JPH02264718A - Aldose reductase inhibitor - Google Patents

Abstract

PURPOSE:To obtain an aldose reductase inhibitor comprising an active substance separated from an extract of dried core material of Sappan lignum of crude drug with methanol as an active ingredient. CONSTITUTION:An aldose reductase inhibitor comprising compounds shown by formula I to formula VII obtained from separated from an extract of dried core material of Sappan lignum of crude drug with methanol as an active ingredient. A dose of the compound is 50mg-5g per adult daily in the case of oral drug and preferably 0.1mg-1g in the case of parenteral drug. Methanol essence is obtained by distilling away a solvent under reduced pressure from an extracted solution of Sappan lignum with methanol, then dissolved in water, extracted with ethyl acetate three times, prepared ethyl acetate essence is chromatographed by silica gel column, extracted with chloroform, then a mixed solvent of chloroform and methanol (methanol content: 5, 10, 15, 20%), elution part of 5 % methanol-chloroform is chromatographed by Sephadex LH-20 column and fractionated to give compounds shown by formula I to formula VII.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to an aldose reductase inhibitor.

[Prior Art and Section H] In recent years, the accumulation of intracellular sorbitol via the polyol pathway, which is a glucose metabolic pathway, has attracted attention as a cause of various complications in diabetes such as cataracts, retinopathy, and renal damage. The polyol pathway is a metabolic pathway in which aldoses such as glucose and galactose are converted to ketoses such as fructose via polyols such as sorbitol and galactitol, and it has been shown by immunohistological techniques that it exists widely in various organs of the body. It has become.

The first step in this pathway, the enzyme that catalyzes the conversion between aldose and polyol, is called aldose reductase.
This enzyme is considered to be the rate-limiting enzyme in the polyol pathway. It has been reported that inhibiting this aldose reductase and reducing the production and accumulation of sorbitol is effective in treating complications in diabetic patients.

Therefore, it has been desired to develop a drug that has an aldose reductase inhibitory effect.

[Means for Solving the Problems] As a result of intensive research to provide an excellent aldose reductase inhibitor, the present inventors have discovered the crude drug Suou, Caesarpinia, which is widely used clinically.
The present invention has been completed by discovering several substances that have an aldose reductase inhibitory effect from the methanol extract of the dried core material of 5appan L., a leguminous plant.

That is, one aspect of the present invention is as shown below.

(1) An aldose reductase inhibitor containing a compound represented by the following formula ■■ as an active ingredient.

(2) An aldose reductase inhibitor containing a compound represented by the following formula ■■ as an active ingredient.

(3) An aldose reductase inhibitor containing a compound represented by the following formula (■) as an active ingredient.

(4) An aldose reductase inhibitor containing a compound represented by the following formula ■■ as an active ingredient.

(6) An aldose reductase inhibitor containing a compound represented by the following formula ■■ as an active ingredient.

■ An aldose reductase inhibitor containing the compound represented by as an active ingredient.

(7) An aldose reductase inhibitor containing a compound represented by the following formula ■■ as an active ingredient.

Hereinafter, the compounds of formulas I to (2) will be collectively referred to as compounds of the formula.

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

Extract Soki or its congeners with water, alcohol, a mixed solvent of water and alcohols, or a mixed solvent of water and acetone, and remove the solvent from the extract and leave the residue as it is or dissolve it in water as necessary. Then, it was extracted with six solvents such as petroleum ether, needles, and chloroform, and the fat-soluble components that migrated to the resulting organic solvent were removed, and then extracted with n-butanol, and the solvent was removed from the extract. The residue was purified using Diaion HP-20 using at least one elution solvent selected from water, methanol, ethanol, acetic acid, chloroform, ethyl acetate, n-hexane, acetone, and benzene.
, MCI gel CHP 20 P and other porous polymers
At least one column chromatography using Sephadex such as Sephadex LH-20, reversed-phase polymer gel, polymer gel, polyamide, activated carbon, or cellulose as a carrier is carried out, and the target components are confirmed and separated using thin layer chromatography. It can be obtained by drawing.

Further, in some cases, it may be purified by recrystallization using a suitable solvent such as methanol or ethanol.

A specific example of the production of the compound of the formula is shown below.

Specific example 1 10 kg of Soki was extracted with 5 liters of methanol 1-1 The solvent was distilled off under reduced pressure from the obtained extract to obtain a methanol extract. This methanol extract was dissolved in water and extracted three times with ethyl acetate. The obtained ethyl acetate extract was subjected to silica gel column chromatography and eluted with 5 L of chloroform and then with 5 Q of a mixed solvent of chloroform-methanol (methanol content: 5.10° to 15.20%).
After distilling off the solvent from the methanol-chloroform eluate under reduced pressure, the residue was subjected to Sephadex LH-20 column chromatography and eluted with methanol. In order of elution, fraction A (fraction eluted at approximately 1.0 to 1.2 times the void volume), fraction B (1.3 to 1.5 times the void volume), fraction C (
1.6 to 1.8 times). After methanol in fraction C was distilled off under reduced pressure, colorless granular product 49019 was obtained by crystallization from aqueous ethanol. The physical and chemical properties of this colorless granular crystal are described in the literature [T, Shimokawa et al.

Chea+, Pharm, Bull, 33.35
45 (1985)].

Specific example 2 Fraction B of specific example 1 was subjected to high performance liquid chromatography [column: CIG (15φ×300111R1 silica gel 10
7a), Elution solvent; methanol:chloroform = 3:9
7] and further crystallized from aqueous methanol to obtain yellow needle crystals 1,69. The physical and chemical properties of this yellow needle crystal are described in the literature [Masahiro Nagai et al., Pharmaceutical Journal, 104, 93]
5. (1984) agreed with the properties of Sarapanchalcone described in Ko.

Specific example 3 Both portions B of specific example 1 were subjected to high performance liquid chromatography [column: CIG (15φx300xm, 107 g of silica gel).
), elution solvent: methanol:benzene-1:9] to obtain 370 ml of colorless powder. The physicochemical properties of this colorless powder are described in the literature [T, 5aitoh etal, C
hem, Pharm, Bull, 34, 2506 (1
986)] was consistent with the properties of 3-deoxysappanon described.

Specific example 4 Fraction B of specific example 1 was subjected to high performance liquid chromatography [column: CIG (15φX3001JI, silica gel 10/
la), elution solvent: methanol; benzene-1=9, and crystallized from water to obtain 1.49 colorless needle crystals. The physical and chemical properties of this colorless needle crystal are described in the literature [M, Nag
ai et al, chem, Pharm, Bull
, 34.1 (1986)].
consistent with the nature of

Specific Example 5 Both fractions eluted with 20% methanol-chloroform in the silica gel column chromatography of Specific Example 1 were further subjected to μ~Bondapak C+m column chromatography, and eluted first with increasing amounts of water and then methanol. . The solvent of the fraction eluted with a methanol content of about 60% was distilled off under reduced pressure to obtain amorphous powder 29.

The physical and chemical properties of this amorphous powder are described in the literature [C, Fukee
tal,, Phytochemistry, 24.2
403 (1985)].

Specific Example 6 Both fractions eluted with methanol content of approximately 50% in the μm Bondapak C+g column chromatography of Specific Example 5 were further purified by Sephadex LH-20 column chromatography (elution solvent: methanol) to obtain 8 g of amorphous powder. Obtained. The physical and chemical properties of this amorphous powder are described in the literature [C, Fuke et al.

Phytochemistry, 24, 2403 (19
85)].

Specific Example 7 Both fractions eluted with methanol content of approximately 65% in the p-Bondapak C+a column chromatography of Specific Example 5 were further purified by Sephadex LH-20 column chromatography (elution solvent: methanol) to obtain amorphous powder. 151? I got it. The physical and chemical properties of this amorphous powder are described in the literature [C, Fuke et al.

PhytocheIIigtry, 24, 240
3 (1985).

Next, the fact that the compound of the formula has an aldose reductase inhibitory effect will be explained with reference to experimental examples.

Experimental Example 1 Measurement of Aldose Retactase Activity Six-week-old Wistar male rats were sacrificed under ether anesthesia, and the crystalline lenses were immediately removed and stored at -20°C.

The crystalline lens was homogenized in 35 pH sodium-potassium-phosphate buffer (pH 7.0) containing 0.5 JAN phenylmethylsulfonyl fluoride, and
．． Centrifugation was performed at 00 Orpm for 30 minutes. The supernatant was used as an aldose reductase crude enzyme solution. Furthermore, all the following operations were performed at 4°C, and the crude enzyme solution sample was stored at -20°C.

Aldose reductase activity was measured using Dufuran (D
[Biochemical M
edicine, 32°99-1.05 (1984)
]. That is, 1001 lithium sulfate,
0.03JIM NADPH (reduced n1cotina
IIlide adenine dinucleoti
de phosphate), and 1 3531M sodium-potassium-phosphate buffer (pi-+ 7.0) prepared to contain glucose as a substrate.
00uf1. Then, 1.0Xl of the above crude enzyme solution and the compound obtained in the "bipod specific example" were each added to DMSO.
Add a drug solution dissolved at 100°C to a final concentration of O''~4.0xlO-'M, and incubate at 30°C for 3
The reaction was allowed to proceed for 0 minutes. Next, the reaction was stopped by adding 0.3-0.5N hydrochloric acid, and by adding 17d of 6N sodium hydroxide containing IO,M imidazole, the NADP (oxidized form n1cot 1na
IIlideadenine dinucleotide
ephosphate) was converted into a fluorescent substance, and the fluorescence intensity was measured 30 minutes later. Fluorescence intensity was measured at room temperature using a fluorometer F-4000 (manufactured by Hitachi Seisakusho) at an excitation wavelength of 36o, ..., and a fluorescence wavelength of 460r! ! The measurement was carried out under the condition of 1 t. Further, the fluorescence intensity measured by reacting in the same manner as above except that DMSO was added instead of adding the solution of the compound obtained in the specific example was used as a control value.

Aldose reductase is an enzyme that converts DL-glyceraldehyde or glucose into polyol using N A D P H as a coenzyme, and with this reaction, N A D P H changes to N A I) P. Therefore, if NADP is low, aldose reductase is inhibited.

The results are expressed as inhibition rate (%) and 50% inhibitory concentration (I
Cs. ) as shown in Table 1.

Table 1 (Inhibitory effect on aldose reductase) Next, an acute maternal test using oral administration of the compound obtained in Example 7 was carried out by ICR.
When the test was carried out using male mice of the strain, there were no deaths after oral administration of 9 on 29/29.

Thus, the compound of the formula has extremely low toxicity and high safety.

Next, the dosage and formulation of the compound of formula will be described.

The compounds of the formula can be administered to animals and humans neat or with conventional pharmaceutical carriers.

The dosage form is not particularly limited and may be selected and used as required, including tablets, capsules, granules, fine granules,
Examples include oral preparations such as powders, parenteral preparations such as injections, and physiological preparations.

In order to achieve the desired effect as an oral agent, it is usually necessary for an adult to take 50119 to 5 g of the compound of the formula in divided doses several times a day, although this will vary depending on the patient's age, weight, and severity of the disease. It seems appropriate to take it.

Oral agents include, for example, starch, lactose, sucrose, mannitol,
It is manufactured using conventional methods using carboxymethyl cellulose, controls, 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] Starch, dextrin, gum arabic powder, gelatin,
Hydroxypropyl starch, medylcellulose, 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] 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 net shapes may contain flavoring agents and coloring agents.

In order to exert the desired effect as a parenteral agent, the weight of the compound of the formula for an adult is usually 0.1■~ per day, although it varies depending on the age, weight, and severity of the disease of the patient! IV injections up to 9;
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, and a stabilizer may be added as necessary. In addition, from the viewpoint of stability, this parenteral preparation may be filled into a vial or the like, then frozen, water removed by ordinary freeze-drying techniques, and a liquid preparation prepared from the freeze-dried product immediately before use. Furthermore, isotonizing agents, stabilizers, preservatives, soothing agents, etc. may be added as appropriate.

Other parenteral preparations include external solutions, liniments such as ointments, and physiological preparations for intrarectal administration, and are manufactured according to conventional methods.

Next, the present invention will be explained in more detail with reference to Examples.
The present invention is not limited to the number of ships.

Example 1 ■Corn starch 449 ■Crystalline cellulose 409 ■Carboxymethyl cellulose calcium 59 ■Light anhydrous silicic acid 0.59 ■Magnesium stearate 0.5g Compound I in Specific Example 1
O total 1009 According to the above recipe, ① to ② were mixed uniformly and compression molded using a tablet machine to obtain a portion of 200 □ tablets.

These tablets contain the compound 20M9 obtained in "Specific Examples" and are taken by adults in 10 to 25 tablets a day in several doses.

Example 2 ■ Crystalline cellulose 84.59 ■ Magnesium stearate 0.59 ■ Calcium carboxymethyl cellulose 59 ■ Ingredients Compound obtained in 2 109 Total 1 oog Mix ■, ■, and part of ■ uniformly according to the above recipe, After compression molding, pulverize, add the remaining amounts of ① and ②, mix, and compress 1° using a tablet machine to make 200 mg tablets.
tablets were obtained.

This tablet contains 20 mg of the compound obtained in Example 2, and an adult should take 10 to 25 tablets in several doses per day.

Example 3 ■ Crystalline cellulose 49.59 ■ 10% hydroxypropyl cellulose ethanol solution 359 ■ Carboxymethyl cellulose calcium 59 ■ Magnesium stearate 0.59 ■ Compound obtained in specific example 3 109 Total 1009 For the formulation of -I- Step 4: Mix ■, ■, and ■ uniformly, neutralize using a conventional method, granulate using an extrusion granulator, dry and crush, mix ■ and ■, and compress using a tablet machine. Some 200 N tablets were obtained by molding.

This tablet contains 20~ of the compound obtained in Specific Example 3, and contains 1 day of the compound obtained in Example 3! Take 0 to 25 tablets in divided doses.

Example 4 ■Corn starch 34.590 Magnesium stearate 509 ■Calcium carboxymethyl cellulose 59 ■Light silicic anhydride 0.5 g ■Compound obtained in Example 4 109 Total +oog Mix ■~■ uniformly according to the above recipe and compress. After compression molding with a molding machine, it was crushed with a crusher and sieved to obtain granules.

This granule 17 contains compound 100N9 obtained in specific example 4.
It contains 2 to 57 doses per day for adults, divided into several doses.

Example 5 ■ Crystalline cellulose 559 ■ 10% hydroxypropyl cellulose ethanol solution 359 ■ Compound obtained in specific example 5
1009 According to the above recipe, ① to ② were uniformly mixed and then combined. After granulating with a press (7-hole granulator), dry and sieve (7
Granules were obtained.

This granule! 9 contains 100 yt of the compound obtained in Specific Example 5.
It contains Ti, and adults should take 2 to 59 doses a day in several doses.

Example 6 ■Corn starch 89.59■Light silicic anhydride 0.59■Compound obtained in specific example 6
to9 total 100g Mix ■～■ uniformly according to the above recipe, 200 m
y was filled into a No. 2 capsule.

One capsule of this preparation contains the compound 2019 obtained in Specific Example 6, and adults should take 10 to 25 capsules a day in several doses.

Example 7 ■ Soybean oil 59 ■ Distilled water for injection 89.59 ■ Soybean phospholipid 2.59 ■ Glycerin 29 ■ Compound obtained in Example 7 l? Total amount 100g Dissolve ■ in ■ and ■ according to the above recipe, and add ■
The solutions of (1) and (2) were added and emulsified to obtain an injection.

Claims (7)

[Claims] (1) Formula I below ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ An aldose reductase inhibitor whose active ingredient is the compound represented by I. (2) An aldose reductase inhibitor containing a compound represented by the following formula II ▲Mathematical formula, chemical formula, table, etc.▼II as an active ingredient. (3) An aldose reductase inhibitor containing a compound represented by the following formula III ▲Mathematical formula, chemical formula, table, etc.▼III as an active ingredient. (4) An aldose reductase inhibitor containing a compound represented by the following formula IV ▲Mathematical formula, chemical formula, table, etc.▼IV as an active ingredient. (5) An aldose reductase inhibitor containing a compound represented by the following formula V ▲Mathematical formula, chemical formula, table, etc.▼V as an active ingredient. (6) An aldose reductase inhibitor containing a compound represented by the following formula VI ▲Mathematical formula, chemical formula, table, etc.▼VI as an active ingredient. (7) An aldose reductase inhibitor containing a compound represented by the following formula VII ▲Mathematical formulas, chemical formulas, tables, etc.▼VII as an active ingredient.