JPH0329765B2 - - Google Patents

Description

[Detailed description of the invention]

INDUSTRIAL APPLICATION FIELD The present invention relates to a therapeutic agent for liver diseases. BACKGROUND OF THE INVENTION Liver diseases are generally classified into two types: those caused by a decrease in the number of normal cells in the liver, and those caused by a decline in liver cell function. Several liver disease treatments have been known, but the latter hepatotoxicants cause hepatic metabolic disorders, resulting in liver disorders caused by decreased liver cell function, such as acute or chronic hepatitis, Known liver disease therapeutic agents for treating liver diseases such as cirrhosis, fulminant hepatitis, drug-induced hepatitis, viral hepatitis, alcoholic hepatitis, fatty liver, or postoperative liver damage are known for their therapeutic effects,
Due to problems such as side effects, the actual situation is still far from being satisfactory. In particular, for patients with the above-mentioned liver disease who are unable or have difficulty in taking nutritional sources such as food orally for some reason and who are suffering from hypoproteinemia or malnutrition, it is recommended to treat the liver disease itself and to treat the liver disease itself. Until now, there has been no known therapeutic agent for liver diseases that can also maintain nitrogen balance and manage nutrition. The present inventors first focused on the constituent components of nucleic acids that control protein synthesis, and as a result of intensive research into these components, they developed a composition containing specific nucleic acid constituents in the form of an infusion solution, etc. It was discovered that when administered to humans, it promotes protein synthesis and provides excellent nutritional management and nitrogen balance maintenance effects, and an invention based on this knowledge was completed (Japanese Patent Application No. 58-233142). As a result of subsequent research, the present inventors have obtained new knowledge that certain of the nucleic acid component compositions according to the above invention have a therapeutic effect on liver diseases. Problems to be Solved by the Invention The present invention aims to provide a novel therapeutic agent for liver diseases. More specifically, therapeutic agents for liver diseases caused by hepatic metabolic disorders caused by hepatotoxicants, etc., and which are based on a decline in the function of hepatocytes, are especially recommended for patients with liver diseases who are unable to take in oral nutritional sources. In contrast, the present invention provides a therapeutic agent for liver diseases that can be administered in conjunction with nutritional management. Means for Solving the Problems The present invention relates to a therapeutic agent for liver diseases characterized by containing any one selected from the following (1) to (3) as an active ingredient. (1) Inosine, cytidine, GMP, uridine, thymidine or IMP (or inosine) CMP, GMP,
UMP (or uridine), thymidine (2) CMP, UMP, IMP (or inosine), thymidine (3) AMP, GMP, CMP, UMP, thymidine [However, IMP is inosine-n'-monophosphate, CMP
represents cytidine-n'-monophosphate, GMP represents gunian-n'-monophosphate, UMP represents uridine-n'-monophosphate, and AMP represents adenine-n'-monophosphate. ] The above (1) to used as active ingredients of the liver disease therapeutic agent of the present invention
Each of the nucleic acid constituents in (3) belongs to ribonucleosides (inosine, cytidine, and uridine), deoxyribonucleosides (thymidine), and ribonucleotides (AMP, IMP, GMP, CMP, and UMP), These do not need to be in free form, but may be in the form of their non-toxic salts. In addition, the abbreviations for the above nucleic acid components are
It follows the IUPAC-IUB regulations or the common symbols in the field. In addition, the nucleobases of the above-mentioned nucleic acid constituents are nucleosides and nucleotides that correspond to each other. For example, guanine, which is a nucleobase, is a nucleoside, that is, guanosine, which is a glycoside, and a nucleoside in which phosphoric acid is further bonded to the nucleoside. That is,
GMP, GDP or GTP can be used as substitutes for each other, and the same applies to each other component. The blending ratio of the above nucleic acid components containing the therapeutic agent of the present invention as an active ingredient depends on the type of nucleic acid components used,
It can be appropriately selected from a wide range depending on the patient's condition, desired therapeutic effect, etc., and is not particularly limited. For examples of preferred combinations of active ingredients, if the above blending proportions are expressed in molar ratios, for example:
CMP:UMP:IMP:Thymidine=4:3:8:1
or 7.5:4:12.5:1, inosine:cytidine:
GMP: Uridine: Thymidine = 4:4:4:3:
1. AMP:GMP:CMP:UMP:Thymidine=
4:4:4:3:1 or IMP:GMP:CMP:
UMP: Thymidine = 4:4:4:3:1 or 2:2:2:1:1 can be given. Even in the above composition, a part or all of each component can be replaced with the corresponding nucleobase, nucleoside, or nucleotide. The therapeutic agent of the present invention is generally preferably used in the form of an injection for intravenous administration, but
It can be used in the form of powders, solutions, suspensions, emulsions, granules, etc. for oral and enteral administration via a tube, etc., and the dosage unit form can be appropriately selected depending on the purpose. Injections as a preferred administration form of the therapeutic agent of the present invention are suitable for liver diseases caused by hepatic metabolic disorders caused by hepatotoxicants, etc., and caused by decreased function of hepatocytes, such as acute or chronic hepatitis, cirrhosis, fulminant hepatitis, etc. It is effective for patients with liver diseases such as drug-induced hepatitis, viral hepatitis, alcoholic hepatitis, fatty liver, and postoperative liver damage. In particular, it is administered to patients who are unable or have a relatively difficult time ingesting nutritional sources such as food, thereby achieving the desired therapeutic effect and increasing protein turnover, maintaining nitrogen balance, and providing nutritional support. It can also be managed. Although the therapeutic agent of the present invention is sufficiently effective when administered alone, when nutritional management is also taken into consideration, even better effects can be achieved by using it in combination with a conventional amino acid preparation. In addition, in order to double the utilization rate of the amino acids co-administered as described above, to help synthesize these amino acids into proteins in the body, and to suppress their consumption as an energy source, for example, glucose, fructose, xylitol, sorbitol, etc. Carbohydrates such as maltose and the like can also be added and blended, and in addition to these sugars, various ingredients that are known to be commonly added and blended in this type of preparations, such as lipids, vitamins, electrolytes,
Further effects can be obtained by optionally adding and blending trace elements and the like. Examples of the above-mentioned lipids include soybean oil, cottonseed oil, sesame oil, egg yolk lecithin, soybean lecithin, etc., and vitamins such as vitamin A, vitamin B, and vitamin B.
_B2 , vitamin _B6 , nicotinic acid, pantothenic acid,
Vitamin C, vitamin D, vitamin E, biotin, folic acid, etc., and sodium chloride as an electrolyte.
Sodium acetate, potassium chloride, magnesium sulfate, magnesium chloride, calcium chloride, dipotassium phosphate, sodium phosphate, etc., and trace elements such as iron, zinc, manganese, copper, iodine,
Examples include cobalt and the like. When the therapeutic agent of the present invention is used as an injection, the preparation method is not substantially different from that of ordinary infusion preparations such as amino acid preparations and electrolyte preparations. The components or their non-toxic salts are mixed and dissolved, and if necessary, stabilizers such as sodium sulfite, sodium bisulfite, sodium pyrosulfite, sodium thiosulfate, etc., and PH regulators such as hydrochloric acid, acetic acid, lactic acid, malic acid, etc. , citric acid or sodium hydroxide, and other additives, and the resulting aqueous solution is sterilized by heat sterilization or aseptic filtration. Alternatively, it can be made into an unpowdered preparation that is dissolved before use, and in that case, it can be easily prepared by appropriate means such as freeze-drying, with or without the addition of various additives, according to conventional methods. be able to. The PH of the therapeutic agent of the present invention thus prepared can be the same as that of ordinary injections such as infusions, and is preferably 3.0 to 9.0, particularly preferably 5.0 to 8.0. In addition, the concentration of nucleic acid components is 0.5~
It is preferable to set it to 10 W/V%, or preferably 2 to 8 W/V%. The therapeutic agent of the present invention in the form of an injection is prepared in the form of a sterile aqueous solution and administered intravenously, such as in a peripheral vein or central vein, or enterally. The dosage of the therapeutic agent of the present invention in the form of an injection is generally about 20 to 500 ml per adult per day, preferably about 30 ml per day.
The amount can be increased or decreased as appropriate depending on the pathological condition, nutritional status, age, weight, concomitant drugs, etc. of the patient to be administered, with the approximate amount being ~200 ml. On the other hand, when the therapeutic agent of the present invention is administered enterally, the above-mentioned injection may be used as is, but it can also be prepared and used in the form of a powder, solution, suspension, emulsion, granule, or the like. In that case, the active ingredient is processed and used in the form of a desired preparation along with conventional additives. Additives include diluents, fillers, and
Bulking agents, binders, suspending agents, disintegrants, surfactants,
Examples include lubricants and excipients. In addition, the above preparations include usual solubilizing agents, buffering agents,
Soothing agents, preservatives, etc., and coloring agents if necessary.
Flavors, flavoring agents, sweeteners, etc. may also be included. These can be appropriately selected depending on the dosage unit form of the therapeutic agent of the present invention, and can produce excellent effects similar to those of injections. Furthermore, when preparing solutions, suspensions, and emulsions, a sufficient amount of common salt, glucose, or glycerin may be included in the above-mentioned preparation form if an isotonic solution is to be prepared. Furthermore, the above preparations can be administered alone like injections, but if nutritional management is also considered, they may be mixed with normal fluid replacement fluids such as glucose and amino acids, or commercially available enteral nutrients, etc., and administered enterally. It is even more suitable if The dosage unit form of the therapeutic agent of the present invention can be appropriately selected depending on its purpose. In addition, the dosage of the preparation for enteral administration should be approximately the same as the active ingredient content administered in the above-mentioned injection, and should be determined based on the pathological condition, nutritional status, age, body weight, or concomitant drugs of the patient to whom it should be administered. The dosage may be increased or decreased as appropriate depending on the situation. Furthermore, depending on the pathology of the patient, the therapeutic agent of the present invention can also be used in combination with conventionally known therapeutic agents for rod disease, thereby significantly shortening the treatment period until cure that would be seen when the above therapeutic agent was administered alone. This is more preferable because it also allows the dosage of the drug to be reduced. Examples Representative examples of the therapeutic agents of the present invention will be described below. In addition, the molar ratio in the following production examples shows approximate values. Manufacturing example 1

【table】

[Table] Pure crystals of each nucleic acid component in amounts having the above composition were added to distilled water for injection, stirred and dissolved, and the pH was adjusted to approximately 7.4 using hydrochloric acid as a pH regulator. Next, the obtained nucleic acid component aqueous solution was filtered aseptically, filled into an injection container, and the container was closed after purging with nitrogen.The container was then sterilized in an autoclave at 105°C for 40 minutes to obtain an injection (total free nucleic acid component concentration). 8W/V%) to obtain the therapeutic agent of the present invention. Manufacturing example 2

[Table] Pure crystals of each nucleic acid component in amounts having the above composition were added to distilled water for injection, stirred and dissolved, and the pH was adjusted to approximately 7.3 using acetic acid as a pH regulator. Next, the obtained nucleic acid component aqueous solution was filtered aseptically, filled into an injection container, the container was closed after purging with nitrogen, and the container was sterilized in an autoclave at 110°C for 40 minutes to obtain an injection (total free nucleic acid component concentration). 8W/V%) to obtain the therapeutic agent of the present invention. Manufacturing example 3

[Table] Pure crystals of each nucleic acid component in amounts having the above composition were added to distilled water for injection, stirred and dissolved, and the pH was adjusted to approximately 6.4 using hydrochloric acid as a pH regulator. Next, the obtained nucleic acid component aqueous solution was filtered aseptically, filled into an injection container, and the container was closed after purging with nitrogen.The container was then sterilized in an autoclave at 105°C for 40 minutes to obtain an injection (total free nucleic acid component concentration). 8W/V%) to obtain the therapeutic agent of the present invention. Manufacturing example 4

[Table] Pure crystals of each nucleic acid component in amounts having the above composition were added to distilled water for injection, stirred and dissolved, and the pH was adjusted to approximately 8.0 using sodium hydroxide as a pH regulator. Next, the obtained nucleic acid component aqueous solution was filtered aseptically, filled into an injection container, and the container was closed after purging with nitrogen.The container was then sterilized in an autoclave at 105°C for 40 minutes to obtain an injection (total free nucleic acid component concentration). 3.4W/Y%) to obtain the therapeutic agent of the present invention. Production Example 5 In the same manner as in Production Example 1, a therapeutic agent of the present invention as an injection having the following composition (total free nucleic acid component concentration: 8 W/V%) is obtained.

[Table] Production Example 6 In the same manner as in Production Example 1, a therapeutic agent of the present invention as an injection having the following composition (total free nucleic acid component concentration 8W/V%) was obtained.

[Table] Production Example 7 In the same manner as Production Example 1, a therapeutic agent of the present invention as an injection having the following composition (total free nucleic acid component concentration 5W/V%) was obtained.

[Table] Production Example 8 A therapeutic agent of the present invention as an injection having the following composition (total free nucleic acid component concentration 7W/V%) was obtained in the same manner as Production Example 4.

[Table] Production Example 9 A therapeutic agent of the present invention as an injection having the following composition (total free nucleic acid component concentration: 8.2 W/V%) was obtained in the same manner as in Production Example 4.

【table】

[Table] Representative examples of the preparation of the therapeutic agent of the present invention in a form for enteral administration are listed below. Production example 10 5'-AMP-2Na 2.34g 5'-CMP-2Na 2.20g 5'-GMP-2Na 2.44g 5'-UMP-2Na 1.65g Thymidine 0.36g Starch 91.01g Each nucleic acid component in amounts to achieve the above composition After the pure crystals are sieved through a 60-mesh sieve, starch as an excipient is added and mixed evenly to obtain the therapeutic agent (powder) of the present invention. Nucleic acid component 5′-AMP-2Na in this therapeutic agent,
5′−CMP−2Na, 5′−GMP−2Na, 5′−UMP−
The molar ratio of 2Na and thymidine is approximately 4:4:
The ratio is 4:3:1. Moreover, the content is 8W/V% as a total free nucleic acid component. Production example 11 Inosine 1.06W/V% Cytidine 1.46 5'-GMP-2Na 2.44 Uridine 1.10 Thymidine 0.36 Refined white sugar 20.00 Ethyl paraoxybenzoate 0.009 Butyl paraoxybenzoate 0.006 Purified water is heated, and refined white sugar is added as a sweetening agent to it. Add and stir to dissolve. After cooling, each nucleic acid component in the amount of the above composition and further ethyl paraoxybenzoate and butyl paraoxybenzoate as preservatives dissolved in a small amount of ethanol are added and dissolved with stirring. The liquid volume is adjusted with purified water, filtered, and filled into a glass container. After replacing with nitrogen, the container is closed, and then heat sterilized to obtain the therapeutic agent (liquid) of the present invention. The molar ratio of the nucleic acid components inosine:cytidine:5'-GMP-2Na:uridine:thymidine in this therapeutic agent is 4:4:4:3:1, and the content is 6.8W/V% as a total free nucleic acid component. be. Production Example 12 In the same manner as Production Example 11, the therapeutic agent of the present invention as a liquid containing each of the following nucleic acid components is obtained. 5'-CMP-2Na 1.12W/V% 5'-GMP-2Na 1.24 5'-UMP-2Na 0.56 5'-IMP-2Na 1.20 Thymidine 0.37 Nucleic acid component 5'-CMP- in this therapeutic agent
2Na:5′−GMP−2Na:5′−UMP−2Na:5′−
The molar ratio of IMP-2Na:thymidine is 2:2:1:
2:1, and the content is as the total free nucleic acid component.
4W/V%. Hereinafter, the therapeutic agent (injection) of the present invention prepared in each of the above production examples will be taken as an example, and the results of animal experiments will be described in detail. Example 1 Effect on galactosamine-induced liver damage Wistar, 8 weeks old, weighing around 250 g
After the male rats were fasted overnight, a catheter was placed at the origin of the right superior vena cava to enable high-calorie infusion in a metabolic cage. The rats were maintained at a constant rate of 150 ml/kg of physiological saline for 24 hours, and then 250 ml/kg of D-galactosamine hydrochloride was administered to the rats.
Kg was administered intraperitoneally to induce liver damage. Immediately after this, high-calorie infusion was administered for 24 hours. The experimental group was a normal high-calorie infusion group containing amino acids (12% commercially available amino acid infusion prescribed by FAO/WHO), glucose, electrolytes, and vitamins as a control group (group C), and in addition to this, A group to which the therapeutic agent of the present invention (hereinafter referred to as "OG-") was added (EO group), a group to which uridine was added in the same amount as OG- (EU group), and aderabine (Sanwa Kagaku Kenkyusho Co., Ltd.) A group (EA group) was established. The dose of OG- was such that the weight ratio of the nucleic acid component to the administered amino acid was 1/10. In addition, the amount of amino acids administered in normal high-calorie infusions (group C) is 8g/Kg/day, the total water content is 200ml/Kg/day,
The total amount of calories administered was 200Kcal/Kg/day.
Its composition and EO group infusion composition are as follows.

[Table] *2 Manufactured by Shionogi & Co., Ltd. *3 Manufactured by Takeda Pharmaceutical Co., Ltd. Blood and liver were collected 30 minutes after the completion of the above infusion, and plasma GTO, GPT activity, and liver glycogen content were used as indicators to determine galactosamine liver. The degree of disability was estimated. The results are shown in Tables 1 and 2.

【table】

[Table] As shown in Tables 1 and 2 above, in the therapeutic agent group of the present invention (EO group), a decrease in plasma GOT and GPT values and an increase in hepatic glycogen level were observed, clearly demonstrating the therapeutic effect on liver damage. It was hot. On the other hand, the above effects were not observed in the EU group or the EA group, which suggests that the therapeutic agents of the present invention exerted the above excellent effects due to the comprehensive action of each nucleic acid component contained therein. Conceivable. In addition, 8-week-old Wistar male rats (weight 250-280 g) were fasted for 2 days in the same manner as above.
The test animals prepared by intraperitoneally administering 250 mg/Kg of D-galactosamine hydrochloride were divided into the following three groups, and each test drug was intraperitoneally administered 3 hours after D-galactosamine administration. (1) Group of the present invention: 25 mg/Kg administration group of the therapeutic agent of the present invention obtained in Production Example 4 (2) Comparative group: Composition based on Formulation Example 4 of French Pharmaceutical Patent No. 5084 [AMP:ATP:GMP:Xanthosin -5-phosphoric acid = 100mg: 100mg: 100mg: 100mg/25ml/
[Kg] Administration group (3) Control group: 0.9% saline (25ml/Kg) administration group Blood was collected from the abdominal aorta of each group of rats 24 hours after administration of each of the above test reagents, and plasma was separated.
GOT, GPT value and LDH value were measured. In addition, a group that only fasted for 3 days was established as a normal group. The above measured values for each group are shown in Table 3 below as mean±standard deviation.

[Table] As is clear from Table 3 above, in the present invention group, the therapeutic agent of the present invention increased GOT due to liver damage.
It can be seen that the therapeutic agent composition of the present invention has the effect of normalizing the GPT value, GPT value, and LDH value, but such an effect was hardly seen in the comparison group. It is clear that it has therapeutic and preventive effects on liver diseases. The reason for this is thought to be that, while the comparative composition contains only purine-based nucleic acid components, the composition of the present invention also contains a pyrimidine-based nucleic acid component such as uridine as an essential active ingredient. Test Example 2 Effect on carbon tetrachloride-induced liver damage Male Wistar rats weighing approximately 250 g were fasted overnight, and a catheter was placed at the origin of the right superior vena cava so that high-calorie infusions could be administered in a metabolic cage. performed the technique. The rats were maintained at a constant rate of 150 ml/Kg of physiological saline over 24 hours, and then 1 ml/Kg of 20% carbon tetrachloride olive oil solution was administered intraperitoneally and subcutaneously in half doses to induce liver damage. . Immediately after this, high-calorie infusion was administered for 48 hours. The experimental group was a normal high-calorie infusion group (C' group, control group) containing amino acids (12% commercially available amino acid infusion prescribed by FAO/WHO), glucose, electrolytes, and vitamins, to which OG- was added. A group (EO' group) was established. The amount of amino acids administered in the normal high-calorie infusion group (Group C') was 8 g/Kg/day, and the total amount of calories administered was 200 Kcal/Kg/day.
The dose of OG in the EO' group was such that the weight ratio of the nucleic acid component to the administered amino acid was 1/10. The compositions of Group C' and Group EO' are the same as Group C and Group EO of Test Example 1, respectively. Thirty minutes after the completion of the above infusion, blood was collected from the inferior aorta and plasma GTO and GPT activities were measured. The results are shown in Table 4.

[Table] As shown in Table 4 above, in the therapeutic agent group of the present invention (EO' group), plasma GTO and GPT values decreased, and a clear therapeutic effect on liver damage was observed.

Claims (1)

[Scope of Claims] 1. A therapeutic agent for liver disease characterized by containing any one selected from the following (1) to (3) as an active ingredient. (1) Inosine, cytidine, GMP, uridine, thymidine or IMP (or inosine) CMP, GMP,
UMP (or uridine), thymidine (2) CMP, UMP, IMP (or inosine), thymidine (3) AMP, GMP, CMP, UMP, thymidine [However, IMP is inosine-n'-monophosphate, CMP
represents cytidine-n'-monophosphate, GMP represents guanine-n'-monophosphate, UMP represents uridine-n'-monophosphate, and AMP represents adenine-n'-monophosphate. 2. The therapeutic agent for liver disease according to claim 1, which is in the form of an injection. 3. The therapeutic agent for liver disease according to claim 1, which is in a form for enteral administration.