TGF-β signal transduction inhibitor

Abstract

To provide a novel TGF-β signal transduction inhibitor.SOLUTION: An extract of Oenothera biennis L. is used as an active ingredient. The extract of Oenothera biennis L. has a TGF-β signal transduction inhibitory effect and thus suppresses TGF-β-induced promotion of fibrosis in tissues. Therefore, it can be expected to have preventive and therapeutic effects on fibrotic diseases, and preventive and improving effects on cellulite.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a TGF-β signaling inhibitor. [Background technology]

[0002] TGF-β (transforming growth factor β) is a multifunctional cytokine that regulates cell proliferation and differentiation. TGF-β is also involved in various pathological diseases. For example, because it promotes fibrosis in tissues, it is closely related to the onset and progression of fibrotic diseases such as pulmonary fibrosis. Furthermore, although it is not a disease, TGF-β has also been suggested to be involved in cellulite, a condition in which the skin becomes uneven due to the degeneration of adipose tissue caused by fibrosis.

[0003] It is known that TGF-β signals are transmitted via the TGF-β / Smad pathway. When TGF-β binds to a receptor present on the cell surface, Smad, a signaling molecule present in the cell, is phosphorylated, and the phosphorylated Smad translocates to the nucleus and regulates the transcription of target genes. In recent years, methods have been attracting attention for preventing and treating fibrotic diseases and preventing and ameliorating cellulite by inhibiting TGF-β signaling, which inhibits the promotion of fibrosis in tissues by TGF-β. For example, Patent Document 1 discloses that compounds with specific chemical structures can be used as active ingredients in TGF-β signaling inhibitors. However, the search for components with TGF-β signaling inhibitory activity remains significant. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 5846527 Summary of the Invention [Problem to be solved by the invention]

[0005] Therefore, an object of the present invention is to provide a novel TGF-β signaling inhibitor. [Means for solving the problem]

[0006] In view of the above, the present inventors have conducted extensive research and found that an extract of evening primrose (Oenothera biennis L.) has an inhibitory effect on TGF-β signaling.

[0007] The TGF-β signaling inhibitor of the present invention, which was developed based on the above findings, contains an extract of evening primrose as an active ingredient, as set forth in claim 1. [Effects of the Invention]

[0008] According to the present invention, a novel TGF-β signaling inhibitor can be provided. [Brief explanation of the drawings]

[0009] [Figure 1] 1 is a graph showing the TGF-β signaling inhibitory effect of an evening primrose extract in an example. DETAILED DESCRIPTION OF THE INVENTION

[0010] The TGF-β signaling inhibitor of the present invention contains an extract of evening primrose as an active ingredient.

[0011] The evening primrose used in the present invention is a biennial plant of the genus Oenothera in the family Onagraceae, also known as evening primrose. In the present invention, either naturally growing plants or plants cultivated by producers may be used.

[0012] In the present invention, an extract of evening primrose can be obtained by, for example, adding water or an organic solvent (such as monoalcohols (e.g., methanol, ethanol, and isopropanol) or polyhydric alcohols (e.g., ether, hexane, ethyl acetate, acetonitrile, etc.), which may contain water, as an extraction solvent to the seeds (which may be dried or crushed) or to compressed seeds, leaving the mixture at room temperature for one hour to one week to extract components, or refluxing the mixture to extract components, followed by filtration to obtain a filtrate. The extraction solvent may be a single solvent or a mixture of multiple solvents. The obtained filtrate may be used as a liquid extract as is, or may be converted into a solid or powder by vacuum concentration or lyophilization. The filtrate may be further purified by ammonium sulfate fractionation, gel filtration, ion chromatography, or the like, and the resulting fraction may be used as a liquid extract. Alternatively, for example, extraction may be performed first using hexane, followed by extraction using solvents of gradually increasing polarity, such as ethyl acetate, acetone, and water, and the filtrate obtained by using any of the solvents may be used as the liquid extract.

[0013] The TGF-β signaling inhibitor of the present invention can be formulated into various dosage forms and taken orally as a pharmaceutical or health food, or can be incorporated into various foods and beverages for ingestion. The TGF-β signaling inhibitor of the present invention can also be applied to the skin in the form of an external preparation such as an ointment, cream, or lotion. The dosage, intake, or application amount can be determined appropriately based on the age, sex, weight, and severity of symptoms of the subject. By taking, ingesting, or applying an appropriate amount, preventive and therapeutic effects against fibrotic diseases and preventive and ameliorative effects against cellulite can be expected based on its TGF-β signaling inhibitory activity. [Example]

[0014] The present invention will be described in detail below with reference to examples, but the present invention should not be construed as being limited to the following description.

[0015] Experimental Example 1: Inhibitory effect of evening primrose extract on TGF-β signal transduction (Experimental Method) The inhibitory effect of evening primrose extract on TGF-β signaling was evaluated using an SBE (Smad binding element)-luciferase reporter assay, a well-known monitoring system for the TGF-β / Smad pathway. Specifically, mouse embryonic fibroblast NIH3T3 (BRC, Japan) cells were electroporated with the pNL[NlucP / SBE / Hygro] Vector (Promega, USA) using the Cell Line Nucleofector Kit V (Lonza, Switzerland). After 24 hours, drug-resistant cells were selected by adding hygromycin at a final concentration of 200 μg / mL, and a hygromycin-resistant NIH3T3 cell line stably expressing the SBE-luciferase reporter was established. Next, approximately 10,000 NIH3T3 cells stably expressing the SBE-luciferase reporter were seeded per well in a 96-well plate in 100 μL of medium (low-glucose DMEM (Sigma, USA) supplemented with 10% FBS (Gibco, USA) and 200 mmol / L L-glutamine solution (x100) (Wako, Japan)). After incubation at 37°C for 16–24 hours, 1 μL of evening primrose extract (final concentration 1%) was added and the cells were incubated for an additional hour. After 1 hour, human TGF-β1 (Wako, Japan) was added at a final concentration of 20 ng / mL and the cells were incubated at 37°C for 4 hours. After 4 hours, the inhibitory effect of the evening primrose extract on TGF-β signaling was evaluated by measuring luciferase activity using the Nano-Glo Luciferase Assay System (Promega, USA) according to the protocol. The evening primrose extract used as the sample was a commercially available product, and its composition (wt%) was evening primrose seed extract (extraction solvent was 50% aqueous 1,3-butylene glycol): 1.0, 1,3-butylene glycol: 49.5, water: 49.5.

[0016] (Experimental results) As a control, 1,3-butylene glycol was added to a final concentration of 0.5% instead of the evening primrose extract. The luciferase activity without human TGF-β1 was set to 1, and the relative values ​​are shown in Figure 1. As is clear from Figure 1, adding evening primrose extract before adding human TGF-β1 significantly reduced the increase in luciferase activity that occurs with the addition of human TGF-β1, indicating activation of the TGF-β / Smad pathway. These results confirmed the inhibitory effect of evening primrose extract on TGF-β signaling.

[0017] Formulation Example 1: Tablets Tablets having the following component composition and inhibitory effect on TGF-β signaling were produced by a method known per se (unit: % by weight). Evening primrose extract powder used in Experimental Example 1 1 lactose 80 Magnesium stearate 19

[0018] Formulation example 2: Biscuits Biscuits having the following component composition and having TGF-β signaling inhibitory activity were produced by a method known per se (unit: weight %). Evening primrose extract used in Experimental Example 1 1 32g plain flour 16 whole eggs Butter 16 Sugar 24 water 10 1 teaspoon baking powder

[0019] Formulation example 3: Jelly A jelly having the following component composition and having TGF-β signaling inhibitory activity was produced by a method known per se (unit: % by weight). Evening primrose extract used in Experimental Example 1 0.01 Gelling Agent 1.3 sugar 20 pH adjuster 2.5 Wednesday 76.19

[0020] Formulation Example 4: Ointment An ointment having the following component composition and having a TGF-β signaling inhibitory effect was produced by a method known per se (unit: % by weight). Evening primrose extract powder used in Experimental Example 1 5 White Vaseline 95

[0021] Formulation Example 5: Cream A cream having the following component composition and having a TGF-β signaling inhibitory effect was produced by a method known per se (unit: % by weight). Evening primrose extract powder used in Experimental Example 1 5 White Vaseline 30 Cetyl alcohol 15 Sodium lauryl sulfate 1 Ethyl parahydroxybenzoate 0.1 Butyl parahydroxybenzoate 0.1 Water 48.8

[0022] Formulation Example 6: Lotion A lotion having the following component composition and having a TGF-β signaling inhibitory effect was produced by a method known per se (unit: % by weight). Evening primrose extract used in Experimental Example 1 10 Polyoxyethylene (40) hydrogenated castor oil 0.5 Ethanol 5 Parabens 0.2 Water 84.3 [Industrial Applicability]

[0023] INDUSTRIAL APPLICABILITY The present invention has industrial applicability in that it can provide a novel TGF-β signaling inhibitor.

Claims

[Claim 1] A TGF-β signaling inhibitor containing an extract of evening primrose as an active ingredient.

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