Anti-inflammatory compositions, uses and skin care products

Abstract

The application belongs to the technical field of skin care, and discloses an anti-inflammatory composition, application and skin care product.The anti-inflammatory composition comprises a kuding tea extract and a litchi extract, and the weight ratio of the kuding tea extract to the litchi extract is 1-3:1-3.The application provides an anti-inflammatory composition, and the kuding tea extract and the litchi extract are compounded, so that the anti-inflammatory composition has excellent skin anti-inflammatory effect.

Description

Technical Field

[0001] This application relates to the field of skin care technology, specifically to an anti-inflammatory composition, its uses, and skin care products. Background Technology

[0002] Anti-inflammatory ingredients significantly improve skin health through multiple mechanisms. Their core value lies in inhibiting the release of pro-inflammatory factors and quickly relieving acute inflammatory reactions such as redness and stinging. At the same time, they regulate the immune response and have a long-term stabilizing effect on chronic inflammatory diseases such as eczema and rosacea. In acne treatment, these products control acne formation from the root and accelerate the fading of acne scars by inhibiting the proliferation of Propionibacterium acnes and regulating sebum secretion. In addition, anti-inflammatory ingredients can block the breakdown of collagen by matrix metalloproteinases, delay the process of "inflammatory aging," and, combined with an antioxidant system, can further reduce UV-induced photodamage.

[0003] Therefore, anti-inflammation is a very important core for skin care. For example, prior art 1: KR20130096621A discloses a composition for skin anti-oxidation, anti-inflammation and skin whitening. Anthocyanin 3-O-glucoside inhibits the production of free radicals, thus having an antioxidant effect. In addition, anthocyanin 3-O-glucoside can also inhibit the production of NO, thereby having an anti-inflammatory effect.

[0004] The specification of prior art 1 states: "The compound of formula (1) can be isolated or synthesized from plant extracts, or a commercially available compound can be used."

[0005] In other words, existing technology 1 provides technical inspiration for using plant extracts for anti-inflammatory purposes. However, due to the safety concerns of cosmetics, the amount of plant extracts used is limited. Therefore, the anti-inflammatory effect that can be achieved by using a single plant extract is also relatively limited. It is necessary to develop a composition of plant extracts to further enhance the anti-inflammatory effect on the skin. Summary of the Invention

[0006] One of the objectives of this application is to provide an anti-inflammatory composition obtained by combining bitter tea extract and lychee extract, which effectively solves the problem of insufficient anti-inflammatory effect on the skin due to the limitation of the amount of single plant extract.

[0007] Another object of this application is to provide the use of an anti-inflammatory composition for the preparation of skin care products.

[0008] In addition, this application also provides a skin care product containing the anti-inflammatory composition of this application, which has excellent anti-inflammatory effects.

[0009] To achieve the above objectives, this application provides an anti-inflammatory composition comprising bitter tea extract and lychee extract, wherein the weight ratio of bitter tea extract to lychee extract is 1-3:1-3.

[0010] Preferably, the weight ratio of the bitter tea extract to the lychee extract is 1:1.

[0011] This application also provides the use of the above-described anti-inflammatory composition in the preparation of skin care products.

[0012] This application also provides a skin care product containing 1-3 wt% of the above-mentioned anti-inflammatory composition.

[0013] Beneficial effects

[0014] Compared with the prior art, this application provides an anti-inflammatory composition that combines bitter tea extract and lychee extract, and has excellent anti-inflammatory effects on the skin. Detailed Implementation

[0015] The present application will be further described below with reference to embodiments, but this does not constitute any limitation on the present application. Any limited modifications made within the scope of the claims of the present application shall still be within the scope of the claims of the present application.

[0016] To illustrate the technical content of this application in detail, the following description is provided in conjunction with the embodiments.

[0017] In the following embodiments and comparative examples, unless otherwise specified, the parts are parts by weight and the percentage is a weight percentage.

[0018] Example 1

[0019] An anti-inflammatory composition comprising 1 part bitter tea extract and 1 part lychee extract.

[0020] Example 2

[0021] An anti-inflammatory composition comprising 3 parts bitter tea extract and 1 part lychee extract.

[0022] Example 3

[0023] An anti-inflammatory composition comprising 1 part bitter tea extract and 3 parts lychee extract.

[0024] Comparative Example 1

[0025] 1 part bitter tea extract.

[0026] Comparative Example 2

[0027] 1 part lychee extract.

[0028] Efficacy test

[0029] I. Capsaicin (Cap) induces keratinocyte-inflammatory factor (IL-8)

[0030] Keratinocytes are a direct target of capsaicin (CAP) and can activate TRPV1. Subcutaneous injection of capsaicin induces neurogenic inflammation, which is related to the release of inflammatory mediators through TRPV1 sensitization of peripheral nerves. TRPV1 activation also participates in the release of inflammatory mediators, mediating the upregulation of IL-8 gene and protein expression levels in cells. The levels of the inflammatory cytokine IL-8 in the negative control and experimental sample groups were measured to evaluate the efficacy of the experimental sample in inhibiting IL-8 levels. IL-8 levels were measured using enzyme-linked immunosorbent assay (ELISA). The results of cellular IL-8 inflammatory cytokine level measurements were compared between the experimental sample and the negative control. If the cellular IL-8 inflammatory cytokine level in the experimental sample was significantly lower than that in the negative control, the experimental sample was considered to have a soothing effect.

[0031] (1) Instruments and equipment: RT-6100 enzyme labeling analyzer.

[0032] (2) Cells used in the experiment: keratinocytes-HaCaT cells.

[0033] (3) Experimental grouping:

[0034] Negative control group (NC): Cells (+), CAP (+), Samples (-);

[0035] Sample group (S): Cells (+) CAP (+) Samples (+).

[0036] Cell viability assay: Keratinocytes (HaCaT cells) were seeded into 96-well plates. After 24 hours, the culture medium was discarded, and basal culture medium containing different concentrations of the test samples was added. After 24 hours, the OD490nm was measured by MTT assay, and the effect of the test samples on cell viability was analyzed by t-test. Cell viability was calculated using the following formula:

[0037] Cell viability (%) = (sample OD) 490nm ) / (Negative control OD 490nm )*100

[0038] IL-8 inflammatory factor assay: Keratinocytes were seeded into 96-well plates and cultured for 24 h. Samples were added according to the groups, and after incubation for 30 min, the supernatant was discarded. Except for blank wells, CAP was added to each well to a final concentration of 8 μmol / L. The plates were incubated in a CO2 incubator for 24 h. The cell supernatant was collected and centrifuged. The supernatant was then used to determine the IL-8 content in the cell culture supernatant using an ELISA kit. The relative IL-8 content was calculated using the following formula:

[0039] IL-8 relative content (%) = (IL-8 content in sample) / (IL-8 content in negative control) * 100

[0040] Before testing the IL-8 inflammatory factor content in the sample cells, cell viability screening was performed to determine the optimal test concentration.

[0041] The results of different concentration tests of Examples 1-3 and Comparative Examples 1-2 were performed according to the above method. The cytotoxicity results are shown in Table 1, and the relative IL-8 content test results are shown in Table 2.

[0042] Table 1. Cytotoxicity results of different concentrations in Examples 1-3 and Comparative Examples 1-2

[0043]

[0044] According to the results in Table 1:

[0045] For those skilled in the art, when the average cytotoxicity value is >90, the toxicity level is non-cytotoxic, while when the average cytotoxicity value is ≤90, it already has different degrees of cytotoxicity.

[0046] According to the data from Comparative Example 1, 0.5% of the bitter tea extract is non-cytotoxic, but when the concentration of the bitter tea extract reaches 1% or even higher, it is cytotoxic.

[0047] According to the data from Comparative Example 2, 0.5-1.5% litchi extract has no cytotoxicity. However, when the concentration of litchi extract reaches 1.8% and 2%, it already has a certain degree of cytotoxicity.

[0048] According to the results of Example 1, when the bitter tea extract and lychee extract were compounded in a weight ratio of 1:1, the concentrations of 0.5-2% were all non-cytotoxic. At the 2% concentration, the concentration of bitter tea extract had reached 1%. According to the data of Comparative Example 1, 1% bitter tea extract had already produced cytotoxicity. However, by compounding with lychee extract in a weight ratio of 1:1, 2% of Example 1 reached the safety level. It is speculated that the active ingredients contained in lychee extract can effectively reduce the cytotoxicity of bitter tea extract.

[0049] However, based on the data from Examples 2 and 3, it was found that the above speculation is not entirely accurate:

[0050] Example 2: When bitter tea extract and lychee extract were combined in a weight ratio of 3:1, they still exhibited cytotoxicity even at a concentration of 0.5%. At this time, the concentration of bitter tea extract in the composition was 0.375% and the concentration of lychee extract was 0.125%. Neither of them had cytotoxicity when they were alone, but the combination showed low cytotoxicity.

[0051] In Example 3, when bitter tea extract and lychee extract were combined in a weight ratio of 1:3, although there was no cytotoxicity at a concentration of 0.5%, low cytotoxicity was observed when the concentration reached 1% or higher. Specifically, at a concentration of 2%, the bitter tea extract concentration in the composition was 0.5% and the lychee extract concentration was 1.5%. Neither of them had cytotoxicity when they existed alone, but the combination showed low cytotoxicity.

[0052] Based on the above experimental data, the following conclusions can be drawn: The composition formed by combining bitter tea extract and lychee extract in a 1:1 weight ratio can significantly reduce the toxicity of both bitter tea extract and lychee extract. Even when the concentration of the composition reaches 2%, its toxicity level is still non-cytotoxic. However, when the weight ratio of bitter tea extract and lychee extract changes, whether the weight proportion of bitter tea extract, which has relatively higher cytotoxicity, is higher, or the weight proportion of lychee extract, which has relatively lower cytotoxicity, is higher, the composition exhibits cytotoxicity that damages cells when the concentration exceeds 1%.

[0053] Table 2. Results of relative IL-8 content tests in Examples 1-3 and Comparative Examples 1-2 at different concentrations

[0054] Comparative Example 1 0.50% 74.79±0.20 Comparative Example 2 1.50% 72.10±0.18 Example 1 2% 32.05±0.09

[0055] As shown in Table 1, the higher concentrations of Examples 1-3 and Comparative Examples 1-2 were cytotoxic and could not be used for relative IL-8 content testing. Therefore, specific concentrations of Examples and Comparative Examples without cytotoxicity were selected for relative IL-8 content testing, and the results are shown in Table 2.

[0056] Based on the results in Table 2, and combined with the results in Table 1, the following analysis was conducted:

[0057] Example 1 of this application combines bitter tea extract and lychee extract in a weight ratio of 1:1, which can break through the upper limit of the amount of bitter tea extract. The resulting composition can reach a concentration of 2% without cytotoxicity. Compared with Comparative Example 1 and Comparative Example 2, the relative content of IL-8 in Example 1 at 2% is significantly reduced, showing a better anti-inflammatory effect.

[0058] II. In vitro PGE2 content determination

[0059] Macrophages can be used as a cell model to study the inhibition of PGE2 levels by cosmetics. Through stimulation with bacterial lipopolysaccharide (LPS), the PGE2 levels in the negative control and experimental sample groups were measured to evaluate the efficacy of the experimental samples in inhibiting PGE2 levels. PGE2 levels were determined using enzyme-linked immunosorbent assay (ELISA). The results of cellular PGE2 level measurements were compared between the experimental sample and the negative control. If the cellular PGE2 level in the experimental sample was significantly lower than that in the negative control, the experimental sample was considered to have a soothing effect.

[0060] (1) Instruments and equipment: RT-6100 enzyme labeling analyzer.

[0061] (2) Cells used in the experiment: macrophages - RAW264.7 cells.

[0062] (3) Sample preparation

[0063] Sample group: Dilute the sample with pure water to the required concentration;

[0064] Negative control group: basal culture medium (cell viability assay); basal culture medium containing LPS (PGE2 content assay).

[0065] Cell viability assay: RAW264.7 cells were seeded in 96-well plates. After 24 hours, the culture medium was discarded, and basal culture medium containing different concentrations of the test samples was added. After 24 hours, the OD490nm was measured by MTT assay, and the effect of the test samples on the viability of RAW264.7 cells was analyzed by t-test. Cell viability was calculated using the following formula:

[0066] Cell viability (%) = (sample OD) 490nm ) / (Negative control OD 490nm )*100

[0067] PGE2 content determination: RAW264.7 cells were seeded into 96-well plates and cultured for 24 h. The culture medium was then replaced with basal medium containing different concentrations of the experimental samples. After 24 h, the cell supernatant was collected and centrifuged. The supernatant was then aspirated, and the PGE2 content in the cell culture supernatant was determined using an ELISA kit to determine the effect of the experimental samples on the PGE2 content of macrophages. The relative PGE2 content was calculated using the following formula:

[0068] PEG2 relative content (%) = Sample PEG2 content / Negative control PEG2 content * 100

[0069] The cytotoxicity results of Examples 1-3 and Comparative Examples 1-2 were tested at different concentrations according to the above method. The results are shown in Table 3. The results of the PGE2 relative content test are shown in Table 4.

[0070] Table 3. Cytotoxicity results of different concentrations in Example 1 and Comparative Examples 1-2

[0071]

[0072] According to the results in Table 3:

[0073] According to the data from Comparative Example 1, when the concentration of bitter tea extract reaches 1.8% or higher, it can also produce cytotoxicity to macrophages and cause damage to them.

[0074] According to the data from Comparative Example 2, the litchi extract showed no cytotoxicity in the concentration range of 0.5-2%.

[0075] Meanwhile, the composition of Example 1 showed no cytotoxicity in the concentration range of 0.5-2%.

[0076] Table 4. Results of relative PGE2 content tests in Examples 1-3 and Comparative Examples 1-2 at different concentrations

[0077]

[0078] According to the results in Tables 3 and 4:

[0079] Kuding tea extract itself has a good anti-inflammatory effect and can significantly reduce the relative content of PGE2; while lychee extract does not have the effect of reducing the relative content of PGE2; however, since kuding tea extract exhibits cytotoxicity after a concentration of 1.80%, the concentration of kuding tea extract should not be too high.

[0080] In Example 1 of this application, it was found that when the bitter tea extract and lychee extract were combined in a weight ratio of 1:1, the combination did not reduce the relative content of PGE2 at concentrations of 1% and 1.5%. Instead, it showed a certain effect of reducing the relative content of PGE2 at a concentration of 1.1%. Furthermore, at a concentration of 2%, it produced a very good effect of reducing the relative content of PGE2. At this point, the concentration of bitter tea extract in the composition was only half, i.e., 1%. It is speculated that lychee extract can effectively enhance the efficacy of bitter tea extract at a certain concentration, thus playing a synergistic role.

[0081] Similarly, it was found that Examples 2 and 3 did not reduce the relative content of PGE2 at concentrations of 1% and 1.5%; however, based on the changes in the relative content of PGE2 at concentrations of 1% and 1.5%, it can be inferred that as the concentration increases, it gradually shows a certain trend of reducing the relative content of PGE2.

[0082] The embodiments presented herein are merely selected implementations based on combinations of all possible embodiments. The appended claims should not be limited to the embodiments described herein. Some numerical ranges used in the claims include sub-ranges within them, and variations within these ranges should also be covered by the appended claims.

Claims

1. An anti-inflammatory composition, characterized in that, It includes bitter tea extract and lychee extract, wherein the weight ratio of bitter tea extract to lychee extract is 1-3:1-3.

2. The anti-inflammatory composition according to claim 1, characterized in that, The weight ratio of the bitter tea extract to the lychee extract is 1:

1.

3. Use of the anti-inflammatory composition as described in claim 1 or 2 to prepare skin care products.

4. A skincare product, characterized in that, Contains 1-3 wt% of the anti-inflammatory composition as described in claim 1 or 2.

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