JPH0788304B2 - Hyperammonemia treatment - Google Patents

Description

TECHNICAL FIELD The present invention relates to a therapeutic agent for hyperammonemia. More specifically, the present invention relates to a therapeutic agent for hyperammonemia, which comprises biotin as an active ingredient.

[Prior art]

Currently, therapeutic agents containing biotin as an active ingredient are used for skin diseases such as acute / chronic eczema, childhood eczema, and contact dermatitis.

For the treatment of hyperammonemia, non-absorbable antibiotics such as furadiomaicin, neomycin (neomysin) have been orally administered, and synthetic disaccharide preparations have been given orally and enema. Are listed.

[Problems to be solved by the invention]

Ammonia in blood is absorbed from the intestinal tract, produced by the coupling reaction of amino acid transferase and glutamate dehydrogenase in protein metabolism in the body, and directly released from amino acids in somatic cells. Ammonia in these blood exists in the state of ammonium ion produced by dissociation equilibrium with free ammonia. Ammonia is toxic in the living body, and therefore, in a healthy person, free ammonia is treated at a urea cycle and the like, and free ammonia is maintained at a very low constant level both in blood and tissues.

On the other hand, in the case of severe liver damage (fulminant hepatitis, cirrhosis, etc.), in addition to an increase in ammonia production, the ammonia processing capacity of the liver decreases, resulting in hyperammonemia, which is one of the main causes of hepatic encephalopathy. It is said to be one. In such cases, the above-mentioned conventional treatment methods often cause diarrhea in patients.
It has the drawback of causing side effects such as bacterial dysplasia, changes in intestinal 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 above-mentioned drawbacks of the conventional therapeutic methods.

Other objects of the present invention will be apparent from the following detailed description and examples.

[Means for Solving the Problems]

The present inventors have observed a significant decrease in blood ammonia by administering biotin in hyperammonemia rats. Further, they have found that the blood ammonia level of a patient with chronic liver failure accompanied by hyperammonemia is lowered, and at the same time, the clinical symptoms of the patient are remarkably improved, and the present invention was completed.

Biotin used as an active ingredient in the present invention is represented by the following formula. : The compound is white crystals or crystalline powder, odorless and tasteless. It has a melting point of about 231 ° C. (decomposition), is hardly soluble in glacial acetic acid, is extremely insoluble in water, ethanol, or n-butanol, and is hardly soluble in ether or chloroform. Dissolve in sodium hydroxide test solution.

Usual pharmaceutical dosage forms containing biotin of the present invention include tablets, coated tablets, capsules, syrups and the like as oral agents, and injections and the like as parenteral agents. Dosage unit forms may contain from about 0.1 to 50 mg biotin.

Solid formulations may include conventional pharmaceutical carriers such as lactose, sucrose, sorbitol, mannitol, starches, cellulose derivatives, calcium and magnesium stearate, carbowax, PVP and the like. The aqueous oral preparation may optionally contain a solubilizing agent such as nicotinic acid amide, a flavoring such as sucrose, or a stabilizer, a flavor and a coloring agent. The injectable aqueous preparation consists of biotin and distilled water for injection, and may optionally contain other components such as a solubilizer, a stabilizer and a soothing agent. These pharmaceutical formulations can be readily prepared according to techniques well known in the art.

The amount of biotin used in the treatment of the present invention varies depending on the symptoms of the patient, but in the case of an adult, the amount of biotin per day is 0.
It is 5 to 100 mg, preferably 1 to 30 mg. It is recommended to administer the drug once or twice a day for injections, and several times a day for oral administration, preferably 3 or 4 times a day.

[Work]

The mechanism by which biotin lowers blood ammonia levels has not yet been established, but it is possible that biotin may be involved in carbonic acid fixation of carbamoylphosphate synthase, which is an early stage of the urea cycle.

[Effect]

The excellent effect of suppressing increase in blood ammonia level of the biotin of the present invention is demonstrated from the results of the test using rats shown below. : Biotin ip administration experiment In this experiment, Wistar rats (male, average weight 20
0 g) was used in the biotin-administered group, 9 animals in the control group, and 9 animals in the control group, for a total of 18 animals. In the biotin administration experiment, urease (concentration 25 units / ml, Sigma) dissolved in physiological saline was administered at 25 units / kg in order to raise the blood ammonia level. Twelve hours before the administration of urease, biotin 1 mg / animal (concentration 1 mg / 2 ml) was intraperitoneally administered using the injection prepared according to the following Example 4, and blood was collected immediately before the administration of urease. After intraperitoneal administration of urease, 2 hours, 4 hours, 6 hours,
Blood was collected at the 9th hour, and the ammonia values of these blood samples were determined by the method of Okuda and Fujii [Shinjitsu Kagaku 21,622-627 (196
6)], that is, the deproteinized supernatant was directly added with the indophenol reagent to develop color, and the amount of ammonia was measured (main experiment). In the control group experiment, 2 ml of physiological saline was intraperitoneally administered instead of biotin, and the experiment was performed in exactly the same manner as the above experiment. The results are shown in Tables 1 and 2 below.

From Tables 1 and 2 above, in the biotin-administered group, the average increase in blood ammonia level 4 hours and 6 hours after urease administration was 126.4 μg / dl and 104.8 μg / dl, respectively, whereas
The average increase in blood ammonia levels 4 hours and 6 hours after administration of urease in the control group was 257.0 μg / dl and 278.1, respectively.
It is μg / dl, and it is recognized that administration of biotin significantly suppresses an increase in blood ammonia level caused by urease loading for 4 to 6 hours after urease administration. Also, regarding the values in Tables 1 and 2 above,
When the t-test of the difference of the mean value corresponding to each was performed, p <0.02 was obtained at 4 hours and 9 hours after urease administration,
It was shown that the biotin of the present invention lowered blood ammonia level with a clear difference of p <0.01 at 6 hours.

Intraperitoneal administration of biotin in rat model of acute liver failure. In this experiment, Wistar rats (male, average weight 240
20) were used. To make a model of acute liver failure, carbon tetrachloride mixed with olive oil at a ratio of 1: 1 was intraperitoneally administered at 2 ml / kg. The biotin-administered group was intraperitoneally administered with 1 mg (1 mg / 2 ml) of biotin each 48 hours, 24 hours, and 6 hours before the administration of carbon tetrachloride, using the injection solution prepared according to Example 4 below. Blood was collected immediately before administration. After intraperitoneal administration of carbon tetrachloride, 5 animals were collected from the jugular vein at 24 hours and 48 hours, respectively. The ammonia levels of these blood samples were measured according to the method described by Okuda and Fujii (main experiment).

In the control experiment, 2 ml of physiological saline was intraperitoneally administered instead of biotin, and the experiment was performed in exactly the same manner as the above experiment. 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 due to carbon tetrachloride loading. Also, in the control group, 2 hours after the administration of carbon tetrachloride, 2 hours after the administration of rats, 2 hours after the administration.
The animals died one by one, but there was no death in the biotin-administered group. When these mortality rates were tested by the Cupran Meier method, the biotin-administered group clearly showed a life-prolonging effect.

Acute toxicity (LD ₅₀ ) when the biotin of the present invention is administered to mice and rats by various administration routes is shown in Table 5 [Fragrance Journal, 61-64 (1980), Fragrance Journal].

Examples The following examples show examples of useful pharmaceutical formulations for dosing the compounds of the present invention.

Was preformed part with biotin trituration of Example 1 tablet dosage per tablet biotin 2mg Lactose 55 Corn starch 16 Microcrystalline cellulose 30 Polyvinylpyrrolidone k-30 5 Calcium stearate 2 total amount 110mg lactose, corn starch,
Microcrystalline cellulose and the remaining lactose are mixed, and biotin is added and mixed. Next, polyvinylpyrrolidone k-30
The above aqueous solution was added as a binder to prepare granules according to a conventional method, and calcium stearate was added and mixed.
Tablets are compressed into 110 mg tablets.

Example 2 Enteric-coated tablet A coating solution having the following composition was sprayed onto the surface of the tablet obtained according to Example 1 according to a conventional method to give a dried product per tablet.
Form 10 mg of film.

Enteric coating solution 100mg of the formulation Eudragit L30D-55 45.0 mg Macrogol 6000 1.5 Talc 3.0 Purified water 50.5 total amount 100.0mg EXAMPLE 3 Injectable solution 1 tube distillation for injection distilled water qs Total 10.0ml biotin for biotin 2.0mg injection Dissolve in water, filter by a conventional method, and fill an ampoule with 10.0 ml. Then, the ampoule is closed and sterilized.

Warmed injection qs capacity may EXAMPLE 4 Injectable solution 1 tube biotin 5.0 mg Nicotinamide 0.15 Benzyl alcohol 0.2 Water for Injection qs Total 10.0ml biotin and nicotinic acid amide (solubilizer) were dissolved biotin Then, the solution is added to distilled water for injection, then dissolved by stirring, cooled to room temperature, benzyl alcohol (soothing agent) is added, and distilled water for injection is added up to a specified amount.

After that, after filtering according to the usual method, fill the ampoule with 10.0 ml,
Sterilize after sealing.

Example 5 Biotin 10 mg in 100 ml of oral syrup agent Nicotinic acid amide 300 D-sorbit 2000 Distilled water proper amount 100 ml Biotin and nicotinic acid amide were added to warm distilled water of about 3/4 volume, and D-sorbit was gradually dissolved after stirring. Add to. After dissolution, cool to room temperature and add the remaining distilled water to the specified amount.

Example 6 Biotin 2 mg lactose in one capsule 298 Total amount 300 mg Biotin was added to about 1/10 of lactose and mixed, and then the remaining lactose was added and mixed uniformly. This is filled into a gelatin hard capsule.

〔Example of use〕

The clinical results of administration of the biotin-containing preparation of the present invention mainly to patients with hyperammonemia based on cirrhosis, that is, the effect of improving blood ammonia level and hepatic coma that accompanies hyperammonemia are shown below. . Incidentally, these patients are the cases in which no improvement was observed even when the other therapeutic agents for hyperammonemia were administered.

Patient 1 Male 67 years Cirrhosis + Gastric cancer 2 Female 65 years Cirrhosis Patient 3 Male 51 years Cirrhosis Biotin was an injection solution prepared according to Example 4 (0.5 mg / m
l) was intravenously administered at 5 mg to 10 mg daily. The administration period varies depending on the condition of each patient.

Blood samples of each patient are collected once or several times a day at approximately the same time, and the blood ammonia level is measured by the microdiffusion method (patient 1, patient 2) and the method of Okuda / Fujii (patient 3), We also examined the degree of hepatic coma.

The degree of hepatic coma was determined based on Table 6 Coma degree classification table]. The classification table, which is the diagnostic criterion for this, is published in Chugai Medical Co., Ltd., page 78 and 12th Inuyama Symposium 110, 1982, Japan Medical Association Vol. 92, No. 7 (issued October 1, 1984). Has been done.

The intervals of biotin administration, blood ammonia level before and after administration, and the degree of hepatic coma of each patient are shown in Tables 7 to 9, respectively.

The results shown in Tables 7 to 9 above show that administration of the biotin of the present invention in all of the examples improves blood ammonia level and hepatic coma.

Claims (1)

[Claims] 1. A therapeutic agent for hyperammonemia, which comprises biotin as an active ingredient.