JP5312780B2 - Food / drink and pharmaceutical composition for reducing blood ammonia concentration - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a blood ammonia concentration reducing agent which is effective, small in adverse reaction and safe. <P>SOLUTION: A food or drink and a medicine composition for reducing blood ammonia concentration contains collagen peptide. In the food or drink and the medicine composition, the number-average molecular weight of collagen peptide is preferably 500 to 2,000. <P>COPYRIGHT: (C)2009,JPO&amp;INPIT

Description

  The present invention relates to a blood ammonia concentration lowering agent. In detail, this invention relates to the food-drinks and pharmaceutical composition which reduce the ammonia concentration in blood containing a collagen peptide.

  Ammonia in the body is produced during protein metabolism, and is metabolized and detoxified not only by the urea pathway but also by pathways such as the production of glutamate, glutamine, and creatine. However, when the ability to metabolize ammonia is reduced, hyperammonemia occurs, neurological symptoms are caused, and so-called hepatic encephalopathy develops. In addition, a decline in the ability to metabolize ammonia causes intellectual developmental deficiencies and liver dysfunction. Currently, as treatment for hyperammonemia, arginine is orally administered, lactulose is orally administered, sodium benzoate is injected, and the like. However, administration of arginine is an activation of the ornithine cycle in the liver, and the effect is incomplete in patients with liver damage. Lactulose is a substance that suppresses the ability to produce ammonia in enteric bacteria, and does not promote the metabolism of ammonia in the body. Sodium benzoate is intended to improve liver function and suppress bacterial growth in the intestinal tract, and does not directly promote ammonia metabolism.

Collagen peptides, on the other hand, are marketed as health foods for the purpose of beautifying the skin (see Non-Patent Document 1), and subsequently proved to be effective for osteoarthritis (see Non-Patent Document 2), increasing the commercial value. ing. These effects are explained as the effect of promoting proteoglycan synthesis in the body from the effect of supplying proline and hydroxyproline, which are extremely poor in general foods. However, it has not been known that collagen peptides reduce blood ammonia concentration.
Kazuaki Kikuchi, Yoshiharu Matahira: “Usefulness of Beverages Containing Marine Collagen Peptide for Dry and Rough Skin of Humans” FRAGRANCE JOURNAL 31, 97-102 (2003) Hashida, M., Miyatake, K., Okamoto, Y., FUJITA, K., MATUMOTO, T., MORIMATSU, F., SAKAMOTO, K, and MINAMI, S. Synergistic effects of D-glucosamine and collagen peptides on healing experimental cartilage injury. Macromol. Biosci., 3: 596-603, 2003

  An object of the present invention is to find a safe and effective substance having a blood ammonia concentration lowering effect and to use it for the treatment of hyperammonemia.

  The present inventors have intensively studied to solve the above problems, and found that the collagen peptide has a blood ammonia concentration lowering action, and completed the present invention.

Thus, the present invention provides:
(1) A food or drink or a pharmaceutical composition for oral administration which contains a collagen peptide and reduces blood ammonia concentration;
(2) The food or beverage or the pharmaceutical composition for oral administration according to (1), wherein the number average molecular weight of the collagen peptide is 500 to 2000;
(3) The food or drink or pharmaceutical composition for oral administration according to (2), wherein the number average molecular weight of the collagen peptide is 600 to 1600;
(4) The food or beverage or the pharmaceutical composition for oral administration according to (3), wherein the number average molecular weight of the collagen peptide is 700 to 1200;
(5) The food or beverage or the pharmaceutical composition for oral administration according to any one of (1) to (4), wherein the collagen peptide is obtained by hydrolysis with a proteolytic enzyme;
(6) The food or drink or the pharmaceutical composition for oral administration according to any one of (1) to (5), wherein the collagen peptide is derived from a fish scale;
(7) A non-human animal feed or a non-human animal pharmaceutical composition containing a collagen peptide, which lowers blood ammonia concentration;
(8) A food or drink or pharmaceutical composition for oral administration according to (1) to (6) or a non-human animal feed or non-human animal according to (7), characterized in that it contains a collagen peptide. A method for producing a pharmaceutical composition is provided.

  According to the present invention, by using a collagen peptide, the blood ammonia concentration can be reduced safely and effectively. Therefore, an effective therapeutic agent for hyperammonemia containing a collagen peptide and an auxiliary therapeutic agent for liver dysfunction associated with hyperammonemia are provided.

  In one aspect, the present invention provides a food or drink containing a collagen peptide that lowers blood ammonia concentration. Collagen is widely distributed in the animal kingdom, and is a protein that constitutes bone, connective tissue, and dermis. In the present invention, a collagen peptide (detailed later) is used as an active ingredient. The number average molecular weight of the collagen peptide used in the present invention is about 500 to about 2000, preferably about 600 to about 1600, more preferably about 700 to about 1200, such as about 700 to about 900, about 800 to about 1000. , About 900 to about 1100, about 1000 to about 1200. The weight average molecular weight of the collagen peptide used in the present invention is about 1500 to about 3000, preferably about 1700 to about 2700, more preferably about 1900 to about 2400, for example, about 1900 to about 2100, about 2000 to about 2200. , About 2100 to about 2300, about 2200 to about 2400. The number average molecular weight and the weight average molecular weight of the collagen peptide can be easily determined by means known to those skilled in the art such as gel filtration and high performance liquid chromatography and known methods.

  The term “collagen peptide” in the present specification refers to a peptide mixture obtained by hydrolyzing collagen by a method as described below. Furthermore, the term “collagen peptide” may refer to individual peptides in a peptide mixture obtained by hydrolyzing collagen.

  The collagen used as the raw material for the collagen peptide of the present invention may be derived from any organism, for example, derived from bones, connective tissue, dermis, etc. of terrestrial animals such as cattle, horses, and sheep, and birds. It may be derived from bones, connective tissue, fins, scales, etc. of aquatic animals such as fish. Collagen which is a raw material of the preferred collagen peptide of the present invention is derived from fish scales. Preferred fish are Thailand, tilapia and the like. Extraction of collagen from these organisms can also be performed by known methods such as hot water extraction and high pressure extraction. If necessary, the extracted collagen may be purified. As a method for purifying collagen, known protein purification methods can be applied, and examples thereof include ammonium sulfate precipitation, ion exchange chromatography, gel filtration chromatography and the like.

  Methods for preparing the collagen peptide of the present invention may be known to those skilled in the art, and general protein hydrolysis methods can be applied to collagen. Examples of the protein hydrolysis method include a method using an enzyme and hydrolysis with an acid or an alkali. Of these, hydrolysis by a proteolytic enzyme is preferable. Various proteolytic enzymes such as plants, animals, and microorganisms are known, and examples include, but are not limited to, papain, bromelain, actinidine, collagenase, trypsin, and pepsin. Since a wide variety of hydrolases are commercially available, these may be used. By appropriately selecting various conditions in the hydrolysis method, for example, the type of enzyme, the amount of enzyme, the concentration of acid or alkali, the type of coexisting buffer, the reaction temperature, the pH of the reaction, the reaction time, etc., the desired molecular weight ( For example, a collagen peptide having a number average molecular weight, a weight average molecular weight) and a desired effectiveness can be obtained. Moreover, if the peptide is isolated from the collagen hydrolyzate and its amino acid sequence is determined, the collagen peptide of the present invention can be produced also by chemical synthesis or genetic engineering techniques.

  As described above, when the blood ammonia concentration rises, neurological symptoms are caused, and not only so-called hepatic encephalopathy develops, but also intellectual failure or liver dysfunction occurs. In order to treat or ameliorate such symptoms, the effect of lowering the blood ammonia concentration of collagen peptides can be used.

  The food and drink of the present invention may use the collagen peptide as it is by a known method, or a drinkable liquid (juice, drink etc.), semi-solid (paste etc.), solid (eg granule, powder (lyophilized powder etc.) )) And the like. Moreover, the food / beverage of this invention can be manufactured also by adding a collagen peptide to food / beverage. Furthermore, the food and drink of the present invention may be in the form of sprinkles or seasonings. The food and drink of the present invention can be a health food, a nutritional functional food, or a food for specified health use (including so-called “Tokuho”).

  A preferred form of the food or drink of the present invention is a supplement containing a collagen peptide. The supplement may have any shape as long as it can be taken orally. For example, it may be a general food, tablet, capsule, granule, powder (eg, freeze-dried powder, etc.) , Suspensions, drinks, elixirs, chewable forms, jelly forms, and the like. These supplements can be produced according to processes used in the food and pharmaceutical fields. For example, when manufacturing a tablet-shaped supplement, general processes such as mixing, drying, and tableting used in the pharmaceutical field can be used. Further, for example, in the case of a capsule form, a general process such as mixing and encapsulation can be used. Soft capsules and hard capsules can also be appropriately selected according to the purpose. To produce a liquid supplement, the extract can be dissolved or suspended in a less toxic medium such as ethanol. In order to produce powder and granule supplements, processes such as usual mixing, drying, grinding and sieving can also be used. When a carrier or excipient is used in the production of supplements, the type and amount thereof can be selected according to the practice of the pharmaceutical field. Examples of solid carriers or excipients include talc, cellulose powder, sugar, starch, and wheat flour. Examples of the liquid carrier include water, ethanol, glycerin, and edible fats and oils. Mixing these carriers and excipients with collagen peptides can also be performed by known methods.

  In another aspect, the present invention provides a pharmaceutical composition for oral administration containing a collagen peptide for reducing blood ammonia concentration. The active ingredient in the pharmaceutical composition of the present invention is the aforementioned collagen peptide. In addition to the collagen peptide, the pharmaceutical composition of the present invention may contain one or more therapeutic agents for ammoniacal disease such as oral administration of arginine, lactulose, sodium benzoate and the like.

  The pharmaceutical composition of the present invention can be formulated into various oral dosage forms. Examples of oral dosage forms include tablets, capsules, granules, powders, and drinks. These dosage forms can be produced by a method known in the pharmaceutical field as described in the supplement, for example.

  The daily intake or dose of the collagen peptide in the food or drink or pharmaceutical composition of the present invention is generally about 5 g to about 10 g per day for an adult (70 kg body weight), It is appropriate to take about 6 g to about 8 g. These amounts can be appropriately changed depending on the severity of the subject's symptoms, the weight and health of the subject, the amount and type of meal, the type and degree of hyperammonemia treatment being received, and the like.

  In addition, since the active ingredient of foods and drinks and pharmaceutical compositions containing the collagen peptide of the present invention is a collagen peptide, there are no problems of safety and side effects, and not only the decrease in blood ammonia concentration but also the overall health condition of the subject. The improvement effect can be expected. For example, by the food and drink and the pharmaceutical composition of the present invention, secondary effects such as improvement of liver function and reduction of arthritis can be expected.

  Foods and beverages and pharmaceutical compositions containing the collagen peptide of the present invention are used in combination with known hyperammonemia treatments or drugs such as oral administration of arginine, oral administration of lactulose, injection of sodium benzoate, etc. It can also be used as an auxiliary food or drink or an auxiliary therapeutic agent.

  In a further aspect, the present invention provides an animal feed containing a collagen peptide that reduces blood ammonia concentration. Hyperammonemia and accompanying liver dysfunction can be treated or improved by lowering the blood ammonia concentration of animals by ingesting the feed of the present invention to animals (except humans).

  In a further aspect, the present invention provides a pharmaceutical composition for oral administration for animals that contains a collagen peptide and reduces blood ammonia concentration. Treating or improving hyperammonemia and the accompanying liver dysfunction by ingesting the animal (except human) pharmaceutical composition for oral administration of the present invention to lower the blood ammonia concentration of the animal be able to. These animal feeds and animal pharmaceutical compositions for oral administration can be used in combination with other hyperammonemia treatments and therapeutic agents.

  Collagen peptide intake or dose in the animal feed or animal oral pharmaceutical composition of the present invention varies depending on factors such as animal type, animal weight, age, health condition, treatment being received, etc. Can be done. Such intake or dose can be easily determined by those skilled in the art.

  In a further aspect, the present invention provides a method for treating hyperammonemia, comprising administering a collagen peptide to a subject. In this treatment, other hyperammonemia treatments and therapeutic agents may be used in combination.

  The invention further provides, in another aspect, the use of a collagen peptide in the manufacture of a medicament for the treatment of hyperammonemia, as well as the use of a collagen peptide for the treatment of hyperammonemia. In this use, other therapeutic agents for hyperammonemia may be used in combination.

  Furthermore, the present invention provides a method for producing a food, beverage, pharmaceutical composition, animal feed (not for humans) or animal drug (not for humans) that lowers the blood ammonia concentration, characterized in that it contains a collagen peptide. Also provide.

  EXAMPLES Hereinafter, the present invention will be described more specifically and in detail with reference to examples, but the present invention is not limited to those described in the examples.

Example 1: Production of collagen peptide (1)
Add approximately 300 liters of water and 30 kg of hydrochloric acid (35%) to tilapia scales (100 kg), react for 3 hours, wash the scales with water, add about 500 liters of water and sodium hydroxide to adjust the pH, and then wash with water and remove. An ash scale was obtained. Add about 300 liters of water to demineralized scale, boil for 2 hours and 30 minutes, drop the temperature to 60 ° C, add 120 g of papain (manufactured by Asahi Food and Healthcare) and inactivate after 1 hour and 40 minutes of reaction. It was. The solution thus obtained was placed in a pressure vessel and treated at 120 ° C. for 4 hours. After the treatment, the temperature was lowered to 80 ° C. and filtered through a membrane filter to obtain a collagen peptide.

The molecular weight of the collagen peptide was measured using high performance liquid chromatography. The column was Asahipak GS-520HQ + GS320HQ + GS220HQ, the eluent was 10 mM Tris-HCl, 0.15 M NaCl, 5 mM CaCl ₂ , the detector was a refractive index detector, and the column temperature was 40 ° C. A pullulan with a known molecular weight was used as a standard sample. The measurement results of the molecular weight of the collagen peptide are shown in FIG. The number average molecular weight (Mn) was 800 and the weight average molecular weight (Mw) was 2200.

Example 2: Production of collagen peptide (2)
Add approximately 300 liters of water and 30 kg of hydrochloric acid (35%) to tilapia scales (100 kg), react for 3 hours, wash the scales with water, add about 500 liters of water and sodium hydroxide to adjust the pH, and then wash with water and remove. An ash scale was obtained. About 300 l of water was added to the demineralized scale, boiled for 2 hours and 30 minutes, the temperature was lowered to 60 ° C., 60 g of papain and 10 g of Amano protease M were added, and the reaction was inactivated after 1 hour and 40 minutes. In addition, 300 l of water was added to the residue (scale) of the resulting solution, boiled for 3 hours, the temperature was lowered to 60 ° C., and 60 g of papain (purified papain F manufactured by Asahi Food and Healthcare) and 10 g of Amano protease M were added. After reacting for 1 hour and 40 minutes, the mixture was deactivated, mixed with the initially obtained solution, adjusted to a temperature of 80 ° C. and then filtered through a membrane filter to obtain a collagen peptide. By this production method, a collagen peptide with little bitterness was obtained.

Example 3 Changes in Blood Amino Acids by Oral Administration of Collagen Peptides to Rabbits Rabbits (Japanese white species, female, body weight 2 kg, 12 weeks old 3 mice) were treated with the collagen peptides obtained in Example 1 (1 g / animal). ) Was orally administered, blood was collected before administration, 2 hours after administration and 24 hours after administration, and the amino acid concentration in the blood was measured. Qualitative and quantitative determination of amino acids was performed using an amino acid analyzer (JLC-500 / V2 fully automatic amino acid analyzer (manufactured by JEOL Ltd.)). The amino acids whose blood concentrations have increased after administration of the collagen peptide are shown in FIGS. The blood glutamine concentration increased 2 times before administration at 2 hours and 2.2 times at 24 hours. The blood aspartic acid concentration increased by 25% at 2 hours after administration and 50% at 24 hours. The blood arginine concentration increased by 25% at 2 hours after administration and 50% at 24 hours, and the blood citrulline concentration increased by 25% at 2 hours after administration, and returned to the value before administration at 24 hours. The blood asparagine concentration increased 15-fold at 2 hours and 28-fold at 24 hours. On the other hand, glutamic acid decreased to 20% before administration at 2 hours after administration, and decreased to 20% or less before administration at 24 hours (not shown). Blood alanine and β-alanine concentrations decreased to about 30% of pre-dose values at 2 hours after administration, and these values were maintained at 24 hours (not shown). Other amino acids whose blood concentration decreased after administration of collagen peptide are threonine, serine, glycine, valine, methionine, cystathionine, isoleucine, leucine, phenylalanine, 1M-histidine, histidine, lysine, tryptophan, hydroxyproline, proline, tyrosine and taurine (Graph shown). Of these, the decrease in blood concentration of taurine was similar to that of alanine and β-alanine, but the blood concentrations of other amino acids were not decreased compared to alanine and β-alanine.

  Arginine and citrulline are basic substances of the ornithine cycle related to ammonia metabolism, and it is thought that the increase in the blood concentration of these substances activates the ornithine cycle in the living body, particularly the urea metabolism in the liver. It is done. Further, since glutamine is synthesized from glutamic acid and urea, an increase in glutamine is considered to mean an increase in urea metabolism. In addition, glutamic acid and aspartic acid combine with blood ammonia to form glutamine and asparagine, respectively, neutralizing the toxicity of ammonia. From the above, it is considered that oral administration of collagen peptide promotes ammonia metabolism in the body and lowers the blood ammonia concentration.

  In addition, glutamine exerts a protective effect on hepatocytes, and asparagine promotes the production of oxaloacetate and promotes the citric acid cycle in the liver. Alanine plays an important role in sugar synthesis in the acetic acid cycle. β-alanine is abundant in muscles. Decreased blood levels of alanine and β-alanine suggest that these substances have been taken up by the liver and muscle. From these, oral administration of the collagen peptide is expected to protect the liver and muscles, and systemic effects can also be expected.

  The amino acids that are mostly contained in collagen peptides are glycine (33.6%), alanine (12.6%), and proline (11%). Surprisingly, these amino acid concentrations in blood are 2 after oral administration of collagen peptide. Decreased in time. In contrast, amino acids whose blood concentrations were increased by oral administration of collagen peptides were glutamine, citrulline, arginine, aspartic acid, and asparagine. An increase in the blood concentration of these amino acids is considered to be a bioactive effect by oral administration of collagen peptide. Moreover, all of these amino acids are directly related to the metabolism of ammonia in the body, and it was strongly suggested that administration of a collagen peptide alone promotes ammonia metabolism in vivo. Based on these points, the following experiment was conducted.

Example 4: Blood ammonia reduction effect of collagen peptide on congenital portal circulatory shunt dog (PSS dog) PSS causes metabolic abnormalities because amino acids absorbed from the intestine bypass the liver, especially high ammonia Blood pressure causes neurological symptoms and causes so-called hepatic encephalopathy. In this experiment, the collagen peptide obtained in Example 1 was given to PSS dogs (beagle, female, 5.5 kg, 8 months old).

Experimental Procedure PSS dogs were fed a normal diet (Kaynes Dogmeal, Cainz Co., Ltd., crude protein content of 21.0% or more) for 1 month, and blood was collected irregularly throughout the period. Thereafter, a low protein diet (liver support, Waltham, crude protein content of 14.0% or more) was given for half a year, and blood was collected irregularly throughout the period. Next, collagen peptide (1 g / meal) was mixed with the low protein diet and given for 2 weeks, and blood was collected three times a week during the period. All meals were given once a day. All blood samples were taken on an empty stomach. The ammonia concentration in the blood sample was measured by the BPB indicator method (Fuji Dry Chemistry System 3500i).

Experimental Results As can be seen from FIG. 4, the blood ammonia concentrations were 290 ± 30 μg / ml at the time of normal diet administration, 220 ± 5 μg / ml at the time of low protein diet administration, and at the time of administration of a diet in which collagen peptide was mixed with the low protein diet The concentration was 135 ± 20 μg / ml, and administration of the collagen peptide showed a significant blood ammonia concentration lowering effect. Moreover, the effect lasted during the administration period, and was effective for 24 hours with a single administration. In addition, no side effects were observed in dogs during the period when the collagen peptide mixed diet was given.

  As can be seen from the results of Examples 2 and 3, it was found that the collagen peptide is effective in treating and improving hyperammonemia by reducing blood ammonia concentration. In addition, it was found that the collagen peptide exerts an effect of lowering blood ammonia concentration even when used in combination with other therapeutic agents for hyperammonemia or dietary therapy therefor. Moreover, the collagen peptide has no confirmed side effects on animals. Collagen peptides are also suggested to be effective for the treatment and improvement of liver dysfunction associated with hyperammonemia. For example, it is considered that collagen peptides can be used as an adjuvant treatment effective for liver dysfunction associated with hyperammonemia. It is done.

  The present invention can be used in the field of foods and animal feeds, particularly health foods. Furthermore, the present invention can also be used in the field of human and animal medicine.

The molecular weight measurement result by the gel filtration chromatography of the collagen peptide obtained in Example 1 is shown. The horizontal axis is the number average molecular weight, and the vertical axis is the refractive intensity. The time course of amino acids whose blood concentration increased after oral administration of collagen peptides before administration (pre), 2 hours after administration (2 hr) and 24 hours after administration (24 hr) is shown. The horizontal axis represents time (hour), and the vertical axis represents the blood concentration at each time relative to the blood concentration before administration (100). The time course of amino acids whose blood concentration increased after oral administration of collagen peptides before administration (pre), 2 hours after administration (2 hr) and 24 hours after administration (24 hr) is shown. The horizontal axis represents time (hour), and the vertical axis represents the blood concentration at each time relative to the blood concentration before administration (100). Shows fasting blood ammonia concentration when a PSS dog is fed a normal diet, a low protein diet (labeled liver support), and a diet mixed with a collagen peptide in a low protein diet (labeled liver support-SCCP) (each blood sample) Sample mean and standard deviation).

Claims (8)

  A pharmaceutical composition for oral administration containing a collagen peptide for reducing blood ammonia concentration, which is used in combination with a low protein diet.   The pharmaceutical composition for oral administration according to claim 1, wherein the number average molecular weight of the collagen peptide is 500 to 2,000.   The pharmaceutical composition for oral administration according to claim 2, wherein the number average molecular weight of the collagen peptide is 600-1600.   The pharmaceutical composition for oral administration according to claim 3, wherein the number average molecular weight of the collagen peptide is 700 to 1200.   The pharmaceutical composition for oral administration according to any one of claims 1 to 4, wherein the collagen peptide is obtained by hydrolysis with a proteolytic enzyme.   The pharmaceutical composition for oral administration according to any one of claims 1 to 5, wherein the collagen peptide is derived from a fish scale. Containing collagen peptide, a veterinary pharmaceutical composition other than humans for lowering the blood ammonia concentrations, except Der Ruhi preparative those to be used in combination with a low protein diet veterinary pharmaceutical composition. Characterized by mixing collagen peptide The method of claims 1 to pharmaceutical compositions for oral administration according to any one of 6 or claim 7 veterinary composition other than human description.

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