JPH0699305B2 - Antidiabetic agent - Google Patents

Description

TECHNICAL FIELD The present invention relates to a therapeutic agent for diabetes, more specifically, a therapeutic agent for non-insulin-dependent type II diabetes.

Conventional technology and its problems The number of diabetic patients is increasing due to the increase in total energy intake due to the westernization of eating habits in Japan in recent years, obesity, and the increase in stress due to sophistication and complexity of society. It is following the path of.

By the way, diabetes can be roughly classified into insulin-dependent type I and insulin-independent type II.

Type I diabetes can control the symptoms of diabetes by the administration of insulin to some extent, but in the case of type II diabetes, the administration of insulin usually cannot directly control the symptoms.

As a treatment method for type II diabetes, in addition to exercise therapy and diet therapy, administration of oral hypoglycemic agents such as sulfonylurea agents can be mentioned as drug therapy, and these drugs promote the secretion of insulin in the body. Since the main action is
It is hard to say that a therapeutic agent for type II diabetes is compatible with its mechanism of action.

Therefore, there is a need in the art for a drug with a completely new mechanism of action that is effective in treating type II diabetes.

Means for Solving the Problems As a result of intensive studies on the above problems, the present inventors have found that at least one selected from the sesquiterpene analogs represented by the following formulas and salts of these compounds is type II. They have found that they are effective as an active ingredient compound of a therapeutic agent for diabetes, and have completed the present invention.

According to the invention, the formula And expression There is provided a therapeutic agent for diabetes comprising at least one selected from the sesquiterpene analogues represented by and a salt of the above compound as an active ingredient.

The compounds represented by the above formulas (1) and (2) used as the active ingredient in the therapeutic agent of the present invention are the compounds previously developed by the applicant of the present invention, for example, West German Patent No. 2821403 (Japanese Patent Laid-Open No. 54-36255). And JP-A-54-37832) and JP-A-64-6214, they are known to be useful for certain pharmaceutical applications including therapeutic agents for ulcerative colorectal diseases. . However, as a result of a separate study, the present inventors have found that the compound has a novel medicinal use which is completely unexpected from the medicinal uses described in the above-mentioned publications, that is, for the treatment of diabetes, particularly insulin-independent type II diabetes. It was found to be effective, and the present invention was completed based on this finding.

When the compound (crystal) represented by the above formula (1) is dissolved in a solvent, particularly a basic solvent, it becomes an equilibrium mixture with the tautomer of the compound represented by the above formula (2). . That is, after the compound (1) was dissolved in a dimethylsulfoxide solvent, according to NMR spectrum analysis measured over time, after 20 minutes of dissolution, in addition to 6.36 ppm showing a characteristic signal of lactol, an aldehyde proton (-C H O )
A slight peak was observed at 9.89 ppm, which is a characteristic signal of the above, and the integration ratio of the two was about 73 for the former to about 27 for the latter, and the peak at 9.89 ppm increased slightly after 2 hours, and the integration ratio was 70. The ratio reached to 30, and this integral ratio did not change in the NMR analysis chart after 63 hours, and the compound (2) exists in the above solvent in a ratio of 3 to 7 with the compound (1).

The therapeutic agent for diabetes of the present invention can contain the compound (1) and the compound (2) in the form of such an equilibrium mixture as active ingredients. The compound (2), which is a tautomer of the compound (1), is not produced when methanol or pyridine is used as the solvent.

The antidiabetic agent of the present invention can also contain a basic compound salt of each compound as an active ingredient, like the above-mentioned compound and the mixture of each compound. The above salt is obtained by reacting at least one functional group of the acidic groups of the compound (1) and the compound (2), that is, the phenolic hydroxyl group and the carboxyl group of the lactol ring of the compound (2) with the basic compound. Can be obtained. Specific examples of the basic compound that can be used for producing such a salt include alkali metal or alkaline earth metal such as sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate. Examples thereof include hydroxides and carbonates. Also, for example, methylamine, ethylamine, isopropylamine, morpholine,
Organic amines such as piperazine, piperidine, and 3,4-dimethoxyphenethylamine can also be used as the basic compound.

Each compound as an active ingredient of the therapeutic agent of the present invention also has a stereoisomer, and the present invention naturally includes utilizing such a stereoisomer as an active ingredient.

The antidiabetic agent of the present invention is usually put into practical use in the form of a pharmaceutical composition using a pharmaceutical carrier together with an active ingredient compound. The above-mentioned pharmaceutical carrier is a diluent such as a filler, a bulking agent, a binder, a moisturizing agent, a disintegrating agent, a surface-active agent, a lubricant and the like, which is usually used for preparing a pharmaceutical preparation according to the usage form. Alternatively, an excipient can be exemplified, and the one or the like is appropriately selected according to the dosage unit form of the obtained preparation.

As the dosage unit form of the therapeutic agent of the present invention, various forms can be selected according to the purpose of treatment, and as a typical example thereof,
Examples thereof include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, injections (solutions, suspensions, etc.) and the like. When forming into tablets, various carriers commonly used in this field can be widely used as the pharmaceutical carrier. Examples thereof include excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, potassium phosphate, water, ethanol, propanol, simple syrup, and glucose solution. , Starch liquid, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone and other binders, sodium carboxymethylcellulose,
Carboxymethyl cellulose calcium, low-substituted hydroxypropyl cellulose, dry starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, and other disintegrants, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, etc. Surfactants, sucrose, stearin, cocoa butter, disintegration inhibitors such as hydrogenated oils, quaternary ammonium bases, absorption promoters such as sodium lauryl sulfate, humectants such as glycerin and starch,
Adsorbents such as starch, lactose, kaolin, bentonite, colloidal silicic acid, refined talc, stearates, boric acid powders, and lubricants such as polyethylene glycol can be used. Further, the tablet is a tablet which is coated with a usual coating as necessary, for example, sugar-coated tablet, gelatin-coated tablet, enteric-coated tablet,
It can be a film-coated tablet, a double-layered tablet, or a multi-layered tablet. When molded into the form of pills, various substances usually used in this field as a pharmaceutical carrier, such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil,
Excipients such as kaolin and talc, gum arabic powder, tragacanth powder, binders such as gelatin and ethanol, disintegrating agents such as laminaran and agar can be widely used. When molding in the form of suppositories, various types of carriers commonly used in this field, such as polyethylene glycol, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semisynthetic glycerides, etc., are used as pharmaceutical carriers. be able to. Capsules can be prepared by mixing the active ingredient compound of the present invention or a salt thereof with the various pharmaceutical carriers exemplified above in a conventional manner and filling, for example, hard gelatin capsules or soft capsules. When the therapeutic agent of the present invention is prepared as an injection such as a solution, emulsion or suspension, it is preferably sterilized and isotonic with blood. When molding into other forms,
Various publicly known diluents commonly used, such as water, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, and polyoxyethylene sorbitan fatty acid esters can be used. In this case, a sufficient amount of sodium chloride, glucose or glycerin for preparing an isotonic solution may be contained in the antidiabetic agent of the present invention, and it is usually well known in the formulations mentioned above. Various additives such as solubilizers, buffers, soothing agents and the like can be added. Further, in the therapeutic agent of the present invention, if necessary, it is possible to include a colorant, a preservative, a flavor, a flavoring agent, a sweetening agent and the like, which are known to be able to be added and blended in this kind of preparation, and other pharmaceuticals. It is possible.

The above formula (1) to be contained in the therapeutic agent for diabetes of the present invention.
The amount of the active ingredient compound represented by the formula (2) and / or the salt thereof is not particularly limited and is appropriately selected from a wide range, but is usually about 0.5 in the whole pharmaceutical preparation composition.
It is suitable that the content is about 30% by weight.

The administration method of the antidiabetic agent of the present invention is not particularly limited and can be determined according to various dosage forms, patient's age, sex and other conditions, degree of disease and the like. For example, tablets, pills, solutions, suspensions, emulsions, granules and capsules can be administered orally. The injections can be administered intravenously alone or in a mixture with a normal replenisher such as glucose, amino acid and the like, and can also be administered intramuscularly, intracutaneously, subcutaneously or intraperitoneally alone if necessary. Suppositories can be administered rectally.

The dose of the therapeutic agent of the present invention is appropriately selected depending on the usage, age of patient, sex and other conditions, degree of disease, etc., and is not particularly limited.
It is suitable to be selected from an amount of about 0.1 mg to 50 mg to be administered, and the preparation can be administered in 1 to 4 divided doses per day.

Examples Hereinafter, production examples of active ingredient compounds of the therapeutic agent for diabetes of the present invention and formulation examples of the therapeutic agent of the present invention using the same will be described. It should be noted that the following is an example of manufacturing and is not restricted by this.

Production example 1 Stachybotrys sp. K76 (Microtech Lab.
No.) was cultured in a 500 ml Sakaguchi flask containing 100 ml of the following composition at 28 ° C. and pH 6 for 4 days with reciprocal shaking.

Glycerin 0.5% Starch 1.0% Lactose 0.2% Soybean flour 0.5% Yeast extract 0.1% Malt extract 0.2% CaCO ₃ 0.3% MgSO ₄ 0.05% One seed culture obtained above was placed in medium 20 of the above composition 30 In a jar fermenter, culture temperature 28 ℃,
The culture was carried out for 5 days at an aeration rate of 1 / medium / minute and a stirring rate of 300 revolutions / minute.

The obtained culture broth was centrifuged at 8000 rpm to remove bacterial cells, methanol of 5 was added to the supernatant, the mixture was stirred and left for 3 hours, and then centrifuged to remove precipitates, Extracted with an equal volume of ethyl acetate. After distilling off the solvent of the ethyl acetate layer under reduced pressure, the residue was dissolved in methanol, passed through an activated carbon column, the eluate was evaporated to dryness under reduced pressure, and then dissolved in chloroform: ethyl acetate = 1: 1 (v / v), The gel was passed through a Sephadex LH-20 column and subjected to thin-layer chromatography [ethyl acetate-chloroform-acetic acid (volume ratio 50: 50: 2) was used as the developing solvent] to obtain an anti-promoting agent corresponding to Rf = 0.34. In the physical activity fraction or similar thin layer chromatography [using a mixed solution of benzene-butanol-acetic acid (volume ratio 60: 15: 5) as a developing solvent], an anti-complement active fraction corresponding to Rf = 0.58 was obtained. gather,
By distilling off the solvent from this, 6,7-dihydroxy-2,5, which is a pale yellow weak acidic substance having anti-complement activity,
5,8a-Tetramethyl-1,2,3,4,4a, 5,6,7,8,8a-decahydronaphthalene-1-spiro-2 '-(6', 7'-diformyl-4'- 2.0 g of hydroxy-2 ', 3'-dihydrobenzofuran) was obtained.

The formation of the above compound is confirmed by the following physicochemical properties.

[Alpha ▲] ²⁰ _D ▼ = -48 DEG (c = 2.5, methanol) Elemental analysis (C ₂₃ H ₃₀ as O ₆₎ Calculated (%) C68.64 H7.51 Found (%) C68.58 H7 .55 Ultraviolet absorption spectrum (UV) analysis Production Example 2 2.1 g of silver nitrate was dissolved in 1 ml of water, 3.5 ml of a 5.8 molar sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 20 minutes,
6,7-Dihydroxy-2,5,5,8a-tetramethyl-1,2,, 4,4a, 5,6,7,8,8a-decahydrophthalene-obtained in Preparation Example 1
A solution prepared by dissolving 1.0 g of 1-spiro-2 '-(6', 7'-diformyl-4'-hydroxy-2 ', 3'-dihydrobenzofuran) in 3 ml of ethanol was added.

After stirring the reaction medium at room temperature for 1.5 hours, the pH was adjusted to about 2 with 2N hydrochloric acid, the reaction solution was extracted with the same amount of ethyl acetate, the extraction solvent was evaporated under reduced pressure, and the obtained residue was purified by a silica gel column. Chromatography [silica gel "Wako C-200",
Wako Pure Chemical Industries' product, eluent: chloroform-ethyl acetate-
Purified with acetic acid (volume ratio 100: 502). Thin layer chromatography [Ethyl acetate-chloroform-acetic acid (volume ratio 5
0: 50: 2) as the developing solvent], and a fraction corresponding to Rf = 0.37 or a similar thin layer chromatography [benzene-butanol-acetic acid (volume ratio 60: 15: 5) mixture As a developing solvent], a fraction corresponding to Rf = 0.71 was collected, and the solvent was distilled off from the fraction to give a pale yellow amorphous crystal.
8-dihydroxy-6-oxo-2,3,6,8-tetrahydro-furo [3,4-g] benzofuran-2-spiro-1'-
(6 ', 7'-dihydroxy-2', 5 ', 5', 8'a-tetramethyl-1 ', 2', 3 ', 4', 4'a, 5 ', 6', 7 ', 8 ′,
700 mg of 8'a-decahydronaphthalene) [compound of formula (1) above] was obtained.

The above compound showed the following physicochemical properties, which confirmed its formation.

[Α ▲] ²⁰ _D ▼ = −44.8 ° (c = 0.9, methanol) Elemental analysis value (as C ₂₃ H ₃₀ O ₇ ) Calculated value (%) C66.03 H7.18 Measured value (%) C65.93 H7 .21 Production Example 3 0.4 ml of 0.4N sodium hydroxide aqueous solution and 5 ml of ethanol
In addition, 4,8-dihydroxy-6-oxo-2,3,6,8-tetrahydro-furo [3,4-g] benzofuran-2-spiro-
1 '-(6', 7'-dihydroxy-2 ', 5', 5 ', 8'a
-Tetramethyl-1 ', 2', 3 ', 4', 4'a, 5 ', 6', 7 ',
418 mg of 8 ', 8'a-decahydronaphthalene) was added, and the mixture was stirred under a nitrogen stream at 30 to 40 ° C for 30 minutes. After completion of the reaction, the solvent was distilled off under reduced pressure and the residue was dried to dryness, and 10 ml of acetone was added to the residue to remove the dissolved portion.

The obtained crude crystals were recrystallized from water-acetone to give pale yellow amorphous crystals, disodium 6,7-dihydroxy-2,
5,5,8a-Tetramethyl-1,2,3,4,4a, 5,6,7,8,8a-decahydronaphthalene-1-spiro-2 '-(6'-carboxylate-7'- Formyl-4'-oxide-2 ',
342 mg of 3'-dihirobenzofuran) [a disodium salt of the compound of the above formula (2)] was obtained.

The obtained compound had the following physicochemical properties, which confirmed its formation.

[Α ▲] ²⁰ _D ▼ = −44.2 ° (c = 1.25, H ₂ O) Elemental analysis value (as C ₂₃ H ₂₈ O ₇ Na ₂ ) Calculated value (%) C59.74 H6.10 Measured value (%) C59.48 H5.91 UV absorption spectrum (UV) analysis Further, in the above, an equimolar amount with respect to the raw material compound, usually using sodium hydroxide in an amount of about 1.2 times the molar amount, sodium 6,7-dihydroxy-2,5,
5,8a Tetramethyl-1,2,3,4,4a, 5,6,7,8,8a-decahydronaphthalene-1-spiro-2 '-(6'-carboxylate-7'-formyl-4 ′ -Oxide-2 ′, 3′-
Dihydrobenzofuran) [a monosodium salt of the carboxyl group of the compound of the above formula (2)] was obtained.

Formulation Example 1 Compound obtained in Production Example 3 [2Na salt of compound of formula (2)] 5 mg Glucose 250 mg Distilled water for injection Appropriate amount 5 ml Dissolve compound and glucose obtained in Production Example 3 in distilled water for injection Then, pour it into a 5 ml ampoule and replace it with nitrogen.
After autoclaving at 15 ° C. for 15 minutes, the antidiabetic agent of the present invention in the form of an injection was obtained.

Formulation Example 2 Compound obtained in Production Example 2 [Compound of Formula (1)] 5 mg Semi-synthetic glyceride base Appropriate amount 500 mg To the semi-synthetic glyceride base, the compound obtained in Production Example 2 was added, and the mixture was heated at 50 ° C. After mixing and suspending, the mixture was poured into a molding mold, naturally cooled, and then taken out to obtain a therapeutic agent for diabetes of the present invention in the form of a suppository.

Formulation Example 3 Compound obtained in Production Example 3 [2Na salt of compound of formula (2)] 5g Avicel (trade name, manufactured by Asahi Kasei Corporation) 120g Constarch 90g Magnesium stearate 6g TC-5 (trade name, hydroxypropylmethylcellulose) 10g Polyethylene glycol-6000 3g Castor oil 40g Ethanol 40g The compound obtained in Production Example 3, Avicel, corn starch and magnesium stearate were taken and after mixed polishing, sugar coating R1
Tableted with 0 mm kine.

The obtained tablets were coated with a film coating agent consisting of TC-5 (hydroxypropyl methylcellulose), polyethylene glycol-6000, castor oil and ethanol to obtain the antidiabetic agent of the present invention in the form of a film coated tablet.

Hereinafter, the pharmacological test of the active ingredient compound of the therapeutic agent for diabetes of the present invention and the result thereof will be described in detail.

The animals used in the following pharmacological tests are male KK mice (Kiwa experimental animals).

The above-mentioned KK mouse is an inbred mouse that was found to develop diabetes in 1962 and has been established and maintained since the origin of the mouse that had been bred in Japan for a long time, and it is usually mild obesity (maximum body weight). 40-50g) is known. The KK mice were individually housed in individual cages in order to prevent conflict in this experiment. As a sample, a special breeding sample (CMF, manufactured by CLEA Japan, Inc.) was freely ingested.

<Pharmacological Test Example 1> Inhibition Test of Weight Gain by Administration of Compound of Active Ingredient for Treatment of Diabetes of the Present Invention The above-mentioned KK mice are divided into two groups, one of which is a compound of active ingredient for treatment of diabetes (sesquiterpene analogue, production example) Monosodium salt of compound of formula (2) obtained in 3) 10
mg / kg administration group (group I) and the other group as control group (II
Group, non-administration group of active ingredient compound of the present invention for treating diabetes.

The compound of the present invention, which is an active ingredient of the therapeutic agent for diabetes in Group I, was dissolved in pure water, stored at 4 ° C., protected from light, and diluted at the time of administration according to the dose. The dilution ratio was adjusted according to the body weight of the mouse.

This experiment was carried out on 6 animals after birth for 8 group I animals and 9 group II animals.
Administration of each drug was started from the age of weeks. The drug was administered by diluting it with the drinking water of mice, and exchanging the drug 3 times a week.
The mice were dosed on the assumption that the water intake was 5 ml / day.

The body weight of each group of mice was measured every week.

As a result, the results shown in Table 1 below were obtained at the following measurement times.

<Pharmacological Test Example 2> Inhibition test of urine sugar by administration of active ingredient compound of the therapeutic agent for diabetes of the present invention The same KK mice as used in the above-mentioned Pharmacological Test Example 1 were divided into 3 groups, and group I (8 mice) was treated for diabetes of the present invention Active ingredient compound (sesquiterpene analog, monosodium salt of compound of formula (2) obtained in Production Example 3) 10 mg / kg administration group, II group (8
The same active ingredient compound 2 mg / kg administration group, group III (9 animals)
Was used as a control group (non-drug administration group).

The administration method of the active ingredient compound of the therapeutic agent for diabetes of the present invention is as described above.
According to the pharmacological test example 1>.

The urine sugar of the mouse was inspected every week using a urine glucose test paper (Diastic II, manufactured by Miles Sankyo Co., Ltd.).

The results were converted into points according to the following criteria, and the onset rate was calculated by multiplying the average points by the number of affected individuals.

That is, the results of the above-mentioned urine sugar test paper are traces: 5, +: 12.5, ++: 2
The average points of each group were calculated and accumulated as 5, +++: 50, +++++: 100.

The results are shown in FIG. 1 for each group.

In FIG. 1, the vertical axis represents the degree of urinary glucose (average point cumulative value) and the horizontal axis represents the age of the experimental mice (weeks). In the figure, (1) shows the results of group III (control group), (2)
Shows group II (the present invention, 2 mg / kg administration group), and (3) shows group I (the present invention, 10 mg / kg administration group).

From FIG. 1 above, the active ingredient compound of the therapeutic agent for diabetes of the present invention
It can be seen that both the 2 mg / kg administration group (Group II) and the 10 mg / kg administration group (Group I) show significantly lower values than the control group (Group III).

<Pharmacological Test Example 3> Effect of administration of the compound of the present invention as an active ingredient for diabetes on blood insulin level After the completion of <Pharmacological Test Example 2>, mice were sacrificed, and the insulin level in blood collected at the time of slaughter was measured. .

The results are shown in Table 2 below.

As shown in Table 2 above, the KK mice (control group (group I), diabetes mellitus of the present invention used in this experiment were compared with the blood insulin levels of normal mice (ICR) separately measured in the same manner. The insulin level of the therapeutic agent active ingredient compound administration group) was extremely high.

On the contrary, the active ingredient compound of the therapeutic agent for diabetes of the present invention is 10
The insulin level of the group administered with mg / kg (group III) is significantly lower than that of the control group (group I), and the administration of the diabetic active ingredient compound of the present invention is effective in lowering the blood insulin level. It turned out that it gives a good result.

In addition, the insulin level in blood of each group of experimental mice in this experiment was measured by the enzyme immunoassay (EIA) (glazyme insulin-EIA TEST) by measuring the insulin level of the serum separated from the whole blood of each experimental mouse according to the standard method. did.

<Pharmacological Test Example 4> Acute Toxicity Test An acute toxicity test of the sesquiterpene analog (the compound of formula (1), the active ingredient compound of the therapeutic agent for diabetes of the present invention) obtained in Production Example 2 was conducted on IV rats.

As a result, the LD ₅₀ value of the compound was 50 mg / kg.

[Brief description of drawings]

FIG. 1 is a graph showing the results of the inhibitory action of urinary sugar according to Pharmacological Test Example 2 of the present invention.

Claims (1)

[Claims] 1. A formula And expression A therapeutic agent for diabetes which comprises, as an active ingredient, at least one selected from the sesquiterpene analogues represented by and the salts of the above compounds.