JPH09157206A - Novel catechol derivative and 5-lipoxygenase inhibitor containing the same as effective component - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new catechol derivative having an inhibiting action against 5-lipoxygenase and useful for curing or prevention of allergic disease such as bronchial asthma or circulatory disease such as edema.

SOLUTION: As this new catechol derivative, the one expressed by the formula (R is H, a 1-10C alkyl or a 2-20C aliphatic, etc.; exhibits C-C single bond or double bond) e.g. 1,7-bis-(3,4-dihydroxyphenyl)-hepta-4,6-diene-3-on is exemplified. The derivative of the formula is obtained by drying whole herb of a native plant at Szechwan Province in China, Aletris glabra Bur. et Franch. (Liliaceae), crushing, extracting with 95% ethanol, distilling off the solvent, separating and purifying the resultant crude extract by using a separation method such as column chromatography. The compound of the formula suppresses generation of a lipoxygenase metabolite such as leukotrienes.

COPYRIGHT: (C)1997,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel catechol derivative and a salt thereof and an active ingredient thereof.
It relates to lipoxygenase inhibitors.

[0002]

2. Description of the Related Art In vivo, 5-lipoxygenase produces 5-hydroperoxyeicosatetraenoic acid from arachidonic acid, and various leukotrienes are biosynthesized using this compound as an intermediate. Are known. Among them, it has been elucidated that leukotriene B4 has a strong leukocyte chemotactic activity and is a mediator of inflammation, and that leukotriene C4 and leukotriene D4 are allergic factors.

Accordingly, compounds having an inhibitory action on 5-lipoxygenase are various diseases caused by lipoxygenase metabolites such as leukotrienes, for example, allergic diseases such as bronchial asthma, nasal allergy, ocular inflammation and atopic dermatitis. In addition, the effects of preventing and treating cardiovascular diseases such as edema, ischemic disease, hypertension, and ischemic encephalopathy are expected.

So far, the welcome company BW-755C
And Merck L-651896, Ono Yakuhin ONO-L
Nearly 200 5-lipoxygenase inhibitors, such as P-219 and Terumo TMK-688, have been developed, but all have problems with respect to toxicity, metabolism, absorption, etc., and most of them have stopped development. .

[0005]

In view of the above circumstances, the present inventors have proceeded with the extraction, separation and purification of Chinese medicinal plants using the 5-lipoxygenase inhibitory action as an index to develop a novel catechol derivative having a strong 5-lipoxygenase inhibitory action. The present invention has been completed and the present invention has been completed.

[0006]

DISCLOSURE OF THE INVENTION It is an object of the present invention to provide a novel catechol derivative and its salt, and a 5-lipoxygenase inhibitor containing these as active ingredients.

That is, according to the present invention, the formula (I)

Embedded image (R is hydrogen, a linear or branched alkyl group having 1 to 10 carbon atoms, preferably a lower alkyl group having 1 to 4 carbon atoms such as methyl or ethyl, or 2 to 1 carbon atoms.
0 aliphatic or aromatic acyl group, preferably an acetyl, a lower aliphatic acyl group such as propionyl, ...
Represents a carbon-carbon single bond or a double bond).

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The compound of the present invention is a plant that grows mainly in Sichuan Province, China.
Bur. Et Franch. (Liliaceae)]. The whole plant is dried, crushed, then extracted with 95% ethanol, the solvent is distilled off, and the obtained crude extract is separated and purified by an appropriate separation method such as column chromatography to obtain the formula ( The compounds of I) can be obtained. The structure of the thus obtained novel catechol derivative of the formula (I), which is the physiologically active substance of the present invention, was determined by the analysis results such as spectroscopy.

In order to use the compound of the present invention as a medicine, the compound of the present invention is administered as it is or as a pharmaceutical preparation to which a pharmaceutically acceptable additive is added. Alternatively, it is administered as a mixture thereof, an extract or extract of hairless powdered sorghum containing them (extracts from which the extraction solvent has been removed).

Examples of the pharmacologically acceptable salt in the present invention include salts with inorganic bases, salts with organic bases, salts with amino acids and the like. Inorganic bases include alkali metals such as lithium, sodium, potassium and the like and alkaline earths such as magnesium, calcium and the like. However, other metals such as aluminum, zinc, iron and the like are also included in the present invention. Examples of the grouped base include amines such as ammonia, primary amine, secondary amine, and 3
And secondary amines and nitrogen-containing heterocycles (including aliphatic and aromatic). More specifically, methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, propylamine, dipropylamine, isopropylamine, diisopropylamine,
Butylamine, dibutylamine, isobutylamine, t
-Butylamine, monoethanolamine, diethanolamine, triethanolamine, pyrrolidine, piperidine, morpholine, pyrrole, pyridine and the like. Amino acids include lysine, arginine, histidine and the like.

Since the present invention has an inhibitory action on 5-lipoxygenase, it suppresses the production of lipoxygenase metabolites such as leukotrienes, and specifically, it is specifically mammalian (eg, human, dog, cat, rabbit, rat). , Mice, etc.), allergic diseases such as bronchial asthma, nasal allergy, ocular inflammation, atopic dermatitis, etc. caused by these metabolites, and circulatory organs such as edema, ischemic disease, hypertension, ischemic brain injury, etc. It can be used for the treatment and prevention of system diseases.

The 5-lipoxygenase inhibitor of the present invention is
It can be administered orally or parenterally. Dosage forms include tablets, pellets, capsules, granules,
Emulsifiers, suspensions, injections and the like can be mentioned. These dosage forms are produced according to a standard method using appropriate carriers, excipients and other additives as necessary.

The daily dose of the catechol derivative represented by the formula (I) depends on the condition and weight of the patient, the kind of the derivative,
For example, parenterally, subcutaneously, intravenously, intramuscularly or intrarectally from about 0.01 to
20 mg / kg / day, preferably about 0.1-10 mg /
Administer kg / day. As an oral agent, about 0.1-100
It is desirable to administer mg / kg / day, preferably about 1-10 mg / kg / day.

[0014]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.

Example 1 1,7-Bis- (3,4-dihydroxyphenyl) -hepta-4,6-dien-3-one:

Embedded image Hairless powdered sugar beet collected in Sichuan, China [Aletris glab
ra Bur. et Franch. (Liliaceae)] is dried, crushed, extracted with 95% ethanol at room temperature, and the obtained extract is concentrated under reduced pressure at 60 ° C. or lower. 8.0 g of this extract was subjected to centrifugal liquid-liquid partition chromatography (CPC) (manufactured by Miki Engineering Co., Ltd.), and the composition ratio of chloroform-methanol-isopropanol-water was 7: 6 :.
Separation was performed using a mixed biphasic solvent system of 2: 5. Merck thin-phase chromatography plate (Silica Gel 6
Each component was fractionated by 0 F 254) and the inhibitory activity was evaluated for each. Active fraction yield 164.7m
g, RF = 0.3-1.0 (chloroform-methanol-isopropanol-water, 4: 3: 1: 2).

This fraction was further added with 22 g of silica gel,
Eluent chloroform-methanol-water, 100: 10:
Separation by chromatography step gradient from 1 to 40: 10: 1 to 20: 10: 1 to methanol. Active fraction yield 26.0 mg, RF = 0.4 (chloroform-methanol-water, 70: 10: 1).

This fraction was further chromatographed on 16 grams of silica gel with chloroform-methanol-water, 20: 1: 0 to 10: 1: 0 to 70: 1.
Separation with a 0: 1 step gradient isolated 16.6 mg pure 5-lipoxygenase inhibitory active ingredient. Yield 16.6 mg, RF = 0.36 (chloroform-methanol, 10: 1).

The active ingredient was found to be a novel catechol derivative having the title structure based on the following spectroscopy. Proton Nuclear Magnetic Resonance Spectra The proton nuclear magnetic resonance spectra measured in deuterated chloroform at 500 MHz using tetramethylsilane as an internal standard are as follows. d 2.78 (m, 2H), 2.86 (m, 2H),
6.22 (d, 1H, J = 15.6), 6.53 (dd, 1H, J = 8.1, 2.1Hz),
6.65 (d, 1H, J = 2.1Hz), 6.67 (d, 1H, J = 8.1Hz), 6.75
(d, 1H, J = 7.9Hz), 6.75 (dd, 1H, J = 15.6, 11.0Hz), 6.
88 (dd, 1H, J = 7.9, 2.1Hz), 6.89 (d, 1H, J = 15.6Hz),
6.99 (d, 1H, J = 2.1Hz), 7.35 (dd, 1H, J = 15.6, 11.0Hz)
ppm carbon-13 nuclear magnetic resonance spectrum The carbon-13 nuclear magnetic resonance spectrum measured at 125.8 MHz using tetramethylsilane as an internal standard in deuterated chloroform is as follows.

D 31.2 (t), 43.2 (t), 114.7 (d), 116.4
(d), 116.6 (d), 116.6 (d), 120.7 (d), 121.9 (d), 125.0
(d), 128.8 (d), 129.9 (s), 134.2 (s), 144.0 (d), 144.6
(s), 146.2 (d), 146.2 (s), 146.7 (s), 148.5 (s), 203.0
(s), ppm

Example 2 1,7-Bis- (3,4-dihydroxyphenyl) -hepta
-1, 4, 6- Triene-3-on:

Embedded image Using 39.2 g of the hairless powdered sugar beet extract, crude fractionation was carried out by CPC under the same conditions as in Example 1 to obtain 7 fractions A to C and a to d, and each fraction was evaluated. . Active fraction (Fr.B)
Yield 1.32g, Rf = 0.3 ~ 1.0 (chloroform-methanol
-Isopropanol-water, 4: 3: 1: 2, organic layer).

This fraction was treated with 75 g of silica gel, eluent chloroform-methanol-water, 100: 10: 1 to 60: 10: 1 to 40:10:
Separation was performed by chromatography with a step gradient of 1 to 20: 10: 1, and 15 fractions of A to O were separated and evaluated. Yield of active fraction (Fr.F): 78.2 mg, Rf = 0.3 (chloroform-methanol-water, 70: 10: 1).

This fraction was chromatographed on 16 g of silica gel and eluted with a step gradient of hexane-ethyl acetate 1: 1 to 1: 2 to 0: 1 to methanol.
It was separated into four D fractions and evaluated. Yield of active fraction (Fr.D) 43.1 mg, Rf = 0.35 (chloroform-methanol-water, 70: 10: 1).

This fraction was further purified by a medium pressure column (size B) with a step gradient of eluent methanol-water, 1: 1 to 1: 0, and as fraction D, 5-lipoxygenase inhibitory active ingredient was 11.9 mg pure. Isolated on.
Yield of active ingredient (Fr.D) 11.9mg, Rf = 0.38 (reverse phase, methanol-water, 3: 2)

The chemical structure of this active ingredient is the novel catechol derivative 1,7-bis-
It was revealed to be (3,4-dihydroxyphenyl) -hepta-1,4,6-trien-3-one. Mass spectrometry (EI-MS method) calcd for C19H16O5; 324.0998 found 324.1024 Proton nuclear magnetic resonance spectrum The proton nuclear magnetic resonance spectrum measured at 500 MHz in tetrachlorosilane as an internal standard in deuterated chloroform is as follows. d 6.64 (d, 1H, J = 15.1Hz), 6.7
6 (d, 1H, J = 8.4Hz), 6.80 (d, 1H, J = 8.3Hz), 6.86 (dd,
1H, J = 15.3, 10.7Hz), 6.91 (d, 1H, J = 15.8Hz), 6.91 (d
d, 1H, J = 8.4, 2.1Hz), 6.95 (d, 1H, J = 15.3Hz), 7.02
(d, 1H, J = 2.1Hz), 7.04 (dd, 1H, J = 8.3, 2.1Hz), 7.12
(d, 1H, J = 2.1Hz), 7.52 (dd, 1H, J = 15.1, 10.1Hz), 7.
59 (d, 1H, J = 15.8Hz) ppm Carbon-13 nuclear magnetic resonance spectrum The carbon-13 nuclear magnetic resonance spectrum measured at 125.8 MHz using tetramethylsilane as an internal standard in deuterated chloroform is as follows. is there. d 114.8 (d), 115.7 (d), 116.
7 (d), 116.8 (d), 122.0 (d), 123.6 (d), 123.7 (d), 125.5
(d), 128.5 (s), 128.5 (d), 130.1 (s), 144.2 (d), 145.7
(d), 146.1 (d), 146.9 (s), 147.1 (s), 148.6 (s), 150.2
(s), 191.9 (s) ppm

Example 3 5-Lipoxygenase Inhibition Experiment: This test was carried out to investigate the 5-lipoxygenase inhibitory action of the compounds of the present invention. 1) Preparation of enzyme solution Rat basophilic leukemia cells (Rat Basophil)
ic Leukemia Cell: RBL-1. ATC
C CRL 1378) was cultured in Eagle medium containing 10% inactivated fetal bovine serum. After washing the cells 3 times with PBS, 0.1 M HEPES containing 1 mM EDTA,
The cells were suspended in pH 7.0 at a rate of 1 × 10 7 cells / ml. The cells were disrupted by ultrasonic waves and then centrifuged at 13,000 rpm for 60 minutes, and the supernatant was used as an enzyme solution. 2) Method for measuring enzyme activity The enzyme solution (54 μl) was added with 1 mM EDTA, 2 mM calcium chloride, and a test substance (dissolved in DMSO).
Add to 1M HEPES, pH 7.0 and bring the total volume to 12
0 μl was used. 0.4m after standing at 37 ℃ for 10 minutes
6 μl of g / ml arachidonic acid (methanol solution) was added and reacted at 37 ° C. for 60 minutes. Acetonitrile 1
The reaction was stopped by adding 20 μl, 13,000 rpm
It was centrifuged for 10 minutes. The 5-hydroxyeicosatetraenoic acid contained in the supernatant was detected by reverse phase high performance liquid chromatography.

As a result, the concentration at which the action of 5-lipoxygenase was inhibited by 50% (IC ₅₀ ) was 1,7-bis-.
(3,4-dihydroxyphenyl) -hepta-4,6-
Dien-3-one with 70 nM 1,7-bis- (3,4-
Dihydroxyphenyl) -hepta-1,4,6-triene-
3-on was 43 nM.

[0027]

The novel catechol derivative of the present invention is
It has a lipoxygenase inhibitory action and is useful as an anti-allergic agent, anti-asthma agent, anti-inflammatory agent and the like.

Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C07C 49/255 9049-4H C07C 49/255 B 69/22 69/22 69/76 69/76 Z (72) Inventor Norio Kawabe 1111 Tehiro, Kamakura City, Kanagawa Prefecture, In the Basic Research Laboratory, Toray Co., Ltd. (72) Inventor Yasuhiko Togami, 1111 Tehiro, Kamakura City, Kanagawa Prefecture, In the Basic Research Laboratory, Toray Co., Ltd. (72) Inventor, Chihiro Ishizuka Kamakura, Kanagawa 1111 Tohiro, Toray Co., Ltd. Basic Research Institute (72) Inventor Yu Wen-Sheng, 4th Section, Chengdu Road, Chengdu, Sichuan Province, China No. 9 (72) Inventor, Hu Xiao-Huang, 4th Section, Chengdu City, Sichuan Province, China

Claims (4)

[Claims] 1. A compound of the formula (I) (R represents hydrogen, an alkyl group having 1 to 10 carbon atoms which may be branched or branched, or an aliphatic or aromatic acyl group having 2 to 10 carbon atoms, and ... Is a carbon-carbon single bond or a double bond. Represents a heavy bond), or a pharmaceutically acceptable salt thereof. 2. A medicine comprising the novel catechol derivative according to claim 1 or a pharmacologically acceptable salt thereof. 3. A 5-lipoxygenase inhibitor comprising the novel catechol derivative according to claim 1 or a pharmacologically acceptable salt thereof as an active ingredient. 4. The therapeutic agent according to claim 3, wherein the 5-lipoxygenase inhibitor is an anti-allergic drug, an anti-asthma drug or an anti-inflammatory drug.