Patent Application
20180185313
The present invention relates to a composition containing an amino acid and a cyclic dipeptide. More specifically, the present invention relates to a composition containing an amino acid or a salt thereof and a cyclic dipeptide or a salt thereof, wherein the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof falls within a specific range, use of the composition for inhibiting angiotensin-converting enzyme, and a method for inhibiting angiotensin-converting enzyme.
It is widely known that lifestyle-related diseases such as hypertension or hyperlipidemia are caused by change in eating habits, a lack of exercise, stress, smoking, genetic factors, etc. Particularly, hypertension is characterized in that its symptoms progress without being subjectively noticed, and exhibits a very high morbidity, also including hypertension suspects. Also, hypertension is known to lead to serious diseases such as stroke (cerebral infarction, cerebral hemorrhage, subarachnoid hemorrhage, etc.), heart-disease (angina pectoris, myocardial infarction, cardiomegaly, heart failure, etc.), and renal failure (hypertensive nephropathy, etc.). Therefore, there is a social demand for prevention and early amelioration of hypertension.
As one of the solutions of hypertension, it is known that inhibition of angiotensin-converting enzyme (also called angiotensin I-converting enzyme) in the renin-angiotensin system known as a mechanism to elevate blood pressure is effective. The angiotensin-converting enzyme is mainly present in the lung, vascular endothelial cells, and the renal proximal tubule and has the property of converting the substrate angiotensin I to angiotensin II serving as a pressor hormone. The angiotensin-converting enzyme also has an action of decomposing bradykinin having a vasodilating action, in addition to the action of yielding angiotensin II. Therefore, angiotensin-converting enzyme inhibitors can also exert a blood pressure-lowering action through the mechanism of a bradykinin decomposition--inhibiting action. Thus, the angiotensin-converting enzyme inhibitors are clinically effective for treatment or prevention of hypertension.
Against this backdrop, amino acids have been found to have a blood pressure-lowering activity or an angiotensin-converting enzyme-inhibiting activity. For example, NPL 1 discloses that decrease in mean arterial pressure is observed by the oral administration of histidine to spontaneously hypertensive rats. However, it is difficult to achieve a sufficient blood pressure-lowering activity by the angiotensin-converting enzyme-inhibiting activity brought about by an amino acid alone.
NPL 2 discloses that a Maillard reaction product obtained from the combination of a specific amino acid and glucose or xylose has an angiotensin-converting enzyme-inhibiting activity. However, a Maillard reaction product, such as acrylamide, formed through the reaction of asparagine with the reducing sugar is suspected of having biological toxicity.
It has been further reported that a composition containing a linear dipeptide or tripeptide including proline, and a free amino acid exhibits an angiotensin-converting enzyme-inhibiting activity (PTL 1). However, the linear dipeptide exposes an amino group and a carboxyl group, which are polar groups, at its ends and is therefore disadvantageous that it has low lipid solubility and is less bioabsorbable.
PTL 1: Japanese Patent Laid-Open No. 2011-102327
NPL 1: Clin Exp Pharmacol Physiol., 37(1), 62-68 (2010)
NPL 2: J. Brew. Soc. Japan., 96(3), 199-206(2001)
An object of the present invention is, for example, to provide a composition which has high bioabsorbability and excellent biological safety and can enhance an angiotensin-converting enzyme-inhibiting activity ascribable to an amino acid.
The present inventors have conducted diligent studies on the object and consequently focused on utilization of cyclic dipeptides or salts thereof. Cyclic dipeptides are dipeptides having a cyclic structure generated through dehydration condensation of an amino group and a carboxyl group each present in the end of a linear dipeptide. In recent years, their various physiological activities have received attention. The present inventors have found that the ratio of the total amount of an amino acid or a salt thereof to the total amount of a cyclic dipeptide or a salt thereof in a composition is set to within a specific range to thereby enhance an angiotensin-converting enzyme-inhibiting activity brought about by the amino acid alone, and arrived at the completion of the present invention.
Specifically, the present invention relates to, but is not limited to, the following aspects:
The present invention can provide a composition which enhances an angiotensin-converting enzyme-inhibiting activity brought about by an amino acid alone. A blood pressure elevation-suppressing action or a blood pressure-lowering action is obtained through inhibition of angiotensin-converting enzyme. Hence, a hypertension-preventing or -ameliorating effect or a preventing or ameliorating effect on various diseases caused by hypertension, such as stroke, heart disease, and renal failure, is exerted by ingestion of the composition according to the present invention.
The amino acid contained in the composition according to the present invention is a naturally derived component, and the cyclic dipeptide is also contained in a heat-treated product of a plant-derived peptide, or the like. Thus, these components are highly safe. Therefore, the composition according to the present invention is considered to have a very low risk of manifesting serious side effects. Cyclic dipeptides have higher lipid solubility than that of linear dipeptides and are not mere dipeptides composed of only a peptide linkage. Thus, the cyclic dipeptide or salt thereof contained in the composition according to the present invention has resistance to the actions of various peptidases secreted into the gastrointestinal tract and can also be expected to have high gastrointestinal absorbability.
1. Amino Acid or Salt Thereof
Throughout the present specification, “amino acid” means a compound having both of an amino group and a carboxyl group as functional groups. Amino acids in the present specification may be essential amino acids or non-essential amino acids or may be amino acids formed through modification after protein synthesis. Throughout the present specification, “amino acid or salt thereof” is occasionally referred to as “amino acid” in a collective and simple manner.
The amino acid contained in the composition according to the present invention is one or two or more members selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, arginine, lysine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, and hydroxyproline. More preferably, three or more members selected from these amino acids are used as an active ingredient. Among the amino acids described above or salts thereof, one or two or more members selected from the group consisting of leucine, threonine, arginine, tyrosine, and tryptophan are preferred, and three or more members selected from these amino acids or salts thereof are more preferably contained.
Throughout the present specification, “salt of an amino acid” refers to any pharmacologically acceptable salt (including inorganic salts and organic salts) of the amino acid. Examples thereof include, but not limited to, sodium salts, potassium salts, calcium salts, magnesium salts, ammonium salts, hydrochlorides, sulfates, nitrates, phosphates, and organic acid salts (acetates, citrates, maleates, malates, oxalates, lactates, succinates, fumarates, propionates, formates, benzoates, picrates, benzenesulfonates, trifluoroacetates, etc.) of the amino acid.
Amino acids or salts thereof used in the present invention may be self-made by using a known method, or commercially available products may be used. For example, an amino acid or a salt, thereof can be produced by chemical synthesis, extraction from a natural product, an enzymatic method, or microbial fermentation. For example, a treatment product obtained by subjecting a plant-derived peptide, for example, a soybean peptide, a tea peptide, or a malt peptide, to enzyme treatment or the like can be suitably used. Alternatively, an amino acid or a salt thereof contained in an existing composition may be used as the amino acid or salt thereof according to the present invention. If the content of a specific amino acid or a salt thereof in an existing composition is less than the desired content, the shortfall of the specific amino acid or salt thereof may be appropriately supplied by using a commercially available product, a synthesized product, or the like.
In the present invention, the amino acid or salt thereof may be obtained from a plant-derived peptide or may be obtained from a heat-treated product of a plant-derived peptide.
2. Cyclic Dipeptide or Salt Thereof
Throughout the present specification, “cyclic dipeptide” refers to a cyclic dipeptide including amino acids as constituent units and having a diketopiperazine structure generated through dehydration condensation of an amino group of an amino acid and a carboxyl group of another amino acid. Throughout the present specification, “cyclic dipeptide or salt thereof” is occasionally referred to as “cyclic dipeptide” in a collective and simple manner. Throughout the present specification, the order of amino acids in a cyclic dipeptide may be inverse as long as the constitution is unchanged, and, for example, Cyclo(Glu-His) and Cyclo(His-Glu) represent an identical cyclic dipeptide.
In a cyclic dipeptide, end portions of two amino acids bond together via an amide linkage (i.e., a cyclic dipeptide has a cyclic structure formed by the amino linkage between an amino end and a carboxy end), and thus cyclic dipeptides are characterized by higher lipid solubility than that of linear dipeptides (particularly, linear dipeptides consisting of the same type of amino acid composition), in which a carboxyl group and an amino group as polar groups are exposed at molecular end portions. Accordingly, cyclic dipeptides are superior in gastrointestinal permeability and membrane permeability to linear dipeptides. This is also evident from previously reported results of compound permeation tests using everted rat gut sacs (J. Pharmacol, 1998, 50:167-172). Also, cyclic dipeptides are considered to also have higher resistance to various peptidases because of their specific structures.
The cyclic dipeptide or salt thereof included in the present invention is one or two or more members selected from the group consisting of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)]. More preferably, all of these cyclic dipeptides or salts thereof are contained.
Throughout the present specification, “salt of a cyclic dipeptide” refers to any pharmacologically acceptable salt (including inorganic salts and organic salts) of the cyclic dipeptide. Examples thereof include, but not limited to, sodium salts, potassium salts, calcium salts, magnesium salts, ammonium salts, hydrochlorides, sulfates, nitrates, phosphates, and organic acid salts (acetates, citrates, maleates, malates, oxalates, lactates, succinates, fumarates, propionates, formates, benzoates, picrates, benzenesulfonates, trifluoroacetates, etc.) of the cyclic dipeptide. Those skilled in the art can easily prepare a salt of a cyclic dipeptide by using any method known in the art.
Cyclic dipeptides used in the present invention can be prepared in accordance with a method known in the art. For example, a cyclic dipeptide may be produced by using chemical synthesis, an enzymatic method, or microbial fermentation, or may be synthesized by using dehydration and cyclization of a linear peptide, or may be prepared in accordance with a method described in Japanese Patent Laid-Open No. 2003-252896, Journal of Peptide Science, 10, 737-737, 2004, International Publication No. WO 2014/200000, etc. For example, a heat-treated product of a plant-derived peptide rich in cyclic dipeptides can be obtained by subjecting a plant-derived peptide obtained through enzyme treatment or heat treatment of a raw material including a plant-derived protein to further high-temperature heat treatment. As desired, the heat-treated product of the plant-derived peptide obtained may be subjected to a process such as filtration, centrifugation, concentration, ultrafiltration, freeze-drying, and pulverization. If the content of a specific cyclic dipeptide in a heat-treated product of a plant-derived peptide is less than the desired content, the shortage of the specific cyclic dipeptide may be appropriately supplied by using other plant-derived peptides, a commercially available product, or a synthesized product.
In the present invention, the cyclic dipeptide or salt thereof may be obtained from a plant-derived peptide or may be obtained from a heat-treated product of a plant-derived peptide.
3. Plant-Derived Peptide
Throughout the present specification, “plant-derived peptide” is not particularly limited, and, for example, a soybean peptide, a tea peptide, or a malt peptide can be used. A plant-derived peptide may be prepared from a plant-derived protein or a raw material including the protein, or a commercially available product may be used.
3-1. Soybean Peptide
Throughout the present specification, “soybean peptide” refers to a low-molecular peptide obtained by lowering the molecular weight of a protein through enzyme treatment or heat treatment of a soybean protein. The soybean (scientific name: Glycine max) as the raw material may be any species and may be produced in any area. Soybean in a stage of processing, such as crushed soybean, can also be used.
3-2. Tea Peptide
Throughout the present specification, “tea peptide” refers to a tea-derived low-molecular peptide obtained by lowering the molecular weight of a protein through enzyme treatment or heat treatment of a tea (including tea leaves and used tea leaves) extract. The tea leaves of the extraction raw material are drinkable parts by extraction of a tea plant (scientific name: Camellia sinensis), such as leaves and stems of tea leaves. In addition, the tea leaves may be in any form, such as a macrophyll or powder form. The harvest time of tea leaves may be any time and is appropriately selected to obtain a desired flavor.
3-3. Malt Peptide
Through the present specification, “malt peptide” refers to a malt-derived low-molecular peptide obtained by lowering the molecular weight of a protein through enzyme treatment or heat treatment of an extract obtained from malt or a crushed product thereof. The malt peptide as the raw material may be any species and may be produced in any area. In particular, barley malt, which is germinated seeds of barley, is suitably used. Throughout the present specification, barley malt is also referred to as “malt” in a simple manner.
4. Composition
One aspect of the present invention is a composition containing an amino acid or a salt thereof and a cyclic dipeptide or a salt thereof, wherein the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof falls within a specific range.
For the present invention, it is important that the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition should fall within a specific range. Specifically, the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition (the total amount of the amino acid or salt thereof in the composition:the total amount of the cyclic dipeptide or salt thereof in the composition) is 1:0.02 or more, preferably 1:0.05 or more, more preferably 1:0.1 or more, and 1:20 or less, preferably 1:10 or less, 1:5 or less, 1:2 or less, 1:1.5 or less, or 1:0.5 or less. Typically, the range of the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition is 1:0. 02 to 1:20, preferably 1:0.02 to 1:5, 1:0.02 to 1:2, more preferably 1:0.02 to 1:1.5.
The amino acid or salt thereof contained in the present invention is one or two or more members selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, arginine, lysine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, and hydroxyproline. Preferably, three or more members selected from these amino acids or salts thereof are contained.
Among the amino acids described above or salts thereof, one or two or more members selected from the group consisting of leucine, threonine, arginine, tyrosine, and tryptophan are preferred, and three or more members selected from these amino acids or salts thereof are more preferably contained.
The ratio of the total amount of the leucine or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition (the total amount of the leucine or salt thereof in the composition:the total amount of the cyclic dipeptide or salt thereof in the composition) is preferably 1:0.02 to 1:1.5 or 1:0.05 to 1:1.5, more preferably 1:0.10 to 1:1.5, still more preferably 1:0.10 to 1:0.50.
The ratio of the total amount of the threonine or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition (the total amount of the threonine or salt thereof in the composition:the total amount of the cyclic dipeptide or salt thereof in the composition) is preferably 1:0.02 to 1:1.5, more preferably 1:0.10 to 1:1.5, still more preferably 1:0.10 to 1:0.50.
The ratio of the total amount of the arginine or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition (the total amount of the arginine or salt thereof in the composition:the total amount of the cyclic dipeptide or salt thereof in the composition) is preferably 1:0.02 to 1:1.5, more preferably 1:0.10 to 1:1.5, still more preferably 1:0.10 to 1:0.50.
The ratio of the total amount of the tyrosine or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition (the total amount of the tyrosine or salt thereof in the composition:the total amount of the cyclic dipeptide or salt thereof in the composition) is preferably 1:0.10 to 1:1.5, more preferably 1:0.10 to 1:0.50.
The ratio of the total amount of the tryptophan or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition (the total amount of the tryptophan or salt thereof in the composition:the total amount of the cyclic dipeptide or salt thereof in the composition) is preferably 1:0.02 to 1:1.5 or 1:0.05 to 1:1.5, more preferably 1:0.10 to 1:1.5, still more preferably 1:0.10 to 1:0.50.
The cyclic dipeptide or salt thereof included in the present invention is one or two or more members selected from the group consisting of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)]. Preferably, three or more selected from these cyclic dipeptides or salts thereof are contained.
The total amount of the amino acid or salt thereof in the composition according to the present invention is not particularly limited as long as the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof in the composition satisfies the range described above, in view of form of administration, an administration method, etc. For example, in the case of using a plant-derived peptide such as a soybean peptide, a tea peptide, or a malt peptide as a raw material, the total amount of the amino acid or salt thereof in the composition according to the present invention is, in terms of a content per Brix, 1.0 ppm/Brix or more, preferably 10 ppm/Brix or more, more preferably 1.0×102 ppm/Brix or more, and 1.0×104 ppm/Brix or less, preferably 5.0×103 ppm/Brix or less, more preferably 1.0×103 ppm/Brix or less, and is typically, 1.0 ppm/Brix to 1.0×104 ppm/Brix, preferably 10 ppm/Brix to 5.0×103 ppm/Brix, more preferably 1.0×102 ppm/Brix to 1.0×103 ppm/Brix. In the case of using a plant-derived peptide such as a soybean peptide, a tea peptide, or a malt peptide as a raw material, the individual content of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, arginine, lysine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, hydroxyproline, or each corresponding salt in the composition according to the present invention is, in terms of a content per Brix, 0.1 ppm/Brix or more, preferably 1.0 ppm/Brix or more, more preferably 5.0 ppm/Brix or more, and 1.0×103 ppm/Brix or less, preferably 5.0×102 ppm/Brix or less, more preferably 1.0×102 ppm/Brix or less, and is typically 0.1 ppm/Brix to 1.0×103 ppm/Brix, preferably 1.0 ppm/Brix to 5.0×102 ppm/Brix, more preferably 5.0 ppm/Brix to 1.0×102 ppm/Brix.
In the present invention, the content of an amino acid or a salt thereof can be indicated by an amount per Brix as described above. Throughout the present specification, “amount per Brix” means an amount defined by a value corresponding to mass percentage of a sucrose solution (an aqueous solution containing only sucrose as a solute) of 20° C. “ppm” used in the present specification refers to ppm in weight/volume (w/v), unless otherwise specified, and 1.0 ppm/Brix can be converted into 0.1 mg/mL or 0.01% by weight when the specific gravity of the solvent is 1.
The content of the amino acid or salt thereof added to the composition according to the present invention can be measured in accordance with a known method. The content can be measured by using, for example, LC-MS/MS or a saccharimeter.
In the case of using a synthesized product or a purified product as the amino acid or salt thereof, the total content of amino acids or salts thereof in the composition according to the present invention is not particularly limited and is, for example, in terms of a content per Brix, 1.0 ppm/Brix or more, preferably 10 ppm/Brix or more, more preferably 10×102 ppm/Brix or more, and 1.0×104 ppm/Brix or less, preferably 5.0×103 ppm/Brix or less, more preferably 1.0×103 ppm/Brix or less, and is typically 1.0 ppm/Brix to 1.0×104 ppm/Brix, preferably 10 ppm/Brix to 5.0×103 ppm/Brix, more preferably 1.0×102 ppm/Brix to 1.0×103 ppm/Brix. In the case of using a synthesized product or a purified product as the amino acid or salt thereof, the individual content of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, arginine, lysine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, hydroxyproline, or each corresponding salt in the composition according to the present invention is, in terms of a content per Brix, 0.1 ppm/Brix or more, preferably 1.0 ppm/Brix or more, more preferably 5.0 ppm/Brix or more, and 1.0×103 ppm/Brix or less, preferably 5.0×102 ppm/Brix or less, more preferably 1.0×102 ppm/Brix or less, and is typically 0.1 ppm/Brix to 1.0×103 ppm/Brix, preferably 1.0 ppm/Brix to 5.0×102 ppm/Brix, more preferably 5.0 ppm/Brix to 1.0×102ppm/Brix.
The total amount of the cyclic dipeptide or salt thereof in the composition according to the present invention is not particularly limited as long as the ratio of the total amount of the amino acid or salt thereof in the composition satisfies the range described above, in view of form of administration, an administration method, etc. For example, in the case of using a plant-derived peptide as a raw material, the total amount of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)], and their corresponding salts in the composition according to the present invention is, in terms of a content per Brix, 1.0 ppm/Brix or more, preferably 5.0 ppm/Brix or more, more preferably 10 ppm/Brix or more, and 1.0×104 ppm/Brix or less, preferably 5.0×103 ppm/Brix or less, more preferably 1.0×103 ppm/Brix or less, and is typically 1.0 ppm/Brix to 1.0×104 ppm/Brix, preferably 5.0 ppm/Brix to 1.0×104 ppm/Brix, more preferably 5.0 ppm/Brix to 5.0×103 ppm/Brix, still more preferably 10 ppm/Brix to 1.0×103 ppm/Brix. In the case of using a plant-derived peptide as a raw material, the individual content of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], or cyclo-isoleucyl-serine [Cyclo(Ile-Ser)], or its corresponding salt in the composition according to the present invention is, in terms of a content per Brix, 0.2 ppm/Brix or more, preferably 1.0 ppm/Brix or more, more preferably 2.0 ppm/Brix or more, and 1.0×104 ppm/Brix or less, preferably 5.0×103 ppm/Brix or less, more preferably 1.0×103 ppm/Brix or less, and is typically 0.2 ppm/Brix to 1.0×104 ppm/Brix, preferably 1.0 ppm/Brix to 5.0×103ppm/Brix, more preferably 2.0 ppm/Brix to 1.0×103 ppm/Brix.
In the case of using a synthesized product or a purified product as the cyclic dipeptide or salt thereof, the total amount of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)], and their corresponding salts in the composition according to the present invention is not particularly limited and is, for example, in terms of a content per Brix, 1.0 ppm/Brix or more, preferably 5.0 ppm/Brix or more, more preferably 10 ppm/Brix or more, and 1.0×104 ppm/Brix or less, preferably 5.0×103 ppm/Brix or less, more preferably 1.0×103 ppm/Brix or less, and is typically 1.0 ppm/Brix to 1.0×104 ppm/Brix, preferably 5.0 ppm/Brix to 1.0×104 ppm/Brix, more preferably 5.0 ppm/Brix to 5.0×103 ppm/Brix, still more preferably 10 ppm/Brix to 1.0×103 ppm/Brix. In the case of using a synthesized product or a purified product as the cyclic dipeptide or salt thereof, the individual content of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], or cyclo-isoleucyl-serine [Cyclo(Ile-Ser)], or each corresponding salt in the composition according to the present invention is, in terms of a content per Brix, 0.2 ppm/Brix or more, preferably 1.0 ppm/Brix or more, more preferably 2.0 ppm/Brix or more, and 1.0×104 ppm/Brix or less, preferably 5.0×103 ppm/Brix or less, more preferably 1.0×103 ppm/Brix or less, and is typically 0.2 ppm/Brix to 1.0×104 ppm/Brix, preferably 1.0 ppm/Brix to 5.0×103 ppm/Brix, more preferably 2.0 ppm/Brix to 1.0×103 ppm/Brix.
In the present invention, the content of a cyclic dipeptide or a salt thereof can be indicated by an amount per Brix as described above. “ppm” used in the present specification refers to ppm in weight/volume (w/v), unless otherwise specified, and 1.0 ppm/Brix can be converted into 0.1 mg/mL or 0.01% by weight when the specific gravity of the solvent is 1.
The content of the cyclic dipeptide or salt thereof added to the composition according to the present invention can be measured in accordance with a known method. The content can be measured by using, for example, LC-MS/MS or a saccharimeter.
The composition according to the present invention may be prepared, for example, by mixing the amino acid or salt thereof and the cyclic dipeptide or salt thereof in a predetermined amount with raw materials of a known composition in accordance with a known production method for a composition, or may be prepared by adding the amino acid or salt thereof and the cyclic dipeptide or salt thereof to a known ready-made composition so as to reach the predetermined amount followed by dissolving and/or suspending. A known composition originally containing the amino acid or salt thereof or the cyclic dipeptide or salt thereof may be used, and it may be appropriately mixed for preparation as long as the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof according to the present invention falls within the predetermined range.
4-1. Mechanism of Action
Angiotensin-converting enzyme is mainly present in the lung, vascular endothelial cells, and the renal proximal tubule and has the property of converting the substrate angiotensin I to angiotensin II serving as a pressor hormone. This angiotensin II acts as a potent pressor hormone and promotes vasoconstriction by binding to its receptor present in the vascular vessel. Also, the angiotensin-converting enzyme has an action of decomposing bradykinin having a vasodilating action. Thus, an effect of suppressing production of the pressor hormone angiotensin II and an effect of suppressing decomposition of bradykinin having a vasodilating action are exerted through inhibition of the angiotensin-converting enzyme activity. This can be expected to produce a blood pressure elevation-suppressing action or a blood pressure-lowering action. Accordingly, the inhibition of the angiotensin-converting enzyme is clinically effective for treatment or prevention of hypertension.
4-2. Other Components
The composition according to the present invention can contain any additive or any component commonly used, for example, in addition to the raw material containing the specific amino acid or salt thereof and the specific cyclic dipeptide or salt thereof. Examples of such additives and/or components include, but not limited to, physiologically active components including vitamins, minerals, nutrient components, and fragrances, and in addition, diluents, binders, emulsifiers, tonicity agents (isotonic agents), buffers, solubilizers, antiseptic agents, stabilizers, antioxidants, colorants, coagulants, and coating agents to be mixed in formation.
4-3. Application
The composition according to the present invention can strongly enhance an angiotensin-converting enzyme-inhibiting activity brought about by an amino acid alone, by adjusting the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof to within a specific range. The composition according to the present invention thereby exerts a blood pressure elevation-suppressing action or a blood pressure-lowering action and can be used for prevention or amelioration of hypertension or prevention or amelioration of various diseases caused by hypertension, such as stroke, heart disease, and renal failure. Thus, one embodiment of the present invention is the composition for suppression of elevation of blood pressure, for lowering of blood pressure, for protection of renal functions, or for prevention or amelioration of stroke or heart disease.
Throughout the present specification, examples of the diseases caused by hypertension include, but not particularly limited to, stroke, heart disease, and renal failure. Examples of the stroke include cerebral infarction, cerebral hemorrhage, and subarachnoid hemorrhage. Examples of the heart disease include angina pectoris, myocardial infarction, cardiomegaly, and heart failure.
Examples of the composition according to the present invention include, but not limited to, drugs, drinks, foods, and cosmetics. Specific examples of the drink according to the present invention may include oolong tea drinks, tea drinks, green tea drinks, fruit juice drinks, vegetable juices, sports drinks, isotonic drinks, enhanced waters, mineral waters, near waters, and energy drinks. Examples of the food according to the present invention may include functional foods, dietary supplements, foods with nutrient function claims, foods for special dietary uses, foods for specified health uses, nutritional supplements, foods for diet treatment, health foods, and supplements. The food or drink according to the present invention may be prepared, for example, by mixing the cyclic dipeptide or salt thereof in a predetermined amount with raw materials of a known food or drink in accordance with a known production method for a food, or may be prepared by adding the cyclic dipeptide or salt thereof to a known ready-made food or drink so as to reach the predetermined amount.
The composition according to the present invention is applicable to any of therapeutic applications (medical applications) and non-therapeutic applications (non-medical applications). Specific examples thereof include use for drugs, quasi-drugs, and cosmetics, and include use for compositions which do not belong to drugs, quasi-drugs, or cosmetics under the Pharmaceutical Affairs Act and nevertheless explicitly or implicitly features an effect of suppressing elevation of blood pressure, an effect of slowing elevation of blood pressure, an effect of suppressing rapid elevation of blood pressure upon awakening, a preventing effect on hypertension, an ameliorating effect on hypertension-, an effect of protecting renal functions, or an effect of ameliorating renal functions.
In another embodiment, the present invention relates to the composition provided with a label indicating a function to be exerted through inhibition of angiotensin-converting enzyme. Examples of such an indication or functional indication include, but not particularly limited to, “expecting lowering of blood pressure”, “suppressing elevation of blood pressure”, “slowing elevation of blood pressure”, “suppressing rapid elevation of blood pressure upon awakening”, “preventing hypertension”, “helping ameliorating hypertension”, “protecting renal functions”, and “ameliorating renal functions”. In the present invention, a label with such indication or functional indication may be provided to the composition itself, or provided to a container or package for the composition.
The composition according to the present invention can be ingested by using an appropriate method in accordance with its form. The ingestion method is not particularly limited as long as the amino acid or salt thereof and the cyclic dipeptide or salt thereof contained in the composition according to the present invention can be transferred into circulating blood. Examples of the form may include, but not limited to, solid preparations for oral administration such as tablets, coated tablets, granules, powders, and capsules, liquid preparations for oral administration such as oral solutions and syrups, and preparations for parenteral administration such as injections, external preparations, suppositories, and transdermal preparations. Throughout the present specification, “ingestion” is used to include all forms such as ingestion, administration, and drinking.
The amount of application of the composition according to the present invention is not constant, and is appropriately set in accordance with its form, administration method, intended use, and the age, body weight, and symptoms of a patient or patient animal as a subject to ingest the composition. The effective amount of ingestion of the composition according to the present invention for a human is not constant, and, for example, in the case of a human with a body weight of 50 kg, is preferably 10 mg or more, more preferably 50 mg or more, in terms of the weight of the amino acid or salt thereof according to the present invention per day. Also, in the case of a human with a body weight of 50 kg, it is preferably 1.0 mg or more, more preferably 5.0 mg or more, in terms of the weight of the cyclic dipeptide or salt thereof according to the present invention per day. Administration may be performed in a single administration or multiple administrations in a day, within a desired range of the amount of application. Any duration may be used for the administration. Here, the effective amount of ingestion of the amino acid or salt thereof or the cyclic dipeptide or salt thereof for a human according to the present invention refers to the total amount of ingestion of the amino acid or salt thereof or the cyclic dipeptide or salt thereof which provides an effective action on a human, and is not limited by the type of the amino acid or the cyclic dipeptide.
The subject to apply the composition according to the present invention is preferably a human, but may be a domestic animal such as cattle, a horse, and a goat, a pet animal such as a dog, a cat, and a rabbit, or a laboratory animal such as a mouse, a rat, a guinea pig, and a monkey. In the case of administration for a non-human animal, the amount of use per day for a mouse of approximately 20 g varies depending on conditions including the contents of the amino acid or salt thereof and the cyclic dipeptide or salt thereof in the composition, and the condition, body weight, sex, and age of a subject to be applied, and in common cases is suitably set to such an amount that allows ingestion of preferably 10 mg/kg or more, more preferably 50 mg/kg or more, as the total amount of the amino acid or salt thereof to be added. Also, it is suitably set to such an amount that allows ingestion of preferably 1.0 mg/kg or more, more preferably 5.0 mg/kg or more, as the total amount of the cyclic dipeptide or salt thereof to be added.
5. Use of Composition Containing Amino Acid and Cyclic Dipeptide for Inhibiting Angiotensin-converting Enzyme
One aspect of the present invention is use of a composition containing an amino acid or a salt thereof and a cyclic dipeptide or a salt thereof for inhibiting angiotensin-converting enzyme, wherein the amino acid is one or two or more members selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, arginine, lysine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, and hydroxyproline, the cyclic dipeptide is one or two or more members selected from the group consisting of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)], and the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof is 1:0.02 to 1:20.
Among the amino acids described above or salts thereof, one or two or more members selected from the group consisting of leucine, threonine, arginine, tyrosine, and tryptophan are preferred, and three or more members selected from these amino acids or salts thereof are more preferably contained.
The type and content of the amino acid or salt thereof, the type and content of the cyclic dipeptide or salt thereof, the range of the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof, etc. are as described above about the composition. Specific examples, and amounts of other components that may be added are also as described above about the composition.
The use according to the present invention includes, but not limited to, for example, use for suppression of elevation of blood pressure, lowering of blood pressure, protection of renal functions, or prevention or amelioration of stroke, heart disease, or hypertension. The use is use for a human or a non-human animal, and may be therapeutic use or non-therapeutic use. Here, “non-therapeutic” is a concept excluding medical practice, that is, therapeutic practice for a human body through treatment.
6. Method for Inhibiting Angiotensin-Converting Enzyme
One aspect of the present, invention is a method for inhibiting angiotensin-converting enzyme using a composition containing an amino acid or a salt thereof and a cyclic dipeptide or a salt thereof, wherein the amino acid is one or two or more members selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, arginine, lysine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, and hydroxyproline, the cyclic dipeptide is one or two or more members selected from the group consisting of cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)], and the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof is 1:0.02 to 1:20.
Among the amino acids described above or salts thereof, one or two or more members selected from the group consisting of leucine, threonine, arginine, tyrosine, and tryptophan are preferred, and three or more members selected from these amino acids or salts thereof are more preferably contained.
The type and content of the amino acid or salt thereof, the type and content of the cyclic dipeptide or salt thereof, the range of the ratio of the total amount of the amino acid or salt thereof to the total amount of the cyclic dipeptide or salt thereof, etc. are as described above about the composition. Specific examples and amounts of other components that may be added are also as described above about the composition.
In the method, an individual in need of inhibition of angiotensin-converting enzyme refers to the above subject for application of the composition according to the present invention. Throughout the present specification, a therapeutically effective amount refers to an amount, at which angiotensin-converting enzyme in the individual administered with the composition according to the present invention is inhibited compared to the case of an individual without administration. A specific effective amount is not constant, and is set appropriately in accordance with form of administration, an administration method, intended use, and the age, body weight, and symptoms of an individual.
In the method according to the present invention, the specific amino acid or salt thereof or the specific cyclic dipeptide or salt thereof may be administered as it is, or as a composition containing the specific amino acid or salt thereof or the specific cyclic dipeptide or salt thereof so as to reach the therapeutically effective amount.
The method according to the present invention enables inhibition of angiotensin-converting enzyme without any side effect.
The present invention will now be described with reference to Examples, but the scope of the present invention is not limited thereby. Those skilled in the art could use the method according to the present invention with various variations or modifications, which are also included in the scope of the present invention.
Example 1
The enhancing effect of a cyclic dipeptide on the angiotensin-converting enzyme-inhibiting action of an amino acid was evaluated. Specifically, as for the inhibiting effects of amino acids and cyclic dipeptides, angiotensin I-converting enzyme-inhibiting activities were measured by using ACE Kit-WST (Dojindo Laboratories) according to the instruction manual attached to the kit. The amino acids were studied at a concentration of 1000 μM, 3000 μM, or 5000 μM, and the cyclic dipeptides were studied at a concentration of 20 μM, 50 μM, 100 μM, 250 μM, or 500 μM. Their-inhibiting activities were evaluated as the residual activity of angiotensin-converting enzyme calculated according to expression 1 given below. A control supplemented with distilled water instead of the aqueous cyclic dipeptide solution was used. Chemically synthesized samples were used as the cyclic dipeptides.
Residual activity (%)=(Absorbance at 450 nm from the cyclic dipeptide-added group/Absorbance from 450 nm from the control)×100 (Expression 1)
The evaluation results of various cyclic dipeptides are shown in Tables 1 to 4 below.
The results of Tables 1 to 4 suggested that the angiotensin-converting enzyme-inhibiting action brought about by the amino acid is enhanced by addition of the specific cyclic dipeptide (cyclo-glutamyl-histidine [Cyclo(Glu-His)], cyclo-arginyl-tyrosine [Cyclo(Arg-Tyr)], and cyclo-isoleucyl-serine [Cyclo(Ile-Ser)]) in a given amount to the amino acid.
The present invention relates to a composition containing an amino acid or a salt thereof and a cyclic dipeptide or a salt thereof at a given ratio. This can enhance an angiotensin-converting enzyme-inhibiting activity brought about by the amino acid alone. The amino acid contained in the composition according to the present invention is a naturally derived component, and the cyclic dipeptide is also contained in a heat-treated product of a plant-derived peptide, or the like. Thus, these components are highly safe, and the composition according to the present invention has a very low risk of manifesting Serious side effects. Cyclic dipeptides have high lipid solubility and are efficiently absorbed from the gastrointestinal tract. Thus, the present invention provides a novel means to contribute to suppression of elevation of blood pressure, etc., and thus has high industrial applicability.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2015-132688 | Jul 2015 | JP | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/JP2016/069381 | 6/30/2016 | WO | 00 |
