JP3249124B2 - Blood lipid lowering agent, blood glucose lowering agent and glycation inhibitor based on ascofuranone and ascofuranone derivative - Google Patents

Description

TECHNICAL FIELD The present invention relates to a hypoglycemic agent and a glycation inhibitor comprising ascofuranone represented by the following general formula (I) and a derivative thereof as an active ingredient, and represented by the following general formula (II) The present invention relates to a blood lipid lowering agent containing an ascofuranone derivative as an active ingredient, and an ascofuranone derivative represented by the following general formula (III).

(Wherein R is a hydrogen atom, a lower alkylcarbonyl group, a pyridylcarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group, and one or two lower alkyl groups. R ′ represents a lower alkylcarbonyl group, a pyridylcarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group, Represents a carbamoyl group substituted by one or two lower alkyl groups, or a lower alkoxycarbonyl lower alkyl group, and R ₀ represents a toluenesulfonyl group or a lower alkoxycarbonyl lower alkyl group.) In the compounds represented by (I), R is water The compound of an elementary atom is a compound known as ascofuranone invented by the present inventors. Ascofuranone is a filamentous fungus Ascochyta visiae
Isoprenoid antibiotics produced by
The specific production method is described in JP-B-56-25310. Ascofuranone has been reported to have a serum lipid-lowering effect ("Journal of Antibiotics", Vol. 26, p. 681, 1973; and "Japan Journal of Pharmacology", Vol. 25, p. 35, 1975).

In addition, the hypoglycemic effect of ascochlorin and its derivatives produced from the filamentous fungi during the production of ascofuranone is already known (see Japanese Patent Publication No. 3-6138).

DISCLOSURE OF THE INVENTION The present inventors have conducted intensive studies in an attempt to explore the new usefulness of ascofuranone as a drug, and as a result, have found that ascofuranone has an excellent blood glucose lowering effect. Not only that, the novel ascofuranone derivative represented by the above general formula (II) has been found to have an excellent blood glucose lowering effect similarly to ascofuranone. Further, as one of the causes of diabetic complications, an increase in glycated glycated protein tissues and lipoproteins has been noted, but the compound represented by the above general formula (I) has a strong glycation inhibitory action. Found to show. In addition, they have found that the compound represented by the above general formula (I) containing ascofuranone has lower toxicity than ascochlorin and its derivatives. Furthermore, it has been found that the novel ascofuranone derivative represented by the general formula (II) has an excellent blood lipid lowering action. The present invention has been completed based on these findings.

The present invention provides a compound represented by the general formula (III) Wherein R ₀ represents a toluenesulfonyl group or a lower alkoxycarbonyl lower alkyl group.

Further, the present invention provides a compound represented by the general formula (II): (Wherein R ′ is a lower alkylcarbonyl group, a pyridylcarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group,
A carbamoyl group substituted by one or two lower alkyl groups or a lower alkoxycarbonyl lower alkyl group) as an active ingredient.

Further, the present invention provides a compound represented by the general formula (I): (Wherein R is a hydrogen atom, a lower alkylcarbonyl group, a pyridinecarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group, and one or two lower alkyl groups. A carbamoyl group or a lower alkoxycarbonyl lower alkyl group) as an active ingredient.

In the above substituents of the compound of the present invention, lower alkyl and lower alkoxy mean alkyl having 1 to 6 carbons and alkoxy having 1 to 6 carbons, respectively.

In the pyridylcarbonyl group, the carbonyl substitution position on the pyridine ring may be any of the 2-position (that is, picolinoyl), the 3-position (that is, nicotinoyl), and the 4-position (that is, isonicotinoyl).

The position of the alkyl group or lower alkoxy group on the benzene ring of the benzoyl group may be any of the ortho, meta and para positions.

In the present invention, the toluenesulfonyl group means any one of an orthotoluenesulfonyl group, a metatoluenesulfonyl group, and a paratoluenesulfonyl group.

In the present invention, the compound represented by the general formula (III) is a novel compound. It is produced by reacting the derivative in the presence of a condensing agent (pyridines, tertiary amines such as triethylamine, dimethylaniline, alkali base, etc.) or without adding a condensing agent.

Among the compounds used in the present invention, the following compounds can be exemplified.

4-O-acetylascofuranone (compound of Example-5) 4-O-propionylascofuranone 4-O-butyrylascofuranone 4-O-isonicotinoylascofuranone (Example-
4-O-nicotinoylascofuranone 4-O-picolinoylascofuranone (compound of Example-5) 4-O-methylcarbamoylascofuranone 4-O-ethylcarbamoylascofuranone 4-O-dimethylcarbamoyl Ascofuranone 4-O-diethylcarbamoylascofuranone (compound of Example-2) 4-O-methoxycarbonylmethylascofuranone (compound of Example-4) 4-O-methoxycarbonylethylascofuranone 4-O-methoxycarbonyl Propylascofuranone 4-O-ethoxycarbonylmethylascofuranone 4-O-ethoxycarbonylethylascofuranone 4-O-ethoxycarbonylpropylascofuranone (compound of Example-3) 4-O-paramethylbenzoylascofuranone 4- -Paramethoxybenzoylascofuranone (compound of Example-5) 4-O-paraethylbenzoylascofuranone 4-O-paraethoxybenzoylascofuranone 4-O-orthomethylbenzoylascofuranone 4-O-orthomethoxybenzoylascofuranone 4-O-orthoethylbenzoylascofuranone 4-O-orthoethoxybenzoylascofuranone 4-O-paratoluenesulfonylascofuranone (compound of Example-5) 4-O-orthotoluenesulfonylascofuranone 4-O-methtoluene Sulfonyluscofuranone Ascofuranone is a compound represented by the general formula (I) wherein R is a hydrogen atom.

When the compound of the present invention is used as a drug, it may be used alone, but it is usually desirable to mix it with a suspension, excipient or other adjuvant and formulate it into a dosage form suitable for oral administration. Lactose, sucrose,
It can be produced using various starches, glucose, cellulose, methylcellulose, carboxymethylcellulose, magnesium stearate, lauryl sulfate, talc, vegetable oil, lecithin and the like.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail based on examples,
This does not limit the invention in any way.

Examples Example-1 1.68 g (3.99 mmol) of ascofuranone was dissolved in 50 ml of dry pyridine, and 1.1 g (6.18 mmol) of isonicotinic acid chloride hydrochloride was added thereto with stirring, followed by heating and stirring at 80-90 ° C for 24 hours. The reaction solution was concentrated to dryness under reduced pressure. The residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid and water, dried, and concentrated under reduced pressure. The remaining oil was separated and purified by silica gel chromatography. 1.8 g of the desired product represented by the following formula was obtained as an oil.

Proton nuclear magnetic resonance <400 MHz, CDCl ₃ , internal standard TMS> δ: 1.21 (3H, s), 1.28 (3H, s), 1.54 (3H, s), 1.62
(3H, s), 1.93 (2H, m), 2.17 (2H, m), 2.40 (2H,
m), 2.69 (3H, s), 3.50 (2H, m), 4.52 (1H, -dd, J =
6.2Hz, 9.9Hz), 5.09 (1H, t, J = 7.0Hz), 5.48 (1H, t, J
= 7.0Hz), 8.04 (2H, dd, J = 1.5Hz, 4.4Hz), 8.91 (2H,
dd, J = 1.5Hz, 4.4Hz), 10.34 (1H, s), 12.61 (1H, s) Example 2 2.52 g (5.99 mmol) of ascofuranone was dissolved in 50 ml of dry pyridine, and 1.5 g (10.84 mmol) of N, N-diethylcarbamoyl chloride was added thereto.
The mixture was heated and stirred for hours. The residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid and water, dried, and concentrated under reduced pressure.
The remaining oily substance was separated by silica gel chromatography, and the oily target substance was dissolved in ethanol and left at room temperature to precipitate 1.0 g of the target substance crystal represented by the following formula. A sample recrystallized from ethanol showed a melting point of 69-70 ° C.

Proton nuclear magnetic resonance <400 MHz, CDCl ₃ , internal standard TMS> δ: 1.21 (3H, t, J = 7.0 Hz), 1.21 (3H, s), 1.28 (3H,
s), 1.31 (3H, t, J = 7.0Hz), 1.63 (3H, s), 1.74 (3
H, s), 2.02 (2H, m), 2.13 (2H, m), 2.40 (2H, br,
m), 2.64 (3H, s), 3.24 (1H, br, s), 3.40 (3H, q, J =
7.0Hz), 3.50 (2H, m), 4.51 (1H, dd, J = 6.6Hz, 9.5H
z), −5.15 (1H, dd, J = 5.5Hz, 7.0Hz), 5.51 (1H, t, J
= 7.0Hz), 10.28 (1H, s), 12.53 (1H, s) Example 3 2.52 g (5.99 mmol) of ascofuranone was dissolved in 50 ml of dimethylformamide. 0.2 g of 60% sodium hydride was added little by little to this. 1.8 g (9.3 mmol) of 4-bromobutyric acid ethyl ester was added to the obtained sodium salt solution.
And heated to 90-100 ° C for 3 hours. Then 0.1 g of 60% sodium hydride and methyl 4-bromobutyrate were added.
5 g was added and heated for another 10 hours. The reaction solution was concentrated under reduced pressure to dryness, and the residue was subjected to 40 ml of 1% hydrochloric acid and 40 ml of chloroform.
Separated with ml.

The chloroform solution is concentrated under reduced pressure to dryness. The remaining oil was separated and purified by silica gel chromatography to obtain 1.2 g of the desired product represented by the following formula as an oil.

Proton nuclear magnetic resonance <400 MHz, CDCl ₃ , internal standard TMS> δ: 1.03 (3H, br, s), 1.76 (3H, br, s), 2.03 (2H,
m), 2.17 (4H, m), 2.34 (1H, dd, J = 9.9Hz, 18.3Hz),
2.41 (1H, dd, J = 6.2Hz, 18.3Hz), 2.62 (2H, m), 2.62
(3H, s), 3.35 (2H, d, J = 6.6Hz), 3.97 (2H, t, -J =
6.0Hz), 4.16 (2H, q, J = 7.0Hz), 4.51 (1H, dd, J = 6.2
Hz, 9.9Hz), 5.17 (1H, dd, J = 6.6Hz, 5.5Hz), 5.50 (1
H, t, J = 6.6Hz), 10.24 (1H, s), 12.51 (1H, s) Example-4 2.52 g (5.99 mmol) of ascofuranone was dissolved in 30 ml of dimethylformamide. 0.2 g of 60% sodium hydride was added little by little to this. 1.5 g (9.8 mmol) of bromoacetic acid methyl ester was added to the obtained sodium salt solution. After standing at room temperature overnight, 0.02 g of 60% sodium hydride and 0.15 g of bromoacetic acid methyl ester were further added. After standing overnight, the mixture was concentrated under reduced pressure. 1 for the remaining oil
100% hydrochloric acid and 100 ml of chloroform were added, and the mixture was stirred with a separating funnel, and the chloroform layer was separated and concentrated to dryness.
Methanol was added to the remaining oil, and the mixture was allowed to stand overnight, whereby 1.9 g of crystals of the target product precipitated. A sample recrystallized from methanol showed a melting point of 77 ° C.

Proton nuclear magnetic resonance <400 MHz, CDCl ₃ , internal standard TMS> δ: 1.21 (3H, s), 1.27 (3H, s), 1.63 (3H, s), 1.76
(3H, s), 2.03 (2H, m), 2.14 (2H, m), 2.35 (1H, dd,
J = 10.3Hz, 18.3Hz), 2.43 (1H, dd, -J = 6.2Hz, 18.3H
z), 2.64 (3H, s), 3.45 (2H, d, J = 7.0 Hz), 3.84 (3
H, s), 4.52 (1H, dd, J = 6.2Hz, 10.3Hz), 4.58 (2H,
s), 5.16 (1H, dd, J = 5.5Hz, 7.0Hz), 5.50 (1H, t, J =
7.0Hz), 10.26 (1H, s), 12.52 (1H, s) Example-5 The following compounds were produced from ascofuranone and the corresponding acid chloride or sulfonyl chloride in the same manner as in each of the above Examples.

(A) 4-O-paramethoxybenzoylascofuranone (tan oil) (b) 4-O-acetylascofuranone (tan oil) (c) 4-O-picolinoylascofuranone (tan oil) (D) 4-O-Paratoluenesulfonylascofuranone (brown oil) In vivo and in vitro experiments were performed to examine the glycation inhibitory action and hypoglycemic action of ascofuranone and its derivatives of the present invention. The glycation inhibitory effect is obtained by adding ascofuranone and its derivative to a reaction solution containing bovine serum albumin and glucose, culturing for a long time, measuring the amount of acid hydrolyzate furosine produced of fructose-lysine, ribose, arginine, Ascofuranone and its derivative were added to the reaction solution containing lysine and cultured, and the glycation inhibitory effect of ascofuranone and its derivative was examined by measuring the amount of pentosidine, one of the late-stage compounds of glycation produced. The hypoglycemic effect was investigated by orally administering ascofuranone and its derivatives to hereditary diabetic db / db mice and streptozotocin diabetic mice, and measuring the blood glucose level.

Example-6 0.1, 0.4, 1.6 mg / ml ascofuranone was added to a reaction solution containing 25 mg / ml bovine serum albumin and 400 mM glucose, and cultured at 37 ° C for 14 days. The produced furosine was measured by high performance liquid chromatography (280 nm, 0.7 mM phosphoric acid) and indicated by the area. The results are shown in Table 1.

As shown in Table 1, the addition of ascofuranone 0.4 or 1.6 mg / ml significantly inhibited the production of furosine.

Example-7 To a reaction solution containing 25 mg / ml bovine serum albumin and 400 mM glucose, 1 mg / ml ascofuranone and its derivative were added, and the mixture was cultured at 37 ° C for 12 days. The produced furosine was measured by high performance liquid chromatography and indicated by its area.

 The results are shown in Table 2.

As shown in Table 2, ascofuranone and its derivatives all significantly inhibited the production of furosine.

Example -8 20,100,500,1000 μg / ml of ascofuranone was added to a reaction solution containing 10 mM of ribose, arginine, and lysine.
The cells were cultured at 7 ° C for 4 or 8 days. The generated pentosidine was measured by high performance liquid chromatography (excitation 335, fluorescence 385, 7 mM phosphoric acid) and indicated by its area. The results are shown in Table 3.

The results were as shown in Table 3, ascofuranone 1000,500μ
The addition of g / ml significantly inhibited the production of pentosidine even at 100%, and even at 100 and 20 μg / ml.

Example-9 1 mg / ml of ascofuranone and its derivative were added to a reaction solution containing 10 mM of ribose, arginine, and lysine, respectively.
C. for 8 days. The resulting pentosidine was measured by high performance liquid chromatography and expressed as its area. The results are shown in Table 4.

As shown in Table 4, ascofuranone, 4-O-methoxycarbonylmethylascofuranone, 4-O-ethoxycarbonylpropylascofuranone and 4-O-isonicotinoylascofuranone produced 90% of pentosidine.
% Or more.

Example -10 Eight-week-old male hereditary diabetic db / db mice were fed with ascofuranone or a derivative thereof containing 0.3% (Clear Japan, C
E-2) was given for one week, sacrificed on the seventh day, and blood glucose and blood triglyceride were measured. Table 5 shows the results.

As a result, as shown in Table 5, all the compounds used in the experiment showed a blood glucose and blood triglyceride lowering action.

Example-11 ddY to which 160 mg / kg of streptozotocin was intraperitoneally administered
8 mg / kg of ascofuranone and its derivative were orally administered to the mice for 7 days, and the blood glucose level was measured.

As shown in Table 6, ascofuranone and its derivatives significantly reduced the blood glucose level of streptozotocin-administered mice.

Example-12 Eight-week-old male hereditary diabetic db / db mice and their normal littermates contained a diet containing 0.3% ascofuranone (CLEA Japan, C
E-2) was given for one week, sacrificed on the seventh day, and blood glucose and blood triglyceride were measured. The results are shown in Table 7.

As shown in Table 7, the blood sugar level and the blood triglyceride level were significantly reduced by feeding db / db mice with ascofuranone in the diet.

Example -13 Ascofuranone 391,156,63 mg / kg suspended in 2% gum arabic was administered to an 8-week-old male hereditary diabetic db / db mouse.
Oral administration for days. The results are shown in Table 8.

As shown in Table 8, the oral administration of the suspension of ascofuranone for 7 days significantly reduced the blood sugar level and blood triglyceride level.

Example -14 Ascofuranone or a derivative thereof was orally administered to ddY mice at 8 mg / kg for 7 days. The results are shown in Table 9.

As shown in Table 9, the 4-O-methoxycarbonylmethylascofuranone-administered group had 26.4% neutral fat and 14.0% cholesterol, and the 4-O-diethylcarbamoylascofuranone-administered group had 17.5% neutral fat. Each showed a significantly lower value than the control group.

INDUSTRIAL APPLICABILITY Ascofuranone and the ascofuranone derivative represented by the formula (II) have excellent hypoglycemic, hypolipidemic and glycation inhibitory effects, and are useful as preventive and therapeutic agents for diabetes, arteriosclerosis, etc. Extremely useful.

Claims (4)

(57) [Claims] 1. A compound of the general formula (III) (Wherein R ₀ represents a toluenesulfonyl group or a lower alkoxycarbonyl lower alkyl group). 2. A compound of the general formula (II) (Wherein R ′ is a lower alkylcarbonyl group, a pyridylcarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group, one or two lower alkyl groups. A carbamoyl group or a lower alkoxycarbonyl lower alkyl group) as an active ingredient. 3. A compound of the general formula (I) (Wherein R is a hydrogen atom, a lower alkylcarbonyl group, a pyridylcarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group, and one or two lower alkyl groups. A carbamoyl group or a lower alkoxycarbonyl lower alkyl group) as an active ingredient. 4. A compound of the formula (I) (Wherein R is a hydrogen atom, a lower alkylcarbonyl group, a pyridylcarbonyl group, a benzoyl group substituted with one lower alkyl group or a lower alkoxy group, a toluenesulfonyl group, and one or two lower alkyl groups. A carbamoyl group or a lower alkoxycarbonyl lower alkyl group) as an active ingredient.