JPH036138B2 - - Google Patents

Description

[Detailed description of the invention]

Diabetes is a typical metabolic disease that develops due to genetic predisposition and the involvement of complex environmental factors. Since the onset of diabetes is directly triggered by the involvement of a wide variety of factors, there is a view that the name diabetes only represents one syndrome. However, in general, there are two types of diabetes: juvenile diabetes, which mainly develops in young people and can lead to serious disorders if exogenous insulin is not supplemented, and adult-type diabetes, which develops in adults and requires little insulin supplementation. It is classified as The latter is known to be closely related to obesity in its onset. Although there are differences in whether insulin injections are necessary or unnecessary for treatment, what all types of diabetes have in common is that the fundamental cause is insufficient insulin action in the body. This is what we do. Insulin is an essential peptide hormone in vertebrates that promotes the synthesis of glycogen, protein, and fat, and exhibits a variety of physiological effects such as suppressing gluconeogenesis in the liver, suppressing lipolysis in adipose tissue, and promoting proliferation in cell culture. It has been known. However, its most essential function is that it is the only hormone that acts to store energy from the food an animal ingests in the body in preparation for the intermeal or fasting period. This is in sharp contrast to other hormones, such as catecholamines, glucagon, ACTH, and glucocorticoids, which act to release energy stored in the body. It is natural that insulin injections have a specific effect on diabetes caused by insufficient insulin action. However, using too much insulin is not a good thing because it disrupts the balance with other hormones. Therefore, the existence of a drug that can enhance insulin action in vivo would be desirable for normalizing metabolism not only in non-insulin-dependent diabetes but even in insulin-dependent diabetes. On the other hand, it is known from various epidemiological studies that ischemic heart disease occurs at a high rate not only in diabetes but also in cases where decreased glucose tolerance and hyperinsulinemia coexist. In this case, deterioration of glucose tolerance is caused by insulin-dependent tissues becoming insulin insensitive. Therefore, the living body is no longer able to maintain homeostasis with the normal amount of insulin secretion, so it tries to compensate for the lack of insulin secretion by increasing insulin secretion from the pancreas. In other words, the seemingly contradictory coexistence of hyperinsulinemia and worsening of glucose tolerance is a compensatory reaction associated with insulin insensitivity. In such pathological conditions, drugs that enhance the action of endogenous and exogenous insulin by improving tissue insulin sensitivity are extremely meaningful for the prevention and treatment of ischemic heart disease and the cause of it, coronary artery sclerosis. I have to say it. As a result of many years of searching for substances that enhance insulin action, the present inventors discovered that ascochlorin 4.
The present inventors discovered that a new derivative obtained by acylating the hydroxyl group at the -position has the property of significantly enhancing insulin action, leading to the completion of the present invention. Ascochlorin is an isobrenoid produced by the filamentous fungus Ascochyta vicia together with several congeners including ascofuranone (Tamura et al. J.
Antibiotics.21:539, 1968). Ascochlorin is also found in some other filamentous fungi, such as Fusarium. sp.
(Fusarium sp.) RHEvans et al., US Patent 3,
546, 073, December 8, 1973). Nectria coccinea (DCAldrich et al., J.
Chem.Soc.Perkin I, page 2136, 1972), Cylindrocladium sp.
Kato et al., J. Antibiotics. 24: 168 pages, 1970)
and Cylindrocladium ilicicola Minato et al., J.
Antibiotics 25:315, 1972) and other filamentous fungi. When ascochlorin is administered to mammals in a form that is rapidly absorbed into the body, severe symptoms of poisoning occur, and in severe cases, death occurs. It is known that a means to specifically weaken the acute toxicity of ascochlorin without impairing its physiological effects is to modify the hydroxyl group at the 4-position of the aromatic ring (Hosokawa et al., Agr. Biol. Chem. 45: 531 page,
(1980, Hosokawa et al., Eur.J.pharmacol.69:429, 1981). However, the 4-0- used here
Methylascochlorin has the disadvantage that it has poor water solubility and is difficult to increase blood concentration when administered systemically or orally. The present inventors investigated with the aim of improving the above-mentioned drawbacks, and found that a new derivative in which the hydroxyl group at the 4-position of the aromatic ring was modified with an acyl group and a carbamoyl group was found to be superior in action and safety, and was able to develop the present invention. Ivy. In other words, the 4-0-acyl derivative of ascochlorin is produced by adding a condensing agent (pyridine, triethylamine, etc.) to ascochlorin, an acid halide, an acid anhydride, or in some cases, a reactive derivative of an acid such as cyanate or isocyanate. amine, dimethylaniline, alkali base, etc.) or without the addition of a condensing agent. The reaction proceeds without solvent, but the yield and
It is usually carried out in a solvent from the viewpoint of operability. A solvent is selected that does not react with the reactive derivative of the acid. Benzene, dimethylformamide, ethers, chloroform, acetone, etc. are usually used. Moreover, there is no problem even if the condensing agent is also used as a solvent. The reaction temperature range can be wide from room temperature to the boiling point of the solvent. However, the use of a large excess of the reactive derivative of the acid should be avoided since it may result in the formation of 2,4-bis-0-acyl derivatives of ascochlorin. However, under the reaction conditions shown in the Examples, the production of bis isomers is almost suppressed. Purification of the desired product can be carried out by common means such as recrystallization, column chromatography, and solvent extraction. The physiological action of the novel compound of the present invention is explained as follows. That is, continuous oral administration for one week to healthy animals significantly lowers blood sugar and blood lipids. This fact is suitable for improving hypercaloremia associated with diabetes and arteriosclerosis and for maintaining normal metabolic homeostasis in the body. In fact, when the compound of the present invention is orally administered to diabetic disease model animals, a marked improving effect is observed. For example, the C57BL/Ksj (db ⁺ /db ⁺ ) strain of genetically obese diabetic mice exhibits hyperglycemia, hyperlipidemia, obesity, hyperinsulinemia, insulin resistance, polydipsia, polyuria, urinary sugar excretion, and vascular disorders in adults. It is said to be a pathological model similar to sexual diabetes. Conventional antidiabetic drugs, such as sulfonylurea and biguanide compounds, are not very effective against these diabetic mice. However, the compound of the present invention significantly improved the abnormalities characteristic of diabetes without reducing food intake. That is, in addition to the reduction in blood sugar and urinary sugar excretion, what is especially noteworthy is that symptoms specific to diabetes such as polydipsia and polyuria were significantly improved. It also showed a reduction in blood sugar and serum lipids and urinary sugar excretion in diabetic animals induced by alloxan and streptozotocin. The results of experiments using the compounds of the present invention to examine their mechanisms of action in vitro have revealed that these compounds enhance the action of insulin and, at the same time, partially act as a substitute for insulin. That is, when the compound of the present invention was applied to normal rat liver sections in the presence of insulin at a low concentration of 10 ^-6 M, glucose oxidation was clearly promoted. Furthermore, when applied to liver sections of diabetic rats in the presence of insulin, fatty acid synthesis was restored to the normal range. On the other hand, when insulin was applied to the epididymal adipose tissue of healthy and diabetic animals in the presence of insulin, glucose uptake and metabolism were enhanced. A similar reaction was observed when added to kidney sections. Furthermore, when the compound of the present invention is applied to the microsomal fraction obtained by grinding epididymal adipose tissue, lipoprotein lipase activity is increased in the same way as when insulin is added, and conversely, neutral fat lipase activity is increased. Activity was strongly inhibited. Effects on diabetic and healthy animals and in
Based on experimental results in vitro, it is presumed that the compound of the present invention not only enhances the action of insulin in vivo, but also partially replaces its action. Such an action has not been found in any conventional drug except insulin. In particular, the compound of the present invention can be said to have a great advantage in terms of clinical treatment in that it exhibits such effects upon oral administration. Although the compound of the present invention may be used alone, it is usually preferable to mix it with a suspending agent, excipient, or other auxiliary agent to formulate a dosage form suitable for parenteral and oral administration. Preferred formulations include, for example, injections, granules, granules, tablets, sugar tablets, pills, capsules, and suppositories. These preparations can be manufactured according to conventional methods using, for example, the following excipients or auxiliaries. Lactose, sucrose, various starches, glucose, cellulose, methylcellulose, carboxymethylcellulose, magnesium stearate, lauryl sulfate, talc, vegetable oil, octyldecyl triglyceride, bicarbonate of soda, various polysorbates,
Polyethylene glycol, lecithin, and mixtures of two or more of these. Preparations for oral administration preferably contain the active ingredient in an amount of 10 to 55% (by weight), and injections preferably contain 1 to 20% (by weight). The medicament of the present invention can be effectively used for the treatment and prevention of diabetes, arteriosclerosis and ischemic heart disease in humans and animals. The toxicity of the compounds of the present invention is rather low, with an acute toxicity LD ₅₀ for rats and mice of 0.5 to 10.0 g/Kg or more for oral administration and 150 to 500 mg/Kg for intraperitoneal administration. The dosage of the drug of the present invention varies depending on the type of pathology, symptoms, etc., but for example, in the case of injection, it is 5 to 1000 mg per adult per day, and in the case of oral administration, it is 30 to 3000 mg per day.
mg, or in the case of suppositories, 5 to 1000 mg can achieve the purpose. Next, formulation examples are shown. 100 mg of 4-0-nicotinoyl ascochlorin is dissolved in a solution consisting of 20 ml of purified corn oil and 200 mg of lecithin, sealed in a light-shielding ampoule filled with nitrogen gas, and sterilized under pressure in a conventional manner. This solution is suspended in an intravenous infusion solution or a sugar solution and injected intravenously. Tablets for oral administration are formulated as follows.
100 parts of 4-0-nicotinyl ascochlorin fine powder (particle size approximately 2μ), 88 parts of lactose, and corn starch
100 copies, HPC-SL 2 copies, L-HPC (PO-30) 50
1 part, crystalline cellulose, 33 parts, calcium stearate, 5 parts, and talc, 10 parts, were added, mixed well, and compressed into tablets with a diameter of 8 mm and a weight of 250 mg using a tablet machine. Example 1 10 g (24.7 mmol) of ascochlorin are dissolved in 50 ml of dry pyridine. Cool the surroundings of the container with ice water and add 6.6 g of nicotinic acid chloride hydrochloride while stirring.
(37.07 mmol) little by little. After the addition is complete, the temperature inside the reaction vessel is gradually returned to room temperature while stirring is continued. After stirring all day and night, the reaction solution was concentrated to dryness under reduced pressure. The residue is separated and extracted with chloroform and water, and the chloroform layer is thoroughly washed with water and then dehydrated by adding anhydrous sodium sulfate. After dehydration, the resulting filtrate is concentrated under reduced pressure, and the remaining oil is separated and purified using silica gel column chromatography.
Fractions of the desired product eluted with chloroform containing 1 to 2% methanol or benzene containing 5% ethyl acetate are collected and concentrated under reduced pressure to obtain a viscous oil. If this is dissolved in ethanol and left to stand, crystals of the desired product will precipitate. Object crystal 9.6
g (76.3%), and the preparation recrystallized from ethanol has a melting point of 159-160°C. The elemental analysis values for this product are C ₂₉ H ₃₂ O ₅ ClN Theoretical values: C68.29%, H6.32%, N2.75% Actual values: C68.23%, H6.36%, N2.80% Structure formula; Example 2 40.5 g of ascochlorin in a 1.0 Erlenmeyer flask
(0.1 mol), dry benzene 50ml, dry pyridine 24
ml (0.297 mol) and shake to dissolve uniformly. While stirring this solution on a magnetic stirrer, 25.22 g of nicotinic acid chloride hydrochloride was added.
(0.142 mol) is added. The mixture was stirred at room temperature for 3 hours, and the precipitated and suspended pyridine hydrochloride was removed by filtration. Add 500 ml of water to the filtrate, shake it, and remove the aqueous layer. Repeat this operation three times. If the mixture becomes turbid during shaking and is difficult to separate, add saline. The benzene layer is dried over anhydrous sodium sulfate and the solvent is distilled off under reduced pressure to obtain a viscous oil. When this is dissolved in 800ml of ethanol and left to stand, 41.0g of target crystals are produced.
(80.4%) was obtained. When recrystallized from ethanol, it shows a melting point of 159-160℃, and the elemental analysis value is
As C ₂₉ H ₃₂ O ₅ ClN Theoretical value (%); C68.29, H6.32, N2.75 Actual value (%); C68.21, H6.32, N2.76 Proton nuclear magnetic resonance δ value < 100MHz, solvent
CDCl ₃ , internal standard TMS > 0.69 (3H, s), 0.80
(3H, d), 0.82 (9H, d), 1.70 (3H,
s), 3.55 (2H, d), 5.37 (1H, d), 5.54
(1H, t), 5.84 (1H, d), 7.49 (1H,
m), 8.55 (1H, d), 8.96 (1H, d), 9.42
(1H, s), 10.34 (1H, s) 12.60 (1H,
s) Structural formula; Example 3 20 g (49.4 mmol) of ascochlorin was dissolved in 100 ml of dry pyridine, and 6.9 g (49.9 mmol) of diethylcarbamoyl chloride was added thereto and heated under reflux. After about 5 hours, 6.9 g of diethylcarbamoyl chloride was added and further heated. Continue reflux. When it is confirmed that ascochlorin has disappeared from the reaction system, the reaction solution is concentrated under reduced pressure to dryness.
The residue is separated between water and benzene, and the benzene layer is thoroughly washed with water, dried over anhydrous sodium sulfate, and the solvent is distilled off. The remaining oil is separated and purified by silica gel column chromatography. A viscous oil is obtained by collecting a fraction of the target product eluted with benzene containing 5% acetic acid ethyl ester and concentrating to dryness. When this was dissolved in ethanol and left in a cold place, 16 g (64%) of the target product precipitated as crystals. When the crude crystals are recrystallized from ethanol, the melting point is 125-127.
℃, elemental analysis values are as C ₂₈ H ₃₈ O ₅ ClN Theoretical value (%); C66.72, H7.60, N2.78 Actual value (%); C66.85, H7.67, N2.80 δ value of proton nuclear magnetic resonance <100MHz, CDCl ₃ , internal standard TMS>: 0.67 (3H, s) 0.79 (3H,
d), 0.82 (3H, d), 1.12~1.36 (6H,
m), 1.86 (3H, s), 2.20~2.50 (3H,
m), 2.63 (3H, s), 3.30~3.60 (6H,
m), 5.35 (1H, d), 5.42 (1H, t), 5.89
(1H, d), 10.28 (1H, s), 12.53 (1H,
s) Structural formula of the target object; Example 4 10 g (24.7 mmol) of ascochlorin is dissolved in 100 ml of dry pyridine, 8.0 g (39.0 mmol) of parachlorophenoxyacetyl chloride is added, and the mixture is heated and stirred at 60-70°C for 5 hours. Next, parachlorophenoxyacetyl chloride 1.0
g (4.9 mmol) and stirred while heating at 60-70°C for 5 hours. Next, the reaction solution was concentrated to dryness under reduced pressure and treated in the same manner as in Example 3. The oily target product separated and purified by silica gel column chromatography was dissolved in ethanol and left at room temperature to precipitate 4.7 g (33.2%) of the target product as crystals. The sample recrystallized from methanol has a melting point of 122-124°C, and the elemental analysis value is C ₃₁ H ₃₄ O ₆ Cl ₂ Theoretical value (%): C64.92, H5.98 Actual value (%): C64. 86, H5.95 δ value of proton nuclear magnetic resonance <100MHz, CDCl ₃ , internal standard TMS>: 0.70 (3H, s), 0.80 (3H,
d), 0.83 (3H, d), 1.87 (3H, s) 2.66
(3H, s), 3.40 (2H, d), 4.93 (2H,
s), 5.20 (1H, t), 5.38 (1H, d), 5.84
(1H, d), 6.92 (2H, d), 7.30 (2H,
d), 10.30 (1H, s), 12.56 (1H, s) Structural formula of target object: Example 5 7 g (17.3 mmol) of ascochlorin is dissolved in 70 ml of dry pyridine, 8.0 g (46.9 mmol) of para-methoxybenzoyl chloride is added thereto, and the mixture is heated under reflux for 5 hours. Thereafter, the desired product obtained by treatment in the same manner as in Example 3 was dissolved in ethanol and allowed to stand at room temperature to precipitate 3.8 g (40.8%) of crystals of the desired product. When this crude crystal is recrystallized with ethanol, it shows a melting point of 155-156℃, and the elemental analysis value is C ₃₁ H ₃₅ O ₆ Cl. Theoretical value (%): C69.07, H6.54 Actual value (%): C68 .82, H6.56 δ value of proton nuclear magnetic resonance <100MHz, CDCl ₃ , internal standard TMS>: 0.68 (3H, s), 0.79 (3H,
d), 0.82 (3H, d), 1.70 (3H, s), 2.67
(3H, s), 3.55 (2H, d), 3.91 (3H,
s), 5.35 (1H, d), 5.50 (1H, t), 5.86
(1H, d), 7.01 (2H, d), 8.16 (2H,
d), 10.32 (1H, s), 12.57 (1H, s) Structural formula of target object: Example 6 10 g (24.7 mmol) of ascochlorin was dissolved in 100 ml of dry pyridine, 5.9 g (29.6 mmol) of para-methoxycarbonylbenzoyl chloride was added thereto, and the mixture was heated and stirred at 60 to 70°C for 7 hours, and further diluted with para-methoxycarbonylbenzoyl chloride. Added 5.9 g (29.6 mmol) of chloride and heated for 7 hours at 60~
Heat and stir at 70℃. Thereafter, the oily target product obtained by the same treatment as in Example 3 was dissolved in ethanol and left at room temperature. Target crystal 3.3g (23.6
%) is precipitated. When the crude crystals are recrystallized from ethanol, they show a melting point of 147-148℃, and the elemental analysis values are
As C ₃₂ H ₃₅ O ₇ Cl Theoretical value (%); C67.78, H6.22 Actual value (%); C67.78, H6.30 δ of proton nuclear magnetic resonance <100MHz, CDCl ₃ , internal standard TMS> Value: 0.68 (3H, s), 0.78 (3H,
d), 0.81 (3H, d), 1.59 (3H, s), 2.69
(3H, s), 3.55 (2H, d), 3.99 (3H, s)
5.35 (1H, d), 5.53 (1H, t), 5.84 (1H,
d), 10.34 (1H, s), 12.60 (1H, s)
8.14 (4H, m) Structural formula of target object: Example 7 20 g (49.4 mmol) of ascochlorin was dissolved in 100 ml of dry pyridine, and 13.2 g (73.7 mmol) of isonicotinic acid chloride hydrochloride was added thereto.
After heating and stirring at ℃ for 4 hours, 5 g (28.1 mmol) of isonicotinic acid chloride hydrochloride was further added and heating and stirring was continued for 7 hours. Then Example 3
The target product oily residue obtained by the same treatment as above is dissolved in ethanol and left in a cold place to obtain 7.2 g (28.6%) of the target product crystals. When the crude crystals are recrystallized from ethanol, they show a melting point of 111-113°C, and the elemental analysis values are as C ₂₉ H ₃₂ O ₅ ClN Theoretical value (%): C68.30, H6.28 Actual value (%): C68.27 , H6.31 Proton nuclear magnetic resonance <100MHz, CDCl ₃ , internal standard TMS> δ value: 0.68 (1H, s) 0.78 (3H,
d), 0.81 (3H, d), 1.67 (3H, s), 2.69
(3H, s), 3.45 (2H, d), 5.28 (1H, d)
5.35 (1H, t), 5.83 (1H, d), 800 (2H,
d), 8.80 (2H, d), 10.34 (1H, s)
12.61 (1H, s) Structural formula of target object: Example 8 5 g (12.35 mmol) of ascochlorin was dissolved in 50 ml of dry pyridine, 4.4 g (24.7 mmol) of picolinic acid chloride hydrochloride was added thereto, and the mixture was heated at 60°C.
After heating and stirring for 3 hours at 60° C., 1.5 g (8.43 mmol) of picolinic acid chloride hydrochloride was added and stirring at 60° C. for 7 hours. Then Example 3
Dissolve the target product in oil form in ethanol and leave it at room temperature to form crystals of the target product.
Obtain 2.0g (32%). When this crude crystal is recrystallized from ethanol, it shows a melting point of 150-152℃,
Elemental analysis values are as C ₂₉ H ₃₂ O ₅ ClN Theoretical value (%); C68.30, H6.28 Actual value (%); C68.30, H6.25 Proton nuclear magnetic resonance <100MHz, CDCl ₃ , internal standard δ value of TMS>: 0.66 (3H, s), 0.77 (3H,
d), 0.79 (3H, d), 1.76 (3H, s), 2.68
(3H, s), 3.55 (2H, d), 5.26 (1H,
d), 5.41 (1H, t), 5.83 (1H, d), 7.62
(1H, m), 7.96 (1H, m), 8.26 (1H,
d), 8.87 (1H, d), 10.33 (1H, s),
12.59 (1H, s) Structural formula of target object: Example 9 Genetic obese diabetic mouse C57BL/Ksj (db ⁺ /
db ⁺ ) to 4-0-nicotinoyl ascochlorin
1 feed containing 0.05% (Japan Clea, CE-2)
I gave it a week. Food intake, water consumption, urine volume, and urinary sugar excretion were measured daily for one week immediately before drug administration and one week after drug administration to examine the effects of the drug.

[Table] As is clear from the table, 4-0-nicotinoyl ascochlorine significantly inhibited polydipsia and polyuria and urinary sugar excretion in genetically obese diabetic mice. Example 10 Five-week-old ddY male mice were fed a diet containing 0.05% of the novel compound of the present invention (Clea Japan, CE-2) for one week, sacrificed on the seventh day, and blood lipids and blood sugar were measured. . The results are shown in Table 2.

[Table] As shown in the table above, all compounds have hypoglycemic effects. Furthermore, 4-0-nicotinoyl ascochlorin, 4-0-(P-chlorophenoxy)acetylar ascochlorin, and 4-0-isonicotinoyl ascochlorin exhibited a serum lipid-lowering effect. Example 11 Streptozotocin 150 mg/Kg was intraperitoneally administered to 5-week-old ddY male mice (n = 7), and 24 hours later, 4-0-nicotinoyl ascochlorine 0.05 was administered intraperitoneally.
% was fed for one week. The control group was given the same drug-free diet (Clea Japan, CE-2). Animals were sacrificed on day 7 and blood glucose and plasma lipids were measured. The results are shown in Table 3.

[Table] As shown in the table, 4-0-nicotinoyl ascochlorine significantly suppressed increases in blood sugar and plasma triglycerides in the streptozotocin diabetic model. Example 12 Genetic obese diabetic mouse C57BL/Ksj (db ⁺ /
db ⁺ ) to 4-0-nicotinoyl ascochlorin
Feed containing 0.05% was given for one week. Control group mice of the same age were given the same drug-free diet (Clea Japan,
CE-2) was given. The animals were sacrificed one week later and blood sugar and blood lipids were measured, and the results are shown in the table below.

【table】

[Table] As shown in the table, 4-0-nicotinoyl ascochlorine significantly improved hyperlipidemia and hyperglycemia in these obese diabetic mice.

Claims (1)

[Claims] 1. General formula (In the formula, R means a pyridyl group, an amino group substituted with a lower alkyl group, a phenoxyalkyl group having a halogen atom in the nucleus, or a phenyl group having a lower alkoxy group or a lower alkoxycarbonyl group in the nucleus. ) ascochlorin derivatives.