JPH0416473B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention discloses a novel compound 4,5-dihydroxy-1,3-dithiolane-2 which has liver function-activating effect.
-This invention relates to a method for producing ylidene malonic acid derivatives. More specifically, the present invention relates to the general formula (): (In the formula, R ¹ and R ² are the same or different and represent an alkyl group, a cyclohexyl group, a benzyl group, an alkenyl group, an alkynyl group, or an alkoxyalkyl group, and M represents an alkali metal atom or an ammonium group.) The compound represented and the following formula: General formula () characterized by reacting glyoxal represented by in the presence of an acid: The present invention provides a method for producing a 4,5-dihydroxy-1,3-dithiolan-2-ylidenemalonic acid derivative represented by the formula (wherein R ¹ and R ² are the same as above). That is, according to the present invention, the 4,5-dihydroxy-1,3-dithiolan-2-ylidenemalonic acid derivative represented by the general formula () is obtained by inactivating the compound represented by the general formula () in the presence of an acid. It can be obtained by reaction with glyoxal in a solvent. Next, the reaction route of the method of the present invention is schematically shown. (In the formula, R ¹ and R ² are the same as above, and M represents an alkali metal atom or an ammonium group.) In the method of the present invention, examples of acids that can be used include inorganic acids such as sulfuric acid and hydrochloric acid, and organic acids such as acetic acid. Mention may be made of acids, of which acetic acid is particularly preferred. Although water can be used as the solvent, it is generally preferable to use an inert organic solvent. Any inert organic solvent may be used as long as it does not interfere with this type of reaction, such as aromatic hydrocarbons such as benzene and toluene; halogenated hydrocarbons such as carbon tetrachloride, chloroform, and dichloromethane; diethyl Examples include ethers such as ether, tetrahydrofuran and dioxane; esters such as ethyl acetate; ketones such as acetone; dimethylformamide and dimethyl sulfoxide. The reaction temperature is preferably 50°C or lower, particularly preferably a low temperature around 15°C. The reaction molar ratio is desirably selected from the range of equimolar to slightly excess molar of glyoxal to the compound represented by the general formula (). The compound represented by the general formula () used in the present invention can be synthesized according to the following scheme. (In the formula, R ¹ , R ² and M are the same as above.) That is, by reacting the malonic acid ester represented by the general formula () with carbon disulfide in the presence of a base, The represented compounds can be obtained. Examples of the base include alkali metal hydroxides such as sodium hydroxide and potassium hydroxide, aqueous ammonia, and the like. The compound represented by the general formula () synthesized by the present invention is a new compound that has not been described in any literature, and has, for example, a liver function-activating effect, and is therefore useful as a liver function-activating agent for humans or animals, or as a therapeutic agent for liver failure. . Representative examples of the compound represented by the general formula () are as follows. General formula ():

【formula】 in

[Table] The compound represented by the general formula () has low toxicity to warm-blooded animals, and acute oral toxicity to rats (male).
For example, the LD ₅₀ value is 1000-3000 mg/Kg for compound No. 1,
3000-5000mg/Kg for compound No. 2, compound No. 3
500mg/Kg or more, 5000mg/Kg or more for compound No. 8,
Compound No. 12 is 1000-3000mg/Kg. The compound represented by the general formula () is particularly useful as a therapeutic agent for liver diseases. For example, it is known that various drugs such as carbon tetrachloride can be administered to healthy test animals to experimentally induce liver damage in the animals (for example, Japanese Patent Publication No. 18579/1983). In addition, as disease models of cholestasis seen in human clinical practice, common bile duct ligation is used for extrahepatic cholestasis, and cholangitis induced by α-naphthyl isothiocyanate is used for intrahepatic cholestasis. It is well known that it is made (Nippon Rinsai, Volume 30, 1).
No. 216 (1972)), and morphologically it is said to be extremely similar to the liver findings of cholestasis in humans (Clinical Studies of Drug-induced Liver Injury, edited by Yuo Yamamoto, p. 234). Therefore, using α-naphthyl isothiocyanate-induced liver damage as a pathological model to search for compounds that have an inhibitory effect will be part of the evaluation of drugs that can be used in human clinical practice in the future. The compound represented by the general formula () can be administered orally or parenterally (e.g., by injection) to test animals with various experimentally created pathological models of liver damage, resulting in significant liver function. It has been found that this has the effect of suppressing or improving the decrease in Therefore, the compounds represented by the general formula () are useful as human and veterinary medicines for the treatment or prevention of liver diseases. In other words, acute or chronic liver diseases in humans and animals caused by various causes, such as fatty liver, alcoholic hepatitis, hepatitis, toxic liver disease, hemorrhagic liver disease, cholestatic liver disease, or their terminal symptoms. It can be used as a treatment for certain liver cirrhosis. The compound represented by the general formula () can be used as a treatment for liver diseases as it is, or it can be formulated as a mixture with a pharmaceutically acceptable diluent and/or other pharmacologically active substances in accordance with pharmaceutical practice. You can also
They may also be formulated in dosage unit form. Possible pharmaceutical forms include the following forms: powders, granules, tablets, dragees, capsules, pills, suspensions, solutions, emulsions, ampoules, injections, isotonic solutions, and the like. In particular, the compounds obtained by the method of the present invention have high water solubility; for example, compound No. 23 has a water solubility of 2000 ppm (200 ppm).
℃), Compound No. 12 is 2200 ppm (20℃), Compound No. 1
Since it is 2% soluble in a 2% gum arabic solution, it is advantageous when used in the form of an injection or drip. The diluent used when preparing a drug from the compound obtained by the method of the present invention is one that is pharmaceutically acceptable. means,
The capsules may be solid, semi-solid, liquid or ingestible and include a variety of agents, such as excipients, fillers, binders, wetting agents, disintegrants, etc.
Surfactants, lubricants, dispersants, buffers, flavoring agents,
Flavoring agents, pigments, fragrances, preservatives, solubilizing agents, solvents, coating agents, sugar coating agents, etc. However, it is not limited to this. Moreover, these may be used alone or as a mixture of more than one. Such pharmaceutically acceptable diluents may also be used in mixtures with other pharmacologically active substances. A medicament based on a compound obtained by the method of the present invention may be produced by any known method. For example, the active ingredient is mixed with a diluent, eg, into granules, and the composition is then shaped, eg, into tablets. Parenterally administered preparations should be sterile. It should also be made isotonic with blood if necessary. Since the compound represented by the general formula () can itself be a therapeutic agent for liver diseases, the active ingredient is generally contained in the composition in an amount of 0.01 to 100% (by weight). In the case of pharmaceutical dosage units, the individual formulation parts forming the formulation may be in mutually different forms or may be the same; for example, the following forms are often employed: tablets, Granules, pills, powders, dragees, capsules, ampoules, etc. The therapeutic agent for liver diseases using the compound obtained by the method of the present invention can be applied to humans and animals for the treatment of liver diseases by methods commonly used in the field. It is administered orally or parenterally.
Oral administration includes sublingual administration. Parenteral administration includes administration by injection (including, for example, subcutaneous, intramuscular, intravenous, and infusion). The pharmaceutical dosage of the compound obtained by the method of the present invention depends on whether the subject is an animal or a human, sensitivity differences, age, sex, body weight, administration method, administration timing, interval, medical condition, physical condition, etc. It varies depending on various factors such as the nature of the pharmaceutical preparation, the type of preparation, and the type of active ingredient. Therefore, in some cases, it may be sufficient to use a dose lower than the lowest dose shown below, and in some cases, it may be necessary to administer a dose exceeding the upper limit dose shown below. In addition, in the case of large-dose administration, it is preferable to divide the administration into several times a day. In order to obtain effective results in animals, the active ingredient should be administered orally in the range of 0.1 to 500 mg per kilogram of body weight per day, preferably in the range of 0.1 to 25 mg per kilogram of body weight per day in the case of parenteral administration. 0.01～
A range of 250 mg, preferably 0.1 to 25 mg is advantageous. Regarding the dosage for obtaining an effective result in humans, the following dosage range is advantageous, for example, taking into account sensitivity differences, safety, etc. from the effective dosage in animals. For oral administration, per kilogram of body weight per day
0.1 to 250 mg, preferably 0.5 to 50 mg, and in the case of parenteral administration, 0.01 to 100 mg per kg of body weight per day, preferably 0.1 to 25 mg. Next, examples of the present invention will be shown. Example 1 Diisopropyl-4,5-dihydroxy-1,
Synthesis of 3-dithiolane-2-ylidene-malonate (Compound No. 23). A mixture of 56.8 g (0.3 mol) of diisopropyl malonate and 22.8 g (0.3 mol) of carbon disulfide is cooled with cold water and stirred. 84 g of a separately prepared 40% caustic potassium aqueous solution (33.6 g of KOH dissolved in 50.4 g of water) was added to this mixture so that the temperature did not exceed 20°C. ) Obtain an aqueous solution of ethylene-1,1-dithiolate. 40% of this dithiolate aqueous solution
% glyoxal 44g, acetic acid 36g, benzene 200
of the mixture and react. During this time, stir the reaction contents well and prepare a solution to keep the reaction temperature below 15°C. After the addition of the mixture was completed, stirring was continued at room temperature, and the color of the reaction mixture changed from reddish brown to light yellow, and the reaction was completed in 1 hour. The benzene layer is separated, washed with water, and the benzene is distilled off to obtain crude crystals.
After drying, it is recrystallized from a mixed solvent of ether and n-hexane. Yield 58g (yield 60%), mp.132.6℃,
NMRδ ^CDCl3 _TMS ppm: 2.25 (12H, d, J=7Hz),
3.80 (2H, broad s), 5.10 (2H, m, J=7Hz),
5.70 (2H, s). Example 2 Diethyl 4,5-dihydroxy-1,3-dithiolane-2-ylidene-malonate (Compound No.
2) Synthesis. Diethyl malonate 16g (0.1mol), carbon disulfide
Cool 7.6 g of the mixture with ice water and stir. Add 24.9 g of 45% KOH aqueous solution to this mixture and set the reaction temperature to 20
When the mixture is added dropwise and reacted while being careful not to exceed the temperature, an aqueous potassium salt solution of 2,2-bis(ethoxycarbonyl)ethylene-1,1-dithiolate is obtained. This aqueous dithiolate solution is added dropwise to a mixture consisting of 14.5 g of 40% glyoxal, 12 g of acetic acid, and 60 ml of benzene for reaction. During this time, stir the reaction contents well and adjust the dropwise addition so as to keep the reaction temperature below 15°C. After the dropwise addition was completed, stirring was continued at room temperature, and the reaction mixture turned from reddish brown to pale yellow, and the reaction was completed for 1 hour. The benzene layer is separated, washed with water, and the benzene is distilled off to obtain crude crystals. After drying, it is recrystallized from a mixed solvent of ethyl acetate, dichloromethane, and n-hexane. Yield 17.8g, Yield 60.5
%. mp87.3℃. Example 3 Diallyl 4,5-dihydroxy-1,3-dithiolane-2-ylidenemalonate (Compound No.
12) Synthesis of A dithiolate salt aqueous solution is prepared from 6.14 g of diallyl malonate in the same manner as in Example 2 using a 45% KOH aqueous solution. Add this to 40% glyoxal 4.8
g, acetic acid 4.0 g, and benzene 30 ml, reacted in the same manner as in Example 2, post-treated, and recrystallized from dichloromethane-n-hexane to give 5.22 g of crystals with an mp. of 81.2°C [yield 49%]. Example 4 Bis(3-methoxypropan-2-yl)4,
5-dihydroxy-1,3-dithiolane-2-
Synthesis of ylidene malonate (compound No. 15) Synthesis was carried out in the same manner as in Example 2 from 8.28 g of bis(3-methoxypropan-2-yl) malonate to obtain the desired product in the form of an oil. This was purified by dry column chromatography using silica gel (developing solvent: ethyl acetate-n-hexane = 1:1) to obtain 3.5 g of an oil with n _D ²⁰ =1.5490. Yield 27.4%. In the following formulation examples, all parts are by weight. The types and proportions of the ingredients can be varied. Formulation Example 1 Compound 22 10 parts heavy magnesium oxide 10 parts lactose 80 parts are uniformly mixed to make a powder or fine granules. Formulation Example 2 Compound 10 10 parts Synthetic aluminum silicate 10 parts Calcium hydrogen phosphate 5 parts Lactose 75 parts were used to prepare a powder according to Formulation Example 1. Formulation Example 3 Compound 3 50 parts Starch 10 parts Lactose 15 parts Crystalline cellulose 20 parts Polyvinyl alcohol 5 parts Water 30 parts After homogeneous mixing, crush, granulate, dry and sieve to obtain granules. Formulation Example 4 1 part of calcium stearate was mixed with 99 parts of the granules obtained in Formulation Example 3, and compression molded to form a product with a diameter of 10 mm.
tablets. Formulation Example 5 Granules were prepared in the same manner as in Formulation Example 3 using 95 parts of Compound 2, 5 parts of polyvinyl alcohol, and 30 parts of water. 10 parts of crystalline cellulose is added to 90 parts of the obtained granules and compressed to form tablets with a diameter of 8 mm. Further, appropriate amounts of syrup gelatin, a mixed suspension of precipitated calcium carbonate, and a coloring agent are added to the tablets to form sugar-coated tablets. Formulation Example 6 Chemical No. 21 0.5 parts Nonionic surfactant 2.5 parts Physiological saline 97 parts are heated and mixed, then sterilized to prepare an injection. Formulation Example 7 The acid agent obtained in Formulation Example 1 is filled into a commercially available capsule container to form a capsule. Test example 1 Carbon tetrachloride liver damage inhibitory effect test method The test compound was dissolved or suspended in olive oil and orally administered to mice (DD male for 6 weeks) at a rate of 250 mg/Kg, and 6 hours later, carbon tetrachloride 0.05
Orally administered at the rate of ml/Kg, 24% of carbon tetrachloride administration
After a period of time, the animals were sacrificed and the degree of liver damage was examined by visual observation. On the other hand, blood was collected at the time of slaughter, plasma was obtained by centrifugation, and plasma glutamic-pyruvic transaminase (GPT) activity was measured according to the Reitman-Frankel method, and the activity was expressed in carmen units (KU). expressed. The liver damage index is as follows. Liver damage index Liver symptoms 0 Healthy liver 2 Slightly affected 4 Obvious damage 6 Severe damage Five mice were used per group, and the average values are shown. Furthermore, for GPT activities exceeding 1000 units, no further measurements were performed, but the average value was calculated as 1000 units for convenience. The results are shown in Table 1.

[Table] As shown in Table 1, the compound obtained by the present invention significantly improves the liver damage index and p-GTP compared to the group administered with carbon tetrachloride alone, indicating that it has a liver damage suppressing effect. ing. Test Example 2 α-naphthyl isothiocyanate liver damage inhibitory effect α-naphthyl isothiocyanate (hereinafter referred to as α-
It's called NIT. ) is known to be able to experimentally induce cholestatic liver damage by administering it to healthy test animals, and it is also used as a model for human pathology. Regarding the α-NIT liver injury model in this test example and the carbon tetrachloride liver injury model in Test Example 1,
Although both cases are accompanied by an increase in plasma GPT activity,
There are considerable differences in the degree of necrosis, and the cause of liver necrosis is that in the former case, small necrosis foci are formed in the liver parenchyma around Gleason's sheath, whereas in the latter case, small necrotic foci are formed in the centrilobular zone. There is a report that coagulative necrosis of hepatocytes is observed (Nippon Rinsai Vol. 30, No. 1, 216
Page (1972)). As can be seen from these facts, it can be seen that the increase in plasma GPT in both pathological models has different characteristics. Measurement of plasma glutamic-pyruvic transaminase (GPT) activity and plasma alkaline phosphatase activity is an appropriate method as an indicator of the degree of liver damage caused by α-NIT and the preventive effect of drugs on that damage. Test method: The test compound was dissolved or suspended in olive oil and orally administered to mice (6 to 7 weeks old, DD male) at a rate of 250 mg/Kg twice at 3 hour intervals. After an hour, 35 mg/Kg of α-naphthyl isothiocyanate (hereinafter referred to as α-NIT) was orally administered. Mice were sacrificed 24 hours after α-NIT administration and blood was collected. Plasma was obtained by centrifuging the blood, and plasma GPT activity was measured according to the Reitman-Frankel method and alkaline phosphatase activity was measured according to the Betsey-Lowry method, and the activity was expressed in Carmen units and Betsey-Lowry units. The results are shown in Table 2.

【table】

【table】

【table】

[Table] ｜
C＝C
／ ＼
CH−S
COC ₃ H ₇ -i
‖
O
As shown in Table 2-1 and 2, the compound obtained by the present invention was found in the α-NIT alone administration group (α-
Compared to NIT control), it significantly improved plasma GPT activity and plasma alkaline phosphatase activity, indicating that it has a liver damage suppressive effect.

Claims (1)

[Claims] 1 General formula (): (In the formula, R ¹ and R ² are the same or different and represent an alkyl group, a cyclohexyl group, a benzyl group, an alkenyl group, an alkynyl group, or an alkoxyalkyl group, and M represents an alkali metal atom or an ammonium group.) The compound represented and the following formula: General formula () characterized by reacting glyoxal represented by in the presence of an acid: (In the formula, R ¹ and R ² are the same as above.) A method for producing a 4,5-dihydroxy-1,3-dithiolan-2-ylidenemalonic acid derivative represented by the following.