Blood pressure suppressant

Abstract

To provide a useful blood pressure elevation suppressant. [Solution] A blood pressure elevation inhibitor containing as an active ingredient one or more compounds selected from the group consisting of saclipin A (a compound represented by the following formula (2)), saclipin B, and derivatives thereof, isolated from an extract of Suizenji Nori. TIFF2026115124000007.tif68169

Description

[Technical Field]

[0001] This disclosure relates to an agent for suppressing blood pressure elevation. [Background technology]

[0002] Patent Document 1 describes a blood pressure-lowering agent containing limonoid as an active ingredient. [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2021-138642 [Overview of the Initiative] [Problems that the invention aims to solve]

[0004] The inventors of this invention have identified a drought-stress-inducing ultraviolet-absorbing substance from the cyanobacterial strain Suizenji-nori. The inventors have been exploring the useful physiological activity of this substance and developing technologies for its effective utilization.

[0005] This disclosure is made in view of the above circumstances and aims to provide a novel blood pressure elevation suppressant. [Means for solving the problem]

[0006] [1] A blood pressure-lowering agent containing as an active ingredient one or more compounds selected from the group consisting of compounds represented by the following formula (1), compounds represented by the following formula (2), and derivatives thereof. [ka] [ka] [2] An oral antihypertensive agent as described in [1]. [Brief explanation of the drawing]

[0007] [Figure 1] It is a graph showing the results of measuring angiotensin converting enzyme inhibitory activity. [Figure 2] It is a graph showing the results of measuring collagenase inhibitory activity. [Figure 3] It is a graph showing the results of measuring hyaluronidase inhibitory activity.

MODE FOR CARRYING OUT THE INVENTION

[0008] Hereinafter, embodiments embodying the present disclosure will be described. In this specification, for a description using "~" for a numerical range, unless otherwise specified, it includes the lower limit value and the upper limit value. For example, in the description "10~20", it includes both the lower limit value "10" and the upper limit value "20". That is, "10~20" has the same meaning as "10 or more and 20 or less". Also, in this specification, the upper limit value and the lower limit value of each numerical range can be arbitrarily combined.

[0009] 1. Blood pressure increasing inhibitor The blood pressure increasing inhibitor of this embodiment contains, as an active ingredient, one or more compounds selected from the group consisting of a compound represented by the following formula (1), a compound represented by the following formula (2), and derivatives thereof.

CHEM.

CHEM.

[0010] Hereinafter, the compound represented by formula (1) above will also be referred to as saclipin B, and the compound represented by formula (2) above will also be referred to as saclipin A. When saclipin A and saclipin B are not distinguished, they will simply be referred to as saclipin. A derivative of the compound represented by formula (1) above will also be referred to as a saclipin B derivative, and a derivative of the compound represented by formula (2) above will also be referred to as a saclipin A derivative. The above "one or more compounds" will also be referred to as compound S.

[0011] (1) saclipin A, saclipin B Aphanothece sacrum, also known as Suizenji-nori, is a cyanobacterium endemic to Japan, and its habitat has only been confirmed in parts of Kyushu. Currently, it grows naturally only in aquaculture farms utilizing the Kogane River in Asakura City, Fukuoka Prefecture, and is distributed as a food product.

[0012] The inventors of this application analyzed an extract of Suizenji-nori and discovered the presence of a substance that strongly absorbs in the ultraviolet region. The substance with strong absorption in the ultraviolet region was isolated and purified by preparative HPLC, and structural analysis of the purified product revealed that it was the compound of formula (1) and the compound of formula (2) above.

[0013] The compound represented by formula (1) is (10E,12Z,14E)-9,16-Dioxooctadeca-10,12,14-trienoic acid in IUPAC nomenclature. The molecular weight of this compound is 306. The inventors of this application have named this compound saclipin B.

[0014] The compound represented by formula (2) is (10E,12E,14E)-9,16-Dioxooctadeca-10,12,14-trienoic acid in IUPAC nomenclature. The molecular weight of this compound is 306. The inventors of this application have named this compound saclipin A. Saclipin A and saclipin B are geometric isomers (cis-trans isomers).

[0015] Thermodynamically, organic compounds with carbon-carbon double bonds are generally considered more stable in the trans (E) form than in the cis (Z) form. It is surprising that both trans (E) saclipin A and cis (Z) saclipin B exist in Suizenji Nori. While the reason for the stable existence of saclipin B is unclear, it is possible that hydrogen bonding stabilizes saclipin B in its enol form. From a stability standpoint, saclipin B is a useful compound for various applications.

[0016] Saclipin A and / or saclipin B may be used in the form of an extract from cyanobacteria or in the form of a purified product. The purification method is not particularly limited and can be carried out by chromatography, such as HPLC. Alternatively, saclipin A and / or saclipin B may be used in the form of the cyanobacteria themselves.

[0017] Saclipin A and / or saclipin B exhibit high stability against treatments such as light irradiation and heat treatment. Saclipin A and / or saclipin B are suitable for oral formulations.

[0018] (2) saclipin A derivatives, saclipin B derivatives In this disclosure, saclipin B derivative means a derivative obtained by modifying one or more functional groups present in saclipin B, and having functionality equivalent to or better than saclipin B. saclipin A derivative means a derivative obtained by modifying one or more functional groups present in saclipin A, and having functionality equivalent to or better than saclipin A. The above "functionality" is not particularly limited and includes, for example, angiotensin-converting enzyme (ACE) inhibitory activity, collagenase inhibitory activity, hyaluronidase inhibitory activity, etc.

[0019] The saclipin B derivative is preferably at least one selected from the group consisting of an ester derivative obtained by reacting a carboxyl group present at the terminal of saclipin B with an alcohol, an amide derivative obtained by reacting a carboxyl group present at the terminal of saclipin B with an amine, and an ammonium salt obtained by reacting a carboxyl group present at the terminal of saclipin B with ammonium. The saclipin A derivative is preferably at least one selected from the group consisting of an ester derivative obtained by reacting a carboxyl group present at the terminal of saclipin A with an alcohol, an amide derivative obtained by reacting a carboxyl group present at the terminal of saclipin A with an amine, and an ammonium salt obtained by reacting a carboxyl group present at the terminal of saclipin A with ammonium. The type of the carboxyl group derivative group in the saclipin B derivative and / or saclipin A derivative can be appropriately selected according to the use, dosage form when used, and the like.

[0020] The carboxyl group derivative group can be represented, for example, as -COOR 1 , -CONH2, -CONHR 2 , -CONR 3 R 4 , -COONH4, -COONH3R 5 . Here, R 1 , R 2 , R 3 , R 4 , R 5Each of these is independently a hydrocarbon group having 1 to 10 carbon atoms, which may have a functional group. Among the carboxyl group derivatives, saclipinB and saclipinA having -COOCH3 are also called methyl-esterified saclipinB and methyl-esterified saclipinA, respectively. Methyl-esterified saclipinB and methyl-esterified saclipinA can enhance hydrophobicity while ensuring UV absorption equivalent to that of saclipinB and saclipinA. Such saclipinB derivatives and / or saclipinA derivatives are more suitable for use in mixtures with hydrophobic components such as oils compared to saclipinB and / or saclipinA.

[0021] (3) Isomer ratio The ratio of saclipin A and its derivatives to saclipin B and its derivatives in a blood pressure-lowering agent is not particularly limited. The ratio of saclipin A and its derivatives to saclipin B and its derivatives (saclipin A and its derivatives:saclipin B and its derivatives, mass ratio) in a blood pressure-lowering agent can be, for example, 70:30 to 5:95, and may also be 50:50 to 8:92, 40:60 to 8:92, or 30:70 to 10:90. In Suizenji Nori, the ratio of saclipin A to saclipin B is usually around 75:35 to 90:10.

[0022] 2. Action and Uses of Blood Pressure Inhibitors (1) Angiotensin-converting enzyme (ACE) inhibitory effect Angiotensin-converting enzyme (ACE) inhibitors are known to be effective in lowering blood pressure by suppressing the production of angiotensin II, which has a blood pressure-raising effect. Angiotensin-converting enzyme (ACE) inhibitors suppress the production of angiotensin II, which triggers an increase in blood pressure, by inducing vasoconstriction and the release of aldosterone.

[0023] Saclipin A has angiotensin-converting enzyme (ACE) inhibitory activity. Saclipin B also has angiotensin-converting enzyme (ACE) inhibitory activity. The trans isomer, saclipin A, exhibits higher angiotensin-converting enzyme (ACE) inhibitory activity than the cis isomer, saclipin B. Similarly, it is presumed that saclipin A derivatives and saclipin B derivatives also have angiotensin-converting enzyme (ACE) inhibitory activity. It is presumed that the trans isomer, saclipin A derivative, exhibits higher angiotensin-converting enzyme (ACE) activity than the cis isomer, saclipin B derivative.

[0024] Based on the above, a blood pressure-lowering agent containing compound S as an active ingredient can be suitably used to suppress blood pressure elevation. A blood pressure-lowering agent containing compound S as an active ingredient is also useful as an angiotensin-converting enzyme (ACE) inhibitor.

[0025] (2) Collagenase inhibitory effect, hyaluronidase inhibitory effect Saclipin A has collagenase inhibitory activity. Saclipin B also has collagenase inhibitory activity. The trans isomer, saclipin A, exhibits higher collagenase inhibitory activity than the cis isomer, saclipin B. Similarly, it is presumed that saclipin A derivatives and saclipin B derivatives also possess collagenase inhibitory activity. It is presumed that the trans isomer, saclipin A derivative, exhibits higher collagenase inhibitory activity than the cis isomer, saclipin B derivative.

[0026] Collagenase is a proteolytic enzyme that breaks down collagen. The accumulation of collagen in dermal fibroblasts is said to be important for maintaining skin elasticity. Blood pressure-lowering agents containing compound S as an active ingredient are also useful as collagenase inhibitors. Furthermore, saclipin is known to have the effect of promoting collagen production and inhibiting collagen glycation. Blood pressure-lowering agents containing compound S as an active ingredient are also useful as anti-aging agents for the skin.

[0027] Saclipin A has hyaluronidase inhibitory activity. Saclipin B also has hyaluronidase inhibitory activity. The hyaluronidase inhibitory activity of saclipin A and saclipin B is approximately equivalent. Similarly, it is presumed that saclipin A derivatives and saclipin B derivatives also have hyaluronidase inhibitory activity.

[0028] Hyaluronidase is a degrading enzyme that breaks down hyaluronic acid. A blood pressure-lowering agent containing compound S as an active ingredient is also useful as a hyaluronidase inhibitor. The accumulation of hyaluronic acid in dermal fibroblasts is said to be important for maintaining skin elasticity. Furthermore, saclipin B is known to promote hyaluronic acid production. A blood pressure-lowering agent containing compound S as an active ingredient is also useful as an anti-aging agent for the skin.

[0029] (3) Other useful effects Saclipin A exhibits an absorption maximum at 316 nm, with a molar extinction coefficient of 26,454 M. -1 cm -1 Saclipin B exhibits an absorption maximum at 319 nm, and its molar extinction coefficient is 30,555 M. -1 cm -1 The blood pressure-lowering agent containing the above compound S as an active ingredient may exert one or more of the following effects (see PCT / JP2024 / 20712 and Japanese Patent Application No. 2024-137649 by the same applicant). UV absorption effect Antioxidant activity; radical scavenging activity, singlet oxygen scavenging activity, etc. Anti-glycation effect; activity that inhibits the glycation reaction of elastin, activity that inhibits the glycation reaction of collagen Anti-aging effects; effects that inhibit elastase activity, promote collagen production in dermal fibroblasts, promote hyaluronic acid production in dermal fibroblasts, etc. Whitening effect; effect of inhibiting tyrosinase activity, effect of suppressing melanin production, etc.

[0030] (4) Oral preparations The blood pressure-lowering agent of this disclosure is suitable, for example, as an oral preparation. Suizenji-nori is a material with a long history of consumption as an edible cyanobacterium. The blood pressure-lowering agent of this disclosure is preferable in terms of safety because it uses a component derived from an edible material as its active ingredient. Furthermore, angiotensin-converting enzyme (ACE) is an enzyme that is widely distributed in the body and is abundant in vascular endothelial cells of the lungs. When compound S is taken orally, it is thought that its efficacy against angiotensin-converting enzyme (ACE) is exerted more efficiently. In addition to the above component, the oral preparation of this disclosure may appropriately contain components that are commonly used in oral preparations, etc., within a quantitative and qualitative range that does not impair the effects of this disclosure. The properties of the oral preparation are not particularly limited and it can be added to or enclosed in any form such as tablets, capsules, granules, powders, tablets, jellies, gummies, gums, drinks, PET bottles, etc., or added to any food or beverage. The oral preparation is suitable as an oral preparation such as a pharmaceutical, a functional food, a supplement, a food additive, etc.

[0031] The amount of compound S in the oral preparation can be appropriately set according to the application and is not particularly limited. From the viewpoint of storage, ease of ingestion, and obtaining a skin-improving effect, the amount of compound S in the oral preparation is preferably 0.01% to 10%, and more preferably 0.1% to 5%, when the total mass of the oral preparation is considered to be 100%. The dosage of the oral preparation can be appropriately adjusted depending on factors such as the age, weight, sex, and condition of the person taking it. For example, the daily intake of the oral preparation is 0.001 mg to 100 mg per day for an adult weighing 60 kg, preferably 0.005 mg to 70 mg, more preferably 0.01 mg to 50 mg, and even more preferably 0.1 mg to 20 mg, based on the above-mentioned compound S, but is not limited to this range. If necessary, the above intake can be divided into one dose or several doses per day, for example, two to three times. [Examples]

[0032] The present disclosure will be further described below with reference to examples. However, the scope of the present disclosure is not limited to these examples.

[0033] 1. Sample preparation (1) Sample of saclipin A, sample of saclipin B Samples of saclipin A and saclipin B were extracted from dried Suizenji-nori (product name "Jusen-ko," distributor "Endo Kanagawa-do") using the method described in Uchida Y, Honda M, Waditee-Sirisattha R, et al. (2024), ACS Agricultural Science & Technology 4: 1260-1270. The saclipin A sample contained saclipin A and saclipin B in a ratio of 0.92:0.08. The saclipin B sample contained saclipin A and saclipin B in a ratio of 0.09:0.91.

[0034] 2. Experiment 1 (ACE inhibitory activity) (1) Measurement of ACE inhibitory activity ACE inhibitory activity was measured using samples of saclipin A and saclipin B. The saclipin concentrations in each sample were 0.4 mM, 2 mM, and 9.8 mM. Three samples were used for each concentration (N=3). The measurements were performed by the Kanagawa Prefectural Institute of Industrial Technology. ACE inhibitory activity was calculated using the ACE Kit-WST (manufactured by Dojin Chemical Laboratories) according to the manufacturer's protocol.

[0035] (2) Results of Experiment 1 (ACE inhibitory activity) The results of Experiment 1 are shown in Figure 1. The horizontal axis of the graph in Figure 1 shows the concentration (mM) of saclipin A or saclipin B. The dark graph on the left for each concentration shows the results for saclipin A, and the white graph on the right shows the results for saclipin B. The vertical axis shows the angiotensin-converting enzyme (ACE) inhibition rate (%). The error bars show the standard deviation of the three measurements.

[0036] As shown in Figure 1, both saclipin A and saclipin B exhibited angiotensin-converting enzyme (ACE) inhibitory activity. IC of saclipin A 50 The value was 0.5 mM. (Saclipin B IC) 50 The value was 1.2 mM. saclipin A showed higher activity than saclipin B. These results indicate that saclipin is useful as a blood pressure elevation suppressant. Furthermore, saclipin was shown to be useful as an angiotensin-converting enzyme (ACE) inhibitor.

[0037] 3. Experiment 2 (Collagenase inhibitory activity) (1) Measurement of collagenase inhibitory activity Collagenase inhibitory activity was measured using samples of saclipin A and saclipin B. The saclipin concentrations in each sample were 0.4 mM, 2 mM, and 9.8 mM. Three samples were used for each concentration (N=3). The measurements were performed by the Kanagawa Prefectural Institute of Industrial Technology. Collagenase inhibitory activity was calculated using the following procedure. First, collagenase was dissolved in 50 mM Tris buffer (pH 7.3) to prepare a 0.02 mg / mL enzyme solution. MOCAc-Pro-Leu-Gly-Leu-A2pr (Dnp)-Ala-Arg-NH2 was dissolved in DMSO to a concentration of 1 mM, and immediately before use, the solution was diluted 200-fold with 50 mM Tris buffer (pH 7.3) to prepare the substrate solution. Measurements were performed using a 96-well microplate. 50 μL of each sample was added to 100 μL of enzyme solution, incubated at 37°C for 10 minutes, then 50 μL of substrate solution was added and stirred, followed by incubation at 37°C for 60 minutes. Fluorescence intensity (excitation wavelength 320 nm, fluorescence wavelength 405 nm) was then measured. Collagenase inhibitory activity was calculated from the change in fluorescence intensity for each sample.

[0038] (2) Results of Experiment 2 (Collagenase Inhibitory Activity) The results of Experiment 2 are shown in Figure 2. The horizontal axis of the graph in Figure 2 shows the concentration (mM) of saclipin A or saclipin B. The dark graph on the left for each concentration shows the results for saclipin A, and the white graph on the right shows the results for saclipin B. The vertical axis shows the collagenase inhibition rate (%). The error bars show the standard deviation of the three measurements.

[0039] As shown in Figure 2, both saclipin A and saclipin B exhibited collagenase inhibitory activity. IC of saclipin A 50 The value was 0.5 mM. (Saclipin B IC) 50 The value was 1.2 mM. Saclipin A was more active than saclipin B. IC25 of saclipin A and saclipin B 50 The values ​​were both 0.4 mM (400 μM) or less. 50The value is the IC of mycosporine-like amino acids (MAAs), known as UV-absorbing compounds in cyanobacteria. 50 The values ​​are sufficient when compared to other values. These results demonstrate that saclipin is also effective as a collagenase activity inhibitor.

[0040] 4. Experiment 3 (Hyaluronidase inhibitory activity) (1) Measurement of hyaluronidase inhibitory activity Hyaluronidase inhibitory activity was measured using samples of saclipin A and saclipin B. The saclipin concentrations in each sample were 0.4 mM, 2 mM, and 9.8 mM. Three samples were used for each concentration (N=3). The measurements were performed by the Kanagawa Prefectural Institute of Industrial Technology. Hyaluronidase inhibitory activity was calculated using the following procedure. First, bovine hyaluronidase was dissolved in 0.1 M acetate buffer (pH 4.0) to prepare a 4 mg / mL enzyme solution. Potassium hyaluronate was dissolved in acetate buffer (pH 4.0) to a concentration of 0.8 mg / mL to prepare a substrate solution. Compound 48 / 80 was dissolved in acetate buffer (pH 4.0) to a concentration of 0.5 mg / mL to prepare an enzyme activation solution. The reaction was carried out in a microcentrifuge tube. 50 μL of sample and 25 μL of enzyme solution were added to a microcentrifuge tube and incubated at 37°C for 20 minutes. Next, 50 μL of enzyme activation solution was added and incubated at 37°C for 20 minutes. Then, 125 μL of substrate solution was added and incubated at 37°C for 40 minutes. Next, 50 μL of 0.4N sodium hydroxide solution was added to stop the reaction. Then, 50 μL of 0.8M borate buffer (pH 9.0) was added and heated on a heat block at 105°C for 5 minutes, after which it was thoroughly cooled on ice. 50 μL of this mixture was transferred to a 96-well microplate, 200 μL of chromogenic solution was added, and incubated at 37°C for 30 minutes, after which the absorbance at 585 nm was measured. Hyaluronidase inhibitory activity was calculated from the change in absorbance in each sample.

[0041] (2) Results of Experiment 3 (Hyaluronidase inhibitory activity) The results of Experiment 3 are shown in Figure 3. The horizontal axis of the graph in Figure 3 shows the concentration (mM) of saclipin A or saclipin B. The dark graph on the left for each concentration shows the results for saclipin A, and the white graph on the right shows the results for saclipin B. The vertical axis shows the hyaluronidase inhibition rate (%). The error bars show the standard deviation of the three measurements.

[0042] As shown in Figure 3, both saclipin A and saclipin B exhibited hyaluronidase inhibitory activity. The hyaluronidase inhibitory activity of saclipin A and saclipin B was approximately equivalent. IC of saclipin A and saclipin B 50 Both values ​​were below 0.4 mM (400 μM). These results indicate that saclipin is also effective as a hyaluronidase activity inhibitor.

[0043] 5. Effects of the Examples This embodiment demonstrates that a useful blood pressure elevation inhibitor can be provided using compound S as the active ingredient.

[0044] The following inventions can also be understood from the above embodiments. The explanations of the specific features of the following inventions will be based on the above explanations as appropriate. An enzyme activity inhibitor that inhibits angiotensin-converting enzyme activity, comprising as an active ingredient one or more compounds selected from the group consisting of the compound represented by formula (1) above, the compound represented by formula (2) above, and derivatives thereof. An enzyme activity inhibitor that inhibits collagenase activity, comprising as an active ingredient one or more compounds selected from the group consisting of the compound represented by formula (1) above, the compound represented by formula (2) above, and derivatives thereof. An enzyme activity inhibitor that inhibits hyaluronidase activity, comprising as an active ingredient one or more compounds selected from the group consisting of the compound represented by formula (1) above, the compound represented by formula (2) above, and derivatives thereof. A food composition containing Suizenji-nori (a type of seaweed) for suppressing blood pressure increases. • A food composition for improving skin quality, containing Suizenji-nori (a type of seaweed).

[0045] This disclosure is not limited to the embodiments detailed above, and various modifications or changes are possible within the scope of the claims set forth herein.

Claims

1. A blood pressure elevation inhibitor containing as an active ingredient one or more compounds selected from the group consisting of compounds represented by the following formula (1), compounds represented by the following formula (2), and derivatives thereof. 【Chemistry 1】 【Chemistry 2】

2. The blood pressure elevation suppressant according to claim 1, which is an oral preparation.

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