WO2013015288A1 - Lipase activity inhibitor - Google Patents

Abstract

Provided are: a lipase activity inhibitor having excellent lipase activity inhibitory action; a cosmetic containing said lipase activity inhibitor; use of a phospholipid for manufacturing said cosmetic; use of a phospholipid for manufacturing a drug, anti-acne agent, anti-obesity agent or anti-inflammatory agent containing said lipase activity inhibitor; a phospholipid for use in inhibiting lipase activity; a phospholipid for use in preventing or treating acne, obesity or inflammation; and a method for preventing or treating acne, obesity or inflammation, including administration of said phospholipids. The present invention pertains to: a lipase activity inhibitor containing a phospholipid as the active ingredient, and a cosmetic, drug, anti-acne agent, anti-obesity agent or anti-inflammatory agent containing said lipase activity inhibitor; use of a phospholipid for manufacturing said cosmetic, drug, anti-acne agent, anti-obesity agent or anti-inflammatory agent; a phospholipid for use in inhibiting lipase activity; a phospholipid for use in preventing or treating acne, obesity or inflammation; and a method for preventing or treating acne, obesity or inflammation, including administration of said phospholipids.

Description

Lipase activity inhibitor

The present invention relates to a lipase activity inhibitor, a cosmetic containing the lipase activity inhibitor, use of a phospholipid for producing the cosmetic, a medicament containing the lipase activity inhibitor, and an anti-acne agent, anti-obesity It relates to the use of phospholipids for producing an agent or an anti-inflammatory agent.

Lipase is a general term for enzymes (including phospholipases A and B) that hydrolyze fatty acid esters. It is found in animal pancreatic juice, intestinal wall / liver, etc., or plant seeds, molds, bacteria, etc., and is involved in lipid metabolism in cells of many organisms.

In the skin, fatty acids are produced from sebum by the action of lipase produced by microorganisms present in the skin or lipase of epidermal keratinocytes, and the resulting fatty acids are one of the causes of acne and dermatitis. It is considered. For example, bacterial lipases in the human body include lipases produced by microorganisms that are resident in the skin surface (for example, Propionibacterium acnes (P. acnes), etc.). The above lipase decomposes triglycerides contained in sebum and produces free fatty acids. This free fatty acid is considered as one of the factors causing an irritating inflammatory reaction to the skin and causing skin diseases such as acne, dermatitis and dandruff. In particular, there is a correlation between the number of P. acnes bacteria that are responsible for acne and the amount of free fatty acids produced, and the lipase produced by P. acnes acts as a stimulating inflammatory reaction against the hair follicle wall. It is thought that the accompanying keratinization abnormality and the formation of comed are caused (Non-patent Document 1).

Conventionally, abnormal fat deposition has occurred in the adipose tissue and various organs of the body, and the resulting obesity has been closely related to the development of various lifestyle-related diseases such as hypertension, arteriosclerosis and diabetes. It is believed that. Obesity is said to be caused by constitutional factors, dietary factors, mental factors, central factors, metabolic factors, lack of exercise, etc., resulting in caloric intake exceeding calorie consumption and fat accumulation. ing. Lipase, which is a kind of digestive enzyme, degrades fats and oils in food and drink that are eaten and consumed. Therefore, treating and preventing obesity by inhibiting the action of lipase and various lifestyle-related diseases (eg, hypertension, diabetes, hyperlipidemia, atherosclerosis or arteriosclerosis) caused by obesity. Will be possible.

However, development of a drug that inhibits bacterial lipase to suppress or prevent diseases or symptoms has not been sufficiently developed yet, and loquat extract (Patent Document 2) as a metal salt (Patent Document 1) or a plant extract. ) Or Kola de Cavallo extract (Patent Document 3) has been reported, but cannot exhibit lipase inhibitory action due to the relationship with other compounding ingredients, and is not necessarily in terms of safety and effectiveness in topical application. It is not satisfactory. Further, triterpenoid saponin compounds have been disclosed as lipase inhibitors (Patent Document 4).

JP 59-186919 A JP-A-10-265364 JP-A-11-228338 JP 2004-43373 A

McGinley, K. J., et al., J. Clin. Microbiol. 12, 672-675 (1980)

Therefore, a lipase activity inhibitor having an excellent lipase activity inhibitory action, a cosmetic containing the lipase activity inhibitor, use of a phospholipid for producing the cosmetic, a medicament containing the lipase activity inhibitor, and It has been desired to use phospholipids to produce anti-acne, anti-obesity or anti-inflammatory agents. Also, phospholipids for use in inhibiting lipase activity, acne, phospholipids for use in the prevention or treatment of obesity or inflammation, and methods for preventing or treating acne, obesity or inflammation including administration of the phospholipids Development was desired.

The present invention relates to a lipase activity inhibitor having an excellent lipase activity inhibitory activity, a cosmetic containing the lipase activity inhibitor, use of a phospholipid for producing the cosmetic, a medicament containing the lipase activity inhibitor And the use of phospholipids for producing anti-acne, anti-obesity or anti-inflammatory agents. Another subject of the present invention is a phospholipid for use in inhibiting lipase activity, acne, phospholipid for use in the prevention or treatment of obesity or inflammation, and acne, obesity or administration comprising administration of the phospholipid. It is to provide a method for preventing or treating inflammation.

As a result of intensive studies, the present inventor has found that a specific phospholipid has a lipase activity inhibitory action and the like, and has completed the present invention.

That is, the present invention relates to the following.
(1) A lipase activity inhibitor containing phospholipid as an active ingredient.
(2) The phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine and phosphatidylglycerol ( 1) The inhibitor according to the above.
(3) The phospholipid is a mixture of two or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. The inhibitor according to (1) above.
(4) A cosmetic comprising the lipase activity inhibitor according to any one of (1) to (3).
(5) Use of a phospholipid for producing the cosmetic according to (4).
(6) The phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol ( 5) Use as described.
(7) The phospholipid is a mixture of two or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Use of said (5) description.
(8) A medicament comprising the lipase activity inhibitor according to any one of (1) to (3).
(9) The medicament according to (8) above, which is an anti-acne agent, anti-obesity agent or anti-inflammatory agent.
(10) The medicament according to (8) or (9) above, which is an external preparation for skin.
(11) Use of a phospholipid for producing an anti-acne agent, an anti-obesity agent or an anti-inflammatory agent.
(12) The phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine and phosphatidylglycerol ( 11) Use as described.
(13) The phospholipid is a mixture of two or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Use of said (11) description.
(14) A phospholipid for use in inhibiting lipase activity.
(15) Phosphorus according to (14), which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Lipids.
(16) The phospholipid according to (14), which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol.
(17) A phospholipid for use in the prevention or treatment of acne, obesity or inflammation.
(18) Phosphorus according to (17), which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Lipids.
(19) The phospholipid according to (17), which is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol.
(20) A method for preventing or treating acne, obesity or inflammation, which comprises administering an effective amount of phospholipid to a person or animal to be administered.
(21) The method according to (20) above, which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. .
(22) The method according to (20), wherein the method is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol.

According to the present invention, a lipase activity inhibitor having an excellent lipase activity inhibitory activity, a cosmetic containing the lipase activity inhibitor, use of a phospholipid for producing the cosmetic, and containing the lipase activity inhibitor And the use of phospholipids for the manufacture of anti-acne, anti-obesity or anti-inflammatory agents. Furthermore, according to the present invention, phospholipids for use in inhibiting lipase activity, acne, phospholipids for use in the prevention or treatment of obesity or inflammation, and acne, obesity or inflammation comprising administration of the phospholipid. Prophylactic or therapeutic methods can be provided.

The present invention relates to a lipase activity inhibitor containing phospholipid as an active ingredient. Another preferred embodiment of the present invention relates to a phospholipid for use in inhibiting lipase activity. In the present invention, “phospholipid” means glycerophospholipid. The glycerophospholipid is a phospholipid having a glycerol skeleton, and means a compound in which a fatty acid is linked to the 1-position and / or 2-position of glycerin, and phosphoric acid is linked to the 3-position. Fatty acid, glycerin, phosphoric acid Etc. as constituent elements.

In the present invention, the phospholipids may be used alone or in combination of two or more. Preferably lysophosphatidylglycerol (LPG), lysophosphatidic acid (LPA), lysophosphatidylcholine (LPC), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylserine (PS) ) And phosphatidylglycerol (PG), or lysophosphatidylglycerol (LPG), lysophosphatidic acid (LPA), lysophosphatidylcholine (LPC), phosphatidylcholine (PC), phosphatidylethanolamine (PE) ), Phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylserine (PS) and phosphatidylglycerol (PG). Among these, from the viewpoint of inhibiting lipase activity, the phospholipid is more preferably lysophosphatidylglycerol (LPG), lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylglycerol (PG). Or at least one selected from the group consisting of lysophosphatidylglycerol (LPG), lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylglycerol (PG) A mixture of two or more. Furthermore, one or more selected from the group consisting of lysophosphatidylcholine (LPC), lysophosphatidylglycerol (LPG) and phosphatidylglycerol (PG) is more preferable, and lysophosphatidylcholine (LPC) and / or lysophosphatidylglycerol (LPG) Particularly preferred.

The fatty acid constituting the phospholipid used in the present invention may be a saturated fatty acid, an unsaturated fatty acid, or may contain both a saturated fatty acid and an unsaturated fatty acid.

The saturated fatty acid is not particularly limited. For example, butanoic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid (eg, palmitic acid) , Heptadecanoic acid, octadecanoic acid (eg stearic acid), nonadecanoic acid, eicosanoic acid (eg arachidic acid), docosanoic acid, tetracosanoic acid, hexacosanoic acid, octacosanoic acid, triacontanoic acid and the like. Preferred saturated fatty acids are tetradecanoic acid, hexadecanoic acid, octadecanoic acid, eicosanoic acid and docosanoic acid, and more preferred saturated fatty acids are tetradecanoic acid, hexadecanoic acid, octadecanoic acid and eicosanoic acid.

Although it does not specifically limit as said unsaturated fatty acid, For example, butenoic acid (for example, crotonic acid, isocrotonic acid etc.), pentenoic acid, hexenoic acid, heptenoic acid, octenoic acid, nonenoic acid, decenoic acid, undecenoic acid, dodecenoic acid ( For example, lauroleic acid etc.), tridecenoic acid, tetradecenoic acid (eg myristoleic acid, myristaleic acid etc.), pentadecenoic acid, hexadecenoic acid (eg palmitoleic acid, palmitelaidic acid etc.), heptadecenoic acid, octadecenoic acid (eg petroseric acid) , Petroceridic acid, oleic acid, elaidic acid, vaccenic acid, etc.), nonadecenoic acid, eicosenoic acid (eg, gadoleic acid, gondrenic acid, etc.), docosenoic acid (eg, erucic acid, brassicic acid, cetreic acid, etc.), tetracosenoic acid ( For example, nervonic acid) Sacosenoic acid, octacosenoic acid, triacontenoic acid, pentadienoic acid, hexadienoic acid (such as sorbic acid), peptadienoic acid, octadienoic acid, nonadienoic acid, decadienoic acid, undecadienoic acid, dodecadienoic acid, tridecadienoic acid, tetradecadienoic acid Dienoic acid, hexadecadienoic acid, heptadecadienoic acid, octadecadienoic acid (for example, linoleic acid, linoelaidic acid, etc.), eicosadienoic acid, docosadienoic acid, tetracosadenoic acid, hexacosadeienic acid, octacosadenoic acid, triacatadienoic acid, hexadecatrienoic acid Octadecatrienoic acid (eg α-linolenic acid, γ-linolenic acid etc.), eicosatrienoic acid (eg dihomo-γ-linolenic acid, mead acid etc.), docosatrienoic acid, tetracosatrienoic acid, hexaco Trienoic acid, octacosatrienoic acid, triacontatrienoic acid, octadecatetraenoic acid (eg stearidonic acid etc.), eicosatetraenoic acid (eg arachidonic acid etc.), docosatetraenoic acid (eg adrenic acid etc.), tetracosa Tetraenoic acid, hexacosatetraenoic acid, octacosatetraenoic acid, triacontatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid (such as crupadononic acid), tetracosapentaenoic acid, docosahexaenoic acid, tetracosahexaenoic acid An acid (for example, nisic acid etc.) etc. are mentioned. Preferred unsaturated fatty acids are octadecenoic acid (eg oleic acid etc.), eicosenoic acid, octadecadienoic acid (eg linoleic acid etc.), octadecatrienoic acid (eg α-linolenic acid, γ-linolenic acid etc.) and more Preferred unsaturated fatty acids are octadecenoic acid (such as oleic acid), octadecadienoic acid (such as linoleic acid), and octadecatrienoic acid (such as α-linolenic acid and γ-linolenic acid).

As the phospholipid used in the present invention, both naturally-derived phospholipids and non-naturally-derived phospholipids can be suitably used. The naturally-derived phospholipid can be obtained by extracting from a raw material containing phospholipid with water or an organic solvent. The naturally-derived phospholipids are, for example, naturally-derived phospholipids (natural lecithin, etc.) derived from plant raw materials such as soybeans and rapeseed, derived from animal raw materials such as egg yolk and fish, fungi or bacteria. May be further subjected to purification, chemical treatment, enzyme treatment, or the like. Examples of the non-naturally-derived phospholipid include chemically synthesized products and / or enzyme synthesized products. In the present invention, “lecithin” is a general term for a mixture mainly composed of various phospholipids obtained from animals or plants, and the phospholipid may be the same as described above.

The enzyme treatment is not particularly limited and can be performed according to a conventional method. For example, there is a method in which natural lecithin is reacted with an enzyme. The enzyme preferably includes lipase, phospholipase D, phospholipase A1, phospholipase A2, and the like, and these can be used alone or in combination of two or more. The enzyme reaction conditions can be appropriately determined by those skilled in the art depending on the enzyme used.

The organic solvent used for the enzyme treatment is not particularly limited, and examples thereof include hydrocarbon solvents such as hexane, heptane, and cyclohexane, ethanol, acetonitrile, acetate, petroleum ether, THF, and acetone. In the extraction, the organic solvents can be used alone or in admixture of two or more.

In addition, phospholipids can be extracted according to a conventional method. The solvent used for extraction is not particularly limited, and examples thereof include hydrocarbon solvents such as hexane, heptane, and cyclohexane, ethanol, acetonitrile, acetate ester, petroleum ether, THF, acetone, and the like. These extraction solvents are used alone. Or 2 or more types can be mixed and used. The amount of the solvent used for the extraction is not particularly limited, and can be appropriately selected according to the weight of the raw material containing the phospholipid. The temperature and extraction time for extraction are not particularly limited. In addition, extraction efficiency can be increased by standing, stirring or shaking.

In the present invention, the phospholipid obtained by the method as described above may be directly used as an active ingredient of the lipase activity inhibitor of the present invention, and the phospholipid is concentrated as long as the effect of the present invention is not impaired. You may use it, performing desalting, distribution, powdering, etc. Examples of the treated phospholipid include those obtained by concentrating under reduced pressure to evaporate the solvent to increase the solid content, those obtained by removing the coloring component by activated carbon treatment, or the liquid / liquid of the aqueous layer and the organic solvent layer. Examples thereof include those obtained by removing water-soluble components by distribution. The phospholipid used in the present invention may be purified by various normal phase or reverse phase chromatography. Furthermore, those obtained by adding an excipient such as dextrin or lactose to the extract or processed product and powdered are also preferably used.

The phospholipid used in the present invention includes lysophospholipid. The lysophospholipid is a phospholipid in which a fatty acid is ester-bonded only at either the 1-position or 2-position of the glycerol skeleton. In the present invention, the method for procuring lysophospholipid is not particularly limited, and natural (animal or plant, microorganism) -derived, synthetic or commercially available products can be used. The lysophospholipid can be obtained by a known method. For example, a method obtained by allowing phospholipase A1 (PLA1) or A2 (PLA2) to act on lecithin; a method described in JP-A-2005-261388; phospholipase extracted from natural products such as soybean lecithin and egg yolk lecithin A method of producing LPG by reacting D (PLD) and glycerol to obtain PG, and then causing PLA2 to act on the obtained PG to hydrolyze the fatty acid ester bond at the 2-position of PG; or lecithin Examples include a method of extracting LPG from enzymatically decomposed lecithin obtained by allowing PLA2 to act. The reaction system for reacting PLD or PLA2 with lecithin is not particularly limited, and may be an aqueous system or a two-phase system. The generated LPG can be analyzed by high performance liquid chromatography (HPLC) or the like. The fatty acid in LPG is not particularly limited in its type, but is preferably a fatty acid having 14 to 24 carbon atoms, more preferably 16 to 20 carbon atoms. Such a fatty acid may be linear or may have a branched chain.

The LPG can be obtained, for example, from Nagase ChemteX Corporation (trade name Nagase Riso PG). The LPG may have various properties, such as emulsifying properties, thermal stability of emulsions, surface activity, foaming properties, binding properties with proteins or starches, antibacterial properties, moisturizing properties, anti-resistance properties, and the like. It may have oxidation property, emulsion storage stability, thermal stability in aqueous solution, pH stability, or the like. Further, the LPG preferably has an action of inhibiting lipase activity, an anti-inflammatory action by inhibiting lipase activity, particularly an anti-acne action. LPG preferably has an anti-obesity effect by inhibiting lipid absorption by inhibiting lipase activity and / or an inhibitory effect on growth of P. acnes.

In the present invention, regarding phospholipids other than LPG, for example, PS is Sigma (product name: L-α-Phosphatidyl-L-serine (soybean); purity is 98% by weight or more), PE is Sigma ( Product name: L-α-Phosphatidylethanolamine (soybean); purity 98 wt% or more), LPC is Sigma (product name: L-α-Lysophosphatidylcholine (soybean); purity 99 wt% or more), PG is Avanti Polar Lipids ( Product name: L-α-Phosphatidyl-DL-glycerol (soybean); purity 99% by weight or more). Furthermore, each of LPA, PA, and PI can be produced by a conventionally known method, and can also be obtained from, for example, Nagase ChemteX Corporation.

The amount of phospholipid in the lipase activity inhibitor can be appropriately selected according to the type of the active ingredient or the shape of the lipase activity inhibitor, and is not particularly limited, but preferably in the total amount of the lipase activity inhibitor. It is 0.0002% by weight or more, more preferably 0.002% by weight or more, further preferably 0.02% by weight or more, and preferably 99.9% by weight or less. Even when two or more kinds of phospholipids are used, they can be used within the above range.

The ratio of various fatty acids in the phospholipid in the lipase activity inhibitor is preferably (1) 1% to 30% by weight of palmitic acid, (2) 1% to 30% by weight of stearic acid, and (3) oleic acid 1 % To 40% by weight, (4) linoleic acid 30% by weight or more, (5) linolenic acid 20% by weight or less, (6) arachidic acid 1% by weight or less, (7) eicosenoic acid 1% by weight or less, more preferably (1) palmitic acid 1 wt% to 20 wt%, (2) stearic acid 1 wt% to 25 wt%, (3) oleic acid 1 wt% to 35 wt%, (4) linoleic acid 35 wt% or more, (5) 15% by weight or less of linolenic acid.

When the lipase activity inhibitor includes a mixture of phospholipids, for example, when PS and PI are included, PS is preferably 0.0006% by weight or more, more preferably 0.006% by weight or more in the total amount of the lipase activity inhibitor. It is 006 weight% or more, Preferably it is 99.9 weight% or less. PI is preferably 0.0004% by weight or more, more preferably 0.004% by weight or more, and preferably 99.9% by weight or less in the total amount of the lipase activity inhibitor.

The lipase activity inhibitor containing the above phospholipid as an active ingredient has an effect of inhibiting P. acnes-derived lipase activity, which is said to cause acne. P. acnes is a bacterium that is resident in the skin and is one of the causes of acne. P.acnes proliferates, P.acnes-derived lipase breaks down triglycerides, and free fatty acids destroy epidermal cells, causing skin inflammation. In the present invention, when a lipase activity inhibitor containing a phospholipid as an active ingredient is used, the activity of lipase of P. acnes can be inhibited, and at the same time, the growth thereof can be suppressed, and the inflammation of the skin is enhanced. Can be suppressed.

The lipase activity inhibitor used in the present invention has the ability to inhibit lipase activity derived from any animal, for example, mammals such as humans, mice, rats, horses, dogs, cats, rabbits, pigs, cattle pancreas, etc. It is preferable to have lipase activity inhibiting ability derived from any bacteria such as P. acnes. As a method for measuring the inhibition of lipase activity, for example, a fluorescence measurement method using, for example, 4-methylumbelliferone oleate as a substrate (for example, fluorescence intensity measurement device (automatic fluorescence measurement system Cyto Fluor II manufactured by Nihon Perceptive Co., Ltd.) excitation wavelength) 360 nm: fluorescence wavelength 460 nm or excitation wavelength 355 nm: fluorescence wavelength 460 nm). The inhibitory activity of the test sample is the amount of test sample (μg / ml) that gives 50% inhibition against the activity of an enzyme solution not containing the test sample (for example, porcine pancreatic lipase 2 U / ml, pH 8), that is, IC ₅₀ Defined. In the present invention, protamine is used as a control, and a phospholipid having a lipase activity inhibitory activity equal to or higher than that of protamine is preferable.

As a method for measuring the growth inhibitory ability of P. acnes and the like by the lipase activity inhibitor of the present invention, for example, a specimen (phospholipid) solution is prepared to 2500 μg / ml using purified water, and the phospholipid is twice as much from the solution. Prepare a diluted series solution, dissolve after sterilization, add 1/9 volume of each diluted series solution of phospholipid to GAM agar kept at 50 ° C and mix well, then dispense into a petri dish and plate Separately, P.acnes was anaerobically cultured in GAM broth medium at 37 ° C. for 18-20 hours to prepare about 10 ⁶ / ml, and the bacterial solution was streaked on the plate. Examples include a method of culturing for ˜20 hours and setting the minimum concentration at which the growth of P. acnes is inhibited to the minimum growth inhibition concentration.

The lipase activity inhibitor of the present invention is mixed with a pharmaceutically acceptable carrier according to a known method, and tablets, granules, capsules, solutions, suppositories, sustained-release agents, pellets, pastes, creams, powders, etc. can do. Examples of the pharmaceutically acceptable carrier include organic carriers or inorganic carriers such as lubricants, binders, disintegrants, excipients, suspending agents, isotonic agents, buffering agents and the like. The mixing ratio of the carrier in the lipase activity inhibitor can be appropriately selected depending on the administration subject, administration route and the like. Since these preparation techniques have been well established, these various known additives and preparation techniques may also be used in the present invention.

The lipase activity inhibitor of the present invention may contain any additive as long as the effects of the present invention are not impaired. Examples of the additive include, but are not particularly limited to, for example, a humectant, a touch improver, a surfactant, a polymer, a thickener, a gelling agent, a solvent, a propellant, an antioxidant, a reducing agent, an oxidizing agent, Preservatives, antibacterial agents, chelating agents, pH adjusters, acids, alkalis, inorganic salts, UV absorbers, whitening agents, vitamins, vitamin derivatives, hair growth agents, blood circulation promoters, stimulants, hormones, anti-wrinkles Agent, anti-aging agent, attractant, astringent, cooling sensation, warming sensation, wound healing promoter, irritation relieving agent, analgesic, cell activator, plant extract, animal extract, microbial extract, antipruritic agent, exfoliating Agents, keratolytic agents, peeling agents, antiperspirants, refreshing agents, enzymes, nucleic acids, fragrances, pigments (for example, colorants, dyes, pigments, etc.), water and the like. These may be added alone or in combination. Since techniques for adding these additives have been well established in the past, these known addition techniques may also be used in the present invention.

The lipase activity inhibitor of the present invention is administered, for example, orally, parenterally, such as subcutaneous, intravenous, intraarterial, intranodal, intramedullary, intrathecal, intraventricular, intranasal, intrabronchial, transdermal, intranodal. Intrarectal, intraperitoneal, intramuscular, intrapulmonary, intravaginal, intrarectal or intraocular administration can be used.

The lipase activity inhibitor of the present invention can be contained in cosmetics or medicines. The form of the product containing the lipase activity inhibitor is not particularly limited, and specifically, for example, face wash, cleansing foam, powder wash, face wash powder, cleansing lotion, cleansing gel, cleansing cream, cleansing milk, cleansing oil, cleansing Mask, lotion, soft lotion, astringent lotion, cleaning lotion, multi-layer lotion, emulsion, emollient lotion, moisture lotion, milky lotion, nourishing lotion, nourishing milk, sun protect, sun protector, ultraviolet ray ( UV) Care Milk, Sunscreen, Makeup Lotion, Makeup Cream, Hand Lotion, Hand Cream, Body Lotion, Body Cream, Emollient Cream, Moisture Cream, Sakae Cream, base cream, pre-makeup cream, sunscreen cream, suntan cream, hair removal cream, deodorant cream, shaving cream, soap, cosmetic soap, medicated soap, liquid soap, shaving soap, synthetic soap, body shampoo, body rinse, body Powder, Pack, Mask, Essence, Moisturizing Essence, Whitening Essence, UV Protection Essence, Beauty Liquid, Basic Cosmetics, White Powder, Dusts, Foundations, Lipsticks, Lip Balm, Lip Gloss, Blusher, Eyeliner, Mascara, Examples include eye shadow, eyebrow, eyebrow, nail enamel, enamel remover, nail treatment, deodorant cosmetic, insect repellent spray, ointment, patch, lotion, and bath preparation.

In a preferred embodiment of the present invention, the lipase activity inhibitor is used as a cosmetic or a skin external preparation. The lipase activity inhibitor itself or a cosmetic or medicine containing the lipase activity inhibitor may be directly applied to or sprayed onto the skin. After being used as soap or shampoo, it may be washed away or left on for a while like makeup cosmetics. It may also be worn by impregnating or spraying clothes.

The cosmetic is not particularly limited, for example, lotion, cosmetic emulsion, cosmetic liquid, cosmetic cream, cosmetic gel, cosmetic lotion, oil, pack agent, facial cleanser, cleansing agent, Basic cosmetics such as soaps, body soaps, body creams, body lotions, hand creams, ointments; hair cosmetics such as shampoos, rinses, hair treatments, hair preparations, permanents, hair nicks, hair dyes, hair growth / hair restorations; Makeup cosmetics such as foundation, lipstick, blusher, eye shadow, eyeliner, mascara, teak; shaving agent, skin cleanser, perfume, skin deodorant, antiperspirant, dentifrice, mouthwash Oral odor prevention agents, oral odor eliminating agents, mouth fresheners, mouthwashes such as gargles, bath agents, sunscreens and the like. The cosmetic of the present invention includes medicinal cosmetics. Among these, skin lotion, cosmetic emulsion, cosmetic lotion, cosmetic cream, cosmetic liquid, cosmetic gel, oil, pack agent, facial cleanser, soap, cleansing agent, body soap, body cream, body Lotion, hand cream, ointment, sunscreen, etc.

Preferred embodiments of the present invention relate to cosmetics (including medicinal cosmetics) containing the lipase activity inhibitor, and preferably to anti-acne cosmetics. The amount of lipase activity inhibitor is preferably adjusted according to the amount of phospholipid contained. The phospholipid used in the cosmetic and the amount used thereof may be the same as described above. The cosmetic of the present invention may contain any of the above additives in addition to the lipase activity inhibitor.

The cosmetic of the present invention can be mixed with a carrier acceptable for cosmetics according to a known method to form a liquid, pellets, paste, cream, powder or the like. Examples of carriers acceptable for cosmetics include organic carriers or inorganic carriers such as lubricants, binders, disintegrants, excipients, suspending agents, isotonic agents, buffers and the like. The mixing ratio of the carrier in the cosmetic can be appropriately selected depending on the administration subject, administration route and the like. Since these preparation techniques have been well established, these various known additives and preparation techniques may also be used in the present invention.

The amount of the lipase activity inhibitor contained in the cosmetic can be appropriately selected according to the type or form of the cosmetic, and is preferably adjusted according to the content of the phospholipid in the lipase activity inhibitor. Phospholipid is used as an active ingredient of the cosmetic of the present invention. The phospholipid used may be the same as the lipase activity inhibitor. The content of the phospholipid in the cosmetic containing the lipase activity inhibitor is not particularly limited, but is preferably 0.0002% by weight or more in the cosmetic, more preferably from the viewpoint of the lipase activity inhibitory effect. It is 002% by weight or more, more preferably 0.02% by weight or more. When the cosmetic contains a mixture of two or more phospholipids, for example, when PS and PI are contained, the PS in the cosmetic is preferably 0.0006% by weight or more, more preferably 0. 0.006% by weight or more, preferably 99.9% by weight or less. PI in cosmetics is preferably 0.0004 wt% or more, more preferably 0.004 wt% or more, and preferably 99.9 wt% or less. The ratio of various fatty acids of phospholipids in the cosmetic is preferably (1) 1% to 30% by weight of palmitic acid, (2) 1% to 30% by weight of stearic acid, and (3) oleic acid 1 % To 40% by weight, (4) linoleic acid 30% by weight or more, (5) linolenic acid 20% by weight or less, (6) arachidic acid 1% by weight or less, (7) eicosenoic acid 1% by weight or less, more preferably (1) palmitic acid 1 wt% to 20 wt%, (2) stearic acid 1 wt% to 25 wt%, (3) oleic acid 1 wt% to 35 wt%, (4) linoleic acid 35 wt% or more, (5) 15% by weight or less of linolenic acid. The cosmetic dosage form and administration method may be the same as described above. Furthermore, in the application of the cosmetic of the present invention, the unit application amount of phospholipid is not particularly limited and can be appropriately selected depending on the age, sex, application location, etc. of the application target. or animal body surface area 1 cm ² per are at 0.001 [mu] g / day or more, from the viewpoint of lipase activity inhibitory effect, preferably 0.004 / day or more, more preferably 0.025 / day or more.

Another preferred embodiment of the present invention relates to the use of a phospholipid for producing the cosmetic. For details regarding the use, reference is made to the description regarding the cosmetic.

A preferred embodiment of the present invention relates to a medicine (including quasi-drug) containing the lipase activity inhibitor. The medicament is preferably an anti-acne agent, an anti-obesity agent or an anti-inflammatory agent. The method for applying the medicine is not particularly limited, and any of oral administration and parenteral administration may be used.

The medicine to be applied by parenteral administration is not particularly limited, but an external preparation is preferable. The external preparation is preferably a skin external preparation. The dosage form of the external preparation for skin is not particularly limited, but is preferably an ointment, cream, lotion, spray, fat, powder, liquid, suspension, emulsion, gel, liniment, poultice, pack. Etc. The amount of the lipase activity inhibitor in the drug can be appropriately selected depending on the form of the drug, and is preferably adjusted according to the content of phospholipid in the lipase activity inhibitor. The phospholipid is used as an active ingredient of the medicament of the present invention. The phospholipid used may be the same as the lipase activity inhibitor. The content of the phospholipid in the medicament containing the lipase activity inhibitor is preferably 0.0002% by weight or more, more preferably 0.002% by weight or more, and still more preferably 0.002% by weight or more from the viewpoint of the lipase activity inhibitory effect. 02% by weight or more. Furthermore, when the medicine contains a mixture of two or more phospholipids, for example, when PS and PI are contained, the PS in the medicine is preferably 0.0006% by weight or more, more preferably 0. 0.006% by weight or more, preferably 99.9% by weight or less. PI in medicine is preferably 0.0004% by weight or more, more preferably 0.004% by weight or more, and preferably 99.9% by weight or less. The proportion of various fatty acids in the phospholipid in the medicament is preferably (1) 1% to 30% by weight of palmitic acid, (2) 1% to 30% by weight of stearic acid, and (3) 1% of oleic acid. % To 40% by weight, (4) linoleic acid 30% by weight or more, (5) linolenic acid 20% by weight or less, (6) arachidic acid 1% by weight or less, (7) eicosenoic acid 1% by weight or less, more preferably (1) palmitic acid 1 wt% to 20 wt%, (2) stearic acid 1 wt% to 25 wt%, (3) oleic acid 1 wt% to 35 wt%, (4) linoleic acid 35 wt% or more, ( 5) Linolenic acid is not more than 15% by weight.

In the application of the external preparation of the present invention, the unit application amount of the phospholipid is not particularly limited and can be appropriately selected according to the age, sex, application state, etc. of the application target. The amount is 0.001 μg / day or more per 1 cm ² of the body surface area, and is preferably 0.004 μg / day or more, more preferably 0.025 μg / day or more from the viewpoint of the lipase activity inhibitory effect.

The administration method of the medicine is not particularly limited, and a method of administering the active ingredient by oral administration, rectal administration, injection or the like can be employed. The form of the pharmaceutical agent is not particularly limited, and a form suitable for the administration method is selected, and it can be mixed with a solid or liquid non-toxic pharmaceutical carrier to form a conventional pharmaceutical preparation. Examples of such preparations include solid preparations such as tablets, granules, powders and capsules, liquid preparations such as solutions, suspensions and emulsions, freeze-dried preparations, and the like. It can be prepared by conventional means. Examples of the non-toxic pharmaceutical carrier include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin , Water, physiological saline and the like. Further, if necessary, conventional additives such as a stabilizer, a wetting agent, an emulsifier, a binder, and an isotonic agent can be appropriately added. The dose of the drug is not particularly limited, and may be set according to usage, patient age, sex, disease severity, and other conditions. For example, phospholipid is 25 mg per day for an average adult. It can be administered to a dose of about 2000 mg, preferably about 90 mg to 1800 mg.

Another preferred embodiment of the present invention relates to the use of phospholipids for producing a medicament. For details on the use, reference is made to the above-mentioned description concerning medicine.

Still another aspect of the present invention relates to a phospholipid for use in the prevention and / or treatment of acne, obesity or inflammation. The description regarding the said pharmaceutical is referred for the description regarding this use.

Another aspect of the present invention relates to a method for preventing and / or treating acne, obesity or inflammation, which comprises administering an effective amount of phospholipid to a person or animal to be administered. For details regarding the method, reference is made to the above-mentioned description relating to medicine.

The present invention relates to a method for screening a phospholipid having a lipase activity inhibitory action as compared with protamine by bringing a lipase having a lipase activity inhibitory action into contact with the lipase. Examples of the method for screening for phospholipids as described above include a method for screening for phospholipids having a lipase activity inhibitory activity compared to protamine using the method for measuring lipase activity inhibition as described above. In the present screening method, the above-mentioned phospholipids and lipases may be used.

The application target of the lipase activity inhibitor, cosmetics and pharmaceuticals of the present invention is not particularly limited, and examples thereof include animals such as humans, cows, horses, sheep, goats, dogs, monkeys, cats, bears, rats and rabbits. Can be mentioned.

Test Example 1. (Inhibition of lipase activity by phospholipids)
Test Sample In order to examine lipase activity inhibition of phospholipid, protamine shown in Table 1 and various phospholipids were used as test samples. As protamine, “Protamine sulfate, derived from salmon” manufactured by Wako Pure Chemical Industries, Ltd. was used. This is known as a lipase activity inhibitor derived from fish and is a positive control in this test. In Table 1, “PIPS Nagase” (manufactured by Nagase ChemteX Corp .; PS approx. 30% by weight, PI approx. 20% by weight, PA approx. 13% by weight, PE approx. 9% by weight, PC approx. 2% by weight) ), PS, PE and LPC are Sigma's “L-α-Phosphatidyl-L-serine (soybean)” (purity 98% by weight or more), “L-α-Phosphatidylethanolamine (soybean)” (purity) 98% by weight), “L-α-Lysophosphatidylcholine (soybean)” (purity 99% by weight or more), and PG is “L-α-Phosphatidyl-DL-glycerol (soybean)” (Avanti Polar Lipids) ( LPG used Nagase ChemteX's "trade name Nagaserizo PG" (74% pure), and LPA, PA, and PI were each manufactured by Nagase ChemteX. Phospholipids (purities 92, 85 and 82% by weight, respectively) were used.

Preparation of enzyme solution Porcine pancreatic lipase (“lipase” manufactured by Cosmo Bio) was used as the enzyme. Porcine pancreatic lipase was prepared to 100 U / ml with 13 mM Tris-HCl buffer (pH 8) containing 150 mM sodium chloride and 1.36 mM calcium chloride, and further to 2 U / ml with the same buffer to prepare an enzyme solution.

Preparation of Substrate Solution 4-Methylumbelliferone oleate was dissolved in DMSO to make a 0.1M 4-methylumbelliferone oleate solution, which was diluted 1000 times with the above buffer solution to prepare a substrate solution. .

Preparation of test sample solution Each phospholipid was prepared in DMSO at various concentrations, and further diluted 25-fold with the above buffer solution was used as a test sample solution. Protamine was directly dissolved in the above buffer without using DMSO to prepare a test sample solution.

Measurement of lipase activity inhibition Lipase activity was measured by a fluorescence measurement method (excitation wavelength: 360 nm: fluorescence wavelength: 460 nm) using 4-methylumbelliferone oleate (sigma) as a substrate. 25 μl of the enzyme solution and 25 μl of various test sample solutions were mixed in a 96-well plate, and 50 μl of the substrate solution was added and mixed here, incubated at 37 ° C. for 30 minutes, and then released 4-methylumbelliferone. The fluorescence intensity was measured with a fluorescence intensity measuring device (fluorescence automatic measurement system CytoFluor II manufactured by Nippon Perceptive Co., Ltd.). The lipase activity was evaluated by calculating the amount of free 4-methylumbelliferone per minute from a calibration curve prepared with the fluorescence intensity of 4-methylumbelliferone. The inhibitory activity of the test sample was determined as the amount of test sample (IC ₅₀ ) that gives 50% inhibition to the activity of the enzyme solution not containing the test sample.

Results Table 1 shows the IC ₅₀ values of various phospholipids and protamine, which is known to inhibit lipase activity as a control. Various phospholipids showed inhibitory action equivalent to or better than protamine, and LPC, LPG, and PG, in particular, showed strong lipase activity inhibitory action.

Test Example 2. (Inhibition of P. acnes lipase activity by LPG)
Preparation of enzyme solution Human sebum scraped with a platinum loop was boiled and degassed and then dispersed in 10 mL of cooled physiological saline, and 1 mL of this was applied to an anaerobic bacterial GAM agar medium (Nissui). After application, the cells were cultured at 37 ° C. for 48 hours to obtain a single colony of P. acnes. The obtained colonies were cultured at 37 ° C. for 48 hours in a GAM liquid medium for anaerobic bacteria (manufactured by Nissui). The cells were collected by centrifugation, washed twice with 50 mM Tris-HCl buffer (pH 7.4), and sonicated in the same buffer under ice cooling to obtain an enzyme solution.

Measurement of lipase activity inhibition Excitation wavelength 355 nm: Fluorescence wavelength 460 nm, except that the above enzyme solution of P. acnes lipase was used as the enzyme solution and LPG was used as the phospholipid. The same was done.

Results of LPG IC ₅₀ for P.acnes lipase activity was 8.8μg / mL, inhibitory effect was observed.

Test Example 3. (Inhibition of LPG growth by P. acnes)
Preparation of plate for sensitivity measurement A sample (LPG) solution was prepared to 2500 μg / ml using purified water, and this was used as a 2-fold dilution series solution. After sterilization and dissolution, add 1/9 volume of each dilution series solution to GAM agar medium (Nissui Co., Ltd.) kept at 50 ° C and mix well. Produced.

Preparation of bacterial solution for inoculation P. acnes strains were anaerobically cultured at 37 ° C. for 18-20 hours in a GAM bouillon medium (manufactured by Nissui), and prepared so that the number of bacteria was about 10 ⁶ / mL.

Cultivation The inoculum was applied to the susceptibility measurement plate with a streak of about 2 cm and anaerobically cultured at 37 ° C. for 18-20 hours. After the cultivation, the lowest concentration at which the growth of P. acnes was inhibited was defined as the minimum growth inhibitory concentration.

Minimum growth inhibition of P. acnes by LPG The minimum growth inhibition concentration of P. acnes by LPG was 125 μg / mL, confirming the growth inhibitory action.

According to the above results, the phospholipid has a lipase activity inhibitory action and a P. acnes growth inhibitory action. In addition, since acne is caused by lipase produced by P. acnes, a phospholipid having a lipase activity inhibitory action or a P. acnes growth inhibitory action has an anti-acne action. Furthermore, since lipase activity is involved in inflammation and lipid absorption, phospholipids having a lipase activity inhibitory action have anti-inflammatory action and anti-obesity action.

According to the present invention, a lipase activity inhibitor, a cosmetic containing the lipase activity inhibitor, use of a phospholipid for producing the cosmetic, a medicament containing the lipase activity inhibitor, and an anti-acne agent, It becomes possible to provide the use of phospholipids for producing anti-obesity agents or anti-inflammatory agents. The medicine can be used as, for example, an anti-acne agent, anti-obesity agent, or anti-inflammatory agent, and the cosmetic can be used as, for example, an anti-acne cosmetic. Furthermore, according to the present invention, phospholipids for use in inhibiting lipase activity, acne, phospholipids for use in the prevention or treatment of obesity or inflammation, and acne, obesity or inflammation comprising administration of the phospholipid. Prophylactic or therapeutic methods can be provided.

Claims (22)

A lipase activity inhibitor containing phospholipid as an active ingredient. The phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Inhibitor. The phospholipid is a mixture of two or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. The described inhibitors. Cosmetics containing the lipase activity inhibitor according to any one of claims 1 to 3. Use of a phospholipid for producing the cosmetic according to claim 4. The phospholipid is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. use. 6. The phospholipid is a mixture of two or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Use of description. A medicament comprising the lipase activity inhibitor according to any one of claims 1 to 3. The medicament according to claim 8, which is an anti-acne agent, an anti-obesity agent or an anti-inflammatory agent. The medicament according to claim 8 or 9, which is an external preparation for skin. • Use of phospholipids to produce anti-acne, anti-obesity or anti-inflammatory agents. The phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. use. The phospholipid is a mixture of two or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Use of description. Phospholipid for use in inhibiting lipase activity. The phospholipid according to claim 14, which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. The phospholipid according to claim 14, which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol. Phospholipids for use in the prevention or treatment of acne, obesity or inflammation. The phospholipid according to claim 17, which is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine and phosphatidylglycerol. The phospholipid according to claim 17, wherein the phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol. A method for preventing or treating acne, obesity or inflammation, which comprises administering an effective amount of phospholipid to a person or animal to be administered. The phospholipid is one or more selected from the group consisting of lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, and phosphatidylglycerol. Method. The method according to claim 20, wherein the phospholipid is at least one selected from the group consisting of lysophosphatidylglycerol, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol.