JP7099821B2 - PCSK9 inhibitor and food composition for improving cholesterol metabolism - Google Patents

Description

The present invention relates to a PCSK9 inhibitor and a food composition for improving cholesterol metabolism.

In order to prevent arteriosclerosis, which is one of the main causes of cardiovascular diseases, it is considered important to remove excess bad cholesterol (LDL cholesterol). In order to control the amount of LDL cholesterol, suppression of cholesterol biosynthesis, that is, reduction of cholesterol influx has been performed in ancient times.

However, recent studies have shown that reducing cholesterol influx alone is not sufficient to reduce the risk of cardiovascular disease. Recently, attention has been paid to attempts to eliminate (clearance) bad cholesterol by reverse transfer of cholesterol. Cholesterol biosynthesized in the liver becomes LDL cholesterol and is carried to the periphery of the body. Cholesterol reverse transfer means that cholesterol that is no longer needed after being transported to the periphery is extracted from the periphery by high-density lipoprotein (HDL) and returned to the liver. HDL is responsible for the first step in the reverse transfer of cholesterol, and the ability to extract unwanted cholesterol is called the extraction ability. Extraction ability is one of the indicators of the quality of HDL. It is believed that the reverse transfer pathway of cholesterol can be activated by improving the quality of good cholesterol (HDL cholesterol).

On the other hand, the reverse transfer of cholesterol can also be activated by improving the uptake of LDL cholesterol. Cholesterol extracted from the periphery by HDL travels in the blood as HDL cholesterol, and then undergoes cholesterol transfer action and is carried to the liver by LDL for uptake. If LDL cholesterol uptake in the liver can be increased, cholesterol can be directed to excretion from the body. Therefore, the function of the LDL receptor responsible for the uptake of LDL cholesterol is important.

In recent years, attention has been focused on attempts to increase LDL receptors and promote the uptake of excess LDL cholesterol by inhibiting the proprotein convertase subtilisin / kexin type 9 (PCSK9), which affects the metabolism of LDL receptors. There is. PCSK9 is a proteolytic enzyme that binds to and degrades the LDL receptor expressed on the surface of the liver. PCSK9 inhibitors have the effect of suppressing the degradation of LDL receptors and promoting the uptake of blood LDL cholesterol into hepatocytes through PCSK9 inhibition. PCSK9 inhibitors have also begun to be manufactured and sold as pharmaceuticals in Japan.

Dong B et al., J. Biol. Chem. 290, 4047-4058 (2015) Tai et al., Mol. Nutr. Food Res. 58, 2133-2145 (2014)

However, the PCSK9 inhibitors that have been manufactured and sold are antibody drugs, and examples of reports on low-molecular-weight compounds or natural products relating to the PCSK9 inhibitory action include berberine (Non-Patent Document 1) and curcumin (Non-Patent Document 2). , Slight. Inhibiting PCSK9 inexpensively and gently with food ingredients is a great advantage for consumers.

It is an object of the present invention to provide a new PCSK9 inhibitor.

The present invention provides a PCSK9 inhibitor containing an unsaturated fatty acid having 16 to 24 carbon atoms or a salt or ester thereof as an active ingredient.

The PCSK9 inhibitor according to the present invention has a high PCSK9 inhibitory effect by containing the above compound as an active ingredient.

In the PCSK9 inhibitor, it is preferable that the unsaturated fatty acid is other than eicosapentaenoic acid.

In the PCSK9 inhibitor, the number of unsaturated bonds in the unsaturated fatty acid may be 1 to 7. As a result, the PCSK9 inhibitory action of the PCSK9 inhibitor becomes more excellent.

In the PCSK9 inhibitor, it is preferable that all unsaturated bonds in the unsaturated fatty acid are cis-type. As a result, the PCSK9 inhibitory action of the PCSK9 inhibitor becomes more excellent.

In the PCSK9 inhibitor, the unsaturated fatty acid is preferably at least one selected from the group consisting of ω-3 fatty acid, ω-6 fatty acid, ω-7 fatty acid and ω-9 fatty acid. As a result, the PCSK9 inhibitory action of the PCSK9 inhibitor becomes more excellent.

In the PCSK9 inhibitor, the unsaturated fatty acid or a salt or ester thereof may be derived from coriander.

The present invention also provides a food composition for improving cholesterol metabolism containing the above-mentioned unsaturated fatty acid or a salt or ester thereof as an active ingredient. Since the food composition contains the above-mentioned ingredient as an active ingredient, the cholesterol metabolism of the ingestor can be improved by ingesting the food composition.

The present invention provides a novel PCSK9 inhibitor.

It is a graph which shows the PCSK9 inhibitory action of the test substance in a test example.

Hereinafter, embodiments for carrying out the present invention will be described in detail. The present invention is not limited to the following embodiments.

The PCSK9 inhibitor according to the present embodiment contains an unsaturated fatty acid having 16 to 24 carbon atoms or a salt or ester thereof as an active ingredient.

The unsaturated fatty acid may be a monounsaturated fatty acid or a polyunsaturated fatty acid. The number of unsaturated bonds (double bonds) in the unsaturated fatty acid may be, for example, 1 to 7, 1 to 6, 1 to 5, or 1 to 4. The unsaturated fatty acid preferably has 16 to 22 carbon atoms.

The unsaturated bond in the unsaturated fatty acid may be trans-type or cis-type. The unsaturated fatty acid may be an all-cis type, that is, an unsaturated fatty acid in which all unsaturated bonds are cis type. Since all unsaturated bonds are cis-type, the PCSK9 inhibitory action of the PCSK9 inhibitor can be further enhanced.

The ester of the unsaturated fatty acid may be, for example, a glycerin ester or an alkyl ester of the unsaturated fatty acid. Examples of the alkyl ester include an alkyl ester having 1 to 3 carbon atoms. Examples of the alkyl ester having 1 to 3 carbon atoms include ethyl ester, methyl ester and propyl ester.

The unsaturated fatty acid is preferably non-conjugated. The unsaturated fatty acid may be, for example, at least one selected from the group consisting of ω-3 fatty acid, ω-6 fatty acid, ω-7 fatty acid and ω-9 fatty acid. The unsaturated fatty acid is preferably at least one selected from the group consisting of ω-3 fatty acid, ω-6 fatty acid and ω-7 fatty acid.

Unsaturated fatty acids having 16 to 24 carbon atoms include, for example, palmitoleic acid (16: 1), loganic acid (16: 3), oleic acid (18: 1), baxenoic acid (18: 1), linoleic acid ( 18: 2), γ-linolenic acid (18: 3), α-linolenic acid (18: 3), pinolenic acid (18: 3), stearidonic acid (18: 4), gadrain acid (20: 1), eikosen Acid (20: 1), Eikosazienoic acid (20: 2), Eikosatrienic acid (20: 3), Meadic acid (20: 3), Eikosatetraenoic acid (20: 4), Arachidonic acid (20: 4) ), Eikosapentaenoic acid (20: 5), Elcaic acid (22: 1), Docosadienoic acid (22: 2), Adrenic acid (22: 4), Kurpanodonic acid (22: 5), Docosahexaenoic acid (22: 6) , Nervonic acid (24: 1), tetracosapentaenoic acid (24: 5), hercinic acid (24: 6) and the like. The number in parentheses indicates the number of carbon atoms in the unsaturated fatty acid: the number of double bonds.

The unsaturated fatty acids are other than eicosapentaenoic acid (all cis-5,8,11,14,17-eicosapentaenoic acid) or docosahexaenoic acid (cis-4,7,10,13,16,19-docosahexaenoic acid). It is preferable that it is one.

The unsaturated fatty acid or a salt or ester thereof may be derived from a natural product or synthesized. The active ingredient may be derived from, for example, oils and fats such as vegetable oil and fish oil, and natural raw materials such as marine animals, microorganisms, and plants. The active ingredient is preferably derived from a plant.

Unsaturated fatty acids derived from natural raw materials or salts or esters thereof can be obtained from natural raw materials by subjecting them to treatments such as fractionation and purification by an appropriately known method. The unsaturated fatty acid or a salt or ester thereof may be derived from, for example, coriander. Examples of unsaturated fatty acids having 16 to 24 carbon atoms derived from coriander include loganic acid (16: 3, all-cis-7,10,13-hexadecatorienic acid).

The active ingredient derived from cilantro can be obtained, for example, by fractionating or purifying the cilantro extract. As the coriander extract, a commercially available product may be used.

When extracting from coriander, the tissue of coriander to be extracted may be, for example, coriander leaves or stems, preferably leaves. The coriander may have been subjected to processing such as heating, drying, freezing, processing, crushing, and sorting.

Extraction from cilantro can be carried out, for example, by immersing cilantro in a solvent and filtering it. As the solvent, for example, ethanol and methanol are suitable. As the solvent, one type may be used alone, or two or more types may be used in combination. Sonication may be performed during immersion. Extraction can also be performed using a supercritical fluid (eg, carbon dioxide). The obtained extract may be concentrated or dried by a known method (for example, concentration under reduced pressure, freeze-drying), or may be further subjected to a treatment such as pulverization.

Fractionation and purification of the desired active ingredient from Pakuchi extract can be performed by known methods (eg, silica gel column chromatography, gel filtration chromatography, flash chromatography, HPLC, filter filtration, centrifugation). can. Whether or not the obtained compound is a desired compound can be confirmed by a known method (for example, mass spectrometry, elemental analysis, nuclear magnetic resonance spectroscopy, ultraviolet spectroscopy, infrared spectroscopy).

The PCSK9 inhibitor according to the present embodiment may contain one selected from the group consisting of the above-mentioned unsaturated fatty acid, an ester of an unsaturated fatty acid, and a salt of an unsaturated fatty acid as an active ingredient, or may contain two or more thereof. But it may be. The content of the active ingredient in the PCSK9 inhibitor according to the present embodiment is 0.001 w / w% or more, 0.01 w / w% or more, 0.1 w / w% or more, 1 w / w% or more, 10 w / w%. It may be more than or equal to 20 w / w or more, and may be 100 w / w% or less, 90 w / w% or less, 70 w / w% or less, or 50 w / w% or less.

Since the PCSK9 inhibitor according to the present embodiment contains the above-mentioned active ingredient, the expression of PCSK9 can be suppressed by ingesting the PCSK9 inhibitor, and as a result, the degradation of the LDL receptor is suppressed. It can promote the clearance of LDL cholesterol and regulate LDL cholesterol in the blood to an appropriate range. Therefore, the PCSK9 inhibitor according to the present embodiment can be used for improving cholesterol metabolism, regulating cholesterol level, reducing LDL cholesterol level, improving high LDL cholesterol disease, and the like.

The PCSK9 inhibitor according to the present embodiment may be in any form such as a solid (for example, powder), a liquid (water-soluble or fat-soluble solution or suspension), a paste, or the like, and may be a powder, a granule, a tablet, or the like. Any dosage form such as a capsule, a solution, a suspension, an emulsion, an ointment, or a plaster may be taken. It can also take the form of a release-controlled preparation. The PCSK9 inhibitor according to the present embodiment may consist only of the above-mentioned active ingredient.

The above-mentioned various preparations include the above-mentioned active ingredients and pharmaceutically acceptable additives (excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspensions). It can be prepared by mixing with an agent or the like.

For example, examples of the excipient include lactose, sucrose, starch, dextrin and the like. Examples of the binder include polyvinyl alcohol, gum arabic, tragant, gelatin, hydroxypropylmethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone and the like. Examples of the lubricant include magnesium stearate, calcium stearate, talc and the like. Examples of the disintegrant include crystalline cellulose, agar, gelatin, calcium carbonate, sodium hydrogencarbonate, dextrin and the like. Examples of the emulsifier or surfactant include Tween60, Tween80, Span80, glycerin monostearate and the like. Examples of the base include cetostearyl alcohol, lanolin, polyethylene glycol and the like. Examples of the solubilizing agent include polyethylene glycol, propylene glycol, sodium carbonate, sodium citrate, Tween80 and the like. Examples of the suspending agent include Tween 60, Tween 80, Span80, glycerin monostearate, polyvinyl alcohol, polyvinylpyrrolidone, methyl cellulose, hydroxymethyl cellulose, sodium alginate and the like.

The PCSK9 inhibitor according to the present embodiment can be used as a component of products such as pharmaceuticals, quasi-drugs, food compositions, food additives, feeds, and feed additives. Examples of foods include breads, noodles, rice, tofu, dairy products, soy sauce, miso, confectionery, and beverages. Examples of the beverage include water, soft drinks, fruit juice beverages, milk beverages, alcoholic beverages, non-alcoholic beverages, sports drinks, nutritional drinks and the like. It can also be used as an ingredient involved in health foods, foods with functional claims, special purpose foods, nutritional supplements, supplements, foods for specified health use, and the like. When the PCSK9 inhibitor according to the present embodiment is used as a component of the above product, the content of the active ingredient in the above product is 0.001 w / w% or more and 100 w / w from the viewpoint of more effectively exerting the PCSK9 inhibitory action. It is preferably contained so as to be% or less, and more preferably 0.01% or more and 95% or less.

The product comprising or containing a PCSK9 inhibitor according to the present embodiment may be for improving cholesterol metabolism. The term "improvement of cholesterol metabolism" means, for example, including regulation of cholesterol level, reduction of LDL cholesterol level, improvement of high LDL cholesterol disease and the like. The above products are labeled to increase their ability to metabolize cholesterol, regulate cholesterol levels, reduce LDL cholesterol levels, prevent high LDL cholesterol disease, and increase cholesterol clearance. You may.

The PCSK9 inhibitor according to this embodiment may be ingested by humans or non-human mammals. The dose (intake) of the PCSK9 inhibitor according to the present embodiment may be, for example, 0.1 mg to 1 g or 1 to 500 mg per 1 kg of body weight per adult per day as an active ingredient. The dose can be appropriately determined according to the condition, age, etc. of the individual.

The PCSK9 inhibitor according to the present embodiment may be orally administered (ingested) or parenterally, but is preferably orally administered. The PCSK9 inhibitor may be administered once a day or in multiple divided doses a day as long as the amount of the active ingredient per day is within the above range.

The present invention also provides a food composition for improving cholesterol metabolism containing the above-mentioned unsaturated fatty acid or a salt or ester thereof as an active ingredient. The food composition for improving cholesterol metabolism may contain one of the above active ingredients alone, or may contain two or more of them. The food composition for improving cholesterol metabolism may be one that improves cholesterol metabolism based on the PCSK9 inhibitory action of the above active ingredient. The content of the active ingredient in the above food composition is 0.001 w / w% or more, 0.01 w / w% or more, 0.1 w / w% or more, 1 w / w% or more, 10 w / w% or more, or 20 w /. It may be w or more, and may be 100 w / w% or less, 90 w / w% or less, 70 w / w% or less, or 50 w / w% or less.

The food composition for improving cholesterol metabolism may be a beverage or a food. Examples of the beverage include soft drinks, fruit juice beverages, milk beverages, alcoholic beverages, non-alcoholic beverages, sports drinks, nutritional drinks and the like. Examples of foods include breads, noodles, rice, tofu, dairy products, soy sauce, miso, and confectionery. The food composition may be a health food, a food with a functional claim, a food for special use, a dietary supplement, a supplement or a food for specified health use.

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

[Test Example 1]
The fatty acids shown in Table 1 were used as the test substance. All unsaturated bonds in unsaturated fatty acids are cis-type.

HepG2 cells were cultured in DMEM + 10% FBS medium at 37 ° C. in a 5% CO ₂ incubator. Each of the above test substances was added to the cultured HepG2 cells at a concentration of 40 μM, and the cells were cultured in a CO ₂ incubator for 24 hours. After culturing, the medium was collected, centrifuged, and the clarified solution was subjected to expression analysis (n = 4). As a control, a substance to which a solvent (DMSO) was added instead of the test substance was used.

The amount of PCSK9 protein secreted extracellularly was evaluated by Western blotting. The results are shown in FIG. The horizontal axis shows a relative value with the control (solvent) as 1.0. The graph shows the mean ± standard deviation. In all the test substances, the amount of PCSK9 protein secreted extracellularly was lower than that of the control, confirming the effect of suppressing the expression of PCSK9. All test substances except palmitic acid, which is a saturated fatty acid, showed a significant difference with respect to the control (P <0.05). In addition, among unsaturated fatty acids, test substances other than eicosapentaenoic acid showed a significant difference from the case of targeting palmitic acid (P <0.05).

Claims (5)

An unsaturated fatty acid having 16 to 24 carbon atoms or a salt or ester thereof is contained as an active ingredient, the number of unsaturated bonds in the unsaturated fatty acid is 1 to 4, and the ester is a glycerin ester or an alkyl ester. PCSK9 inhibitor. The PCSK9 inhibitor according to claim 1 , wherein the unsaturated bonds in the unsaturated fatty acid are all cis-type. The PCSK9 inhibitor according to claim 1 or 2 , wherein the unsaturated fatty acid is at least one selected from the group consisting of ω-3 fatty acids, ω-6 fatty acids, ω-7 fatty acids and ω-9 fatty acids. The PCSK9 inhibitor according to any one of claims 1 to 3 , wherein the unsaturated fatty acid or a salt or ester thereof is derived from coriander. A food composition for improving cholesterol metabolism containing the unsaturated fatty acid according to any one of claims 1 to 4 or a salt or ester thereof as an active ingredient.

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