TGF-β signaling inhibitor

Abstract

To provide a novel TGF-β signaling inhibitor. 【Solution means】 It contains an aqueous extract of _Sagina maxima_ as an active ingredient. Since the aqueous extract of _Sagina maxima_ has an inhibitory effect on TGF-β signaling, it is expected to have a preventive and therapeutic effect on fibrotic diseases and a preventive and ameliorating effect on cellulite by suppressing the promotion of fibrosis in tissues by TGF-β.

Description

Technical Field

[0001] The present invention relates to a TGF-β signal transduction inhibitor.

Background Art

[0002] TGF-β (transforming growth factor β) is a multifunctional cytokine that controls cell growth and differentiation. TGF-β is pathologically involved in various diseases. For example, since it has an effect of promoting fibrosis in tissues, it is closely related to the onset and progression of fibrotic diseases such as pulmonary fibrosis. Also, regarding the uneven skin condition (cellulite) that occurs when adipose tissue degenerates due to fibrosis, although it is not a disease, the involvement of TGF-β has been suggested.

[0003] The signal of TGF-β is known to be transmitted through the TGF-β / Smad pathway. When TGF-β binds to a receptor present on the cell surface, Smad, an information-transmitting molecule present inside the cell, is phosphorylated. The phosphorylated Smad translocates to the nucleus and regulates the transcription of target genes. In recent years, a method of preventing or treating fibrotic diseases or preventing or improving cellulite by suppressing the promotion of fibrosis in tissues by TGF-β by inhibiting the signal transduction of TGF-β has attracted attention. For example, in Patent Document 1, it is introduced that a compound having a specific chemical structure can be used as an active ingredient of a TGF-β signal transduction inhibitor. However, the search for components having a TGF-β signal transduction inhibitory action is still in a significant situation.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems 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 Problems

[0006] As a result of intensive studies in view of the above points, the present inventors have found that the water extract of Oenothera biennis L. has an inhibitory effect on TGF-β signaling.

[0007] Based on the above findings, the TGF-β signaling inhibitor of the present invention comprises, as described in claim 1, seeds the water extract of Oenothera biennis L. as an active ingredient.

Effects of the Invention

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

Brief Description of the Drawings

[0009]

Figure 1

Modes for Carrying Out the Invention

[0010] The TGF-β signaling inhibitor of the present invention comprises the water extract of Oenothera biennis L. as an active ingredient.

[0011] The Oenothera biennis L. used in the present invention is an annual herbaceous plant of the genus Oenothera in the family Onagraceae, which has the alias of evening primrose. In the present invention, those growing naturally in the wild may be used, or those cultivated by producers may also be used.

[0012] In the present invention, the water extract of *Marsilea quadrifolia* can be obtained as a filtrate, for example, by adding water (such as tap water, distilled water, ion-exchanged water, sterilized water, warm water at 30°C or higher like hot water) as an extraction solvent to the seeds themselves (which may be dried or crushed) or the pressed product of the seeds, followed by allowing to stand or stirring for 1 hour to 1 week for component extraction, or performing reflux for component extraction and then filtering. The obtained filtrate may be used as a liquid extract as it is, or may be concentrated under reduced pressure or freeze-dried to be used as a solid or powder. In addition, the filtrate may be further purified by ammonium sulfate fractionation, gel filtration, ion chromatography, etc., and the obtained fraction may be used as a liquid extract.

[0013] In addition to being able to be taken or ingested as pharmaceuticals, health foods, etc. by formulating the TGF-β signal transduction inhibitor in the present invention into various dosage forms, it can also be incorporated into various foods and beverages for ingestion. Further, the TGF-β signal transduction inhibitor in the present invention can also be applied to the skin in the form of external preparations such as ointments, creams, and lotions. The dosage, ingestion amount, and application amount can be appropriately determined based on the age, gender, weight, degree of symptoms, etc. of the applicable subject. By taking, ingesting, or applying an appropriate amount, based on its TGF-β signal transduction inhibitory effect, a preventive effect and a therapeutic effect on fibrotic diseases, a preventive effect and an improvement effect on cellulite can be expected.

Examples

[0014] Hereinafter, the present invention will be described in detail by way of examples, but the present invention is not construed as being limited to the following description.

[0015] Experimental Example 1: TGF-β Signal Transduction Inhibitory Effect of the Hot Water Extract of *Marsilea quadrifolia* (Experimental Method) The inhibitory effect of the hot water extract of *Equisetum arvense* L. on TGF-β signal transduction was evaluated by an SBE (Smad binding element)-Luciferase reporter assay known as a monitoring system for the TGF-β / Smad pathway. Specifically, first, pNL[NlucP / SBE / Hygro] Vector (Promega, USA) was introduced into mouse embryonic fibroblasts NIH3T3 (BRC, Japan) by electroporation using Cell Line Nucleofector Kit V (Lonza, Switzerland). After 24 hours, drug-resistant cell selection was performed by adding Hygromycin at a final concentration of 200 μg / mL, and an SBE-Luciferase reporter stable expression NIH3T3 cell line with Hygromycin resistance was established. Next, approximately 10,000 SBE-Luciferase reporter stable expression NIH3T3 cells per well were seeded in a 96-well plate with 100 μL of medium (low glucose DMEM (Sigma, USA) supplemented with 10% FBS (Gibco, USA) and 200 mmol / L L-glutamine solution (×100) (Wako, Japan), the same below). After culturing at 37°C for 16 - 24 hours, 1 μL of the medium in which a commercially available hot water extract (water-soluble powder) of *Equisetum arvense* L. was dissolved to a concentration of 10 mg / mL (final concentration 100 μg / mL) was added, and the cells were further cultured for 1 hour. After 1 hour, human TGF-β1 (Wako, Japan) at a final concentration of 20 ng / mL was added, and the cells were cultured at 37°C for 4 hours. After 4 hours, the inhibitory effect of the hot water extract of *Equisetum arvense* L. on TGF-β signal transduction was evaluated by measuring the Luciferase activity according to the protocol using the Nano-Glo Luciferase Assay System (Promega, USA).

[0016] (Experimental results) As a control, only 1 μL of the medium was added instead of the medium dissolved with the hot water extract of *Memma tsuyoigusa*, and the relative value with the Luciferase activity when human TGF-β1 was not added was set to 1, as shown in Figure 1. As is clear from Figure 1, by adding the hot water extract of *Memma tsuyoigusa* before adding human TGF-β1, the increase in Luciferase activity, which means that the TGF-β / Smad pathway activated by the addition of human TGF-β1, was significantly reduced. The IC50 was 53.7 μg / mL. From the above results, the TGF-β signal transduction inhibitory effect of the hot water extract of *Memma tsuyoigusa* could be confirmed.

[0017] Formulation Example 1: Tablet A tablet having a TGF-β signal transduction inhibitory effect and consisting of the following component composition was produced by a method known per se (unit: weight %). Hot water extract of *Memma tsuyoigusa* used in Experimental Example 1 1 Lactose 80 Magnesium stearate 19

[0018] Formulation Example 2: Biscuit A biscuit having a TGF-β signal transduction inhibitory effect and consisting of the following component composition was produced by a method known per se (unit: weight %). Hot water extract of *Memma tsuyoigusa* used in Experimental Example 1 1 Weak flour 32 Whole egg 16 Butter 16 Sugar 24 Water 10 Baking powder 1

[0019] Formulation Example 3: Jelly A jelly having a TGF-β signal transduction inhibitory effect and consisting of the following component composition was produced by a method known per se (unit: weight %). Hot water extract of *Memma tsuyoigusa* used in Experimental Example 1 0.01 Gelling agent 1.3 Sugar 20 pH adjuster 2.5 Water 76.19

[0020] Formulation Example 4: Ointment An ointment having an inhibitory effect on TGF-β signal transduction and consisting of the following component composition was produced by a method known per se (unit: % by weight). Hot water extract of *Caltha palustris* L. used in Experimental Example 1 5 White petrolatum 95

[0021] Formulation Example 5: Cream A cream having an inhibitory effect on TGF-β signal transduction and consisting of the following component composition was produced by a method known per se (unit: % by weight). Hot water extract of *Caltha palustris* L. used in Experimental Example 1 5 White petrolatum 30 Cetanol 15 Sodium lauryl sulfate 1 Ethyl paraben 0.1 Butyl paraben 0.1 Water 48.8

[0022] Formulation Example 6: Lotion A lotion having an inhibitory effect on TGF-β signal transduction and consisting of the following component composition was produced by a method known per se (unit: % by weight). Hot water extract of *Caltha palustris* L. used in Experimental Example 1 10 Polyoxyethylene (40) hydrogenated castor oil 0.5 Ethanol 5 Paraben 0.2 Water 84.3

Industrial Applicability

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

Claims

【Claim 1】 A TGF-β signaling inhibitor comprising an aqueous extract of seeds of Equisetum arvense as an active ingredient.

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