Composition with PPAR-gamma inhibition effect and application thereof

Abstract

The invention discloses a composition with a PPAR-gamma inhibition effect and application of the composition. The composition comprises glycyrrhizic acid, witch hazel extract, pseudoalteromonas fermentation product extract and hydrolyzed olive leaf extract in a mass ratio of (0.5-15): (5-35): (40-75): (5-25), after the witch hazel extract is compounded with the other three components, the composition has a PPAR-gamma inhibition effect, the components have an extremely remarkable synergistic interaction effect, and the composition has a good application prospect. The lipid inhibition effect and the anti-inflammatory effect of the composition can be remarkably improved, and the skin inflammation reaction can be better inhibited; the effect of the composition is obviously better than that of each component used independently or any three components used in a compounding manner, the effects of inhibiting lipid synthesis, reducing sebum secretion and inhibiting inflammatory reaction of the three components are further improved, and more choices are provided for drugs for treating inflammatory dermatosis caused by excessive lipid secretion.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and more specifically, relates to a composition with PPAR-γ inhibitory effect and application thereof. Background Art

[0002] Skin lipids are composed of lipids produced by sebocytes, keratinocytes, and microorganisms, and can significantly affect skin status through different mechanisms. Lipids are important metabolites with many important cellular functions, including participation in energy metabolism, signal transduction, enzyme activation, transmembrane transport, cell proliferation, development, differentiation, and apoptosis. Changes in lipid metabolism are an important factor in the pathogenesis of various diseases, such as metabolic diseases, cancer, neurodegenerative diseases, and skin diseases. It is well known that skin status is significantly affected by skin lipids, which are one of the most important compounds for maintaining the function of the skin barrier and are important multifunctional mediators that affect epidermal metabolism, inflammatory responses, cell proliferation, and differentiation processes.

[0003] In the human epidermis, peroxisome proliferator-activated receptors (PPARs) are a group of key skin lipid-regulating transcription factors that are involved in controlling lipid metabolism, skin barrier permeability, inflammatory response, cell proliferation and differentiation. Among them, peroxisome proliferator-activated receptor γ (PPAR-γ) belongs to the nuclear hormone receptor superfamily and is a ligand-activated transcription factor with multiple biological functions. It plays an important role in regulating metabolism, controlling inflammation, regulating glucose and lipid metabolism, improving atherosclerosis, inhibiting tumors and regulating immunity. PPAR-γ plays a central role in lipid synthesis. When PPAR-γ is activated, it can promote adipogenesis and lipid synthesis as well as the differentiation of adipocytes, enhance the uptake and storage capacity of adipose tissue for lipids, lead to excessive sebum secretion, and be accompanied by this inflammatory response, which can easily cause inflammatory skin diseases such as acne, seborrheic dermatitis, atopic dermatitis, acute contact dermatitis, psoriasis and rosacea. At the same time, PPAR-γ can also regulate the expression of genes related to lipid synthesis, such as lipoprotein lipase (LPL), fatty acid transport and oxidation genes, thereby controlling the rate and amount of lipid synthesis. Therefore, the study of PPARγ helps to understand different inflammatory skin diseases. By inhibiting the binding of PPARγ to ligands, ligands can inhibit lipid synthesis, reduce sebum secretion, and inhibit inflammatory responses, thereby reducing skin inflammation.

[0004] Inflammatory skin diseases are closely related to factors such as excessive sebum secretion, obstruction of the sebaceous gland ducts, bacterial infection and inflammatory response. At present, the drug treatments for inflammatory skin diseases mainly include oral drugs such as antibiotics, retinoic acid drugs, etc., and topical drugs such as benzoyl peroxide and salicylic acid. Oral drugs can effectively relieve symptoms, but long-term use may cause side effects such as gastrointestinal discomfort, dry skin, and abnormal liver function; topical drugs may also cause adverse reactions such as skin irritation, dryness, and peeling. Therefore, most of the currently used drug treatments for inflammatory skin diseases have adverse reactions. Long-term use of antibiotics may cause bacteria to develop drug resistance, which reduces the efficacy of the drugs, or even has defects and shortcomings such as ineffectiveness.

[0005] So far, there is still a lack of efficient and natural drugs that have PPAR-γ inhibitory effects and can inhibit inflammatory responses to inhibit lipid synthesis, reduce sebum secretion and inhibit inflammatory responses, thereby reducing the occurrence of inflammatory skin diseases. The applicant's previous studies have shown that the composition of glycyrrhetinic acid, hydrolyzed olive leaf extract and Pseudoalteromonas fermentation product extract can regulate lipid secretion disorders and reduce sebum secretion by inhibiting SREBP-1; however, the lipid inhibition effect of the composition needs to be improved, and whether it can be used to treat inflammatory skin diseases remains to be studied. At present, it is necessary to develop and explore more sebum action targets to provide more natural drugs for the treatment of inflammatory skin diseases with better effects, high specificity, few side effects and low prices. Summary of the invention

[0006] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of existing drugs for treating inflammatory skin diseases, and to provide a composition with PPAR-γ inhibitory effect and its application.

[0007] The object of the present invention is to provide a composition having a PPAR-γ inhibitory effect.

[0008] Another object of the present invention is to provide a use of the composition having PPAR-γ inhibitory effect.

[0009] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0010] The invention provides a composition with PPAR-γ inhibitory effect, which comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (0.5-15): (5-35): (40-75): (5-25).

[0011] Based on previous studies, the present invention further compounds the composition obtained by compounding the witch hazel extract on the basis of the three components of glycyrrhizic acid, pseudoalteromonas fermentation product extract, and hydrolyzed olive leaf extract, and has a significant PPAR-γ inhibitory effect. After compounding, the components have a significant synergistic effect, which can significantly reduce the expression of proinflammatory factors TNF-α and IL-1β, reduce the content of triglycerides, inhibit lipid synthesis, reduce sebum secretion, and better inhibit sebum inflammation. Among them, the effects of using witch hazel extract and pseudoalteromonas fermentation product extract alone are similar, and the present invention finds that after compounding the witch hazel extract, under the premise of controlling the total amount of the composition unchanged, the amount of each component is reduced, and the effect of the composition at the same concentration is significantly better than the use of each component alone or the compound use of any three components, which can significantly inhibit lipid synthesis, reduce sebum secretion and inhibit inflammatory response, and is used to prevent and / or treat diseases caused by excessive lipid secretion, and provide more choices for the treatment of inflammatory skin diseases, which is of great significance for the development of natural medicines with high specificity, small side effects and low price.

[0012] Preferably, the composition comprises components in the following mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (1-10): (10-30): (45-70): (10-20).

[0013] More preferably, the composition comprises components in the following mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (5-10): (10-15): (60-70): (10-15).

[0014] The present invention provides a method for preparing the composition, which comprises directly mixing respective amounts of glycyrrhetinic acid, witch hazel extract, pseudoalteromonas fermentation product extract and hydrolyzed olive leaf extract.

[0015] The present invention provides application of the composition in preparing a lipid synthesis inhibitor.

[0016] The present invention provides application of the above composition in preparing a PPAR-γ inhibitor.

[0017] The present invention provides application of the composition in preparing anti-inflammatory products.

[0018] The present invention provides application of the above composition in preparing a product for reducing triglyceriditis.

[0019] Preferably, the product is a medicine, a skin care product or a cosmetic.

[0020] The present invention provides application of the composition in preparing drugs for treating inflammatory skin diseases.

[0021] Preferably, the inflammatory skin disease is acne, seborrheic dermatitis, atopic dermatitis, acute contact dermatitis, psoriasis or rosacea.

[0022] The present invention has the following beneficial effects:

[0023] The invention provides a composition with PPAR-γ inhibitory effect, comprising the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (0.5-15): (5-35): (40-75): (5-25). After the witch hazel extract is further compounded on the basis of the other three groups, the composition has a significant PPAR-γ inhibitory effect, and the components have a very significant synergistic effect, which can significantly improve the lipid inhibitory effect and anti-inflammatory effect of the composition, and better inhibit skin inflammatory reaction; the effect of the composition is significantly better than that of each component used alone or any three components used in combination, and can inhibit lipid synthesis, reduce sebum secretion and inhibit inflammatory reaction, thereby providing more choices for inflammatory skin disease treatment drugs, and can be used for preventing and / or treating diseases caused by excessive lipid secretion, and has important significance for developing natural skin disease drugs with high specificity, small side effects and low price. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is the result graph of SZ95 cell survival rate.

[0025] Figure 2 The results of PPAR-γ expression are shown in Figure 2

[0026] Figure 3 This is the result diagram of TNF-α expression.

[0027] Figure 4 This is the result diagram of IL-1β expression.

[0028] Marks in the figure: compared with the blank group, ####P<0.000; compared with the model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.001. DETAILED DESCRIPTION

[0029] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0030] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0031] The glycyrrhetinic acid used in the examples was purchased from Foshan Kangni Ailun Biotechnology Co., Ltd., the witch hazel extract was purchased from Shanghai Biohobo Biotechnology Co., Ltd., the Pseudoalteromonas fermentation product extract was purchased from Guangzhou Rongdao Chemical Co., Ltd. (manufacturer: LIPOTEC SAU), and the hydrolyzed olive leaf extract was purchased from Shanghai Boshuo Industrial Co., Ltd. (manufacturer: KATAKURA & CO-OP AGRI CORP.).

[0032] Example 1

[0033] A composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = 1:30:49:20.

[0034] The preparation method of the composition is as follows: 0.1 g of glycyrrhetinic acid, 3.0 g of witch hazel extract, 4.9 g of Pseudoalteromonas fermentation product extract and 2 g of hydrolyzed olive leaf extract are directly mixed to obtain the composition.

[0035] Example 2

[0036] A composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = 2.5:15:67.5:15.

[0037] The preparation method of the composition is as follows: 0.25 g of glycyrrhetinic acid, 1.5 g of witch hazel extract, 6.75 g of Pseudoalteromonas fermentation product extract and 1.5 g of hydrolyzed olive leaf extract are directly mixed to obtain the composition.

[0038] Example 3

[0039] A composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = 5:15:70:10.

[0040] The preparation method of the composition is as follows: 0.5 g of glycyrrhetinic acid, 1.5 g of witch hazel extract, 7.0 g of Pseudoalteromonas fermentation product extract and 1.0 g of hydrolyzed olive leaf extract are directly mixed to obtain the composition.

[0041] Example 4

[0042] A composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = 10:10:70:10.

[0043] The preparation method of the composition is as follows: 1.0 g of glycyrrhetinic acid, 1.0 g of witch hazel extract, 7.0 g of Pseudoalteromonas fermentation product extract and 1.0 g of hydrolyzed olive leaf extract are directly mixed to obtain the composition.

[0044] Comparative Example 1

[0045] The difference from Example 1 is that it contains only 10 g of glycyrrhetinic acid.

[0046] Comparative Example 2

[0047] The difference from Example 1 is that only 10 g of witch hazel extract is contained.

[0048] Comparative Example 3

[0049] The difference from Example 1 is that it contains only 10 g of the Pseudoalteromonas fermentation product extract.

[0050] Comparative Example 4

[0051] The difference from Example 1 is that it only contains 10g of hydrolyzed olive leaf extract.

[0052] Comparative Example 5

[0053] The difference from Example 1 is that: the composition does not contain the hydrolyzed olive leaf extract, and comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: Pseudoalteromonas fermentation product extract = 20:10:70.

[0054] The preparation method of the composition is as follows: 2.0 g of glycyrrhetinic acid, 1.0 g of witch hazel extract and 7.0 g of Pseudoalteromonas fermentation product extract are directly mixed to obtain the composition.

[0055] Comparative Example 6

[0056] The difference from Example 1 is that: the extract of the fermentation product of Pseudoalteromonas is not contained, and the composition comprises the following components in the following mass ratio: glycyrrhetinic acid: witch hazel extract: hydrolyzed olive leaf extract = 30:30:40.

[0057] The preparation method of the composition is as follows: 3.0 g of glycyrrhetinic acid, 3.0 g of witch hazel extract and 4.0 g of hydrolyzed olive leaf extract are directly mixed to obtain the composition.

[0058] Comparative Example 7

[0059] The difference from Example 1 is that: the composition does not contain witch hazel extract, and comprises the following components in a mass ratio: glycyrrhetinic acid: hydrolyzed olive leaf extract: Pseudoalteromonas fermentation product extract = 10:10:80.

[0060] The preparation method of the composition is as follows: 1.0 g of glycyrrhetinic acid, 1.0 g of hydrolyzed olive leaf extract and 8.0 g of Pseudoalteromonas fermentation product extract are directly mixed to obtain the composition.

[0061] Comparative Example 8

[0062] The difference from Example 1 is that: glycyrrhetinic acid is not contained, and the composition comprises the following components in a mass ratio: witch hazel extract: hydrolyzed olive leaf extract: Pseudoalteromonas fermentation product extract = 10:10:80.

[0063] The preparation method of the composition is as follows: 1.0 g of witch hazel extract, 1.0 g of hydrolyzed olive leaf extract and 8.0 g of Pseudoalteromonas fermentation product extract are directly mixed to obtain the composition.

[0064] Test Example 1 Cytotoxicity and PPAR-γ Inhibitory Activity of Composition

[0065] (1) Cytotoxicity assay

[0066] Human sebaceous gland cells (SZ95 cells) were inoculated into DMEM medium containing 10% fetal bovine serum and 1% double antibody and cultured at 37°C and 5% CO2. The cultured SZ95 cells were digested and separated with 0.25% insulin-EDTA and plated in a 96-well plate at 5×10 3 Cells were cultured at a concentration of cells / well. After 24 hours of inoculation, complete culture medium was added to the blank group, and complete culture medium was added to the experimental group at a concentration of 0.05% (prepared with deionized water) of the composition of Examples 1 to 4 and Comparative Examples 1 to 8. After 24 hours of incubation, the supernatant was discarded, 100 μL of MTT working solution was added to each well, and the reaction was carried out at 37°C in the dark for 4 hours. After discarding the waste liquid, 150 μL of DMSO solution was added to dissolve the purple formazan crystals, and the fluorescence intensity was detected at 490 nm.

[0067] Cell survival rate = (OD value of drug-treated group / OD value of blank group) × 100%

[0068] The results of SZ95 cell survival rate are as follows Figure 1 As shown, the compositions of Examples 1 to 4 and Comparative Examples 1 to 8 have no cytotoxicity to SZ95 cells and are safe and reliable.

[0069] (2) PPAR-γ expression measurement

[0070] Based on SZ95 human sebaceous gland cells, an in vitro sebum secretion model was established by applying dihydrotestosterone (DHT) as a stimulating factor. DHT is a type of androgen that has a clear stimulating effect on sebaceous gland cells and can promote the secretion of sebum and the expression of inflammatory factors. Human sebaceous gland cells (SZ95 cells) were inoculated into DMEM culture medium containing 10% fetal bovine serum and 1% double antibody and cultured at 37°C and 5% CO2. The cultured SZ95 cells were digested and separated with 0.25% insulin-EDTA and plated in 12-well plates at 2×10 4 Cells were cultured at a concentration of cells / well. Complete medium was added to the blank group, complete medium containing 50 μmol / L DHT was added to the model group, and complete medium containing 50 μmol / L DHT and 0.05% concentration of the composition of Examples 1 to 4 and Comparative Examples 1 to 8 was added to the experimental group. After 48 hours of culture, the original medium of the 12-well plate was removed, 1 mL of PBS was added to wash 3 times, and the plate was placed on ice at the same time. 1 mL of RIPA lysis solution containing PMSF was added to each well, and the cells were scraped in the same direction with a scraper, and lysed on ice for 30 minutes. The lysis solution was pipetted into a 2 mL EP tube with a pipette, and the tube was centrifuged at 12000 rpm for 5 minutes. The supernatant was taken, and the total protein concentration was determined using a BCA kit, and then the expression level of the lipid metabolism regulator PPAR-γ was determined using an ELISA kit.

[0071] The results of PPAR-γ expression were as follows Figure 2 As shown, after DHT modeling, the PPAR-γ expression of SZ95 cells increased significantly. After the administration of Examples 1 to 4 and Comparative Examples 1 to 8, the PPAR-γ expression level of the cells decreased significantly, and the cells had PPAR-γ inhibitory activity; among them, the inhibitory effect of the four components in Examples 1 to 4 was the best, which was significantly better than the comparative examples of each component used alone or three components used in combination. Under the premise that each composition had the same total amount ratio and concentration, the four compositions in Examples 1 to 4 were significantly improved.

[0072] Test Example 2 Anti-inflammatory effect of the composition

[0073] The supernatant in step (2) of Test Example 1 was taken, and the total protein concentration was determined using a BCA kit, and then the expression levels of pro-inflammatory cytokines TNF-α and IL-1β were determined using an ELISA kit.

[0074] TNF-α assay results Figure 3As shown, the expression of TNF-α in SZ95 cells in the model group was significantly increased, and after the administration of Examples 1 to 4 and Comparative Examples 1 to 8, the expression level of TNF-α in the cells was significantly reduced, among which the four components in the composition of Examples 1 to 4 had the most obvious effect on reducing the content of the proinflammatory cytokine TNF-α, and the effect was significantly better than the comparative example in which each component was used alone or the three components were used in combination. The combination of the four components further improved the TNF-α inhibitory effect, which had an unexpected effect, indicating that glycyrrhetinic acid, witch hazel extract, Pseudoalteromonas fermentation product extract and hydrolyzed olive leaf extract had a certain synergistic effect after being combined.

[0075] IL-1β assay results Figure 4 As shown, the IL-1β expression of SZ95 cells in the model group was significantly increased, while after the administration of Examples 1 to 4 and Comparative Examples 1 to 8, the IL-1β expression level of the cells was significantly reduced, among which the four components in the composition of Examples 1 to 4 had the most obvious effect on reducing the content of the pro-inflammatory cytokine IL-1β, and the effect was significantly better than the comparative examples in which each component was used alone or the three components were used in combination.

[0076] Test Example 3 Effect of the composition on lipid levels

[0077] Triglyceride (TG) is one of the components of lipids, which is formed by esterification of glycerol and three fatty acids. The effect of the composition of the embodiment and comparative example on cell lipid synthesis was determined by measuring the expression content of cell TG. The experimental treatment method was the same as step (2) in test example 1, and finally the concentration of TG was determined using an ELISA kit.

[0078] The statistical results of TG content in cells are shown in Table 1. In the model group, the TG expression level of SZ95 cells was significantly increased, and after the administration of Examples 1 to 4 and Comparative Examples 1 to 8, the TG expression level of the cells was significantly reduced. Among them, the four components in the composition of Examples 1 to 4 had the most obvious effect on reducing TG content, and the effect was significantly better than the comparative examples in which each component was used alone or the three components were used in combination.

[0079] Table 1 TG content of SZ95 cells

[0080]

[0081]

[0082] Note: Compared with the blank group, ####P<0.000; compared with the model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.001.

[0083] In summary, the study shows that the effect of using witch hazel as a single component is similar to that of the fermentation product of Pseudoalteromonas, and the present invention significantly enhances the lipid inhibition and anti-inflammatory effects of the composition containing three components: glycyrrhetinic acid, Pseudoalteromonas fermentation product extract and hydrolyzed olive leaf extract after compounding the four components, which has unexpected effects; at the same time, the lipid inhibition and anti-inflammatory effects are further improved after compounding the four components, which has a synergistic effect, not only enhancing the inhibitory effect of PPAR-γ, reducing TG content, but also improving the anti-inflammatory effect and inhibiting skin inflammatory response. It shows that the composition provided by the present invention can better inhibit lipid synthesis and inflammatory response, can be used to prevent and / or treat diseases caused by excessive lipid secretion, and is better used to prepare products for treating inflammatory skin diseases such as acne, seborrheic dermatitis, atopic dermatitis, acute contact dermatitis, psoriasis or rosacea.

[0084] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A composition having a PPAR-γ inhibitory effect, characterized in that: The composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (0.5-15): (5-35): (40-75): (5-25).

2. The composition according to claim 1, characterized in that The composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (1-10): (10-30): (45-70): (10-20).

3. The composition according to claim 2, characterized in that The composition comprises the following components in a mass ratio: glycyrrhetinic acid: witch hazel extract: pseudoalteromonas fermentation product extract: hydrolyzed olive leaf extract = (5-10): (10-15): (60-70): (10-15).

4. The method for preparing the composition according to any one of claims 1 to 3, characterized in that: The glycyrrhetinic acid, witch hazel extract, Pseudoalteromonas fermentation product extract and hydrolyzed olive leaf extract in different amounts are directly mixed.

5. Use of the composition according to any one of claims 1 to 3 in the preparation of a lipid synthesis inhibitor.

6. Use of the composition according to any one of claims 1 to 3 in the preparation of a PPAR-γ inhibitor.

7. Use of the composition according to any one of claims 1 to 3 in the preparation of anti-inflammatory products.

8. Use of the composition according to any one of claims 1 to 3 in the preparation of a product for reducing triglycerides.

9. Use of the composition according to any one of claims 1 to 3 in the preparation of a drug for treating inflammatory skin diseases.

10. The use according to claim 9, characterized in that: The inflammatory skin disease is acne, seborrheic dermatitis, atopic dermatitis, acute contact dermatitis, psoriasis or rosacea.

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