JPH01226814A - Remedy for hyperammonemia - Google Patents

Abstract

PURPOSE:To provide a remedy for hyperammonemia containing biotin as an active component and effective in lowering the ammonia level in blood and ameliorating hepatic coma. CONSTITUTION:The objective remedy can be prepared by mixing the biotin of formula with conventional drug carrier, dissolution assistant, stabilizing agent, distilled water for injection, etc., and forming to a drug preparation by conventional method. It can be used in the form of tablet, capsule, syrup, injection, etc. Biotin is administered at a rate of 0.5-100mg daily for adult in 1-2 divided doses in the case of injection or in several divided doses in the case of oral drug. Biotin of formula is a white crystal or crystalline powder free from taste and odor. It has a melting point of about 231 deg.C and is little soluble in glacial acetic acid, scarcely soluble in water, ethanol and n-butanol and almost insoluble in ether and chloroform. It is soluble in sodium hydroxide reagent.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to a therapeutic agent for hyperammonemia.

More specifically, the present invention relates to a therapeutic agent for hyperammonemia containing biotin as an active ingredient.

[Prior art]

Currently, biotin is used as a therapeutic agent containing biotin as an active ingredient for skin diseases such as acute and chronic eczema, childhood eczema, and contact dermatitis.

Conventionally, non-absorbable antibiotics such as fradiomycin (fura-dioa
+aicin), neomycin, neomycin
Oral administration of synthetic disaccharide preparations and enema administration have been used successfully.

[Problem that the invention is trying to solve]

Ammonia in the blood is absorbed from the intestinal tract, produced by the coupled reaction of amino acid transferase and glutamate dehydrogenase during protein metabolism in the body, and directly separated from amino acids in body cells. Ammonia in the blood exists in the form of ammonium ions generated by dissociation equilibrium with free ammonia. Ammonia is toxic to living bodies, so in healthy people it is processed through the urea cycle, and free ammonia is kept at a constant, extremely low level in both blood and tissues.

On the other hand, in cases of severe liver damage (fulminant hepatitis, cirrhosis, etc.), in addition to increased ammonia production, the liver's ability to process ammonia decreases, resulting in hyperammonemia, which is one of the main causes of hepatic encephalopathy. It is said that there is one. In such cases, the above conventional treatment methods often leave patients with diarrhea,
It has the disadvantage of causing side effects such as bacterial alternation, changes in intestinal bacterial flora, and gastrointestinal disorders.

An object of the present invention is to provide an excellent therapeutic agent for hyperammonemia in warm-blooded animals including humans, which overcomes the drawbacks of the conventional therapeutic methods described above.

Other objects of the invention will become apparent from the detailed description and examples below.

[Means to solve the problem]

The present inventors observed a significant decrease in blood ammonia by administering biotin to hyperammonemic rats. Furthermore, the present invention was completed based on the knowledge that the present invention lowers the blood ammonia level of patients with chronic liver failure accompanied by hyperammonemia, and at the same time significantly improves the clinical symptoms of the patients.

Biotin used as an active ingredient in the present invention is represented by the following formula. : (CHz)9COOH This compound is a white crystal or crystalline powder with no odor or taste. It has a melting point of about 231° C. (decomposed), is soluble in glacial acetic acid, extremely sparingly soluble in water, ethanol, or n-butanol, and almost insoluble in ether or chloroform. Dissolves in sodium hydroxide test solution.

Typical pharmaceutical dosage forms containing the biotin of the present invention include, for example, oral preparations such as tablets, coated tablets, capsules, and syrups, and parenteral preparations such as injections. The dosage unit form may contain about 0.1 to 50 μg of biotin.

Solid formulations may include conventional pharmaceutical carriers such as lactose, sucrose, sorbitol, mannitol, starches, cellulose derivatives, calcium and magnesium stearate, carbowaxes, PvP, and the like. Oral aqueous preparations may optionally contain solubilizing agents such as nicotinamide, flavoring agents such as sucrose, or stabilizers, flavoring agents, coloring agents, and the like.

The aqueous preparation for injection consists of biotin and distilled water for injection, and may contain other ingredients such as solubilizers, stabilizers, and soothing agents, if necessary. These pharmaceutical formulations can be readily manufactured according to techniques well known in the art.

The amount of biotin used in the treatment of the present invention varies depending on the patient's symptoms, but for adults, the amount of biotin used per day is 0.
．． It is 5 to 100 square meters, preferably 1 to 30 square meters. In the case of injections, it is recommended to administer the drug once or twice a day, and in the case of oral preparations, it is recommended to administer the drug several times a day, preferably in 3 or 4 divided doses.

[For production]

Although the mechanism by which biotin lowers blood ammonia levels has not yet been established, it is thought that biotin may be involved in carbon dioxide fixation by carbamoyl phosphate synthase, which is an early step in the urea cycle. The excellent effect of the biotin of the invention on suppressing the rise in blood ammonia levels is demonstrated by the results of the test using rats shown below. : Intraperitoneal administration experiment of biotin This experiment involved Wistar rats (male, average weight 2
00 g) was used for a total of 18 animals, 9 animals in the biotin administration group and 9 animals in the control group. In the biotin administration experiment, in order to increase the blood ammonia level, urease (concentration 25 units/-, Sigma) dissolved in physiological saline was administered at a dose of about 25j#/kg. 12 hours before administering urease, inject biotin 1/2 hours using an injection prepared according to Example 4 below.
The mice (concentration 1/2-) were administered intraperitoneally, and blood was collected immediately before administration of urease. After intraperitoneal administration of urease, blood was collected at 2 hours, 4 hours, 6 hours, and 9 hours, and the ammonia levels of these blood samples were determined using the method of Okuda and Fujii [Modern Medicine 21.622-627 (1966). ), that is, indophenol reagent was directly added to the deproteinized supernatant to develop color, and ammonia was measured according to the method of quantifying it (this experiment).
. A control group experiment was conducted in exactly the same manner as the above experiment, with 2 ml of raw silicon saline being intraperitoneally administered instead of biotin. The results are shown in Tables 1 and 2 below.

From Tables 1 and 2 above, the average increase in blood ammonia levels in the biotin-administered group 4 and 6 hours after urease administration was 126.4 μg/d, respectively! , 104.8μg/d
! On the other hand, the average increase in blood ammonia levels in the control group 4 hours and 6 hours after administration of urease was 257.0 μg/dl and 278.1 μg/d1, respectively; Increase in blood ammonia levels caused by urease loading after urease administration4
Significant suppression was observed for up to 6 hours. In addition, when a t-test was conducted on the difference between the corresponding mean values for the values in Tables 1 and 2 above, it was found that p<0.02.6 hours and p<0.01 at 4 hours and 9 hours after urease administration. It was shown that the biotin of the present invention lowers blood ammonia levels with a clear difference.

Experiment on intraperitoneal administration of biotin in acute liver failure model rats.
0 g) were used. Carbon tetrachloride mixed 1:1 with olive oil was used to create an acute liver failure model.
d/kg was administered i.p. For the biotin-administered group, 48 hours, 24 hours, and 6 hours before the administration of carbon tetrachloride, each dose of biotin (1/2 w11) was intraperitoneally administered using an injection solution prepared according to Example 4 below. Blood was collected immediately before carbon chloride administration.

After intraperitoneal administration of carbon tetrachloride, blood was collected from the jugular vein of five animals each at 24 hours and 48 hours. The ammonia levels of these blood samples were measured according to the method described by Okuda and Fujii (this experiment).

A control experiment was conducted in exactly the same manner as the above experiment, with physiological saline being intraperitoneally administered to 21R1 instead of biotin. The results are shown in Tables 3 and 4 below.

From Tables 3 and 4 above, it is recognized that biotin tends to suppress the increase in blood ammonia level in the acute liver failure model caused by carbon tetrachloride loading. Furthermore, in the control group, two rats each died 24 hours and 48 hours after carbon tetrachloride administration, whereas there were no deaths in the biotin administration group. When these mortality rates were tested using the Kabran-Meier method, the biotin-administered group clearly showed a survival benefit.

The acute toxicity (LD,.) when the biotin of the present invention is administered to mice and rats via various administration routes is shown in Table 5 below [Fragrance Journal, 61-64 (1980),
Fragrance Journal Co.].

Table 5 Acute Toxicity of Biotin Examples Examples of useful pharmaceutical formulations for administering compounds of the invention are provided in the following examples.

Example l Biotin 2 ■ Milk
Sugar 55 Corn starch 16 Microcrystalline cellulose 30 Polyvinylpyrrolidone -3
05 Calcium stearate 2 total amount 110
■ Pre-prepared biotin trichloride with a portion of lactose, cornstarch,
Add the biotin suspension to the mixture of microcrystalline cellulose and remaining lactose and mix. Next, -3 to polyvinylpyrrolidone
An aqueous solution of No. 0 is added as a binder, followed by preparing aggregated granules according to a conventional method. After adding calcium stearate and mixing, the mixture is compressed into 110 square tablets.

Example 2 Enteric Coated Tablets A coating solution having the following composition is sprayed on the surface of the tablets obtained according to Example 1 according to a conventional method to form a coating of 10 cm per tablet as a dry product.

Enteric coating '100■'s Eudragit L30D -5545,0 Macrogol 6000
1.5 talc
3.0 Purified water
50.5 Total amount 100.0 ■ Example 3 Biotin 2.0 ■ Distilled water for injection Total amount 10.0 d Biotin is dissolved in distilled water for injection, filtered according to the usual method, and then filled into ampules with 10.0 d. . The ampoule is then fused and sterilized.

Example 4 Biotin 5.0 ■Nicotinic acid amide 0.15 Benzyl alcohol 0.2 Distilled water for injection
Appropriate amount of whole amount 10. oIlll Add biotin and nicotinic acid amide (solubilizing agent) to an appropriate volume of warmed distilled water for injection in which biotin can be dissolved, then stir and dissolve, cool to room temperature, and add benzyl alcohol (soothing agent) and distilled water for injection. Add water to make the total volume.

Thereafter, after filtering according to the usual method, 10.0Pnl was added to the ampoule.
Fill, seal and sterilize.

Example 5 Kotan Satoyuki IM Yu average 4 main biotin 10 z Nicotinic acid amide 300D-Sorvit 2000 water distilled water
Add biotin and nicotinic acid amide to approximately 374 volumes of hot distilled water (100 M1 total volume), stir and dissolve, and then gradually add D-Sorbi Nod. After dissolving, cool to room temperature and add the remaining distilled water to the specified amount.

Example 6 Biotin in 1 capsule 2 ■Milk
Sugar 298 total amount 3
00■ Add biotin to about 1710 ml of lactose and mix, then add the remaining lactose and mix uniformly. This is filled into hard gelatin capsules.

〔Example of use〕

The clinical results of administering the biotin-containing preparation of the present invention to patients with hyperammonemia, mainly due to liver cirrhosis, are as follows: These patients are examples in which no improvement was seen even after administration of other hyperammonemia therapeutics.

Patient 1: Male, 67 years old, liver cirrhosis + gastric cancer patient 2: Female, 65 years old, liver cirrhosis patient 3: Male, 51 years old Liver cirrhosis biotin was an injection prepared according to Example 4 (0.5
/-) was administered intravenously 5 to 10 times a day.

The administration period varies depending on each patient's condition.

Blood samples from each patient were collected once or several times a day at approximately the same time, and blood ammonia levels were measured using the microdiffusion method (patients 1 and 2) and the method described by Okuda and Fujii (patient 3). The degree of hepatic coma was also determined.

The degree of hepatic coma was determined based on Table 6 [Coma Degree Classification Table]. The classification table that serves as the diagnostic standard for this is published in the Journal of the Japan Medical Association, Vol. 92, No. 7 (published October 1, 1980).
Page 78 and 12th Inuyama Symposium 110°198
2. Published in Chugai Igakusha.

Tables 7 to 9 show the biotin administration interval, blood ammonia level before and after administration, and degree of hepatic coma for each patient.

3) a) Blood ammonia level: Represents the blood ammonia concentration before biotin administration (unit: μg/di) b) Hepatic coma level 2 Represents the hepatic coma level according to Table 6 .

However, ■-■ indicates an intermediate state between m and ■ (the same applies hereinafter).

Indicates an intermediate state close to m<n (hereinafter the same) Mr).

O represents a normal state.

C) Biotin dose: represents the biotin dose per day (24 hours).

The results in Tables 7 to 9 above show that administration of the biotin of the present invention reduces blood ammonia levels and improves the degree of hepatic coma in all cases.

Patent applicant: Sansei Pharmaceutical Co., Ltd.

Claims (1)

[Claims] A therapeutic agent for hyperammonemia containing biotin as an active ingredient.