As the preservative such as oily ester, methyl P-hydroxybenzoate, sorbic acid and the like are used.

When the compound is to be prepared as a preparation for oral administration, it is preferably administered in a form suitable for absorption from the intestinal tract like a capsule.

In capsules for oral administration, binders such as gelatin,
Excipients such as sorbit, for example, lactose, sugar, starch, glycine, etc., stabilizers such as magnesium stearate, ascorbic acid, etc., and potato starch etc. as a disintegrating agent can be appropriately contained. In preparing an orally-administered preparation of the compound, a film can be further applied using a pharmaceutically acceptable enteric film-forming substance. As the film-forming substance, cellulose acetate phthalate (CAP) is used as a carbohydrate derivative such as cellulose and saccharide, and methyl acrylate / methacrylic acid copolymer is used as a polyvinyl derivative such as acrylic acid copolymer and dibasic acid monoester. Examples thereof include polymers and methyl methacrylate / methacrylic acid copolymers.

In order to improve the disintegration and dissolution of the active ingredient of the present invention, one or more kinds of surfactants such as alcohols, esters, polyethylene glycol derivatives, sorbitan fatty acid esters and the like can be added. In addition, as an excipient, for example, one or more kinds of sucrose, lactose starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium aluminometasilicate, calcium carbonate, sodium hydrogen carbonate and the like may be combined. It can be added. As the lubricant, for example, magnesium stearate, talc, hydrogenated oil, etc. may be used alone or in combination.
One or more can be added, and as a corrigent or flavoring agent, salt, saccharin, sugar, mannitol, orange oil,
Sweeteners such as licorice extract, citric acid, glucose, menthol, eucalyptus oil, malic acid, etc., flavors, colorants, preservatives and the like may be contained. Examples of adjuvants such as suspending agents and wetting agents may include coconut oil, olive oil, sesame oil, peanut oil, calcium lactate, safflower oil, soybean phospholipid, and the like.

When the drug of the present invention is orally administered, it is administered as a suspension, soft or hard capsule or tablet, granule, fine granule, powder, or aerosol, and when parenterally administered, it is solid or suspended. It is administered as a viscous liquid so that sustained mucosal absorption can be maintained.

Hereinafter, the present invention will be described with reference to formulation examples and test examples.

Formulation Example 1. (Injection / Drip) 5 g of powdered glucose is added so as to contain 500 mg of the above compound, and the mixture is aseptically distributed to a vial, which is sealed and stored. Before use, 500 ml of 0.85% physiological saline is added to give an intravenous injection, and 10 to 500 ml is administered by intraarterial injection or drip infusion depending on the condition for one day.

Formulation Example 2. (Tablets) 100 adult tablets were prepared with the following composition.

[A]

Compound 10g Lactose 9.94g Hydroxypropyl cellulose 0.06g Magnesium stearate 0.2g [B] Cellulose acetate phthalate 0.6g Hydroxypropylmethylcellulose phthalate 0.6g Take the ingredients of A and mix well and press directly or
Alternatively, after kneading well, granules were formed through a screen of an extrusion type granulator, sufficiently dried, and then pressed to prepare tablets.

Further, if necessary, the formed tablet is coated with a base material B well dissolved to prepare an enteric coated tablet.

Formulation Example 3. (Capsules) 100 adult capsules were prepared with the following ingredients.

[A]

Compound 10g Lactose 2.46g Hydroxypropylcellulose 0.04g [B] Cellulose acetate phthalate 1g Hydroxypropylmethylcellulose phthalate 1g Take the above component A, mix well, shape into granules according to a conventional method, and dry well After sieving to obtain granules suitable for capsules, the granules were filled into capsules to prepare capsules. Further, if necessary, the base material of B dissolved in this granule while floating and flowing is coated, and the granule is prepared as an enteric granule, which is filled in a capsule to form an enteric capsule.

Antiallergic Action Test Example 1. (5-Lipoxygenase Inhibitory Activity) 1) Assay for 5-Lipoxygenase Activity Masttocytoma P in Carcinomatous Mastocytoma
-815 cells were transferred to an in vitro suspension culture system and cloned to obtain a cell line with high 5-lipoxygenase activity, 2-E-6 strain [Kawamura, M. et al: Prostaglandins,
19 : 659 (1980)] and treated with n-butyric acid for 40 hours to give 5
-Remarkably increased lipoxygenase activity and PG synthase activity (Koshihara, Y. et al: J. Biol. Chem., 257, 730).
2, (1982)].

As shown in FIG. 1 (a), in the case of the malignant mastocytoma carcinoma, 12- rather than 5-lipoxygenase (5-HETE) activity was observed.
Has much higher lipoxygenase (12-HETE) activity,
As shown in FIG. 1 (b), the ratio is completely reversed in the 2-E-6 strain.

2) 5-lipoxygenase and PG synthase inhibitory activity: 10,000 × g supernatant of a homogenate of 50 mM phosphate buffer (pH 7.4), 1 mM EDTA and 0.1% gelatin obtained from the 2-E-6 strain treated with n-butyric acid. Was used as an enzyme source, and the drug effect of the compound on its 5-lipoxygenase activity was compared as a control when the compound was not added. The inhibitory effect is the number of counts (cp
m) and the control (no compound added) was set as 100. The results are shown below.

(Effects of the Invention) The antiallergic drug of the present invention exhibits extremely excellent 5-lipoxygenase selective inhibitory activity and has an excellent antiallergic action. That is, it is extremely useful as a powerful anti-allergic drug such as a therapeutic drug for asthma, anti-inflammatory drug, and immunostimulant, together with its safety.

[Brief description of drawings]

FIG. 1 shows mast cell carcinoma mast cytomas P-815.
(A) and its cloned cell culture cells 2-E-6 strain (b)
2 is a thin-layer chromatography radio spectrum showing the lipoxygenase activity of Escherichia coli.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Agustin Smartno, Republic of Indonesia South Jakarta Kuva Yoran Rama Kotak 2 Sinar Passer Jumuad Jalan (No address) (56) Reference JP 62-270544 (JP, A)

Claims (3)

[Claims] 1. A structural formula: An anti-allergic drug containing a dihydrochalcone compound represented by as an active ingredient. 2. The antiallergic drug according to claim 1, which is in a parenteral dosage form. 3. The antiallergic drug according to claim (1) in the form of oral dosage form.