FABP5 production inhibitor

Abstract

To provide a FABP5 production inhibitor.SOLUTION: Provided is a FABP5 production inhibitor containing one or more selected from tranexamic acid, pantothenic acid, and pantothenyl alcohol.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to an FABP5 production inhibitor that inhibits the production of FABP5 in the body. [Background technology]

[0002] A family of proteins called fatty acid-binding proteins (FABPs) has been attracting attention. FABPs are low-molecular-weight proteins of 14 to 15 kDa that are mainly present in the cytoplasm and are thought to be involved in the uptake and transport of fatty acids and in the regulation of gene expression by forming complexes with nuclear receptors (Non-Patent Document 1). Nine types of FABPs are known in mammals, and are generally abundant in tissues with high fatty acid metabolic activity, such as cardiac muscle, skeletal muscle, intestine, liver, and adipocytes. FABPs are named after the tissue or cells from which they were first isolated, such as heart-type (H-), brain-type (B-), liver-type (L-), and intestinal-type (I-). However, because each FABP is expressed in a variety of tissues and cells, they are increasingly being designated by numbers, such as FABP3 (H-FABP), FABP5 (E-FABP), and FABP7 (B-FABP). This numbering designation will be used throughout the present specification.

[0003] FABP5 is the first FABP reported in the epidermis. FABP5 is used as a wrinkle formation marker (Patent Document 1), a marker for evaluating acne-prone skin (Patent Document 2), and a biomarker common to degenerative diseases of the nervous system (Patent Document 3). FABP5 is used as one of many markers for evaluating fatigue and stress states in Patent Document 4.

[0004] FABP5 is thought to be a cause of atherosclerosis and diabetes because the onset of these diseases is suppressed in genetically mutated or knockout mice, but its relationship to these diseases in humans has not yet been clarified.

[0005] Patent Document 5 lists a triazolopyrimidinone derivative (5-((3-chloro-2-methylphenoxy)methyl)-2-phenyl-[1,2,4]triazolo[1,5-α]pyrimidin-7(4H)-one) that has the effect of inhibiting FABP5 expression or activity as an example of a substance that inhibits FABP5, and describes that this compound and its derivatives inhibit FABP5 and have the effect of inhibiting hair growth, and also describes a method for evaluating hair growth inhibitors using FABP5 as an indicator. Patent Document 6 describes an invention in which an aqueous extract of adzuki beans, or a substance obtained by adsorbing this onto an adsorbent or carrier and eluting it with a solvent such as methanol or ethanol, is used as an active ingredient in anticancer drugs such as cancer metastasis inhibitors, FABP5 gene activity inhibitors, and cancer growth inhibitors.

[0006] Patent Document 7 describes that FABP4 and FABP5 are expressed in intestinal cells that secrete GIP, that inhibiting FABP4 / 5 reduces blood GIP levels, and that substances that inhibit FABP4 / 5 are useful as agents for suppressing increases in GIP levels. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Japanese Patent Application Laid-Open No. 2010-164404 [Patent Document 2] Japanese Patent Application Laid-Open No. 2016-95185 [Patent Document 3] Japanese Patent Application Laid-Open No. 2014-071016 [Patent Document 4] Japanese Patent Application Laid-Open No. 2013-150558 [Patent Document 5] Japanese Patent Application Laid-Open No. 2016-214085 [Patent Document 6] Japanese Patent Application Publication No. 2017-155012 [Patent Document 7] Japanese Patent Application Publication No. 2017-215336 [Non-patent literature]

[0008] [Non-Patent Document 1] Nature Reviews Drug Discovery,2008,7:489-503 Summary of the Invention [Problem to be solved by the invention]

[0009] An objective of the present invention is to provide an agent for suppressing FABP5 production. [Means for solving the problem]

[0010] The main configuration of the present invention is as follows. 1. A FABP5 production inhibitor comprising one or more selected from tranexamic acid, pantothenic acid, and pantothenyl alcohol. 2. The FABP5 production inhibitor according to 1., wherein the tranexamic acid concentration is 0.1% by weight or more and 5% by weight or less, the pantothenic acid concentration is 0.001% by weight or more and 0.02% by weight or less, and the pantothenyl alcohol concentration is 0.01% by weight or more and 0.5% by weight or less. [Effects of the Invention]

[0011] The FAPB5 production inhibitor of the present invention can inhibit the production of FABP5 in human cells. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 shows the amount of FABP5 produced in an example. DETAILED DESCRIPTION OF THE INVENTION

[0013] The FABP5 production inhibitor of the present invention comprises one or more selected from tranexamic acid, pantothenic acid, and pantothenyl alcohol. The FABP5 production inhibitor of the present invention may comprise one or more selected from the above-mentioned tranexamic acid, pantothenic acid, and pantothenyl alcohol, and may comprise two or more. As used herein, "FABP5" is an abbreviation for fatty acid-binding protein 5, meaning fatty acid-binding protein 5. Inhibition of FABP5 production means inhibition of FABP5 gene transcription, inhibition of FABP5 gene expression, and inhibition of FABP5 protein amount.

[0014] Tranexamic acid Tranexamic acid is an abbreviation for trans-4-aminomethylcyclohexane-1-carboxylic acid. In the present invention, tranexamic acid may be a salt of tranexamic acid, and examples of the salt include alkali metal salts such as sodium salt, potassium salt, calcium salt, and magnesium salt, alkaline earth metal salts, mineral acid salts such as hydrochloride, nitrate, and sulfate, and organic acid salts such as methanesulfonate. Commercially available products include "Japanese Pharmacopoeia Tranexamic Acid" (Maruzen Pharmaceutical Co., Ltd.).

[0015] Pantothenic acid Pantothenic acid, also known as vitamin B5, is found in meat, wheat germ, kidney, liver, nuts, brewer's yeast, etc. In the present invention, pantothenic acid may be a salt of pantothenic acid, and examples of the salt include alkali metal salts such as sodium salt, potassium salt, calcium salt, and magnesium salt, alkaline earth metal salts, mineral acid salts such as hydrochloride, nitrate, and sulfate, and organic acid salts such as methanesulfonate. Among these, calcium salt is preferred because it is a food additive produced by chemical synthesis, is inexpensive, and is highly safe.

[0016] Pantothenyl alcohol Pantothenyl alcohol is an alcohol derivative of pantothenic acid and is also called provitamin B5 because it is a precursor of pantothenic acid.

[0017] In the FAPB5 production inhibitor of the present invention, the concentration of tranexamic acid is not particularly limited, but is preferably 0.1% by weight or more and 5% by weight or less from the viewpoint of inhibiting the production of FABP5. The tranexamic acid concentration is more preferably 0.2% by weight or more, even more preferably 0.5% by weight or more, more preferably 4% by weight or less, even more preferably 3% by weight or less, and even more preferably 2% by weight or less.

[0018] In the FAPB5 production inhibitor of the present invention, the pantothenic acid concentration is not particularly limited, but from the viewpoint of inhibiting FABP5 production, it is preferably 0.001% by weight or more and 0.02% by weight or less, more preferably 0.0015% by weight or more, even more preferably 0.002% by weight or more, more preferably 0.015% by weight or less, even more preferably 0.012% by weight or less, and even more preferably 0.008% by weight or less.

[0019] In the FAPB5 production inhibitor of the present invention, the pantothenyl alcohol concentration is not particularly limited, but from the viewpoint of inhibiting FABP5 production, it is preferably 0.01% by weight or more and 0.5% by weight or less, more preferably 0.02% by weight or more, even more preferably 0.03% by weight or more, and more preferably 0.4% by weight or less, even more preferably 0.03% by weight or less.

[0020] The FAPB5 production inhibitor of the present invention contains one or more selected from tranexamic acid, pantothenic acid, and pantothenyl alcohol. If two or more are contained, it is preferable that any one of them satisfies the above-mentioned concentration, more preferable that any two of them satisfies the above-mentioned concentration, and even more preferable that all three of them satisfies the above-mentioned concentration.

[0021] The FAPB production inhibitor of the present invention can be used as food, beverage, health food such as supplement, pharmaceutical, cosmetic, and the like. The food may be a normal food, a nutritional supplement, a functional food, a health food, a food for specified health uses, etc. The beverage may be blended with, for example, a soft drink, a fruit drink, a whey drink, an alcoholic drink, etc. When used as a supplement, the form may be, for example, a solid preparation such as a powder, a dusting, a granule, a tablet, or a capsule, or a liquid preparation such as a solution, a suspension, or an emulsion.

[0022] For pharmaceutical use, the active ingredient can be administered in the form of a conventional pharmaceutical preparation by mixing it with a solid or liquid pharmaceutical carrier suitable for oral or parenteral administration (e.g., rectal administration or injection). Examples of such forms include solid preparations such as powders, granules, tablets, and capsules; liquid preparations such as solutions, suspensions, and emulsions; lyophilized preparations; and topical preparations, with topical preparations being preferred. These preparations can be prepared by conventional methods. Examples of such pharmaceutical carriers include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glycerides, polyethylene glycol, hydroxyethylene starch, ethylene glycol, polyoxyethylene sorbitan fatty acid esters, amino acids, gelatin, albumin, water, and physiological saline. If necessary, additives such as stabilizers, wetting agents, emulsifiers, binders, and isotonic agents can be added as appropriate.

[0023] The dosage of the FABP5 production inhibitor of the present invention is appropriately selected and determined depending on the age, weight, symptoms, administration route, administration schedule, formulation, etc. of the subject. For oral administration, 0.001 to 100 g per day is preferred. For external use, a composition containing 0.001 to 1% by mass is applied multiple times per day.

[0024] When the FABP5 production inhibitor of the present invention is used in cosmetics, various ingredients commonly used in ordinary cosmetics can be appropriately blended within a range that does not impair the effects of the present invention. Examples of such other ingredients that can be blended include oils, surfactants, moisturizers, thickeners, colorants, alcohols, UV protection agents, amino acids, vitamins, whitening agents, organic acids, inorganic salts, enzymes, antioxidants, stabilizers, preservatives, disinfectants, skin activators, blood circulation promoters, antiseborrheic agents, anti-inflammatory agents, sequestering agents, pH adjusters, astringents, refreshing agents, fragrances, pigments, and water.

[0025] The form of the cosmetic containing the FABP5 production inhibitor of the present invention is not particularly limited, and can be used, for example, in the form of lotion, emulsion, cream, serum, foundation, etc., as skin care cosmetics, makeup cosmetics, massage cosmetics, and pack cosmetics. [Example]

[0026] The present invention will be specifically described below with reference to test examples.

[0027] FABP5 production inhibition test (1) Test method 1) Cells used in the test Normal human epidermal keratinocytes (NHEK passage 4 / Thermo Fisher Scientific).

[0028] 2) Cell culture conditions Cells were cultured at 37°C under 5% carbon dioxide and 95% CO₂ atmosphere. The medium used was EpiLife Medium with 60μM Calcium (Life Technologies Japan #M-EPI-500-CA) supplemented with HuMedia-KG Growth Additive Set (Kurabo #KK-6150). For standard culture, cells were seeded into 75cm flasks, and when 80-90% confluent, 0.05% Trypsin-EDTA (Sigma-Aldrich) was added to detach the cells, followed by subculture.

[0029] 3) Test sample The test samples (tranexamic acid, calcium pantothenate, pantothenyl alcohol) were all dissolved in 50 μM oleic acid before the test. Untreated medium served as a control. Medium supplemented with 50 μM oleic acid alone was also tested. Note that oleic acid was added to stimulate FABP5 production.

[0030] 4) Stimulation of FABP5 production by LPS and fatty acids 4 × 10 NHEK cells were cultured in a 24-well plate. 4 Cells were seeded at 1000 cells / well and pre-cultured for 24 hours. Then, cells were cultured for 8 hours in a medium supplemented with 40 μg / ml LPS (lipopolysaccharide, derived from Escherichia coli 055:B5: Sigma-Aldrich). The medium was then replaced with a test sample dissolved in 50 μM oleic acid, and the cells were cultured for 20 hours.

[0031] 5) Protein extraction The cells were lysed in Cell Lysis Buffer, shaken for 20 minutes, and then the cell lysate was collected. The protein content of the collected cell lysate was measured using a BCA protein kit according to the protocol provided with the kit.

[0032] 6) Measurement of FABP5 100 μl of anti-FABP5 antibody (Bio Vender) diluted with PBS was added to a 96-well EIA / RIA plate, and the plate was stored at 4° C. to immobilize the antibody. The next day, the cells were washed three times with 300 μl of wash buffer, then excess liquid was removed, and Reagent Diluent (R&D) was added and blocked for 1 hour at 25°C. After washing three times with 300 μl of PBS-T (0.05%) and removing excess liquid, 100 μl each of FABP5 (Funakoshi Co., Ltd.) diluted with cell lysis buffer and the test sample were added. After reacting at 37°C for 1.5 hours, the plate was washed three times with 300 μl of PBS-T (0.05%), and then excess liquid was removed. 100 μl of anti-FABP5 antibody (R&D) diluted with PBS-T (0.05%) was added and reacted at 37°C for 1 hour. After the reaction, the plate was washed three times with 300 μl of PBS-T (0.05%), and excess solution was removed. 100 μl of anti-Streptavidin-HRP2 diluted in PBS-T (0.05%) was added and incubated at 37°C for 30 min. The plate was washed three times with 300 μl of PBS-T (0.05%), and excess solution was removed. 100 μl of TMB One solution (Promega) was added and incubated at room temperature until the color changed. The reaction was then stopped by adding 50 μl of 2N sulfuric acid (Wako Pure Chemical Industries, Ltd.), and the absorbance at 450 nm was measured. The amount of FABP5 produced was measured by comparing it with a previously prepared calibration curve. The resulting FABP5 measurement was normalized by dividing it by the amount of cellular protein to determine the amount of FABP5 produced.

[0033] (2) Test results The results are shown in Figure 1. Tranexamic acid, pantothenic acid, and pantothenyl alcohol were able to suppress the production of FABP5 in cells stimulated with oleic acid.

Claims

1. A FABP5 production inhibitor containing tranexamic acid.

2. The FABP5 production inhibitor according to claim 1, wherein the concentration of the tranexamic acid is 0.1% by weight or more and 5% by weight or less.

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