TIPARP production promoter

Abstract

To provide a novel TIPARP production promoter.SOLUTION: A TIPARP production promoter contains fermented Alteromonas macleodii extract and / or extract from flower buds of Lonicera japonica as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention provides a TIPARP production promoter. [Background technology]

[0002] TIPARP is an important regulator of immunity, stem cell pluripotency, and transcription factor regulation. TIPARP is also a transcription repressor of the aryl hydrocarbon receptor (AhR), a transcription factor that mediates the toxic effects of drugs and environmental hazards. If TIPARP production can be promoted, it is expected that various diseases and conditions caused by a decrease in TIPARP or activation of AhR can be prevented or improved. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication No. 2006-528239 [Patent Document 2] JP 2019-156752 A [Patent Document 3] Japanese Patent Application Publication No. 07-157412 [Patent Document 4] JP 2012-246226 A [Non-patent literature]

[0004] [Non-Patent Document 1] Cancer Management and Research 2019:11 8991-9004 [Non-Patent Document 2] Int. J. Mol. Sci. 2019, 20, 2312; doi:10.3390 / ijms20092312 [Non-Patent Document 3] Sci. STKE, 11 September 2007 Vol. 2007, Issue 403, p. pe49; DOI: 10.1126 / stke.4032007pe49 Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a TIPARP production promoter. [Means for solving the problem]

[0006] As a result of intensive research, the present inventors have found that Alteromonas fermentation extract and Golden-silver-flower extract have a high TIPARP production promoting effect. Based on the above findings, the present inventors have completed the following invention: (1) A TIPARP production promoter containing Alteromonas fermentation extract and / or Gold and Silver Flower extract as active ingredients. (2) A composition containing the TIPARP production promoter described in (1). Effect of the Invention

[0007] According to the present invention, a composition or oral preparation containing a TIPARP production promoter can be provided. [Brief description of the drawings]

[0008] [Figure 1] Figure 1 shows the level of TIPARP expression in normal human epidermal keratinocytes when Alteromonas fermentation extract was added at various concentrations (0.10% by weight, 0.50% by weight) in Experiment 1, expressed as a relative value (ΔΔCt method) with the level when the negative control (1,3-BG only) was added set at 1.00. [Diagram 2] Figure 2 shows the level of TIPARP expression in normal human epidermal keratinocytes when various concentrations (0.10% by weight, 0.50% by weight) of Golden Ginseng extract were added in Experiment 1, expressed as a relative value (ΔΔCt method) with the level when the negative control (ethanol only) was added set at 1.00. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The present invention provides a TIPARP production promoter containing Alteromonas fermentation extract and / or Golden Flower extract as an active ingredient. TIPARP (TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin)-inducible poly-ADP-ribose polymerase), also known as PARP7 and ARTD14, is a transcriptional repressor of mono-ADP-ribosyltransferases and aryl hydrocarbon receptor (AhR). TIPARP is an important regulator in immunity, stem cell pluripotency, transcription factor regulation, etc., and it is known that knockdown of TIPARP causes liver fat accumulation, and in breast cancer, TIPARP methylation occurs and its expression is reduced (Non-Patent Documents 1 and 2). If the production of TIPARP can be promoted by the present invention, it is expected to improve immunity and prevent / ameliorate various diseases and conditions caused by TIPARP reduction.

[0010] In one embodiment of the TIPARP production promoter of the present invention, the activation of AhR is suppressed by promoting TIPARP production. AhR is a transcription factor that mediates the toxic effects of drugs and environmental hazardous factors. It has been reported that AhR binds to aromatic hydrocarbons and dioxins to induce xenobiotic metabolism, and induces growth factors, antioxidant factors, inflammatory substances, and the like (Non-Patent Document 3). It is known that the activation of AhR in the skin causes adverse effects such as the generation of active oxygen, dullness, and melanin production, and these adverse effects are exacerbated by the accumulation of ultraviolet rays. It is expected that the use of the TIPARP production promoter of the present invention can suppress the activation of AhR and prevent and improve various diseases and conditions such as those described above caused by the activation of AhR.

[0011] The promotion of TIPARP production means, for example, that the expression of the TIPARP gene increases the amount of TIPARP protein, for example, that the expression of the TIPARP gene or the amount of TIPARP protein increases when a TIPARP production promoter is applied, compared to a state where nothing is applied (control). The promotion of TIPARP production in the present invention means, for example, that the expression of the TIPARP gene or the amount of TIPARP protein increases by 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 200% or more, 300% or more, 400% or more, or 500% or more when a TIPARP production promoter is applied, compared to a state where nothing is applied (control). The expression amount of the TIPARP gene can be determined by any known technique.

[0012] Such TIPARP production promoting activity can be measured by various methods including in vivo, in vitro, ex vivo, etc. For example, the test substance is administered to cells such as liver cells, mammary gland cells, adipocytes, and skin cells, and the mRNA expression level of TIPARP in the cells is measured by a method such as qPCR, or the protein amount is measured by a method such as Western blotting. However, the measurement method is not limited to the above method, and any other method may be adopted. For example, an in vivo or ex vivo method may be adopted in which the mRNA expression level of TIPARP or the protein amount of TIPARP is measured in samples such as liver, breast, fat, and skin tissue / model after administration to an animal such as a human.

[0013] The present invention provides a TIPARP production promoter comprising Alteromonas fermentation extract and / or rhododendron amplexicaule extract as an active ingredient. The TIPARP production promoter of the present invention may be in any form, such as an oral agent, a parenteral agent such as a transdermal agent or an injection.

[0014] The Alteromonas fermented extract (CAS number: 267233-41-0) used in the present invention is an extract of a fermented product obtained from Alteromonas macleodii, a microorganism that lives in the deep sea, and is a polymer of polysaccharide glucoside. It is known to have anti-inflammatory, skin irritation, and sensitive skin symptom-relieving effects (Patent Documents 1 and 2). It can be easily fermented, purified, extracted, etc. by known methods, and is also easily available as a commercial product such as Abyssine PF by UNIPEXIN NOVATIONS.

[0015] The honeysuckle extract used in the present invention is an extract of the flower buds of honeysuckle (Lonicera japonica). It is known to have anti-inflammatory, antibacterial, and antispasmodic effects (Patent Documents 3 and 4). It can be easily dried, purified, extracted, etc. by known methods, and is also easily available as a commercially available product. It can be used either raw or dried, but it can also be used as an extract, dried product, dried powder, raw material powder, squeezed juice, etc., and the form to be used can be appropriately selected.

[0016] The above extract can be extracted by, for example, solvent extraction. In the case of solvent extraction, the fermented product or plant in whole or in part is dried as necessary, and further chopped or crushed as necessary, and then extracted by using an aqueous extractant, water, for example, cold water, warm water, or hot water at or below the boiling point, or a water-containing organic solvent, an organic solvent, for example, ethanol, methanol, ether, 1,3-butylene glycol, etc., at room temperature or after heating, with a suitable solvent selected according to the properties of the raw material and the use of the composition. However, the extraction method is not limited to solvent extraction, and may be any conventional method known in the art. The extraction method and the form of the extract used in the present invention are arbitrary as long as they do not impair the effects of the present invention. The form of the above extract may be not only the extract itself, but also one that has been appropriately diluted or concentrated by a conventional method, and may further be a powdered or lumpy solid obtained by drying the extract.

[0017] As an example of the aqueous organic solvent, an aqueous lower alcohol such as aqueous ethanol may be used, in which case the water content may be, for example, 0 to 10 v / v%, 10 to 40 v / v%, 20 to 30 v / v%, 30 to 50 v / v%, 50 to 80 v / v%, 80 to 99.5 v / v%, etc.

[0018] The TIPARP production promoter of the present invention can be administered by various administration routes such as transdermal, oral, and local injection, and it is preferable to apply the active ingredient of the present invention in an amount that can fully exert the effect of promoting TIPARP production. The amount of Alteromonas fermentation extract and / or cypress extract to be added can be appropriately determined depending on the type, purpose, form, and method of use thereof.

[0019] When the TIPARP production promoter of the present invention is used as a transdermal or oral preparation, it is preferable to prepare it so that the dry weight of Alteromonas fermentation extract and / or Crown and Silver Flower extract is about 0.00001 to 1.0% by weight, more preferably about 0.001 to 0.8% by weight, and more preferably about 0.10 to 0.5% by weight, of the total amount of the transdermal or oral preparation.

[0020] In addition, the frequency of administration is not limited, but according to one example, it can be administered, for example, once every two weeks, once a week, once every three days, once every two days, once a day, twice a day, three times a day, four times a day, etc. In addition, it may be administered as needed, continuously, or intermittently at intervals of, for example, several months.

[0021] In addition, the TIPARP production promoter of the present invention can be contained in a composition for percutaneous or oral administration, such as a pharmaceutical, cosmetic, or food composition. The form of the TIPARP production promoter and composition of the present invention can be selected from any of a variety of forms, such as powder, liquid, solid such as tablet, granule, granule, paste, gel, cream, and the like.

[0022] The TIPARP production promoter and composition of the present invention can be used in combination with additives selected arbitrarily as necessary. For additives, excipients, colorants, preservatives, thickeners, binders, disintegrants, dispersants, stabilizers, gelling agents, antioxidants, surfactants, preservatives, pH adjusters, etc., known additives can be appropriately selected and used.

[0023] The present invention also provides a method for preventing or improving diseases and conditions caused by inhibition of TIPARP or activation of AhR, such as promotion of TIPARP production in liver tissue including hepatocytes, breast tissue including mammary gland, and skin tissue including epidermal or dermal cells, and fatty liver, breast cancer, skin dullness, and melanin production through inhibition of AhR activation by promoting TIPARP production, by administering Alteromonas fermentation extract and / or Golden Ginseng extract, for example, percutaneously, orally, or locally by injection. The method of the present invention is a method for cosmetic purposes, and may not be a treatment by a doctor or medical professional.

[0024] Furthermore, the present invention provides use of the Alteromonas fermentation extract and / or the Golden Flower extract in the manufacture of a pharmaceutical, such as an oral or enteral preparation, for preventing or ameliorating diseases or conditions caused by inhibition of TIPARP or activation of AhR, such as fatty liver, breast cancer, and skin aging.The present invention also provides the Alteromonas fermentation extract and / or the Golden Flower extract for use in a method for preventing or ameliorating diseases or conditions caused by inhibition of TIPARP, such as fatty liver, breast cancer, skin dullness, and melanin production, by promoting TIPARP production in liver, fat, and skin tissues, for example, by oral or transdermal administration. EXAMPLES

[0025] The present invention will now be described in more detail with reference to examples, although the present invention is not limited thereto.

[0026] Experiment 1: Sample preparation As candidate samples for TIPARP production promoters, Alteromonas fermentation extract and / or Golden Flower extract were prepared as shown in the table below.

[0027] [Table 1]

[0028] A total of 29 types of candidate samples were prepared, including natural ingredients such as animal and plant extracts and synthetic ingredients. The samples were adjusted to a final concentration of 0.10% to 0.5% by weight in the medium using the solvent of each extract (ethanol or 1,3-BG (1,3-butylene glycol)). As a control, the solvent of each extract that did not contain anything was used.

[0029] Experiment 2: TIPARP expression analysis by qPCR In this study, normal human epidermal keratinocytes (Kuraray: KK-4009) were used. Epidermal cells were cultured in a cell culture dish at 37°C and 5% CO2 using a medium (Thermo Fisher Scientific) containing EpiLife Medium M-EPI-500-CA and Human Keratinocyte Growth Supplement (HKGS). 5 cells / cm 2 After the cells were seeded at a density of 100 μg / cm2 and allowed to settle, they were treated with hydrogen peroxide to induce senescence. The senescence state was confirmed by detection of SA-β-gal (Dojindo: SG03 Cellular Senescence Detection Kit - SPiDER-βGal). In addition, ultraviolet light (UVB 400 mJ / cm2) was applied to the cells. 2 ,UVA 5J / cm 2 ) and cultured for 6 hours. Each extract and solvent were added and cultured overnight, after which 10 nM FICZ (Aryl Hydrocarbon Receptor Agonist) was added to activate AhR. Total RNA was collected after 24 hours of culture.

[0030] cDNA was synthesized from total RNA in normal human epidermal keratinocytes cultured using the above method (Takara: PrimeScript RT reagent kit). qPCR was performed using this cDNA with SYBR Green (KAPA SYBR Fast qPCR kit (Universal qPCR kit)). β-actin was used as the internal control to evaluate the ability to increase TIPARP mRNA production. At the same time, gene expression of CYPA1A and CYP1B1, which are reactive oxygen producing genes downstream of AhR, was also evaluated, and it was confirmed that there was no increase in their production. The specific sequences of the primers corresponding to each gene are shown in Table 2 below. [Table 2]

[0031] The results are shown in Figure 2. As shown in Figure 2, when the Alteromonas fermentation extract and the Golden Flower extract were administered, the expression level of TIPARP was significantly increased in a concentration-dependent manner compared to the control. Therefore, it can be seen that the extract of the present invention has an excellent effect of promoting TIPARP production.

Claims

1. A TIPARP production promoter containing Alteromonas fermentation extract and / or Tunga japonica extract as active ingredients.

2. A composition for promoting TIPARP production, comprising the TIPARP production promoter according to claim 1.

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