Traditional Chinese medicine composition and extract for reducing skin matrix metalloproteinase as well as preparation method and application of traditional Chinese medicine composition and extract

Abstract

The invention discloses a traditional Chinese medicine composition and extract for reducing skin matrix metalloproteinase as well as a preparation method and application of the traditional Chinese medicine composition and extract. The traditional Chinese medicine composition comprises 20-30 parts of folium artemisiae argyi, 10-20 parts of honeysuckle and 5-15 parts of coix seeds. All the components in the traditional Chinese medicine composition can achieve a synergistic effect, and increase of fibroblast matrix metalloproteinase caused by UV is reduced. By optimizing the extraction method, the obtained extract has better effective components and better impression, and can be used for preparing ultraviolet-proof skin care products.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine beauty and skin care, and particularly relates to a traditional Chinese medicine composition for reducing skin matrix metalloproteinases, an extract, and a preparation method and application thereof. Background Art

[0002] Matrix Metalloproteinases (MMPs) are a class of zinc-dependent endopeptidases that can degrade various components in the extracellular matrix (ECM). Matrix Metalloproteinases (MMPs) play an important role in the physiological and pathological processes of the skin. Overexpression of MMPs leads to excessive degradation of matrix components, destroys the normal skin tissue structure, and causes wrinkles or decreased elasticity of the skin. This causes tissue damage and loss of function. Therefore, inhibiting the overexpression of MMPs in the skin matrix under physiological or environmental inducements is crucial to maintaining skin health.

[0003] Commonly used MMPs inhibitors include synthetic small molecule inhibitors such as Batimastat (BB-94) and Marimastat. These compounds bind to the catalytic domain of MMPs and prevent them from degrading the matrix. These inhibitors may lack specificity and easily inhibit multiple MMPs, resulting in a wide range of physiological functions being affected and possibly causing side effects. Natural plants show many advantages in inhibiting matrix metalloproteinases (MMPs). For example, compared with synthetic drugs, natural plant extracts generally have lower toxicity and side effects. Natural plant ingredients generally have good biocompatibility and can interact well with other biological molecules in the human body.

[0004] These ingredients often work by regulating the body's natural balance mechanism and are not likely to cause drug resistance. Plants are rich in polyphenols, flavonoids, saponins and alkaloid compounds, which can inhibit MMPs through different pathways. In addition, natural plants have the advantages of mature production technology, stable raw material supply and low price. However, the main active ingredients in a single plant extract may be limited. In order to achieve a significant inhibitory effect, a single plant extract may require a higher dose, and long-term use may still have safety and tolerability issues.

[0005] Therefore, there may be greater application potential through the synergistic enhancement of multiple plant ingredients. The combination of multiple plant ingredients can work together through different pathways and mechanisms to produce a synergistic effect and improve the overall MMPs inhibition effect. Different plant ingredients can target different MMPs targets and provide a more comprehensive inhibitory effect. By combining multiple plant extracts, the dosage of each ingredient can be reduced, the potential risk of side effects can be reduced, and safety can be improved. Summary of the invention

[0006] The primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide a Chinese medicine composition for reducing skin matrix metalloproteinases.

[0007] Another object of the present invention is to provide an extract for reducing skin matrix metalloproteinases.

[0008] Another object of the present invention is to provide a method for preparing the extract for reducing skin matrix metalloproteinases.

[0009] Another object of the present invention is to provide a use of the extract for reducing skin matrix metalloproteinases.

[0010] The purpose of the present invention is achieved through the following technical solutions: A traditional Chinese medicine composition for reducing skin matrix metalloproteinases comprises the following components in parts by mass: 20-30 parts of wormwood, 10-20 parts of honeysuckle, and 5-15 parts of coix seeds; preferably comprises the following components in parts by mass: 20-25 parts of wormwood, 10-15 parts of honeysuckle, and 5-10 parts of coix seeds; preferably comprises the following components in parts by mass: 25 parts of wormwood, 15 parts of honeysuckle, and 10 parts of coix seeds.

[0011] The skin matrix metalloproteinase is preferably at least one of MMP-1 and MMP-3.

[0012] An extract capable of reducing skin matrix metalloproteinase is prepared from the above-mentioned Chinese medicinal composition capable of reducing skin matrix metalloproteinase.

[0013] The method for preparing the extract for reducing skin matrix metalloproteinases comprises the following steps: (1) Soaking wormwood leaves and honeysuckle in an ethanol solution; then adding water, heating and refluxing for extraction, and filtering to obtain a first extract; (2) Powdering coix seeds, adding water, heating under reflux for extraction, and filtering to obtain a second extract; (3) The first extract and the second extract are mixed and clarified to obtain an extract for reducing skin matrix metalloproteinases.

[0014] Step (3) is preferably: mixing the first extract and the second extract, clarifying, and drying to obtain an extract that reduces skin matrix metalloproteinases.

[0015] The ethanol solution described in step (1) is preferably an ethanol solution with a concentration of 70 to 100% by volume.

[0016] The soaking time in step (1) is preferably 0.5 to 2 hours, more preferably 1 hour.

[0017] The amount of the ethanol solution in step (1) is 1 to 2 times the total volume of the wormwood leaf and honeysuckle; more preferably, it is 1 times the total volume of the wormwood leaf and honeysuckle.

[0018] The volume of water used in step (1) is 4 to 9 times the volume of the ethanol solution; preferably 8 to 9 times the volume of the ethanol solution.

[0019] The time of the heating reflux extraction in step (1) is preferably 0.5 to 2 hours; more preferably 0.5 to 1 hour.

[0020] The amount of water used in step (2) is preferably 9 to 11 times the volume of coix seeds; more preferably 9 to 10 times the volume of coix seeds.

[0021] The time of the heating reflux extraction in step (2) is preferably 0.5 to 2 h; more preferably 0.5 to 1 h.

[0022] The clarification method described in step (3) is preferably centrifugation followed by filtration using a 0.22 μm microporous membrane.

[0023] The drying method is preferably vacuum freeze drying or spray drying.

[0024] The use of the extract capable of reducing skin matrix metalloproteinase in the preparation of UV protection skin care products.

[0025] The dosage forms of the anti-ultraviolet skin care products include but are not limited to facial masks, external creams and external patches.

[0026] Compared with the prior art, the present invention has the following advantages and effects: (1) The present invention provides a traditional Chinese medicine composition for reducing skin matrix metalloproteinases. The components in the traditional Chinese medicine composition can synergistically enhance their effectiveness and reduce the increase of fibroblast matrix metalloproteinases caused by UV.

[0027] (2) The present invention has a low effective concentration and is non-toxic and non-irritating.

[0028] (3) The present invention intentionally soaks mugwort and honeysuckle in alcohol during the extraction process, which increases the dissolution rate of the two traditional Chinese medicine ingredients. In addition, extracting coix seed alone prevents the high viscosity of coix seed extract from inhibiting the dissolution of the effective ingredients of mugwort and honeysuckle and the darkening of the color after mixed extraction, thereby preventing the appearance from being affected. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the result of the effect of Chinese herbal extract on fibroblasts on MMP-1 and MMP-3; among them, ** indicates that compared with the model group, P <0.01; *** indicates that compared with the model group, P<0.001; ns means there is no significant difference compared with the model group. DETAILED DESCRIPTION

[0030] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0031] Unless otherwise specified, the present invention has no special requirements on the sources of the components of the traditional Chinese medicine composition, and commercially available products known to those skilled in the art may be used.

[0032] Example 1 A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following parts by mass: 25 parts of wormwood leaves, 15 parts of honeysuckle flowers, and 10 parts of coix seeds.

[0033] (1) Weigh wormwood leaves and honeysuckle and soak them in 1 volume of anhydrous ethanol for 1 hour; add 9 volumes of water to the ethanol-soaked wormwood leaves and honeysuckle, heat and reflux for extraction for 1 hour, and separate with a filter bag to obtain a first extract.

[0034] (2) Weigh coix seeds and grind them into powder. Add 10 times the volume of water and heat under reflux for extraction for 1 hour. Separate with a filter bag to obtain the second extract.

[0035] (3) The first extract and the second extract were mixed at room temperature, centrifuged at 8500 rpm, and then clarified by a 0.22 μm microporous filter membrane to obtain a third extract. The third extract was dried to obtain a Chinese herbal extract for reducing skin matrix metalloproteinases.

[0036] Example 2 The difference between this embodiment and embodiment 1 is that the mass fraction of the Chinese medicine composition is changed.

[0037] A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following parts by mass: 20 parts of wormwood leaves, 10 parts of honeysuckle flowers, and 5 parts of coix seeds.

[0038] (1) Weigh wormwood leaves and honeysuckle and soak them in 1 volume of anhydrous ethanol for 0.5 h; add 8 volumes of water to the wormwood leaves and honeysuckle soaked in anhydrous ethanol, heat and reflux to extract for 0.5 h, and separate with a filter bag to obtain the first extract.

[0039] (2) Weigh coix seeds and grind them into powder. Add 9 times the volume of water and heat under reflux for extraction for 0.5 h. Separate with a filter bag to obtain the second extract.

[0040] (3) The first extract and the second extract were mixed at room temperature, centrifuged at 8500 rpm, and then clarified by a 0.22 μm microporous filter membrane to obtain a third extract. The third extract was dried to obtain a Chinese herbal extract for reducing skin matrix metalloproteinases.

[0041] Example 3 The difference between this embodiment and embodiment 1 is that the mass fraction of the Chinese medicine composition is changed.

[0042] A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following parts by mass: 30 parts of wormwood leaves, 20 parts of honeysuckle flowers, and 15 parts of coix seeds.

[0043] (1) Weigh wormwood leaves and honeysuckle and soak them in 2 times the volume of anhydrous ethanol for 2 hours; add 9 times the volume of water to the wormwood leaves and honeysuckle soaked in anhydrous ethanol, heat and reflux for extraction for 2 hours, and separate with a filter bag to obtain a first extract.

[0044] (2) Weigh coix seeds and grind them into powder. Add 11 times the volume of water and heat under reflux for extraction for 2 h. Separate with a filter bag to obtain the second extract.

[0045] (3) The first extract and the second extract were mixed at room temperature, centrifuged at 8500 rpm, and then clarified by a 0.22 μm microporous filter membrane to obtain a third extract. The third extract was dried to obtain a Chinese herbal extract for reducing skin matrix metalloproteinases.

[0046] Comparative Example 1 The difference between the comparative example and Example 1 is that honeysuckle and coix seed are removed.

[0047] A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following mass parts: 50 parts of mugwort leaves.

[0048] Weighing mugwort leaves and putting them into 1 volume of anhydrous ethanol to soak for 1 hour; adding 9 volumes of water to the mugwort leaves soaked in anhydrous ethanol, heating and refluxing for extraction for 1 hour, and separating through a filter bag to obtain a first extract. The first extract is centrifuged at 8500 rpm, clarified and dried through a 0.22 μm microporous filter membrane to obtain a Chinese medicine extract that reduces skin matrix metalloproteinases.

[0049] Comparative Example 2 The difference between the comparative example and Example 1 is that wormwood leaf and coix seed are removed.

[0050] A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following parts by mass: 50 parts of honeysuckle.

[0051] Weigh honeysuckle and put it into 1 volume of anhydrous ethanol to soak for 1 hour; add 9 volumes of water to the honeysuckle soaked in anhydrous ethanol, heat and reflux for extraction for 1 hour, and separate through a filter bag to obtain a first extract. The first extract is centrifuged at 8500rpm, clarified and dried through a 0.22μm microporous filter membrane to obtain a traditional Chinese medicine extract that reduces skin matrix metalloproteinases.

[0052] Comparative Example 3 The difference between the comparative example and embodiment 1 is that wormwood leaf and honeysuckle are removed.

[0053] A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following parts by mass: 50 parts of coix seeds.

[0054] Weigh coix seeds and grind them into powder, add 10 times the volume of water, heat and reflux to extract for 2 hours, separate the extract with a filter bag, and dry it to obtain the Chinese medicine extract that reduces skin matrix metalloproteinases.

[0055] Comparative Example 4 The difference between the comparative example and Example 1 is that the extraction preparation method is changed.

[0056] A traditional Chinese medicine composition for reducing skin matrix metalloproteinases is prepared according to the following parts by mass: 25 parts of wormwood leaves, 15 parts of honeysuckle flowers, and 10 parts of coix seeds.

[0057] Weigh wormwood leaves, honeysuckle and coix seeds, mix them, add 10 times the volume of water, heat and reflux to extract for 1 hour, and separate with a filter bag to obtain the first extract.

[0058] The first extract was centrifuged at 8500 rpm, clarified by a 0.22 μm microporous filter, and dried to obtain a Chinese herbal extract for reducing skin matrix metalloproteinases. The Chinese herbal extract was darker in color than the extract obtained in Example 1, which should be due to the co-heating reaction of the sugars in coix seeds and wormwood and honeysuckle.

[0059] Test Example 1 (1) Fibroblasts (mouse embryonic fibroblasts NIH-3T3, Beina Biotech) were revived from liquid nitrogen and cultured in a 37°C incubator with 5% CO2. Cells were cultured using DMEM medium (containing 10% v / v FBS) until cell confluence reached 70-80%.

[0060] (2) Wash the cells twice with PBS and digest them with trypsin (0.25%) to prepare a cell suspension. Calculate the cell concentration and adjust it to an appropriate concentration (about 1×10 5 The cell suspension was seeded into a 96-well plate, 200 µL was added to each well, and cultured for 24 hours to allow the cells to adhere.

[0061] (3) Remove the culture medium and wash the cells twice with PBS buffer.

[0062] (4) Place the culture plate under a UV lamp and expose it to 254 nm ultraviolet light (UV) at a dose of 40 mJ / cm 2 .

[0063] (5) After irradiation, 200 µL of fresh culture medium or 200 µL of fresh culture medium containing each Chinese herbal medicine extract (final concentration of 10 μg / mL) was immediately added and cultured for another 24 h.

[0064] (6) After 24 hours, the cell culture supernatant of each group was collected and centrifuged to remove cell debris. The supernatant was used to detect MMP-1 and MMP-3 by ELISA. The test results were as follows: Figure 1 As shown. After UV irradiation, MMP-1 and MMP-3 in the model group were significantly increased, among which Examples 1 to 3 could effectively inhibit the increase of MMP1 and MMP-3, and were significant. Comparative Examples 1, 2 and 3 using wormwood alone, honeysuckle alone and coix seed alone, as well as changing the extraction method, were less effective than Example 1.

[0065] In summary, the Chinese medicine composition provided by the present invention can effectively reduce skin matrix metalloproteinases and produce a synergistic effect. Although the above embodiment has made a detailed description of the present invention, it is only a part of the embodiments of the present invention, rather than all the embodiments. People can also obtain other embodiments based on this embodiment without inventiveness, and these embodiments all belong to the protection scope of the present invention.

Claims

1. A Chinese medicine composition for reducing skin matrix metalloproteinases, characterized in that The invention comprises the following components in parts by mass: 20 to 30 parts of wormwood leaves, 10 to 20 parts of honeysuckle flowers and 5 to 15 parts of coix seeds.

2. The Chinese medicine composition for reducing skin matrix metalloproteinases according to claim 1, characterized in that The invention comprises the following components in parts by mass: 20-25 parts of wormwood leaves, 10-15 parts of honeysuckle flowers and 5-10 parts of coix seeds.

3. An extract for reducing skin matrix metalloproteinases, characterized in that: The composition is prepared from the Chinese medicinal composition for reducing skin matrix metalloproteinases according to claim 1 or 2.

4. The method for preparing the extract for reducing skin matrix metalloproteinases according to claim 3, characterized in that The steps include: (1) Soaking wormwood leaves and honeysuckle in an ethanol solution; then adding water, heating and refluxing for extraction, and filtering to obtain a first extract; (2) Powdering coix seeds, adding water, heating under reflux for extraction, and filtering to obtain a second extract; (3) The first extract and the second extract are mixed and clarified to obtain an extract for reducing skin matrix metalloproteinases.

5. The method for preparing an extract for reducing skin matrix metalloproteinases according to claim 4, characterized in that: Step (3) is: mixing the first extract and the second extract, clarifying, and drying to obtain an extract that reduces skin matrix metalloproteinases.

6. The method for preparing an extract for reducing skin matrix metalloproteinases according to claim 4 or 5, characterized in that: The ethanol solution described in step (1) is an ethanol solution with a concentration of 70 to 100% by volume; The amount of the ethanol solution in step (1) is 1 to 2 times the total volume of wormwood leaves and honeysuckle; The volume of water used in step (1) is 4 to 9 times the volume of the ethanol solution; The amount of water used in step (2) is 9 to 11 times the volume of coix seeds.

7. The method for preparing an extract for reducing skin matrix metalloproteinases according to claim 4 or 5, characterized in that: The soaking time in step (1) is 0.5 to 2 hours; The heating reflux extraction time in step (1) is 0.5 to 2 hours; The time of the heating reflux extraction in step (2) is 0.5 to 2 hours.

8. The method for preparing an extract for reducing skin matrix metalloproteinases according to claim 5, characterized in that: The clarification method is centrifugation; The drying method is vacuum freeze drying or spray drying.

9. Use of the extract for reducing skin matrix metalloproteinases according to claim 3 in the preparation of UV protection skin care products.

10. The use according to claim 9, characterized in that: The dosage form of the anti-ultraviolet skin care product is a facial mask, an external cream or an external patch.

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