JPH07116033B2 - Anti-allergic drug - Google Patents

Description

TECHNICAL FIELD The present invention relates to an antiallergic drug containing caffeic acid ester of quinic acid, caffeic acid sinapinic acid ester of quinic acid, or a mixture thereof 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 seminal vesicles of sheep, and with this discovery, six types of PG, PGE ₁ , E ₂ , E ₃ , F ₁ α, F ₂ α, and F ₃ α were first extracted and isolated. Separated, its chemical structure was also revealed. These PGs are lipid-soluble substances having physiological activities such as actions on smooth muscle contraction and relaxation, actions on platelet aggregation, actions on adenylate cyclase, actions on gastric acid secretion, actions on reproductive physiology, etc. Relatively strong PGE ₁ , E ₂ and F ₂ α etc. have already been formulated,
It is used as a peripheral vasodilator, antithrombotic drug, and parturition promoter.

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 (Hydroperxyeicosatetraenoic acid) (Hipoxygenase (Lipoxygenase)
Hydroxyeicosatetraenoic acid (Hydrox
It is converted to a physiologically active substance called yeicosatetraenoic 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 it actually has leukotriene biosynthetic activity in inflamed tissues. 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 a compound having 5-lipoxygenase inhibitory activity and leukotriene production inhibitory activity, the present inventors have found that caffeic acid ester of quinic acid or caffeic acid sinapinic acid ester of quinic acid is 5 −
The inventors have newly found that they have excellent physiological activity of inhibiting lipoxygenase 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 the Invention) The present invention is an anti-allergic drug containing quinic acid caffeic acid ester, quinic acid caffeic acid sinapinic acid ester, or a mixture thereof as an active ingredient.

Examples of the quinic acid ester which is the active ingredient compound of the present invention include the following compounds.

(1) 3-O-caffeoyl-4-O-sinapoylquinic acid (2) 3,5-O-dicaffeoylquinic acid (3) 4,5-O-dicaffeoylquinic acid (4) 3,4 —O-Dicaffeoylquinic acid The structural formula is:

The compound (1) which is an active ingredient of the present invention is a novel compound which has the physical properties described below and is extracted and isolated from Yamajiko, which is a medicinal plant produced in China, Taiwan and the like. An example of the extraction / isolation method is shown below.

(Extraction / isolation example) Grinded Yamajiko (3.7 kg) with 50% water-containing acetone (7)
Extract at room temperature for 12 hours. After separating the extract, the residue is similarly extracted twice with 50% water-containing acetone (3) twice. After the extracts were combined and concentrated to about 1.5, n-hexane (11 x
3 times), ethyl acetate (1 × 3 times) and then n-butanol (1 × 3 times), and a hexane soluble portion (2.3 g),
Ethyl acetate soluble part (18.9g), butanol soluble part (182.1
g) and a water-soluble part (445.5 g). Dissolve 171 g of butanol-soluble part in 200 ml of water, and then use Diaion H
It was mounted on a column of P-20 (8.5φ × 27 cm) and fractionated by the stepwise gradient elution method of water and methanol as follows.

Fractionation Eluent Volume () Fractional weight (g) 1 Water 3 82.6 2 10% Methanol 3 25.4 3 20% Methanol 3 13.6 4 40% Methanol 3 17.9 Fractionation Eluent Volume () Fractional weight (g) 5 60% Methanol 4 24.3 6 80% Methanol 3 16.8 7 Methanol 4 6.8 80% Methanol Elution fraction (10.1 g) was further analyzed by column chromatography using Diaion HP-40 (4.5φ × 20 cm) as shown below. Was fractionated as follows.

Fractionation Eluent Volume (ml) Fraction weight (g) 1 Water 300 0 2 60% Methanol 400 2.22 3 65% Methanol 400 3.34 4 70% Methanol 400 2.23 5 75% Methanol 400 1.34 6 80% Methanol 400 0.67 7 85% methanol 400 0.25 8 90% methanol 500 0.06 9 methanol 500 0.02 Of these, the fractions eluted with 60% and 65% methanol were Fuji gel.
Preparative high performance liquid chromatography (HPL) using RQ-2
Repeat the fractionation with C) [eluent: acetonitrile (20-3
0%)-water-acetic acid (1%)], (1) 256 mg was isolated.

High performance liquid chromatography (column: TSKODS SOTM
[4.6φ × 250 mm], eluent; acetonitrile: water: formic acid = 32: 68: 1, flow rate 1.2 ml / min, detection 330 nm), as a result of analysis, (1) is Fraction 3 of Diaion HP-40 and acetic acid. Folch partition of ethyl layer (chloroform: methanol: water = 8:
4: 3) was also detected in the water layer. [Physical Properties of Compound (1)] Shape: Light yellow amorphous powder Mass spectrometry: FD-MS m / z 560 (M -) ¹ H-NMR (acetone-d ₆ -D ₂ O, 270 MHz) δ: 7.61 (1H, d, J = 15.9), 7.54 (1H, d, J = 15.9), 7.13
(1H, d, J = 2.0), 7.01 (1H, dd, J = 2.0,8.3), 6.98 (2
H, s), 6.84 (1H, d, J = 8.3), 6.40 (1H, d, J = 15.9),
6.22 (1H, d, J = 15.9), 5.68 (1H, ddd, J = 4.8,9.7,10.
6), 5.16 (1H, dd, J = 2.9,9.7), 4.5 (1H, br, d, J = 2.
9), 3.89 (6H, s), 2.39 (1H, ddd, J = 2.5,4.8,13.
2), 2.29 (2H, m), 2.18 (1H, dd, J = 11.0,13.2). ¹³ C-NMR (acetone-d ₆ -D ₂ O, 22.5 MHz) δ: 167.7, 167.4, 163.9, 155.1, 152.9, 149.4 (2C), 14
7.2, 147.0, 146.8, 127.7, 126.5, 123.1, 116.7, 11
6.2, 115.2 (2C), 107.4 (2C), 78.1, 76.1, 69.7, 6
8.8, 57.2 (2C), 40.3, 38.5 Compounds (2), (3) and (4) are compounds extracted and isolated from coffee beans (Phytochemisery.196).
5, Vol. 4, pp 527-529).

It should be noted that the compounds (1) to (4) used in the present invention have 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 agent 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), infusion preparation, suppository preparation and the like.
The effective amount may vary depending on the symptoms, administration route, dosage form, number of administrations and the like. When used for treating an adult, the effective dose is usually 0.005 to 100 mg / kg of body weight of the above-mentioned compound as an active ingredient, 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. Further, as an excipient, for example, one or more kinds of sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium metasilicate aluminate, calcium carbonate, sodium hydrogen carbonate, etc. are combined. Can be added. As the lubricant, for example, magnesium stearate, talc, hydrogenated oil and the like can be added alone or in combination of two or more, and as a corrigent or flavoring agent, salt, saccharin, sugar, mannitol, orange oil, licorice Sweeteners such as extracts, citric acid, glucose, menthol, eucalyptus oil, malic acid, flavors, colorants,
Preservatives and the like may be included. 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 compound (1) or (2), and the mixture is aseptically distributed to a vial, and 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 intravenous injection or infusion per day depending on the symptoms.

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

[A]

Compound (3) 10 g Lactose 9.94 g Hydroxypropyl cellulose 0.06 g Magnesium stearate 0.2 g [B] Cellulose acetate phthalate 0.6 g Hydroxypropylmethylcellulose phthalate 0.6 g Take the ingredients of A, 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 (4) 10 g Lactose 2.46 g Hydroxypropyl cellulose 0.04 g [B] Cellulose acetate phthalate 1 g Hydroxypropylmethyl cellulose phthalate 1 g Take the above component A, mix well, and mold it into granules according to the usual method. It was thoroughly dried and sieved to give granules suitable for capsules, which 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.

Anti-allergic action Test example 1. (5-lipoxygenase inhibitory activity) 1) Assay of 5-lipoxygenase activity Mastocytoma P of cancerous 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 [Koshihara, Y. et al: Biochim. Biophy.
s. Acta, 712, 42 (1982)] was treated with n-butyric acid for 40 hours to significantly increase 5-lipoxygenase activity and PG synthase activity [Koshihara, Y. et al: J. Biol. Chem., 2
57, 7302, (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 activity Using 10,000 mM 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 as an enzyme source , The efficacy of the compound on its 5-lipoxygenase activity was compared as a control without the addition of the compound. 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 anti-allergic drug of the present invention exhibits extremely excellent 5-lipoxygenase inhibitory activity and has an excellent leukotriene production inhibitory action. That is, it is extremely useful as a powerful asthma therapeutic agent, anti-inflammatory drug, anti-allergic drug such as immunostimulant, etc. in combination 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 (56) References J. Nat. Prod. , 50 (3), p. 392-9 (1987)

Claims (3)

[Claims] 1. An anti-allergic agent comprising a caffeic acid ester of quinic acid or a caffeic acid sinapinic acid ester of quinic acid 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.