JPH0459773A - New chromone and use of the same chromone as medicine - Google Patents

Abstract

NEW MATERIAL:Chromone expressed by the formula. USE:Having 5-lipoxygenase inhibitory action and aldose reductase inhibitory action and useful as a remedy such as an antiallergic agent for various disease conditions based on arachidonic acid dysbolism and as a remedy for complications in diabetes. PREPARATION:A crude drug Bupleurum root, Buplerum falcatum L. which is an original plant thereof or other plants of the same genus are extracted with a solvent such as hexane or diethyl ether and the solvent is subsequently removed from the resultant extract solution. The residue is then subjected to column chromatography, etc., using a solvent such as hexane or diethyl ether as an elution solvent and a porous polymer such as Diaion HP-20(R) or Sephadex(R) such as Sephadex LH-20(R) as a support to afford the objective compound expressed by the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a novel chromone and its use as a medicine.

[Prior art and problems] Bupleurum falcatum L-
is a crude drug that has been used as an antipyretic, sedative, and analgesic since ancient times, and saponins, flavones, etc. are known as its main ingredients. Detailed research has been conducted on saponin, and it has been reported that it has a central depressant effect, anti-inflammatory effect, anti-allergic effect, etc. However, at present, sufficient research has not been conducted regarding other trace components.

[Means for Solving the Problems] The present inventors have been researching the ingredients of saihu, and as a result of intensive examination of its pharmacological effects, the herbal medicine saihu,
The present inventors have discovered that chromones contained in the original plant, Mishimasaiko, or other congenerous plants have 5-lipoxygenase and aldose reductase inhibitory effects, leading to the completion of the present invention.

That is, the present invention provides a chromone represented by the following formula % (hereinafter referred to as the compound of the present invention),
and 5-lipoxygenase inhibitors and aldose reductase inhibitors containing the chromone as an active ingredient.

5-Lipoxygenase is an important enzyme in the arachidonic acid cascade, and inhibitors of this enzyme are thought to be useful as medicines to treat various pathological conditions based on abnormalities in arachidonic acid metabolism, such as inflammation, thrombosis, allergies, and asthma. There is.

Aldose reductase is an enzyme that catalyzes the first step of the polyol pathway in which aldoses such as glucose and galactose are converted to ketoses such as fructose via polyols such as sorbitol and galactitol. It has been reported that reducing the production and accumulation of sorbitol is effective in treating complications in diabetic patients.

Therefore, this aldose reductase inhibitor is considered to be useful as a therapeutic agent for complications in diabetes.

Furthermore, it was revealed for the first time by the present inventors that chromones are contained in saiko.

As mentioned above, an object of the present invention is to provide a therapeutic agent for various pathological conditions based on abnormalities in arachidonic acid metabolism, such as an anti-allergic agent, and a therapeutic agent for complications in diabetes, which have a 5-lipoxygenase inhibitory effect and an aldose reductase inhibitory effect. It's about doing.

For example, the following methods can be used to obtain the compound of the present invention.

The compound of the present invention is obtained by extracting the herbal drug Saigo, its original plant Mishima saiko, or other congenerous plants with at least one solvent selected from hexane, diethyl ether, petroleum ether, ethyl acetate, chloroform, acetone, methanol, and ethanol. Then, the residue obtained by removing the solvent from the obtained extract is treated with at least one solvent selected from hexane, diethyl ether, petroleum ether, ethyl acetate, chloroform, acetone, methanol, ethanol, acetonitrile, tetrahydrofuran, and water. As elution solvent, Diaion HP-20, MCI gel CHP2
Porous polymers such as 0P, Sephadex LH-2
Column chromatography or high-performance liquid chromatography using Sephadex, reversed-phase silica gel, silica gel, polyamide, cellulose, etc. as a carrier is performed once or several times, and the objective is determined by thin-layer chromatography or high-performance liquid chromatography. The compound of the present invention can be obtained by fractionating while checking the components.

Next, the compound of the present invention has an excellent 5-lipoxygenase inhibitory effect and an aldose reductase inhibitory effect,
The usefulness of the present invention as a medicine such as an anti-allergy agent and a treatment for various complications of diabetes will be explained using experimental examples.

Experimental example 1 (5-lipoxygenase inhibitory effect) RBL-1
Cultured cells were 1ff so that it was 5 x 106 cells/mj.
Ljl (suspended in 50rnM phosphate buffer (pH 7,4) containing EDTA and 10% ethylene glycol, after sonication, incubated at 10,000XG for 10 min, then for 10 min.
The upper groove, which was centrifuged at 5.0 OOXG for 60 minutes, was used as a 5-lipoxygenase enzyme preparation.

As a substrate, a dimethyl sulfoxide (DMSO) solution of 10-tone arachidonic acid, the enzyme preparation prepared as described above, and the compound obtained in the example below were used at a final concentration of 1100p.
The mixture was placed in a test tube and reacted at 37°C for 10 minutes. As an internal standard, 10 Jd of 0.25 M butyl-3,5-dinitrobenzoate was added and n-hexane L
Extracted with 8mt. The amount of 5-HETE in this was measured using high performance liquid chromatography [column; TSK gelODS].
-80TMCT (manufactured by TOYO8ODA), mobile phase: acetonitrile:water:acetic acid (60:40:0.02), flow rate: 1 ml/min, detection: ultraviolet light (235,...).

From this result, the inhibition rate was calculated using the following formula, and 5. When the lipoxygenase inhibitory activity was investigated, it was revealed that it had an inhibitory activity of 70.4% at a concentration of 100%.

C: 5-HETE without the compound obtained in the example
Peak area (corrected by internal standard) S: 5-HETE when the compound obtained in the example was added
Peak area (corrected by internal standard) Experimental example (Aldose reductase inhibitory effect) Six-week-old male Wistar rats were sacrificed under ether anesthesia, and the crystalline lenses were immediately removed and stored at -20°C.

The crystalline lens was homogenized in 135d sodium, potassium-phosphate buffer (pH 7.0) containing 0.5rnM phenylmethylsulfonyl fluoride for 30.000r
Centrifuged at pm for 30 minutes. The supernatant was used as a sample for inhibition of aldose reductase activity. Furthermore, all of the above operations were performed at 4°C, and the specimens were stored at -20°C.

Aldose reductase activity was measured using Dufuran (Dufuran).
frane) et al. [Biochemical Me.
dicine, 32, 99-105 (1984)]
This was done by That is, 100d lithium sulfate, 0.
03rnMNADPH (reduced nicotinamide
e adenine dinucleotidepho
135rnM sodium-potassium-phosphate buffer (pH 7,0) prepared to contain 20rnkI glucose as a substrate, 5ooJd&:
, drug solution 1007.1 in which the above sample 10oJd and the compound obtained in the example were dissolved in DMSO to a final concentration of 1.0X10'~i, oxio'M, respectively.
were added and reacted at 30°C for 30 minutes. Next, 0.3 ml of 0.5N hydrochloric acid was added to stop the reaction, and 1
6N sodium hydroxide containing 0WLhI imidazole 1
By adding rn1, NADP (oxidized nicotinamiden) produced by the above reaction is
ine dinucleotide phosphate
e) was converted into a fluorescent substance, and its fluorescence intensity was measured 30 minutes later. Fluorescence intensity was measured using a spectrophotometer F-4000 at room temperature.
(manufactured by Hitachi, Ltd.) with an excitation wavelength of 360,
Measurement was performed under the condition of a fluorescence wavelength of 460ru1s. In addition, the fluorescence intensity measured by reacting with soap was used as a control value in the same manner as above except that DMSO was added instead of adding the solution of the compound obtained in the example.

Aldose reductase uses NADPH as a coenzyme,
DL-An enzyme that converts glyceraldehyde or glucose into polyol, and along with this reaction, NADP
H changes to NADP. Therefore, if NADP is low, aldose reductase is inhibited.

As a result, when the aldose reductase inhibitory activity of the compound of the present invention was examined, it was found that at a concentration of 1100p, it was 55.3%
was found to have inhibitory activity.

Next, an acute toxicity test of the compound of the present invention was conducted using ICR male mice, and there were no deaths after oral administration of 1 g/hg.

Next, the dosage and formulation of the compound of the present invention will be explained.

The compounds of the present invention can be administered to animals and humans either neat or with conventional pharmaceutical carriers. The dosage form is not particularly limited and may be selected and used as necessary, and may include oral dosage forms such as tablets, capsules, granules, fine granules, and powders, parenteral dosage forms such as injections, and full dosage forms. .

In order to exert the desired effect as an oral agent, although it varies depending on the age, weight, and severity of the disease of the patient, it is usual for adults to administer 50 mg to 5 g of the compound of the present invention in divided doses several times a day. It seems appropriate to take it.

Oral agents include, for example, starch, lactose, sucrose, mannitol,
It is manufactured using conventional methods using carboxymethyl cellulose, corn starch, inorganic salts, etc.

In addition to the excipients mentioned above, binders, disintegrants, surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents, fragrances, and the like can be used in this type of preparation as appropriate. Specific examples of each are shown below.

[Binder 1 Starch, dextrin, gum arabic powder, gelatin,
Hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low substituted hydroxypropyl cellulose.

[Surfactant] Sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80° [Lubricant J Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, Polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The compounds of the invention can also be administered as suspensions, emulsions, syrups, elixirs,
These various dosage forms may contain flavoring agents and coloring agents.

In order to exert the desired effect as a parenteral agent, the compound of the present invention should be administered intravenously in an amount of 0.1 rng to 1 g per day for adults, although this will vary depending on the age, weight, and severity of the disease of the patient. , intravenous drip, subcutaneous injection, and intramuscular injection are considered appropriate.

This parenteral preparation is manufactured according to a conventional method, and generally uses distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. as a diluent. be able to. Furthermore, a bactericide, a preservative, a stabilizer, and an emulsifier may be added as necessary.

In addition, from the standpoint of stability, this parenteral drug is frozen after being filled into vials, etc., and the water is removed using normal freeze-drying techniques.
Solutions can also be reconstituted from the lyophilizate immediately before use. Furthermore, isotonizing agents, stabilizers, preservatives, soothing agents, etc. may be added as appropriate.

Other parenteral preparations include liquid preparations for external use, liniments such as ointments, half-open preparations for intrarectal administration, etc., and are manufactured according to conventional methods.

EXAMPLES Next, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited thereto in any way.

Example 1 8.5 kg of ground saiko was extracted with 2711 hexane, followed by chloroform and 257 methanol (1:1).

The solvent of the chloroform-methanol (1:1) extract was distilled off to obtain chloroform-methanol (1:1) extract 1.
470g was obtained. The extract was fractionated into upper and lower layers with a chloroform-methanol-water (8:4:3) mixture, and 935 g of crude fraction A (upper) and 935 g of crude fraction B (lower) were obtained.
) 376g was obtained.

500 g of crude fraction B was further subjected to column chromatography using silica gel, and eluted with n-hexane-chloroform (1:1), chloroform, chloroform containing 2% methanol, and chloroform containing 20% methanol.

14.82 g of the fraction eluted with chloroform was
Further, the product was subjected to column chromatography using silica gel and eluted with acetone-hexane (1:2) to obtain 76.6 IrL,1 of colorless needle crystals. This colorless needle-like crystal has the physical and chemical properties shown below, and is therefore 5-hydroxy-2-hydroxymethyl-7-methoxy4dan-1-
Benzobilan 4-one (5-hydroxy-2-hy
droxymethyl-7-methoxy4H-1
-benzopyran4-one).

Melting point = 185-192°C (acetone-chloroform: 15:85) Mass spectrum C1C11H10 05m/z: 222 (M”), 193HR-MS m
/zH Calculated value 222.05281 Actual value 222.05121 Infrared absorption spectrum 'NQI''cm-1; 330
0.1660, 1635, 1580, 1520 proton nuclear magnetic resonance spectrum (δppm in pyridine-d5): 3.7
3 (3H, d, J = 1.0Hz).

4.75 (2H, s).

6.48 (IH, d, J = 2.0Hz).

6.59 (IH, d, J = 2.0Hz).

6.78 (IH, br, s).

7.84 (IH, br) (δppm in CD013): 3.86 (3H, s), 4.57 (2H, br, d,
J = 5.5Hz).

6.33 (IH, t, J = 1.0Hz).

6.36, 6.37 (each IH, d, J =
2.0Hz) 13C-nuclear magnetic resonance spectrum (δppm in pyridine-d5): 55.
9(q), 60.2(t), 92.8(d), 98.4
(d).

106.1(s), 106.4(d).

158.2, 162.7, 165.8, 171.6, 1
83.0 (each s) Example 2 ■Corn starch 44g ■Crystalline cellulose 40g ■Carboxymethylcellulose calcium 5g ■Light anhydrous silicic acid 0.5g■Magnesium stearate o, sgCompound obtained in Example 1
10 total 100gMix ■~■ uniformly according to the above recipe, compress and mold with a tablet machine, and make one tablet 200g. ．． g tablets were obtained.

This tablet contains the compound 2orrLg obtained in Example 1.
It contains 10 to 25 tablets per day for adults, divided into several doses.

Example 3 ■ Crystalline cellulose 84.5 g ■ Magnesium stearate 0.5 g ■ Calcium carboxymethyl cellulose 5 g Compound listed in Example 1 1゜ Total 100 g According to the above recipe, ■, ■, and part of ■ were uniformly mixed and compressed. After molding, crush it, add the remaining amounts of ■ and ■, and mix.
Compression molding was performed using a tablet machine to obtain a portion of 200 #v tablets.

These tablets contain 2oIng of the compound obtained in Example 1, and are taken by adults in 10 to 25 tablets a day in several doses.

Example 4 ■Crystalline cellulose 49.5g ■10% hydroxypropyl cellulose ethanol solution 35g ■Carboxymethylcellulose calcium 5g ■Magnesium stearate 0.5g Compound 1 in Example 1 Total 100g ■, ■, and ■ were uniformly mixed according to the above recipe. After mixing ① and ②, making a net by a conventional method, granulating it with an extrusion granulator, drying and crushing it, and compressing it with a tablet machine, a portion of 2.
00. ．． g tablets were obtained.

These tablets contain the compound 209 obtained in Example 1, and are taken by adults in 10 to 25 tablets a day in several doses.

Example 5 ■Corn starch 34.5g ■Magnesium stearate 50g ■Carboxymethyl cellulose calcium 5g ■Light anhydrous silicic acid 0.5g Combined again in Example 1 10 Total 100g Mix ■~■ uniformly according to the above recipe, and use a compression molding machine. After compression molding, the mixture was crushed using a crusher and sieved to obtain granules.

1 g of this granule contains 100 g of the compound obtained in Example 1, and adults take 2 to 5 g a day in several doses.

Example 6 ■ Crystalline cellulose 55 g ■ 10% hydroxypropyl cellulose ethanol solution 35 g Compound 1 obtained in Example 1 Total 100 g According to the above recipe, ■ to ■ were uniformly mixed and suspended. After granulation using an extrusion granulator, the mixture was dried and sieved to obtain granules.

1 g of this granule contains 100 # of the compound obtained in Example 1, and adults should take 2 to 5 g in several doses per day.

Example 7 ■Corn starch 89.5g ■Light silicic anhydride o, sg Example 1 and compound 1
0 Total 100g ① to ② were mixed uniformly according to the above recipe, and 200g was filled into No. 2 capsules.

One capsule of this preparation contains 20 mg of the compound obtained in Example 1, and adults should take 10 to 25 capsules a day in several doses.

Example 8 ■ Soybean oil 5g ■ Distilled water for injection 89.5g ■ Soybean phospholipid 2.5g ■ Glycerin
2 g Total amount of hardware used in Example 1 100 g According to the above recipe, ■ was dissolved in ■ and ■, and the solutions of ■ and ■ were added to emulsify to obtain an injection.

Claims (3)

[Claims] (1) The following formula ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ A chromone represented by . (2) A 5-lipoxygenase inhibitor whose active ingredient is a chromone represented by the following formula ▲ Numerical formula, chemical formula, table, etc. ▼. (3) An aldose reductase inhibitor whose active ingredient is a chromone represented by the following formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼.