JPH10251246A - New chromene derivative and 5-lipoxygenase inhibitor containing the same derivative as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new compound having 5-lipoxygenase inhibiting action and useful as an antiallergic agent, an antiasthmatic agent, an anti- inflammatory agent, etc. SOLUTION: This compound is represented by the formula (R1 to R4 are each an alkyl, an alkenyl, an alkynyl, an aryl or an acyl and R4 represents silyl; X and Y are each oxygen, sulfur or nitrogen; Z is H, an alkyl or a silyl; X exists only when X is nitrogen; two lines comprising full line and dotted line are each a single bond or double bond), e.g. 2,7-dimethyl-5-hydroxy-2-(4- methyl-4-hydroperoxypento-2-enyl)chromene. The compound of the formula is contained in Rhododendron cephaluntum which is a plant mainly naturally grown in Shisen Province, China and is obtained by drying and pulverizing the whole grass of the plant, extracting the resultant powder with 95% ethanol, distilling the solvent and separating and purifying the resultant crude extract with column chromatography, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to novel chromene derivatives and salts thereof, and 5-lipoxygenase inhibitors containing these as an active ingredient.

[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. Up to now, Welcome BW-755
C and Merck L-651896, Ono Pharmaceutical ONO-
Nearly 200 types of 5-lipoxygenase inhibitors such as LP-219 and Terumo's TMK-688 have been developed, but all have been discontinued due to toxicity problems, poor absorption, and rapid metabolism. There are only a few compounds under development.

[0004]

DISCLOSURE OF THE INVENTION In view of the above circumstances, the present inventors have promoted the extraction, separation and purification of Chinese medicinal plants using 5-lipoxygenase inhibitory activity as an index, and have developed a novel chromene derivative having a strong 5-lipoxygenase inhibitory activity. And completed the present invention.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel chromene derivative, a salt thereof, and a 5-lipoxygenase inhibitor containing these as an active ingredient. That is, according to the present invention, the following formula (I)

[0006]

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(Wherein, R _{1 to} R ₄ are each independently H, an alkyl group having 1 to 6 carbon atoms having no substituent, an alkyl group having 1 to 6 carbon atoms having a substituent, and a substituent. Alkenyl group having 1 to 6 carbon atoms, alkenyl group having 1 to 12 carbon atoms having a substituent, 1 to 6 carbon atoms having no substituent
6, an alkynyl group having 1 to 12 carbon atoms having a substituent, an aryl group having 1 to 6 carbon atoms having no substituent, an aryl group having 2 to 12 carbon atoms having a substituent, not having a substituent An acyl group having 1 to 6 carbon atoms, an acyl group having 1 to 12 carbon atoms having a substituent, R ₄ also represents a silyl group, and X and Y each independently represent an oxygen atom, a sulfur atom, or a nitrogen atom. , Z represents a hydrogen, an alkyl group, or a silyl group when X is a nitrogen atom, and when X is an oxygen atom or a sulfur atom, Z does not exist, and a double line consisting of a solid line and a dotted line is a carbon-carbon A chromene derivative represented by a single bond or a double bond).

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The compound of the present invention is mainly known as a plant that grows naturally in Sichuan Province, China, such as [Cib035; Rhododendro].
n cephaluntum]. The whole plant is dried, pulverized, extracted with 95% ethanol, and the crude extract obtained by distilling off the solvent is separated and purified using an appropriate separation method such as column chromatography to obtain the following compound. (I)

[0009]

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Wherein R _{1 to} R ₄ are each independently H,
C1-C6 alkyl group having no substituent, C1-C6 alkyl group having a substituent, C1-C6 alkenyl group having no substituent, C1-C6 having a substituent
12 alkenyl groups, unsubstituted C1-6 alkynyl groups, substituted C1-12 alkynyl groups, unsubstituted C1-6 aryl groups,
An aryl group having 2 to 12 carbon atoms having a substituent, an acyl group having 1 to 6 carbon atoms having no substituent, an acyl group having 1 to 12 carbon atoms having a substituent, and R ₄ also represents a silyl group;

As the substituent, halogen, alkoxy,
Hydroxy, alkylthio, amino, nitro, nitrile, formyl, carbamoyl, alkoxylcarbonyl, carboxy; X and Y each independently represent an oxygen atom, a sulfur atom, or a nitrogen atom; When X is an oxygen atom or a sulfur atom, Z does not exist, and a double line consisting of a solid line and a dotted line represents a carbon-carbon single bond or a double bond. The chromene derivative shown can be obtained.

The structure of the thus obtained novel chromene derivative of the formula (I), which is the physiologically active substance of the present invention, was determined by analysis results such as spectroscopy. In order to use the compound of the present invention as a pharmaceutical, 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).

In the present invention, pharmacologically acceptable salts 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. As the organic base, amines such as ammonia, primary amine, secondary amine,
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 inhibits the production of lipoxygenase metabolites such as leukotrienes, and specifically, can be used for mammals (eg, human, dog, cat, rabbit, rat) , Mice, etc.), allergic diseases such as bronchial asthma, nasal allergy, ocular inflammation and atopic dermatitis caused by these metabolites, and circulatory organs such as edema, ischemic disease, hypertension and ischemic encephalopathy It can be used for treatment and prevention of systemic diseases.

The 5-lipoxygenase inhibitor of the present invention comprises
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 chromene derivative represented by the above formula (I) varies depending on the condition and weight of the patient, the type of the derivative, the administration route and the like. About 0.01 in the muscle, rectum
-20 mg / kg / day, preferably about 0.1 / 10 mg / kg / day
Daily. As an oral preparation, about 0.1 to 100 mg / kg
Per day, preferably about 1-10 mg / kg / day.

[0018]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, but it should not be construed that the present invention is limited thereto. Embodiment 1 FIG. 2,7-dimethyl-5-hydroxy-2-
(4-Methyl-4-hydroperoxypent-2-enyl
Le) Chromene (CIB035 (2) -3AAPDEB)
Isolation

[0019]

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The hair throat collected in Sichuan Province, China [CIB0
35; Rhododendron ce phaluntum] is dried, pulverized, extracted with 95% ethanol at room temperature, and the obtained extract is concentrated under reduced pressure at 60 ° C or lower. 25 g of this extract was partitioned between water and ethyl acetate, and the organic layer (Fr. A) was separated by centrifugal liquid-liquid chromatography (CPC) (III) using a solvent system having a chloroform-methanol-water mixture ratio of 4: 3: 2. According to Oni Engineering Co., Ltd.), the composition ratio of chloroform-methanol-isopropanol-water is 7: 6: 2: 5.
Using a mixed two-phase solvent system.

Each component was fractionated using a thin-phase chromatography plate (Silica Gel 60F254) manufactured by Merck, and the inhibitory activity was evaluated for each component. Active fraction (Fr.AA) yield 5.94 g, Rf
= 0.9-1.0 (chloroform-methanol, 7:
3) A medium pressure column (hexane-ethyl acetate, 9: 1-8: 2-1: 1-chloroform-methanol 1: 1) obtained by further packing this fraction in a glass column of 55 mm in diameter and 60 cm in length with silica gel in a dry manner. , A step gradient), and evaluated as 18 fractions A to R.

The active fraction (Fr.P) yield 175.5 mg,
Rf = 0.5-0.6 (hexane-ethyl acetate, 7:
3) Further, this fraction was subjected to a medium pressure column (normal phase, size
Separation was carried out by the eluent of dichloromethane-ethyl acetate 98: 2 according to B) to obtain 7 fractions A to G, which were evaluated. Active fraction (Fr. D) yield 101.0 mg, Rf = ~
0.58 (dichloromethane-ethyl acetate 95: 5) This fraction was further eluted with a medium pressure column (normal phase, size B), and a dichloromethane-ethyl acetate 98: 2-95: 5-methanol step gradient. G was separated into 7 fractions and evaluated.

The active fraction (Fr.E) yield 11.4 mg, R
f = 0.41 (dichloromethane-ethyl acetate, 95:
5) Finally, this fraction was applied to a medium pressure column (normal phase, size
A), purification with an eluate of chloroform-methanol, 99: 1 was repeated three times, and a novel compound, 2,7-dimethyl-5-hydroxy-2-, was used as an active ingredient.
(4-Methyl-4-hydroperoxypent-2-enyl) chromene was isolated. Active ingredient (Fr.B) yield 3.5 mg, Rf = 0.26 (chloroform-methanol, 97: 3)

The results of LC-MS measured by the mass spectrum APCI method are as follows. calcd. for C ₁₇ H ₂₂ O ₄ ; 290.36 m
/ Z = 291 ((M + H) ⁺ , 273 ((MO
H) ⁺ ), 257 ((M-OOH) ⁺ )

Proton Nuclear Magnetic Resonance Spectrum The proton nuclear magnetic resonance spectrum measured in deuterated chloroform at 500 MHz using tetramethylsilane as an internal standard is as follows. ¹ H NMR (CDCl ₃ , 500 MHz) δ 1.26
(S, 3H), 1.26 (s, 3H), 1.40 (s,
3H), 2.19 (s, 3H), 2.41 (m, 2
H), 4.65 (bs, 1H), 5.46 (d, 1H,
J = 10 Hz), 5.56 (d, 1H, J = 16 Hz),
5.76 (dt, 1H, J = 16, 7.5 Hz), 6.1
0 (bs, 1H), 6.22 (bs, 1H), 6.64
(D, 1H, J = 10 Hz), 7.17 (bs, 1H) pp
m

Carbon-13 nuclear magnetic resonance spectrum The carbon-13 nuclear magnetic resonance spectrum measured at 125.8 MHz in heavy chloroform using tetramethylsilane as an internal standard is as follows. ¹³ C NMR (CDCl ₃ , 125.8 MHz) δ 21.
5 (q), 24.2 (q), 24.2 (q), 26.6
(Q), 44.3 (t), 77.8 (s), 82.2
(S), 106.8 (s), 108.5 (d), 10
9.6 (d), 117.2 (d), 126.4 (d),
126.4 (d), 137.0 (d), 139.9
(S), 151.1 (s), 154.2 (s) ppm

Infrared absorption spectrum The infrared absorption spectrum measured by the liquid film method is as follows. 4300, 3350, 3054, 2982, 2932,
1628, 1580, 1512, 1456, 1427,
1379, 1365, 1330, 1267, 1203
1141,1096,1067,992,977,91
3,828,775,739,704cm ^-1

Example 2.5-Lipoxygenase Inhibition Experiment This test was performed to examine the 5-lipoxygenase inhibitory action of the compound of Example 1 of the present invention. 1) Preparation of enzyme solution Rat Basophilic Leukemia cells (Rat Basophilic Leukemia)
Cell: RBL-1. ATCC CRL1378) was cultured in Eagle's medium containing 10% inactivated fetal bovine serum. PB cells
After washing 3 times with S, 0.1 M HE containing 1 mM EDTA
The cells were suspended in PES, pH 7.0 at a rate of 1 × 10 ⁷ cells / ml. After crushing the cells by ultrasound, 13,000 rpm
And the supernatant was used as an enzyme solution.

2) Method for Measuring Enzyme Activity 54 μl of the enzyme solution was mixed with 0.1 mM EDTA containing 2 mM calcium chloride and a test substance (dissolved in DMSO).
Add to HEPES, pH 7.0, and add a total volume of 120 μl
And After standing at 37 ° C. for 10 minutes, 6 μl of 0.4 mg / ml arachidonic acid (methanol solution) was added.
For 60 minutes. The reaction was stopped by adding 120 μl of acetonitrile and centrifuged at 13,000 rpm for 10 minutes. 5-Hydroxyeicosatetraenoic acid contained in the supernatant was detected by reversed-phase high-performance liquid chromatography. As a result, the concentration (IC ₅₀ ) at which the action of 5-lipoxygenase was inhibited by 50% was 0.3 μM.

Embodiment 3 FIG . Arachidone of the compound of Example 1
Measurement of anti-inflammatory effect using acid-induced mouse ear edema model Six groups of ICR mice were used, and the test drug was administered to both ears in 20 mice.
μl / ear was applied. Thirty minutes later, 1 mg / ml arachidonic acid dissolved in 90% ethanol was applied to the right ear at 20 μl / ear. The same volume of 90% ethanol was applied to the left ear. Ear thickness was measured at 30 minutes and 60 minutes after arachidonic acid application. The test drugs were all dissolved in acetone. (Ear thickness at each time point) − (ear thickness before application) was defined as an increase in thickness due to edema. The results are shown in Table 1 below.

[0031]

[Table 1]

Embodiment 4 FIG . RBL-1 of the compound of Example 1
Measurement of Leukotriene Production Amount in Cells Leukotriene B4 was measured as follows. RB
L-1 cells were placed in a 48-well plate at 2 × 10 ⁵ c.
The cells were seeded so as to become cells / well. The test drug was added, and the cells were cultured at 37 ° C. for 15 minutes. A23187 was added (final concentration 10 μM), and the cells were further cultured for 15 minutes. Thereafter, the culture solution was collected, and a supernatant was obtained by centrifugation. The amount of leukotriene B4 contained in this supernatant was determined by EIA k
It was measured in it. Also, leukotrienes C4, D4,
For the measurement of E4, cells were measured at 5 × 10 ⁵ cells / well.
And the other steps were performed in the same manner as for leukotriene B4. The results are shown in Table 2 below.

[0033]

[Table 2]

[0034]

The novel chromene derivative of the present invention has a 5-lipoxygenase inhibitory activity and is useful as an antiallergic agent, an antiasthmatic agent, an antiinflammatory agent and the like.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 35/78 A61K 35/78 C C07D 335/06 C07D 335/06 (72) Inventor Norio Kawabe 1111 Tehiro, Kamakura City, Kanagawa Prefecture Toray (72) Inventor Chihiro Tokuda 1111 Tehiro, Kamakura City, Kanagawa Prefecture Toray Co., Ltd.Basic Research Laboratory (72) Inventor Yasuhiko Togami 1111 Tehiro, Kamakura City, Kanagawa Prefecture Toray Co., Ltd. Basic Research Laboratory (72 Inventor Yu Wen-sheng, 4th ninth, Renmin South Rd.

Claims (4)

[Claims] 1. A compound represented by the following formula (I): (Wherein, R _{1 to} R ₄ are each independently H, an alkyl group having 1 to 6 carbon atoms having no substituent, an alkyl group having 1 to 6 carbon atoms having a substituent, and a carbon number having no substituent. 1-6
Alkenyl group having 1 to 12 carbon atoms having a substituent, alkynyl group having 1 to 6 carbon atoms having no substituent, alkynyl group having 1 to 12 carbon atoms having a substituent, carbon having no substituent An aryl group having 1 to 6 carbon atoms, an aryl group having 2 to 12 carbon atoms having a substituent, an acyl group having 1 to 6 carbon atoms having no substituent, and 1 to 12 carbon atoms having a substituent
And R ₄ also represents a silyl group; X and Y each independently represent an oxygen atom, a sulfur atom, or a nitrogen atom; and when X is a nitrogen atom, Z represents a hydrogen, an alkyl group, or a silyl group. Wherein X is an oxygen atom or a sulfur atom, Z is absent, and a double line consisting of a solid line and a dotted line represents a carbon-carbon single bond or a double bond) or a pharmacology thereof. Acceptable salts. 2. A medicament comprising the novel chromene derivative according to claim 1 or a pharmacologically acceptable salt thereof as an active ingredient. 3. A 5-lipoxygenase inhibitor comprising the novel chromene derivative according to claim 1 or a pharmacologically acceptable salt thereof as an active ingredient. 4. The therapeutic agent according to claim 2, wherein the 5-lipoxygenase inhibitor is an antiallergic drug, an antiasthmatic drug or an antiinflammatory drug.