JPH08769B2 - Drugs for treating or preventing liver diseases - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a drug for treating and preventing kidney and liver diseases, and more particularly, to a renal disease and a liver disease that occur independently and when diseases of both organs also occur. The present invention also relates to a drug for treating and preventing kidney and liver diseases, which exhibits excellent therapeutic and preventive effects.

[0002]

2. Description of the Related Art As is well known, the kidney in a living body, in its renal tubular epithelial cells, collects waste products from the blood and harmful substances, such as drugs or poisons, which are taken up in the living body in the urine by a mechanism of filtration and secretion. Has the function of excreting into.

On the other hand, the liver in the living body mainly has the following three functions. That is, as the first function, the liver acts as a digestive gland and the
Secretes ~ 1000 cc of bile to aid in digestion and absorption of fat in the small intestine. In addition, as a second function, the liver participates in chemical changes, storage and utilization in the body of various nutrients, that is, intermediate metabolism. Furthermore, the third
As a function of, the liver performs a detoxification action, and detoxification treatment such as oxidation, reduction, conjugation, etc. is added to harmful substances from outside the body, such as poisons and harmful substances made in the body, or this is mixed with bile and discarded. Or send it to the kidneys and excrete it in the urine.

As described above, the kidney and the liver have their own respective functions, but the waste products and the drugs are likely to be accumulated locally, and therefore, the kidney and the liver alone are caused by these substances. Not only diseases, but also multiple diseases of both organs are likely to occur. The substances that cause such single or concomitant renal and hepatic diseases are specifically diverse, and they are as follows when the drugs are taken as examples and shown separately for each organ. Among the drugs listed below, drugs marked with a circle on their head cause damage to both the kidney and the liver.

(Drugs prone to renal damage) ○ Analgesics, lower fever, anti-inflammatory agents, anti-rheumatic agents phenacetin, aspirin, indomethacin, mefenamic acid, fenoprofen, gold preparations, D-penicillamine and the like. ○ Antibiotics Aminoglycosides, polypeptides, polyenes, cephalosporins, penicillins, etc. ○ Chemotherapeutic agents such as sulfamides. -Anti-cancer agent mitomycin C, daunomycin, cisplatin, nitrosourea type, etc. -Immunosuppressant such as cyclophosphamide. ○ Anesthetic such as methoxyflurane. ○ Diuretics such as thiazide.・ Sharpener

(Drugs that easily cause liver damage) ○ Analgesics, lower fever, anti-inflammatory agents, anti-rheumatic agents acetaminophen, aspirin, phenylbutazone,
Sulindac, evefenac, gold preparation, etc. ○ Antibiotics Aminoglycosides, polyenes, cephalosporins, penicillins, tetracyclines, etc. ○ Chemotherapeutic agents Sulfa drugs, isoniazid, etc. -Anti-cancer agent mitomycin C, cisplatin, 6-MP, nitrosourea type, etc. ○ Anesthetics such as halothane and methoxyflurane.・ Psychoactive drugs such as chlorpromazine, diazepam, and barbital. ○ Diuretics such as thiazide.

[0007] The examples of drugs causing the above-mentioned kidney and liver diseases are summarized based on the following documents. That is, [Clinical Adult Disease Vol. 16 No. 8 (198
6) 45-62, 85-103], [Latest Medical Library 42 "Medical Treatment of Chronic Hepatitis" by Mikio Nishioka, Shinshoku Publishing Co., Ltd., issued January 26, 1987, pages 29-31] ,
[Clinical and Research Volume 63, Issue 4, April 1986, 38-
39], ["Liver Diseases" Toshiji Oda et al., Chugai Medical Co., Ltd., October 20, 1980, 323-333.
Page], ["Pharmaceutical handbook" edited by Osaka Prefectural Hospital Pharmacists Association, Yakuza Jikhosha, published November 10, 1983]. As mentioned above, kidney and liver diseases have been considered from the viewpoint of the substance metabolism function of those organs. However, in renal diseases, there are other diseases that are thought to be mediated by an immune mechanism.

The renal diseases referred to in the present invention are (a) renal dysfunction caused by abnormal substance metabolism, for example, acute nephritis caused by drugs and the like, chronic nephritis in which such acute nephritis is chronic, and (b) immune system. (1) Acute nephritis mediated by the pathology and chronic nephritis caused by such acute nephritis, (c) acute nephritis caused by bacterial and viral infections and chronic nephritis caused by these chronic kidney diseases, etc., and functions at kidney sites such as glomeruli, renal tubules, and lupus A disease that presents a disorder.

Similarly, the liver disease referred to in the present invention is
(A) Liver dysfunction caused by abnormal substance metabolism and biosynthesis, for example, acute hepatitis caused by a drug or chronic hepatitis in which such acute hepatitis has become chronic, (b)
Fatty liver caused by drinking or the like, chronic hepatitis caused by these chronic diseases, (c) viral hepatitis caused by viral infection, or chronic hepatitis caused by the chronic infection, and diseases exhibiting widespread liver dysfunction such as cirrhosis resulting from the above disorders. Say.

By the way, as a method for treating kidney disease,
Currently, depending on its severity, therapy to maintain renal function,
For example, rest therapy, diet or drug therapy, hemodialysis therapy or kidney transplantation are known. The hemodialysis therapy is considered to be the final therapy for renal diseases, but the renal function is not improved only by removing the waste products accumulated in the body due to renal dysfunction from the blood, and the patient should undergo lifelong dialysis therapy. In the end, in many cases, heart failure, infectious diseases, etc. occur together, resulting in death. On the other hand, as drug therapy, diuretics, immunosuppressive agents, corticosteroid therapy, mannitol, lactulose therapy, etc. are currently performed, but these are all symptomatic treatments, and their therapeutic effects are poor, At present, patients depend on diet and rest therapy. For example, a diuretic is used as a therapeutic agent for renal disease, but its pharmacological action is to supplement the reduced renal function, and does not prevent renal disease or cure renal disease. Further, among the diuretics, most of the thiazide type diuretics and the non-thiazide type diuretic furosemide have a problem that they cause liver dysfunction as a side effect and are not preferable in treatment.

Therefore, at present, it cannot be said that there is a sufficiently satisfactory drug as a drug for treating renal diseases, and there is a strong demand for the development of a more effective drug.

Similarly, as methods for treating liver diseases, there are currently known therapies for maintaining liver function, such as rest therapy, diet therapy and drug therapy. Among these therapies, as drug therapy, specifically, amino acids such as aspartic acid, thioctic acid, methionine, thioproline,
There is a therapy using peptides such as glycine and glutathione, liver extract, liver hydrolyzate, placenta hydrolyzate, glucuronic acid derivative and glycyrrhizin extracted and purified from licorice. Of these drugs, glycyrrhizin is known to be effective in improving liver function caused by chronic hepatitis, but this glycyrrhizin has a problem in administration that it is ineffective by oral administration. .

The mechanism of liver disease has not been clarified except for viral hepatitis and toxic hepatitis, and the treatment methods for liver disease have mainly been rest therapy and diet therapy. However, in recent years, for example, hepatocyte protein synthesis and RNA
By activating the synthesis, malotilate was developed, which has the effect of suppressing the progress of hepatic fibrosis. However, such malotilate is only approved for application in the treatment of cirrhosis. Furthermore, although its mechanism of action is unknown, cyanidanol has been developed as an immunostimulant and is said to be effective in treating type B viral hepatitis. However, the sale of cyanidanol is currently suspended in Japan due to a fatal accident in a patient who received cyanidanol in Italy and Portugal.

Therefore, under the present circumstances, there is a strong demand for the development of a drug which is truly effective in treating and preventing a wide range of liver diseases and has excellent safety. Furthermore, as described above, the kidney and the liver have important roles in the metabolic function of the body and are related to each other. This is because when one of the liver and the kidney is dysfunctional, the metabolic load is increased on the other, resulting in dysfunction of both organs in many cases. As the disorder progresses, renal dysfunction often occurs. Therefore, as a therapeutic drug, a drug that simultaneously improves the functions of both organs is desirable.

[0015]

[Means for Solving the Problems] The inventors of the present invention have conducted extensive studies in order to solve the above-mentioned problems, and as a result of studying a wide variety of substances, as a result of focusing on isoliquiritigenin, I came to obtain such knowledge. It has long been known that isoliquiritigenin exists in nature as a plant licorice component. However, the content of isoliquiritigenin in licorice is very small, most of it exists as glycosides, and many similar homologs coexist, which makes isolation and purification difficult. The pharmacological effect of liquiritigenin was unclear. For example, JP-B-48-8485 reports that a mixture of liquiritin, isoliquiritin, liquiritigenin, isoliquiritigenin and the like extracted from licorice has an antiulcer action.
However, in such a report, there is no clear suggestion that isoliquiritigenin, which coexists in a very small amount, is an active ingredient for antiulcer action.

The purity of isoliquiritigenin used in the same manner is unknown, but isoliquiritigenin is reported to have a relatively strong antispasmodic activity (Pharmaceutical Journal, Vol. 80, 620-624, page 1960). By Shibata et al. In addition, the pharmacological activity of chalcones has been attracting attention in recent years. For example, the carcinogenic promoter TPA by Nakadate et al.
Of the inhibitory effect of dermal hypervascular permeability (“inflammation”)
Vol. 4, pp. 554-556, 1984) and Japanese Patent Application No. 60-1
There is a report on the differentiation-inducing activity of animal tumor cells in 78815 and its usefulness as a carcinostatic agent.

Thus, based on such knowledge, the inventors of the present invention independently conducted research, and previously proposed the usefulness of isoliquiritigenin as an antiallergic agent (Japanese Patent Application No. 61-49530). . Therefore, as a result of further studies based on such findings, new findings were obtained that isoliquiritigenin is effective in treating and preventing kidney and liver diseases. The present invention has been made based on such findings. That is, the present invention is a drug for treating and preventing kidney and liver diseases, which comprises isoliquiritigenin or a pharmaceutically acceptable salt thereof as an active ingredient.

As described above, the term "kidney disease" refers to (a) renal dysfunction caused by abnormal substance metabolism, for example, acute nephritis caused by a drug or the like, and chronic nephritis in which such acute nephritis is chronic ( ) Acute nephritis mediated by an immune mechanism and chronic nephritis in which such acute nephritis becomes chronic, (c)
Acute nephritis caused by bacterial and viral infections and chronic nephritis caused by chronic infection, such as glomeruli, renal tubules, lupus, and other functional disorders of the kidney. Also,
Similarly, the term “liver disease” as used herein means (a) hepatic dysfunction caused by abnormal substance metabolism and biosynthesis, for example, acute hepatitis caused by a drug or chronic hepatitis in which such acute hepatitis is chronic, (b) Fatty liver caused by drinking or the like, chronic hepatitis caused by these chronic diseases, (c) viral hepatitis caused by viral infection, or chronic hepatitis caused by the chronic infection, and diseases exhibiting widespread liver dysfunction such as cirrhosis resulting from the above disorders. Say.

The pharmaceutically acceptable salt of isoliquiritigenin includes non-toxic salts such as alkali metal and alkaline earth metal salts, for example, sodium salt, potassium salt, magnesium salt, calcium salt and the like. Yes, and
There are non-toxic amine salts such as ammonium salts.

The drug for treating and preventing kidney and liver diseases of the present invention can be administered orally or parenterally (for example, intravenous injection, subcutaneous administration, rectal administration, etc.). Can be prepared into different dosage forms. Such agents include tablets, capsules, granules, powders, fine granules, pills, troches, sublingual tablets, suppositories, ointments, injections, emulsions, suspensions, syrups, etc., depending on their use. It can be prepared in any formulation form.

In preparing these, for example, non-toxic excipients, binders, disintegrants, lubricants, preservatives, antioxidants, isotonic agents, buffers, which are commonly used in this type of drug, are used. Agent,
Coating agents, flavoring agents, solubilizing agents, bases, dispersants,
It can be formulated by a known method using additives such as a stabilizer and a colorant. The specific examples of each of the non-toxic additives that can be used are as follows.

First, as excipients, starch and its derivatives (dextrin, carboxymethyl starch, etc.), cellulose and its derivatives (methyl cellulose,
Hydroxypropyl methylcellulose etc.), sugars (lactose, sucrose, glucose etc.), silicic acid and silicates (natural aluminum silicate, magnesium silicate, magnesium aluminometasilicate etc.), carbonates (calcium carbonate, magnesium carbonate, carbonic acid) Sodium hydrogen, etc.), aluminum hydroxide / magnesium, synthetic hydrotalcite, polyoxyethylene derivatives, glyceryl monostearate, sorbitan monooleate and the like.

As the binder, starch and its derivatives (pregelatinized starch, dextrin etc.), cellulose and its derivatives (methyl cellulose, sodium carboxymethyl cellulose, hydroxypropylmethyl cellulose etc.), gum arabic, tragacanth, gelatin,
Examples include sugars (glucose, sucrose, etc.), ethanol, polyvinyl alcohol, polyvinylpyrrolidone, and the like.

As the disintegrant, starch and its derivatives (carboxymethyl starch, hydroxypropyl starch, etc.), cellulose and its derivatives (carboxymethyl cellulose, sodium carboxymethyl cellulose, crystalline cellulose, hydroxypropyl methyl cellulose, etc.), carbonates (calcium carbonate, Sodium bicarbonate, etc.), tragacanth, gelatin, agar and the like.

Lubricants include stearic acid, calcium stearate, magnesium stearate, talc,
Examples thereof include silicic acid and salts thereof (light anhydrous silicic acid, natural aluminum silicate, etc.), titanium oxide, calcium hydrogen phosphate, dried aluminum hydroxide gel, macrogol, and the like.

As preservatives, paraoxybenzoic acid esters, sulfites (sodium sulfite, sodium pyrosulfite, etc.), phosphates (sodium phosphate, calcium polyphosphate, sodium polyphosphate, sodium metaphosphate, etc.), alcohols ( (Chlorobutanol, benzyl alcohol, etc.), benzalkonium chloride, benzethonium chloride, phenol, cresol, chlorocresol, dehydroacetic acid, sodium dehydroacetate, glycerin sorbate, sugars and the like.

As the antioxidant, sulfites (sodium sulfite, sodium hydrogen sulfite, etc.), Rongalit, erythorbic acid, L-ascorbic acid, cysteine, acetylcysteine, thioglycerol, butylhydroxyanisole, dibutylhydroxytoluene, propyl gallate. , Ascorbic acid palmitate, dl-
Examples include α-tocopherol and nordihydroguaiaretic acid.

Examples of the tonicity agent include sodium chloride, potassium nitrate, sodium nitrate, dextran, glycerin, glucose and the like.

As the buffer, sodium carbonate, hydrochloric acid,
Examples thereof include boric acid and phosphates (sodium hydrogen phosphate, etc.).

As coating agents, cellulose derivatives (hydroxypropyl cellulose, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate, etc.), shellac, polyvinylpyrrolidone, polyvinylpyridines (poly-2-vinylpyridine, poly-2-vinyl-5). -Ethylpyridine, etc.), polyvinyl acetal diethylaminoacetate, polyvinyl alcohol phthalate, and a methacrylate / methacrylic acid copolymer.

As the corrigent, sugars (glucose, sucrose,
Lactose, etc.), sodium saccharin, sugar alcohols and the like.

As the solubilizing agent, ethylenediamine,
Nicotinamide, sodium saccharin, citric acid,
Examples thereof include citrates, sodium benzoate, soaps, polyvinylpyrrolidone, polysorbates, sorbitan fatty acid esters, glycerin, propylene glycol, benzyl alcohol, jetilin, sugar esters and the like.

As the base, fats (pork fat etc.), vegetable oils (olive oil, sesame oil etc.), animal oils, lanolin, petrolatum, paraffin, waxes, resins, bentonite, glycerin, glycols, higher alcohols (stearyl alcohol, Cetanol, etc.) Cellulose derivatives and the like can be mentioned.

As the dispersant, gum arabic, tragacanth, cellulose derivative (methyl cellulose etc.), polyoxylstearic acid, sorbitan sesquioleate,
Examples thereof include aluminum monostearate, sodium alginate, polysorbates, sorbitan fatty acid esters and the like.

Finally, examples of the stabilizer include sulfites (sodium hydrogen sulfite, etc.), nitrogen, carbon dioxide and the like.

The content of isoliquiritigenin in such a preparation varies depending on its dosage form, but it is generally desirable to contain it in a concentration of 0.1 to 100% by weight. The dose of the preparation according to the present invention can be widely varied depending on the type of warm-blooded animal including human beings, the severity of symptoms, the diagnosis of a doctor, etc. 0.01 to 300 mg / kg / kg body weight per day, preferably 0.01
˜50 mg / kg, for parenteral administration, 0.01 to 150 mg / kg / kg body weight daily, preferably
It is preferable to administer 0.01 to 20 mg / kg. However, it is possible to change the dosage range according to the severity of the patient's symptoms and the doctor's diagnosis. The above dose can be administered once or several times a day. Examples of the present invention will be shown below. The present invention is not limited to these examples.

[Example 1] "Toxicity test" This example was carried out to confirm the safety of isoliquiritigenin, which is the active ingredient of the present invention. Isoliquiritigenin was orally and intraperitoneally administered to five 5-week-old male ddY mice. As a result, the minimum lethal dose was 3000 mg / kg or more (oral) and 1000 mg / kg or more (intraperitoneal).

Example 2 “Effect on Gentamycin-induced Acute Kidney Injury” In this example, among renal dysfunction caused by abnormal substance metabolism, a model of acute nephritis caused by a drug was used.
Select the aminoglycoside antibiotic gentamicin, which is known to have an extremely strong effect of inducing renal dysfunction,
This was carried out to investigate the action of isoliquiritigenin on acute nephritis caused by this and to confirm the usefulness of the drug of the present invention. Suzuki et al. Method (Journal of the Japanese Pharmacological Society 84
Vol. 463-469, 1984). That is, male Wistar rats weighing 160 to 180 g (Charles River Japan Co., Ltd., 5 animals per group) were subcutaneously administered with gentamicin 80 mg / kg / day once a day for 15 days to induce renal damage to induce renal damage. % 50,150,300 mg / kg of isoliquiritigenin suspended in an aqueous solution of 1% carboxymethyl cellulose (hereinafter abbreviated as 1% CMC)
/ Day was orally administered to 5 ml / kg for 15 consecutive days.
Gentamicin was administered to the solvent control group in the same manner as in the isoliquiritigenin administration group, and then 5 ml of a 1% CMC aqueous solution was added.
/ Kg was orally administered for 15 consecutive days. Collect urine by collecting urine for 24 hours using a metabolic cage from the day before the first administration and the day after the last administration, and after measuring the urine volume, 3000
After centrifugation at rpm for 15 minutes, the lactate dehydrogenase activity (LDH) in urine of the supernatant was analyzed automatically (AU-5
50, Olympus Optical Co., Ltd.). Furthermore, N-acetyl-β-D-glucosamidase activity (NAG) in urine was measured using NAG Test Shionogi (Shionogi Pharmaceutical Co., Ltd.). After the end of urine collection and before the start of the test,
Blood was collected from the tail vein. On the day after the final administration, blood was collected from the abdominal aorta under ether anesthesia, exsanguination was performed, the laparotomy was performed, and the kidney was removed. Blood sampled is 3000 rpm, 1
After centrifugation for 5 minutes, blood urea nitrogen (BUN) of the serum was measured using an automatic analyzer. The excised kidney was divided into two, fixed with 10% buffered formalin, and a hematoxylin / eosin stained specimen was prepared and examined microscopically. The results are shown in Tables 1 and 2 below.

[0039]

[Table 1]

[0040]

[Table 2]

(Results) (1) In the group administered with gentamicin alone, NAG and LDH increased remarkably 15 days after administration of gentamicin, and B
An increase in UN was also recognized, and the onset of renal damage due to gentamicin was clearly recognized. (2) Isoliquiritigenin significantly suppressed the increase in NAG, LDH, and BUN levels in a dose-dependent manner as compared with the gentamicin-induced control group. (3) Also in the histopathological findings of the kidney, isoliquiritigenin 300 mg / kg compared with the gentamicin-induced control group.
In the administration group, marked improvement in necrosis, degeneration, regeneration and division, and invasion of tubular epithelial cells was observed. Further, improvement of necrosis, regeneration and division of tubular epithelial cells was also observed in the isoliquiritigenin 50, 150 mg / kg administration group. From the above test results, it is clear that isoliquiritigenin significantly improves drug-induced acute nephritis, and isoliquiritigenin is considered to be extremely useful for the treatment of drug-induced acute nephritis. .

[Example 3] "Effect of isoliquiritigenin on cisplatin-induced acute renal injury" This Example is a model of acute nephritis caused by a drug among renal dysfunction caused by abnormal substance metabolism. The anticancer drug cisplatin, which is known to have a very strong renal dysfunction-inducing action, was selected, and the action of isoliquiritigenin on the acute nephritis caused by this was investigated, and the usefulness (preventive and therapeutic effects) of the drug of the present invention was confirmed. It was done in order to do.

(1) Preventive test 1% for 20 to 25 g of ddY male mice (10 mice per group)
Isoliquiritigenin 200m suspended in CMC aqueous solution
Cisplatin 1 dissolved in physiological saline at 2 mg / 10 ml on the 5th day after oral administration of g / kg / day for 5 consecutive days
6 mg / kg was subcutaneously administered once, and 5 more days from the next day
Similarly, daily administration of isoliquiritigenin was performed, blood was collected, centrifuged at 3000 rpm for 15 minutes, and then BU in serum was
N was measured.

(2) Prevention test 20 to 25 g of male ddY mice (10 mice per group) were treated with 16 mg of cisplatin 2 mg / 10 ml dissolved in physiological saline.
mg / kg once subcutaneously administered, 1 mg of isoliquiritigenin 200 mg / kg / day suspended in 1% CMC aqueous solution
Orally administered once daily for 5 consecutive days, then blood sampled for 3000 r
After centrifugation at pm for 15 minutes, BUN in serum was measured.

In the tests (1) and (2), 20 to 2
5 g of male ddY mice (10 mice per group), which were continuously orally administered with isoliquiritigenin 200 mg / kg / day, were used as a control group administered with isoliquiritigenin alone,
As a nephritis-induced control group, 20 mg to 25 g of ddY male mice (10 mice per group) were subcutaneously administered with 16 mg / kg of cisplatin, and 1% CMC was used instead of isoliquiritigenin.
Except that the aqueous solution was continuously orally administered, the same treatment was performed as in the cisplatin + isoliquiritigenin administration group, and each was compared with the same administration group. Table 3 shows the results.

[0046]

[Table 3]

(Results) (1) Compared to the untreated control group, the BUN was remarkably increased in the control group inducing cisplatin, which clearly showed the onset of acute renal injury by cisplatin. (2) Isoliquiritigenin significantly suppressed the increase in BUN in both the preventive test and the therapeutic test, as compared with the control in which renal damage was induced. (3) No change in BUN was observed in the control group to which isoliquiritigenin was administered alone, which did not cause renal injury, as compared with the non-treated control group, and isoliquiritigenin increased BUN due to the onset of renal injury. It is clear that it suppresses specifically. From the above results, it is clear that isoliquiritigenin remarkably improves drug-induced acute kidney injury, and is considered to be extremely useful for the prevention and treatment of drug-induced acute kidney injury.

[Example 4] "Effect of isoliquiritigenin on rat anti-GBM nephritis" It is said that 70% of human chronic kidney disease develops through an immune mechanism, and particularly in glomerulonephritis, many In these cases, findings suggesting the involvement of the immune mechanism were recognized, and based on this, various studies have been made on the production of animal experimental models as human chronic nephritis models. Above all, the Masugi nephritis model developed in Japan is widely used worldwide as a nephritis model due to an antibody against the glomerular tissue antigen (GBM).
Therefore, the present inventors chose the Masugi nephritis model as the human chronic nephritis model, and used the method of Suzuki Yoshio et al. (Journal of the Nephrological Society of Japan, Vol. 23, pp. 323-331, 1981) and (Jpn. (1981), this example was performed to confirm the usefulness of the drug of the present invention.

That is, 0.5 ml / rat of anti-rat GBM rabbit serum was added to male SD rats weighing 180 to 200 g (Charles River Japan Co., Ltd., 7 in the nephritis-inducing control group, 6 in the other groups). Inject intravenously and immediately 1% CMC
Isoliquiritigenin 30, 100, suspended in an aqueous solution,
300 mg / kg / day was orally administered to 5 ml / kg daily for 3 weeks continuously in each group of rats. The nephritis-induced control group was intravenously injected with 0.5 ml of anti-rat GBM rabbit serum / rat, and then 1
% CMC aqueous solution was continuously orally administered to 5 mg / kg daily for 3 weeks. Urine was collected by collecting urine for 24 hours using a metabolic cage from the last administration day of the third week of anti-GBM serum administration to the next day, and after measuring the urine volume, centrifugation was performed at 3000 rpm for 15 minutes, and the supernatant was lactate dehydrogenated in urine. Enzyme activity (LD
H), alkaline phosphatase (ALP), and protein content were measured using an automatic analyzer (AU-550, Olympus Optical Co., Ltd.). After completion of urine collection, BUN, creatinine, ALP, total cholesterol, total protein, and albumin were measured in serum collected from the abdominal aorta under ether anesthesia and centrifuged at 3000 rpm for 15 minutes. The results are shown in Table 4 below.
And shown in Table 5.

[0050]

[Table 4]

[0051]

[Table 5]

(Results) (1) In the nephritis-induced control group at the 3rd week, the total protein, total ALP, and total LDH in urine significantly increased as compared with the untreated control group.
Although BUN and creatinine in the blood did not show significant changes, a decrease in ALP and an increase in total cholesterol were observed. Further, although there was no change in the total serum protein amount, a decrease in albumin content was observed, and a corresponding decrease in the A / G ratio was observed. From the above results, it is clear that the renal system is mediated by the immune system. (2) In each dose group of isoliquiritigenin administration group, total protein and total AL in urine were higher than those of the nephritis-induced control group.
The increase in P and total LDH was significantly suppressed. Also, blood AL
Significant improvements in P, total cholesterol and albumin levels were observed. From the above results, it is considered that isoliquiritigenin is extremely useful for the prevention and treatment of immune-mediated renal damage.

[Example 5] "Effect of isoliquiritigenin on d-galactosamine-induced liver injury" d-galactosamine is a compound that induces a lesion similar to the lesion of viral hepatitis in humans,
It is used as a model for viral hepatitis. Therefore, the present inventors have proposed the method of Sakamoto et al. (“Efficacy Screening Method Vol. 1 for New Drug Development” 69-82, 1984).
(Koji Sakamoto, Seisho Shoin), the present example was carried out as follows to confirm the usefulness of the drug of the present invention. That is, 1% CMC was applied to Wistar male rats weighing 180 to 200 g (Charles River Japan, 6 per group).
Isoliquiritigenin 10, 30 mg suspended in aqueous solution
/ Kg / day was orally administered to each rat group for 5 days at 5 ml / kg, and 1 hour after administration of isoliquiritigenin for 4 days, 250 m of d-galactosamine dissolved in physiological saline was administered.
g / 5 ml / kg was intraperitoneally administered, and 48 hours after the administration of d-galactosamine, blood was collected from the abdominal aorta under ether anesthesia, and centrifuged at 3000 rpm for 15 minutes. ,
ALP was measured by an automatic analyzer AU-550.

The hepatopathy-induced control group was treated in the same manner as the isoliquiritigenin administration group except that an 1% CMC aqueous solution was orally administered instead of the isoliquiritigenin 1% CMC suspension. Also, as a positive control group, instead of isoliquiritigenin, malotilate 30 mg / kg / day, 100 mg / kg / da suspended in 1% CMC aqueous solution, respectively.
y, cyanidanol 30mg / kg / day, 100m
Treatment was carried out in the same manner as in the isoliquiritigenin administration group except that orally administered to g / kg / day. The measurement results are shown in Table 6.

[0055]

[Table 6]

(Results) (1) In the liver injury-induced control group, a clear increase in GOT, GPT, and ALP was observed as compared with the untreated control group. (2) In the group administered with isoliquiritigenin, GOT, GPT,
An increase in ALP was suppressed, and a clear improvement in GOT and GPT was observed at 30 mg / kg. (3) 10 for positive control group malotilate and cyanidanol
Isoliquiritigenin 10 in the 0 mg / kg administration group
Almost the same suppression of GOT and GPT increase as in the mg / kg administration group was observed. From the above results, isoliquiritigenin is malotilate,
It is clear that d-galactosamine-induced liver damage is suppressed at a dose lower than that of cyanidanol, and it is considered to be useful for the prevention and treatment of viral hepatitis and the like.

[Examples 6 and 7] Carbon tetrachloride generates a highly reactive free radical by the drug-metabolizing enzyme system in hepatocytes, which binds to a hepatocyte membrane protein to strongly suppress cell activity or It is said that hepatocyte necrosis and hepatic fat accumulation are caused by peroxidation of membrane lipids of internal organs, and it is most widely used as an experimental model of human acute drug hepatitis, fatty liver, chronic hepatitis, and cirrhosis. Has been. Therefore, the present inventors have proposed the method of Imaizumi et al. (Japan Jour
nal of Pharmacology vol.31 15-21 1981
1) and Minoru Kato et al. (Journal of the Japanese Pharmacological Society, 80 volumes, p.
83-91 1982) and the following Examples 6 and 7 were carried out to confirm the usefulness of the drug of the present invention.

[Example 6] "Effect of isoliquiritigenin on carbon tetrachloride (CCl ₄ ) -induced acute liver injury" Wist having a body weight of 180 to 200 g
ar male rats (Charles River Japan, 1
Isoliquiritigenin (10, 30 mg / kg / day) suspended in a 1% CMC aqueous solution was orally administered to 6 animals in a group for 4 consecutive days. For the solvent control group, 5 ml of 1% CMC aqueous solution /
kg / day was continuously administered orally for 4 days, and 30 mg / kg / day of malotilate was added to the positive control group.
100mg / kg / day and Cyanidan
ol) 30 mg / kg / day, 100 mg / kg / da
Y was suspended in a 1% CMC aqueous solution and continuously orally administered for 4 days. For carbon tetrachloride, a precise analysis reagent was used, and rats were orally administered for 3 consecutive days from the second day of drug administration to 0.2 ml /
Subcutaneous administration was performed at kg / day. Twenty-four hours after the final administration of carbon tetrachloride, blood was collected from the abdominal aorta under ether anesthesia. Blood was GOT for serum spun at 3000 rpm for 15 minutes,
The GPT was measured using an automatic analyzer (AU-550, Olympus Optical Co., Ltd.). The results are shown in Table 7.

[0059]

[Table 7]

(Results) (1) The liver damage-inducing control group remarkably G
An increase in OT and GPT was observed, and the occurrence of liver damage due to carbon tetrachloride was clearly observed. (2) In the isoliquiritigenin administration group, GOT and GPT were significantly improved in each dose group as compared with the liver injury-induced control group. (3) 30% for positive control group malotilate and cyanidanol
In the mg / kg administration group or the 100 mg / kg administration group, almost the same suppression of GOT and GPT increase as in the isoliquiritigenin 10 mg / kg administration group was observed. From the above results, isoliquiritigenin is malotilate,
It is considered that at a dose lower than cyanidanol, it suppresses carbon tetrachloride-induced acute liver injury and is useful for the prevention and treatment of acute liver injury.

[Example 7] "Effect of isoliquiritigenin on carbon tetrachloride-induced chronic liver injury" Male Wistar rats weighing 180 to 200 g (Charles River Japan Co., Ltd., 6 animals per group) Iritigenin 30 mg / kg / day 1% CM
It was suspended in a C aqueous solution and continuously orally administered every day for 3 weeks. The solvent control group was orally administered with 1% CMC aqueous solution 5 ml / kg / day continuously for 3 weeks, and the positive control group was malotilate 30 mg / kg / day and cyanidanol 30 mg / day.
kg / day was suspended in a 1% CMC aqueous solution and continuously orally administered every day for 3 weeks. Carbon tetrachloride is 3 from the 3rd day of drug administration
0.5 ml / kg was intraperitoneally administered twice a week for consecutive weeks. On the fourth day after the final administration of carbon tetrachloride, blood was collected from the abdominal aorta under ether anesthesia. Blood was centrifuged at 3000 rpm for 15 minutes, and serum was analyzed by GOT and GPT with an automatic analyzer (A
U-550). The results are shown in Table 8.

[0062]

[Table 8]

(Results) (1) Compared with the non-treated control group, GO control group
A marked increase in T and GPT is seen, and the onset of severe liver damage due to carbon tetrachloride is clear. (2) In the isoliquiritigenin administration group, almost the same suppression of increase in GOT and GPT as in the positive control group cyanidanol was observed. On the other hand, malotilate was significantly suppressed in this test system. From the above results, isoliquiritigenin is considered to be useful for the prevention and treatment of chronic liver damage.

[Example 8] "Effect of isoliquiritigenin on cisplatin-induced acute liver injury" The anticancer agent cisplatin is known to damage the liver as well as the kidney (F.
Cawalli et al Cancer Treatment Reports vol.62 No.1
22125-2126 1978). The present inventors found that the anticancer drug cisplatin damages the liver as well as the kidney,
Moreover, it has been found that even a lower dose of cisplatin that causes kidney damage causes liver damage. In this example,
The usefulness of the drug of the present invention was confirmed by examining the action of isoliquiritigenin on this cisplatin-induced acute liver injury. Wist weighing 150-180g
ar male rats (Charles River Japan, 1
Cisplatin 8.5 mg / kg dissolved in physiological saline 2 mg / 10 ml was subcutaneously administered to 1 group (6 animals) once, and then 1% CM
Isoliquiritigenin 200mg /
After oral administration of kg / day for 10 consecutive days, the animals were dissected,
A hematoxylin-eosin stained specimen of the liver was prepared and examined microscopically. As a control group for inducing liver damage, 150-180 g
1% C instead of isoliquiritigenin in male Wistar rats (Charles River Japan, 6 per group)
The same treatment as in the isoliquiritigenin administration group was used except that the MC aqueous solution alone was orally administered for 10 consecutive days. The results are shown in Table 9.

[0065]

[Table 9]

(Results) (1) Compared with the non-treated control group, it is clear that the liver damage was caused by cisplatin administration in the cisplatin-administered control group. (2) Isoliquiritigenin significantly suppressed cisplatin-induced acute liver injury. From the above results, it is clear that isoliquiritigenin remarkably improves cisplatin-induced acute liver injury, and it is considered that the drug is extremely useful for the treatment of acute liver injury.

[Example 9] "Effect of isoliquiritigenin on cisplatin-induced acute kidney and liver injury" As described above, isoliquiritigenin is used for the prevention and treatment of renal and liver injury. Since it has been clarified that it is useful, the present inventors conducted the following preventive and therapeutic tests using the experimental model of the anticancer drug cisplatin of Example 3 and Example 8 described above to determine the kidney and liver of the drug of the present invention. Was confirmed to be effective against concomitant diseases.

(1) Preventive test 1% CM was applied to male Fisher rats (Charles River Japan, Ltd., 3 animals per group) weighing 215 to 240 g.
20 mg / k of isoliquiritigenin suspended in C aqueous solution
g / day was orally administered to one group, and intraperitoneally administered to one group once a day for 12 consecutive days. From the 6th day after the start of administration, 2 mg / kg of cisplatin dissolved in physiological saline was administered.
g / day was subcutaneously administered once a day for 4 consecutive days, and isoliquiritigenin was administered in the same manner for 3 days on the day of cisplatin administration and after the end of cisplatin administration. However, on the cisplatin administration day, isoliquiritigenin was administered immediately after the administration of cisplatin. Cisplatin was administered to the renal and hepatic disorder-induced control group in the same manner as the isoliquiritigenin administration group, and 1% CM was used instead of isoliquiritigenin 1% CMC suspension.
The treatment was performed in the same manner as in the isoliquiritigenin administration group except that the C aqueous solution was orally administered. 24 hours after the end of administration of isoliquiritigenin, blood was collected from the abdominal aorta under ether anesthesia,
After autopsy, the liver was removed, fixed in 10% buffered formalin solution, stained with hematoxylin-eosin, and PAS.
Staining was performed and microscopic examination was performed. Blood collected is 3000 rp
After centrifuging at m for 15 minutes, BUN of the serum was measured using an automatic analyzer (AU-550, Olympus Optical Co., Ltd.).

(2) Therapeutic test Male Fisher rats weighing 215 to 240 g (Nihon Charles River Co., Ltd., 3 animals per group) were treated with 2 mg / 10 ml of cisplatin dissolved in physiological saline.
It was subcutaneously administered once a day for 4 days at a dose of kg / day to induce renal and hepatic disorders. Immediately after daily cisplatin administration, 1% CMC
20 mg / kg of isoliquiritigenin suspended in an aqueous solution
/ Day was orally administered to one group and intraperitoneally to the other group, and was administered once a day for 4 consecutive days, and after cisplatin administration, was administered once a day for 3 consecutive days. As control for inducing renal and hepatic disorders, cisplatin was administered in the same manner as in the isoliquiritigenin administration group, and 1% CMC aqueous solution was orally administered instead of 1% CMC suspension of isoliquiritigenin. The same treatment was performed as in the tigenin-administered group.

Twenty-four hours after the end of administration of isoliquiritigenin, blood was collected from the abdominal aorta under ether anesthesia, necropsy was performed, the liver was extracted, fixed in 10% buffered formalin solution, and stained with hematoxylin-eosin and PAS. I did a microscopic examination. The collected blood was centrifuged at 3000 rpm for 15 minutes, and then BUN of serum was analyzed by an automatic analyzer (AU-5).
50, Olympus Optical Co., Ltd.). In addition, the untreated control group was a male Fisher rat having a body weight of 215 to 240 g (Japan Charles River Co., Ltd.,
1% CMC aqueous solution was orally administered once a day for 12 days instead of isoliquiritigenin 1% CMC suspension to 3 animals per group). Twenty-four hours after the end of administration, blood collection and autopsy were performed, and BUN measurement and microscopic examination were performed in the same manner as above. The results are shown in Table 10 and Table 11.

[0071]

[Table 10]

[0072]

[Table 11]

(Results) (1) Compared with the untreated control group, the kidney and liver injury-inducing control group had increased BUN and pathological findings of liver tissues and cells, that is,
It is clear that cisplatin causes renal damage and hepatic damage due to oxidative and vacuolar degeneration of hepatocytes. (2) In the preventive test and the therapeutic test, the isoliquiritigenin administration group was compared with the renal and hepatic disorder-induced control group.
Both oral administration and intraperitoneal administration markedly suppressed the increase in BUN and suppressed the onset of liver damage. From the above results and Example 3 and Example 8, it is considered that isoliquiritigenin is extremely useful for the prevention and treatment of complications of renal disorder and liver disorder.

In the above-mentioned Examples, the usefulness of the drug of the present invention in each disease of kidney and liver and in the co-morbidity of both organs was shown. Subsequently, in the examples shown below,
A prescription example in which the dosage form and composition are specifically specified is shown. In addition,
The present invention is not limited to the following examples.

[Example 10] 5 mg and 2 per tablet
An example of a tablet formulation containing 5 mg of active ingredient is as follows. Prescription example 1 (5 mg tablet) Isoliquiritigenin 5 Lactose 137 Starch 45 Carboxymethylcellulose calcium 10 Talc 2 Magnesium stearate 1 200 mg / tablet Prescription example 2 (25 mg tablet) Isoliquiritigenin 25 Lactose 120 Starch 42 Carboxymethylcellulose calcium 10 Talc 2 Magnesium stearate 1 200 mg / tablet The details of the production method are as follows. The crystals of isoliquiritigenin are ground, and lactose and starch are added thereto and mixed. Add 10% starch paste to the above mixture,
Stir to produce granules. After drying, the particle size was adjusted to about 850 μm, and talc and magnesium stearate were mixed and compressed into tablets.

[Example 11] (20 mg capsule) Isoliquiritigenin 20 Lactose 53 Starch 25 Magnesium stearate 2 100 mg / tablet Isoliquiritigenin was crushed well, and starch, lactose and magnesium stearate were sufficiently added. After mixing, it was filled into capsules.

[0077]

INDUSTRIAL APPLICABILITY As described above, the present invention provides a therapeutic and prophylactic drug for kidney and liver diseases, characterized by containing isoliquiritigenin or a pharmaceutically acceptable salt thereof as an active ingredient. According to the present invention, each of a wide range of renal and hepatic diseases that occur independently, and further, when the diseases of both organs coexist, the excellent therapeutic and prophylactic effect is exhibited. A prophylactic drug can be provided.

Claims (1)

[Claims] 1. A therapeutic or prophylactic drug for liver diseases, which comprises isoliquiritigenin or a pharmaceutically acceptable salt thereof as an active ingredient.