Inhibitor containing lecithin as active ingredient

Abstract

To discover new physiological activities of lecithin and to provide new applications thereof.SOLUTION: Lecithin exhibits excellent physiological functions including inhibition of interleukin-1α production, inhibition of cyclooxygenase-2 production, inhibition of endothelin-1 production, and inhibition of DNA damage, and thus can be suitably employed as an active ingredient of an interleukin-1α production inhibitor, a cyclooxygenase-2 production inhibitor, an endothelin-1 production inhibitor, and a DNA damage inhibitor.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an interleukin-1α production inhibitor, a cyclooxygenase-2 production inhibitor, an endothelin-1 production inhibitor, and a DNA damage inhibitor, each of which contains lecithin as an active ingredient. [Background technology]

[0002] Lecithin is a general term for a mixture primarily composed of various phospholipids obtained from animals or plants. It is a naturally derived surfactant that is not irritating to the skin and is widely used in the fields of food, cosmetics, pharmaceuticals, etc. Lecithin is primarily used as an emulsifier, but is also used as a liposome membrane material, moisturizer, texture improver, etc. The physiological effects of lecithin have been reported to include skin whitening (Reference 1) and collagen production promotion (Reference 2). However, despite lecithin's widespread use in a wide range of fields, its physiological effects have not yet been fully studied. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 62-056411 [Patent Document 2] Patent Publication No. 2013-060476 Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to discover new physiological effects of lecithin and to provide new uses thereof. [Means for solving the problem]

[0005] As a result of extensive research conducted by the present inventors to solve the above problems, they discovered that lecithin has various physiological effects, and thus completed the present invention. [Effects of the Invention]

[0006] Lecithin has excellent physiological effects such as an inhibitory effect on interleukin-1α production, an inhibitory effect on cyclooxygenase-2 production, an inhibitory effect on endothelin-1 production, and an inhibitory effect on DNA damage, etc. Therefore, lecithin can be preferably used as an active ingredient of an inhibitor of interleukin-1α production, an inhibitor of cyclooxygenase-2 production, an inhibitor of endothelin-1 production, or an inhibitor of DNA damage. DETAILED DESCRIPTION OF THE INVENTION

[0007] The present invention relates to an interleukin-1α production inhibitor, a cyclooxygenase-2 production inhibitor, an endothelin-1 production inhibitor, and a DNA damage inhibitor, each of which contains lecithin as an active ingredient.

[0008] Specific examples of lecithin used in the present invention include lecithin obtained from plants such as soybean, rapeseed, sunflower, safflower, peanut, cottonseed, corn, rice, and barley, as well as egg yolk, hydrogenated products thereof, and lyso forms thereof. Lecithin with different phosphatidylcholine (hereinafter referred to as PC) contents depending on the degree of purification is commercially available, and lecithin with any PC content can be used in the present invention. Among these, lecithin obtained from soybean or sunflower and hydrogenated products thereof are preferred from the viewpoint of availability.

[0009] As shown in the tests in the Examples described below, lecithin has excellent physiological effects such as an inhibitory effect on interleukin-1α production, an inhibitory effect on cyclooxygenase-2 production, an inhibitory effect on endothelin-1 production, and an inhibitory effect on DNA damage. Therefore, based on these effects, lecithin can be preferably used as an active ingredient in an inhibitor of interleukin-1α production, an inhibitor of cyclooxygenase-2 production, an inhibitor of endothelin-1 production, or an inhibitor of DNA damage.

[0010] The amount of lecithin to be added to the interleukin-1α production inhibitor, cyclooxygenase-2 production inhibitor, endothelin-1 production inhibitor, or DNA damage inhibitor is not particularly limited, and may be added in an amount of 0.01 to 99% by weight, preferably 0.1 to 90% by weight.

[0011] The various inhibitors of the present invention containing lecithin as an active ingredient preferably further contain sterols. Specific examples of sterols include animal-derived sterols such as cholesterol, dihydrocholesterol, lanosterol, dihydrolanosterol, and desmosterol; plant-derived sterols such as stigmasterol, sitosterol, campesterol, brassicasterol, and mixtures thereof such as phytosterols; microbial-derived sterols such as ergosterol; and γ-oryzanol. These may be used alone or in combination of two or more. Among these, cholesterol, phytosterol, and γ-oryzanol are preferred. The combined use of such components can further enhance the effects of the present invention. The amount of sterols to be incorporated into the various inhibitors of the present invention is not particularly limited, and is generally 0.01 to 40% by weight, preferably about 0.05 to 30% by weight.

[0012] When lecithin and sterols are used in combination as the active ingredient of the present invention, a composition can be used in which the active ingredient is composed mainly of lecithin and sterols, which are pre-complexed or premixed to improve the dispersibility and solubility of the lecithin. Such compositions are commercially available from Nippon Fine Chemical Co., Ltd., as listed in Tables 1 and 2. These products are used as liposome-forming agents or emulsifiers, but can also be used as the active ingredient of the present invention. The Presome / Phytopresome / PrimeLipid series use hydrogenated soybean lecithin with a high PC content, while the Composite / Phytocompo series uses hydrogenated soybean lecithin with a low PC content.

[0013] [Table 1]

[0014] [Table 2]

[0015] The various inhibitors of the present invention containing lecithin as an active ingredient can further contain known components having anti-inflammatory effects, such as tranexamic acid, glycyrrhizinic acid and its derivatives, glycyrrhetinic acid and its derivatives, salicylic acid and its derivatives, hinokitiol, guaiazulene, allantoin, indomethacin, zinc oxide, hydrocortisone acetate, prednisone, diphedramine hydrochloride, chlorpheniramine maleate, peach leaf extract, mugwort leaf extract, phytosterol esters such as phytosteryl oleate, phytosteryl isostearate, phytosteryl macadamia nut oil fatty acid, and phytosteryl sunflower seed oil fatty acid. The amount of these components to be added to the various inhibitors of the present invention is not particularly limited, and is generally 0.001 to 30% by weight, preferably about 0.01 to 10% by weight.

[0016] Methods for administering to a living body interleukin-1α production inhibitors, cyclooxygenase-2 production inhibitors, endothelin-1 production inhibitors, and DNA damage inhibitors containing lecithin as an active ingredient include oral administration, injection, and transdermal administration. The dosage is not particularly limited as long as it achieves the effects of the present invention, and may be adjusted appropriately depending on the formulation, application site, age, sex, etc. Lecithin may be used as is, or may be dissolved, dispersed, or mixed in a common base such as water, gel, polyhydric alcohol, petrolatum, paraffin, vegetable oil, or silicone oil. Various additives may also be used as needed. Additives that can be used are not particularly limited as long as they are commonly used to obtain the desired dosage form, and known additives such as excipients, colorants, thickeners, binders, disintegrants, dispersants, stabilizers, gelling agents, antioxidants, surfactants, preservatives, moisturizers, and pH adjusters may be appropriately selected and used. Alternatively, the active ingredient may be incorporated into pharmaceuticals or cosmetics to exert the desired effects. In the case of pharmaceuticals, they can be incorporated into oral preparations such as tablets, capsules, granules, powders, liquids, and suspensions; topical preparations such as dermatological preparations, patches, eye drops, nasal drops, oral preparations, and suppositories; and parenteral preparations such as drip infusions and injections.In the case of cosmetics, they can be incorporated into skin cosmetics such as lotions, lotions, gels, emulsions, serums, creams, packs, face washes, and body washes; makeup cosmetics such as foundations, lipsticks, lip glosses, and mascaras; and hair cosmetics such as shampoos, rinses, treatments, hair mists, hair waxes, setting lotions, color lotions, hair manicures, and hair growth agents.

[0017] The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples in any way.

[0018] <Suppression of interleukin-1α (IL-1α) production> (with ultraviolet irradiation) Each of the various lecithins or lecithin-containing compositions listed in Table 3 was dispersed in water to a lipid concentration of 0.2% by mass to prepare an aqueous dispersion. 1.0 × 10 normal human epidermal keratinocytes were placed on a 24-well flat plate. 5After culturing for 16 hours in HuMedia-KB2 medium (Kurabo), each aqueous dispersion was added to the medium so that the lipid concentration in the medium became 100 ppm, and the cells were cultured for 24 hours. UVB was irradiated at 40 mJ / cm. 2 After irradiation, each aqueous dispersion was added again to the medium so that the lipid concentration was 100 ppm, and the cells were cultured for an additional 24 hours. The amount of IL-1α produced (normalized by total protein amount) was then quantified using ELISA. The amount of production without lecithin was quantified as a control, and the amount of production with 40 ppm dipotassium glycyrrhizinate added was quantified as a comparison control. The results are shown in Table 3 as relative values, with the amount of IL-1α produced by the control set at 100.

[0019] [Table 3]

[0020] <Suppression of interleukin-1α (IL-1α) production> (without UV irradiation) Each of the various lecithins or lecithin-containing compositions listed in Table 4 was dispersed in water to a lipid concentration of 0.2% by mass to prepare an aqueous dispersion. 1.0 × 10 normal human epidermal keratinocytes were placed on a 24-well flat plate. 5 Cells were seeded at a concentration of 100 cells / well and cultured in HuMedia-KB2 medium (Kurabo Industries, Ltd.) for 16 hours. Each aqueous dispersion was then added to the medium to bring the lipid concentration to 100 ppm, and the culture was continued for another 24 hours. The amount of IL-1α produced (normalized by total protein) was then quantified using ELISA. The amount of production without lecithin was used as a control, and the amount of production with 40 ppm dipotassium glycyrrhizinate added was also quantified. The results are shown in Table 4 as relative values, with the amount of IL-1α produced by the control set at 100.

[0021] [Table 4]

[0022] <Suppression of cyclooxygenase-2 (COX-2) production> Each of the various lecithins or lecithin-containing compositions listed in Table 5 was dispersed in water to a lipid concentration of 0.2% by mass to prepare an aqueous dispersion. 1.0 × 10 normal human epidermal keratinocytes were placed on a 24-well flat plate. 5 After culturing for 16 hours in HuMedia-KB2 medium (Kurabo), each aqueous dispersion was added to the medium so that the lipid concentration in the medium became 100 ppm, and the cells were cultured for 24 hours. UVB was irradiated at 40 mJ / cm. 2 After irradiation, each aqueous dispersion was added again to the medium so that the lipid concentration was 100 ppm, and the medium was further cultured for 6 hours. The COX-2 gene expression level (normalized by GAPDH) was then measured using real-time PCR. The expression level was measured in the same manner without lecithin as a control. The results are shown in Table 5 as relative values, with the COX-2 gene expression level of the control set at 100.

[0023] [Table 5]

[0024] <Suppression of endothelin-1 (ET-1) production> Each of the various lecithins or lecithin-containing compositions listed in Table 6 was dispersed in water to a lipid concentration of 0.2% by mass to prepare an aqueous dispersion. 1.0 × 10 normal human epidermal keratinocytes were placed on a 24-well flat plate. 5 After culturing for 16 hours in HuMedia-KB2 medium (Kurabo), each aqueous dispersion was added to the medium so that the lipid concentration in the medium became 100 ppm, and the cells were cultured for 24 hours. UVB was irradiated at 40 mJ / cm. 2After irradiation, each aqueous dispersion was added again so that the lipid concentration in the medium became 100 ppm, and the cells were further cultured for 6 hours. Then, the expression level of the ET-1 gene (normalized by GAPDH) was measured using the real-time PCR method. The expression level in the case of no addition of lecithin was measured in the same manner as a control. The results are shown in Table 6 as relative values when the expression level of the ET-1 gene in the control was set to 100.

[0025]

Table 6

[0026] <DNA damage suppression> For various lecithin-containing compositions described in Table 7, an aqueous dispersion was prepared by dispersing them in water so that the lipid concentration became 0.2% by mass. Human normal epidermal keratinocytes were seeded in a 24-well flat plate at a concentration of 1.0×10 5 cells / well and cultured in HuMedia-KB2 medium (manufactured by Kurabo Industries Ltd.) for 16 hours. Then, each aqueous dispersion was added so that the lipid concentration in the medium became 100 ppm, and the cells were cultured for 24 hours. After irradiation with UVB at 30 mJ / cm 2 each aqueous dispersion was added again so that the lipid concentration in the medium became 100 ppm, and the cells were further cultured for 24 hours. Then, cyclobutane-type pyrimidine dimers, which are generally used as an index of DNA damage, were detected by immunofluorescence staining, and the DNA damage level was evaluated using the fluorescence intensity (normalized by DAPI) as an index. The fluorescence intensity in the case of no addition of lecithin was measured in the same manner as a control. The results are shown in Table 7 as relative values when the fluorescence intensity in the control was set to 100.

[0027]

Table 7

[0028] From the results of Tables 3 to 7, it was confirmed that lecithin has an inhibitory effect on interleukin-1α production, an inhibitory effect on cyclooxygenase-2 production, an inhibitory effect on endothelin-1 production, and an inhibitory effect on DNA damage.

[0029] Interleukin-1α is known as a proinflammatory cytokine involved in various inflammatory responses in the body. In skin cells such as epidermal keratinocytes and fibroblasts, interleukin-1α production is increased by UV irradiation, which is known to be one of the factors that cause UV-induced skin inflammation. Furthermore, it has been shown that interleukin-1α produced by UV irradiation promotes the production of melanocyte-activating factors such as endothelin and prostaglandins, thereby inducing melanocyte activation. Therefore, inhibiting interleukin-1α production in skin cells can alleviate various symptoms caused by inflammatory responses and prevent or improve pigmentation caused by excessive melanin secretion.

[0030] Cyclooxygenase-2 is known to be one of the rate-limiting enzymes that regulates the synthesis of prostaglandins in the body. Prostaglandins are mediators that play a central role in inflammatory responses, and increased production of prostaglandins in the skin is known to induce melanocyte activation. Therefore, inhibiting the production of cyclooxygenase-2 in skin cells can alleviate various symptoms caused by inflammatory responses and prevent and improve pigmentation caused by excessive melanin secretion.

[0031] Endothelin-1 is known as a potent vasoconstrictor peptide produced by vascular endothelial cells, but its production in skin cells is known to be enhanced by UV irradiation, inducing melanocyte activation. Therefore, inhibiting endothelin-1 production in skin cells can prevent and improve pigmentation caused by excessive melanin secretion.

[0032] UV rays are known to be one of the factors that cause DNA damage, and when UV exposure causes DNA damage in skin cells, normal cell differentiation and proliferation are hindered, accelerating cellular aging and causing various problems such as a weakened barrier function, age spots, dullness, wrinkles, and sagging skin. Therefore, suppressing DNA damage in skin cells will help prevent skin aging.

Claims

1. An interleukin-1α production inhibitor whose active ingredient is lecithin.

2. A cyclooxygenase-2 production inhibitor containing lecithin as an active ingredient.

3. An endothelin-1 production inhibitor containing lecithin as an active ingredient.

4. A DNA damage inhibitor whose active ingredient is lecithin.

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