JP2023072797A - Angiogenesis inducing agent - Google Patents

Abstract

To provide a novel angiogenesis inducing agent.SOLUTION: An angiogenesis inducing agent comprises at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof as an active ingredient.SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to an angiogenesis promoting agent and a food composition for promoting angiogenesis.

Examples of angiogenesis-promoting agents include, for example, angiogenesis-promoting agents containing acetylsalicylic acid or a pharmacologically acceptable salt thereof as an active ingredient (Patent Document 1), and cyclobutylpurine derivatives having a specific structure (Patent Document 2). ) have been reported.

U.S. Patent Application Publication No. 2006/252732 U.S. Patent Application Publication No. 2011/230659

One of the objects of the present invention is to provide a novel angiogenesis-promoting agent.

The present inventors have newly discovered that decenedioic acid and sebacic acid, which are abundantly contained in royal jelly metabolites, have excellent angiogenesis-promoting action.

The present invention provides an angiogenesis promoter containing, as an active ingredient, at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof.

The decenedioic acid may be 2-decenedioic acid or may be trans-2-decenedioic acid.

At least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof may be derived from royal jelly metabolites.

Furthermore, the present invention provides a food composition for promoting angiogenesis, containing the angiogenesis-promoting agent.

The food composition may be selected from the group consisting of foods, health foods, foods with function claims, foods with nutrient function claims, nutritional supplements, supplements, and foods for specified health uses.

The present invention provides a novel angiogenesis-promoting agent.

1 is a photograph of a tube formed by HUVEC under the addition of decenedioic acid, observed with a phase-contrast microscope. DMSO is a negative control, and each concentration indicates the concentration of decendioic acid in the medium before the start of culture. 1 is a photograph of a tube formed by HUVECs under the addition of sebacic acid, observed with a phase-contrast microscope. DMSO is a negative control, and each concentration indicates the concentration of sebacic acid in the medium before starting the culture. The results of calculation and comparison of the total length (a), the number of network structures (b), and the number of branch points (c) of HUVECs formed by HUVEC under the addition of decenedioic acid are shown by analysis using ImageJ. Graph (n=6, *p<0.05; **p<0.01). The concentration on the horizontal axis indicates the concentration of decendioic acid in the medium before the start of culture. Graph showing the results of calculation and comparison of the total length (a), the number of network structures (b), and the number of branch points (c) of HUVECs formed by HUVEC under conditions of addition of sebacic acid by analysis using ImageJ. (n=6, *p<0.05; **p<0.01). The concentration on the horizontal axis indicates the concentration of sebacic acid in the medium before the start of culture.

DETAILED DESCRIPTION OF THE INVENTION Embodiments for carrying out the present invention will be described in detail below. However, the present invention is not limited to the following embodiments.

An embodiment of the present invention provides an angiogenesis promoter containing, as an active ingredient, at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof.

As used herein, “promoting angiogenesis” means promoting the formation of new blood vessel structures. The promotion of angiogenesis can be evaluated, for example, by the degree of vascular structure formation after a certain period of time has passed after administration of an angiogenesis-promoting agent. The degree of vascular structure formation can be evaluated, for example, by the length of blood vessels, the number of network structures, the number of branching points of blood vessels, and the like. It can be said that angiogenesis is promoted when one or more, two or more, or three selected from an increase in the number of branch points is confirmed. For example, it can be determined that angiogenesis is promoted when the average blood vessel length after a certain period of time from the administration of an angiogenesis-promoting agent is increased compared to a reference value.

For example, it can be determined that angiogenesis is promoted when the average blood vessel length after a certain period of time from the administration of an angiogenesis-promoting agent is increased compared to a reference value. For example, the average blood vessel length after a certain period of time after administration of an angiogenesis-promoting agent is at least 10%, 15%, 20%, 25%, 30%, 35%, 40% compared to the reference value. , 45%, 50%, 60%, 70%, 80%, or 90% increase angiogenesis.

Further, for example, when the average number of network structures after a certain period of time after administration of an angiogenesis-promoting agent increases compared to the reference value, it can be said that angiogenesis is promoted. . For example, the average number of network structures after a certain period of time after administering an angiogenesis-promoting agent is at least 10%, 15%, 20%, 25%, 30%, 35%, 40% compared to the reference value. , 45%, 50%, 60%, 70%, 80%, or 90% increase in angiogenesis.

In addition, for example, angiogenesis is promoted when the average number of branch points of blood vessels after a certain period of time after administration of an angiogenesis-promoting agent is increased compared to a reference value. can be done. For example, the average branch point of blood vessels after a certain period of time after administration of an angiogenesis-promoting agent is at least 10%, 15%, 20%, 25%, 30%, 35%, 40% compared to the reference value. , 45%, 50%, 60%, 70%, 80%, or 90% increase in angiogenesis.

Promotion of angiogenesis can also be evaluated, for example, by the time from administration of an angiogenesis-promoting agent to the formation of blood vessels that meet the criteria. The reference blood vessel can be set by, for example, the length of the blood vessel, the number of network structures, the number of branch points of the blood vessel, and the like. The time required for 1 or more, 2 or more, or 3 or more selected from the length of blood vessels, the number of network structures, and the number of branching points of blood vessels to reach a reference value is the same as when the angiogenesis-promoting agent is not administered. Angiogenesis can be promoted if it is relatively short.

For example, when the time from administration of an angiogenesis-promoting agent until the average blood vessel length reaches a reference value (length) is shorter than the reference value (time), angiogenesis is promoted. It can be assumed that there is For example, the time for the mean blood vessel length to reach a reference value (length) is at least 10%, 15%, 20%, 25%, 30%, 35% compared to the reference value (time), Angiogenesis can be promoted when shortening by 40%, 45%, 50%, 60%, 70%, 80%, or 90%.

Also, for example, when the time from administration of an angiogenesis-promoting agent until the average number of network structures reaches a reference value (length) is shorter than the reference value (time), angiogenesis is promoted. It can be said that For example, the time for the average number of networks to reach a reference value (length) is at least 10%, 15%, 20%, 25%, 30%, 35% compared to the reference value (time), Angiogenesis can be promoted when shortening by 40%, 45%, 50%, 60%, 70%, 80%, or 90%.

Further, for example, when the time from administration of an angiogenesis-promoting agent until the average number of branch points of blood vessels reaches a reference value (length) is shorter than the reference value (time), angiogenesis can be said to be promoted. For example, if the time to reach a reference value (length) for the average number of vascular branch points is at least 10%, 15%, 20%, 25%, 30%, 35% compared to the reference value (time). %, 40%, 45%, 50%, 60%, 70%, 80%, or 90% shortening can promote angiogenesis.

The reference or reference value is the value when or when the angiogenesis-promoting agent is not administered, the value immediately before or when administering the angiogenesis-promoting agent, the value immediately after or at the time of administering the angiogenesis-promoting agent, An appropriate one can be used in accordance with the evaluation method based on blood vessels in tissue or values based thereon.

When evaluating the promotion of angiogenesis in a living tissue, it can be determined, for example, by an evaluation system using a blood vessel visualization mouse. When evaluating the promotion of angiogenesis in culture of cells such as vascular endothelial cells and stem cells that differentiate into blood vessels, for example, cell masses or tissues produced by culture can be used for determination. The degree of vascular structure formation, for example, the length of blood vessels, the number of network structures, the number of branch points of blood vessels, etc., can be observed with a phase-contrast microscope or the like, and can be analyzed using image analysis software such as ImageJ.

Subjects to whom the angiogenesis-promoting agent is administered may be rodents, ungulates, carnivores, marsupials, and primates, and may be tissues or cells derived therefrom. The subject is preferably a mammal, such as a cynomolgus monkey and a human, more preferably a human. The cells may be, for example, vascular endothelial cells and stem cells that differentiate into blood vessels.

Royal jelly is metabolized in the body of an administration subject to become a royal jelly metabolite. Decenedioic acid and sebacic acid are major constituents of royal jelly metabolites, for example, it has been reported that they are often found in plasma of human subjects after administration of royal jelly (Yamagata et al., RSC Advances (2019), 9, 15392-15401). This time, it was newly found that decenedioic acid and sebacic acid, which are major constituents of royal jelly metabolites, have an action to promote angiogenesis. Therefore, the angiogenesis-promoting agent of the present embodiment contains, as an active ingredient, at least one compound selected from the group consisting of decendioic acid, sebacic acid, and salts thereof, and thus has an action of promoting angiogenesis. .

Salts of decenedioic acid and sebacic acid may be salts acceptable for food use or pharmaceutical use. Salts of decenedioic acid and sebacic acid include, for example, salts with alkali metals, alkaline earth metals, other metals, ammonium, and the like. More specific examples of salts of decenedioic acid and sebacic acid include potassium, sodium, calcium and magnesium salts.

The decenedioic acid may be 2-decenedioic acid. 2-decenedioic acid may be either cis or trans, and is preferably trans-2-decenedioic acid.

Decenedioic acid, sebacic acid and salts thereof may be chemically synthesized products or may be naturally derived products such as royal jelly. Since decenedioic acid, sebacic acid and salts thereof are abundant in royal jelly metabolites, for example, decenedioic acid, sebacic acid and salts thereof may be derived from royal jelly metabolites.

The angiogenesis-promoting agent of the present embodiment may contain decenedioic acid, sebacic acid, and salts thereof singly or in combination of two or more thereof. When a mixture of two or more types is contained, two or more compounds may be simply mixed, or two or more compounds may be simply mixed. Or it may be a mixture obtained by processing (eg, concentrating) its metabolites.

The content of decenedioic acid or a salt thereof in the angiogenesis-promoting agent according to the present embodiment may be 0.1% by mass or more and 99% by mass or less in terms of dry solid content relative to the total amount of the angiogenesis-promoting agent. 1% by mass or more, 1% by mass or more, 3% by mass or more, 3.5% by mass or more, 5% by mass or more, 7% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, 70% by mass or more, 75% by mass 80% by mass or more, 85% by mass or more, 90% by mass or more, 93% by mass or more, 95% by mass or more, 98% by mass or more, 99% by mass or more, or 100% by mass. In addition, the content of decenedioic acid or a salt thereof in the angiogenesis-promoting agent according to the present embodiment is 100% by mass or less, 99% by mass or less, 98% by mass or less, 95% by mass or less in solid content relative to the total amount of the angiogenesis-promoting agent. % by mass or less, 93% by mass or less, 90% by mass or less, 85% by mass or less, 80% by mass or less, 75% by mass or less, 70% by mass or less, 65% by mass or less, 60% by mass or less, 55% by mass or less, 50 % by mass or less, 45% by mass or less, 40% by mass or less, 35% by mass or less, 30% by mass or less, 25% by mass or less, 20% by mass or less, 15% by mass or less, 10% by mass or less, 8% by mass or less, 5 % by mass or less, 3% by mass or less, or 1% by mass or less.

The content of sebacic acid or a salt thereof in the angiogenesis-promoting agent according to the present embodiment may be 0.1% by mass or more and 99% by mass or less in terms of dry solid content relative to the total amount of the angiogenesis-promoting agent. % by mass or more, 1% by mass or more, 3% by mass or more, 3.5% by mass or more, 5% by mass or more, 7% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more , 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, 70% by mass or more, 75% by mass or more , 80% by mass or more, 85% by mass or more, 90% by mass or more, 93% by mass or more, 95% by mass or more, 98% by mass or more, 99% by mass or more, or 100% by mass. In addition, the content of sebacic acid or a salt thereof in the angiogenesis-promoting agent according to the present embodiment is 100% by mass or less, 99% by mass or less, 98% by mass or less, and 95% by mass in solid content relative to the total amount of the angiogenesis-promoting agent. % or less, 93 mass% or less, 90 mass% or less, 85 mass% or less, 80 mass% or less, 75 mass% or less, 70 mass% or less, 65 mass% or less, 60 mass% or less, 55 mass% or less, 50 mass% % or less, 45% by mass or less, 40% by mass or less, 35% by mass or less, 30% by mass or less, 25% by mass or less, 20% by mass or less, 15% by mass or less, 10% by mass or less, 8% by mass or less, 5% by mass % or less, 3 mass % or less, or 1 mass % or less.

When administered to humans, the dose of the angiogenesis-promoting agent can be appropriately set according to factors such as the type of compound, the health condition of the person taking it, the administration method, and the combination with other agents.

The angiogenesis promoter of the present embodiment can be used at a dose of 1 mg or more and 10 g or less, preferably 5 mg or more and 8 g or less, preferably 10 mg or more and 3 g per day for an adult with a body weight of 60 kg in terms of active ingredient amount. It is more preferable to use the following doses, more preferably 15 mg or more and 1.5 g or less, even more preferably 20 mg or more and 1 g or less, and more preferably 22 mg or more and 500 mg or less. It is even more preferred and particularly preferred to use a dose of 24 mg or more and 250 mg or less. When the angiogenesis-promoting agent of the present invention is used in children, the dose is, for example, 3/5 the adult dose for children aged 6 to under 13 years, 2/5 the adult dose for children aged 1 to under 6 years, and A 1/5 dose may be used. The dose can be appropriately set within the above range depending on factors such as the health condition of the person taking the drug, administration method, and combination with other agents.

The angiogenesis-promoting agent of this embodiment may be orally administered (ingested) or parenterally administered (for example, intranasally administered). The angiogenesis-promoting agent of the present invention may be administered once a day, or in multiple doses such as twice a day, three times a day, etc., as long as the amount of the active ingredient per day is within the range described above. may also be administered. Moreover, the angiogenesis-promoting agent of the present invention is preferably administered continuously. By continuous administration, the angiogenesis activating effect is exhibited more remarkably.

The angiogenesis-promoting agent according to this embodiment may contain only the above active ingredients, or may further contain other ingredients as long as the effects of the present invention are not impaired. Other ingredients include, for example, pharmaceutically acceptable ingredients (e.g., excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents). , Food acceptable ingredients (e.g., minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, cooling agents, binders, sweeteners, disintegrants, lubricants, coloring agents , fragrances, stabilizers, preservatives, sustained release modifiers, surfactants, solubilizers, wetting agents).

The angiogenesis-promoting agent of this embodiment may be in any form such as solid, liquid, and paste. The angiogenesis-promoting agent of the present invention may be in the form of, for example, plain tablet, sugar-coated tablet, granule, powder, tablet, capsule (hard capsule, soft capsule, seamless capsule). The angiogenesis-promoting agent of the present invention can be prepared, for example, by mixing the compound of the present invention or a salt thereof as an active ingredient with other ingredients, if necessary, and molding the mixture into the above dosage form. .

The angiogenesis promoting agent of the present embodiment can be used as a pharmaceutical, quasi-drug, or food composition itself, or added to a pharmaceutical, quasi-drug, or food composition. The food composition is preferably a food in which the tertiary function of food (physical conditioning function) is emphasized. Examples of foods in which the tertiary functions of foods are emphasized include health foods, foods with functional claims, foods with nutrient function claims, dietary supplements, supplements, and foods for specified health uses.

A pharmaceutical, quasi-drug, or food composition comprising the angiogenesis-promoting agent of the present embodiment, or a pharmaceutical, quasi-drug, or food composition comprising the angiogenesis-promoting agent of the present embodiment is for promoting angiogenesis. For example, by promoting angiogenesis through angiogenesis-promoting action, vascular damage or abnormality repair, skin healing or reconstruction (e.g., wound / burn healing or improvement), improvement by angiogenesis promotion It can be used for prevention or treatment of angiogenesis-related diseases.

A pharmaceutical, quasi-drug, or food composition comprising the angiogenesis-promoting agent of the present embodiment, or a pharmaceutical, quasi-drug, or food composition comprising the angiogenesis-promoting agent of the present invention also has an angiogenesis-promoting action , can promote improvement or restoration of blood flow.

A pharmaceutical, quasi-drug, or food composition comprising the angiogenesis-promoting agent of the present embodiment, or a pharmaceutical, quasi-drug, or food composition comprising the angiogenesis-promoting agent of the present embodiment is further associated with aging. It can be used to improve or treat diseases or symptoms associated with decreased capillaries.

The content of the angiogenesis-promoting agent of the present embodiment in the pharmaceutical, quasi-drug, and food composition is such that the amount of the active ingredient taken per day is within the range described above. It may be appropriately set according to the type of object.

The food composition containing an angiogenesis-promoting agent according to the present invention may contain the angiogenesis-promoting agent in an amount of, for example, 0.005 to 10.0% by mass. When the compound is 2-decenedioic acid, it may be from 0.1 mg to 20 mg/1 g dry weight of the food composition, preferably from 1 mg to 10 mg/1 g dry weight of the food composition. When the compound is sebacic acid, it may be, for example, 0.05 mg to 10 mg/1 g dry weight of the food composition, preferably 0.5 mg to 5 mg/1 g dry weight of the food composition.

When the angiogenesis-promoting agent of the present embodiment is used as a food composition itself or added to a food composition, the form of the food composition is not particularly limited. Soft drinks, milk drinks, lactic acid drinks, yogurt drinks, carbonated drinks, etc.); Spreads (custard cream, etc.); Pastes (fruit paste, etc.); , cookies, cakes, puddings, etc.); Japanese sweets (daifuku, mochi, steamed buns, castella, anmitsu, yokan, etc.); ice desserts (ice cream, popsicles, sherbet, etc.); , soup, meat sauce, pasta, pickles, jam, etc.);

The angiogenesis-promoting agent of the present embodiment can be used as a food in which the tertiary function of food is emphasized (e.g., health food, food with functional claims, nutritional supplement, supplement, or food for specified health use) itself, or the tertiary function of food. When used by being added to a food in which the tertiary function of the food is emphasized, the form of the food in which the tertiary function of the food is emphasized is, in addition to the forms of the food described above, for example, uncoated tablets, sugar-coated tablets, granules, powders, tablets, capsules (Hard capsule, soft capsule, seamless capsule).

When the angiogenesis promoting agent of the present embodiment is used as a drug or quasi-drug itself, or added to a drug or quasi-drug, the form of the drug or quasi-drug is not particularly limited. Tablets, sugar-coated tablets, granules, powders, tablets, capsules (hard capsules, soft capsules, seamless capsules) may be used.

The method for producing the pharmaceutical, quasi-drug, or food composition to which the angiogenesis-promoting agent of the present embodiment is added is not particularly limited, and a known method can be appropriately followed. For example, by mixing the angiogenesis promoter of the present embodiment with an intermediate product or final product in the manufacturing process of a drug, quasi-drug, or food composition, the drug, quasi-drug, or A food composition can be obtained.

This embodiment described above can also be considered as at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof for use in promoting angiogenesis. This embodiment can also be regarded as an application of at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof in the production of an angiogenesis promoting agent.

In the above-described embodiment, the step of administering an effective amount of an angiogenesis-promoting agent containing at least one compound selected from the group consisting of decenedioic acid, sebacic acid, and salts thereof to a subject in need thereof. It can also be regarded as a method of promoting angiogenesis, including Subjects can be rodents, ungulates, carnivores, marsupials, and primates. The subject is preferably a mammal, such as a cynomolgus monkey and a human, more preferably a human.

EXAMPLES The present invention will be described in more detail below based on examples. However, the present invention is not limited to the following examples.

The angiogenesis-promoting effects of 2-decenedioic acid (Sundia MediTech, hereinafter referred to as "decenedioic acid") and sebacic acid (Nacalai tesque) were examined according to the following method.

1. Experimental method 1-1. Tube Formation assay (evaluation of in vitro angiogenesis ability)
150 μL of Matrigel (Corning) was added dropwise to a 48-well plate, heated in a 37° C. incubator for 30 minutes or longer, and then HUVECs (Lonza, human umbilical vein endothelial cells) were seeded (in the experiment, the number of passages was 5) were cultured in EBM-2 medium (Lonza) supplemented with various concentrations of decenedioic acid and sebacic acid for 8 hours. The cell density before culture was 4.0×10 ⁵ cells/mL, and the culture conditions were 37°C. Six hours post-culturing cultures with good tube formation were used for evaluation of angiogenic potential.

For the evaluation, a phase-contrast microscope was used to record the formation process of tubes formed by HUVECs on the gel, and images were taken at least 4 points per well. Photographs taken with a phase-contrast microscope 6 hours after culture are shown in FIGS. 1 and 2 (n=6, *p<0.05; **p<0.01). FIG. 1 shows the results for decenedioic acid and FIG. 2 for sebacic acid.

3 and 4 show the results of calculating the total length of the tube, the number of network structures, and the number of branch points in the image by analysis using ImageJ. FIG. 3 shows the results for decenedioic acid and FIG. 4 for sebacic acid.

2. Experimental results 2-1. Effect of RJ Metabolites on In Vitro Angiogenic Ability Tubes formed by HUVEC under the conditions of adding decenedioic acid and sebacic acid were compared with the control (DMSO added), respectively. It was found that angiogenesis was promoted by the method (Figs. 1 and 3). In particular, when comparing the dotted line frame (control) and the solid line frame in FIGS. 1 and 3, the difference is remarkable.

A comparison of the total length of tubes, the number of network structures, and the number of branch points formed by HUVEC under the conditions of the addition of decenedioic acid and the addition of sebacic acid revealed that the addition of decenedioic acid reduced the total length of the tubes and the number of network structures. It was confirmed that the number of and the number of branch points increased significantly (Fig. 2). In addition, the addition of sebacic acid also tended to increase the total length of the tube, the number of network structures, and the number of branch points, although not as much as that of decenedioic acid (Fig. 4).

Claims (5)

An angiogenesis promoter containing, as an active ingredient, at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof. The angiogenesis promoter according to claim 1, wherein the decenedioic acid is 2-decenedioic acid. The angiogenesis promoter according to claim 1 or 2, wherein at least one compound selected from the group consisting of decenedioic acid, sebacic acid and salts thereof is derived from royal jelly metabolites. A food composition for promoting angiogenesis, comprising the angiogenesis-promoting agent according to any one of claims 1 to 3. 5. The food composition according to claim 4, which is selected from the group consisting of food, health food, food with function claims, food with nutrient function claims, dietary supplement, supplement and food for specified health use.

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