CN115089645A

CN115089645A - Composition and skin care product for inhibiting TRPV1 pathway and reducing skin irritation

Info

Publication number: CN115089645A
Application number: CN202210598546.6A
Authority: CN
Inventors: 张建华; 郭文姣
Original assignee: Nord Traceability Guangzhou Biotechnology Co ltd
Current assignee: Nord Traceability Guangzhou Biotechnology Co ltd
Priority date: 2022-05-30
Filing date: 2022-05-30
Publication date: 2022-09-23
Anticipated expiration: 2042-05-30
Also published as: CN115089645B

Abstract

The invention provides a composition for inhibiting a TRPV1 pathway and reducing skin irritation, which comprises a camellia extract and a purslane extract, wherein the mass ratio of the camellia extract to the purslane extract is 0.01-10: 0.01-10. The composition for inhibiting the TRPV1 channel and reducing skin irritation comprises a camellia extract and a purslane extract which can inhibit the TRPV1 channel, and the camellia extract and the purslane extract have the combined action, so that the threshold value of a irritation response can be increased, the skin sensitivity response can be reduced, and the skin irritation, pruritus and burning sensation caused by the activation of the TRPV1 channel can be reduced.

Description

Composition and skin care product for inhibiting TRPV1 pathway and reducing skin irritation

Technical Field

The invention belongs to the technical field of daily cosmetics, and particularly relates to a composition for inhibiting a TRPV1 pathway to reduce skin irritation, and a skin care product for inhibiting a TRPV1 pathway to reduce skin irritation.

Background

The skin is an organ covering the surface of the human body, is one of the most important organs, and has the effects of protecting the human body from the external environment and irritation and maintaining moisture balance. When the skin is strongly stimulated, the barrier on the surface of the skin may be damaged to cause a series of immune reactions, and at the same time, the nerve endings on the surface of the skin receive signals of stimulation and transmit the signals to the nerve cells to cause a series of feelings such as stabbing pain, itching, burning and the like. The nerve endings receive the stimulation signals with a certain threshold value, and if the stimulation signals are lower than the threshold value, the corresponding signal conduction can not be generated to generate response. Hypersensitivity may be caused when the nerve response is excessive, and slight stimulation can cause reactions such as stabbing pain, itching, burning and the like. Sensitive skin may cause a stronger response to mild disturbance due to either exposure of nerve endings or hyperactivity of nerve endings due to barrier damage.

Transient receptor potential vanilloid 1 (TRPV1) channel is a non-selective cation channel, and after activation, it can make extracellular calcium ion flow in, resulting in increased intracellular calcium ion concentration, and further mediating a series of physiological or pathological reactions. In recent years, TRPV1 channels were found to have a significant impact on stinging, itching, burning. By exogenously administering the TRPV1 antagonist, the stress response of the skin can be effectively reduced, and the skin barrier can be improved.

Camellia (academic name: Camellia sp.) is also called Camellia, wherein 700 discovered compounds capable of promoting human health comprise flavone, polyphenol and other components, the Camellia has been proved to have various physiological activities, and the Camellia extract can inhibit the generation of tyrosinase, so that the skin color can be evened, the photoaging caused by UVB can be resisted, and the skin damage caused by environmental pollution can be reduced.

Purslane is also named as locust vegetable, is a common traditional Chinese medicine, and tender stems and leaves of the purslane can be used as vegetables. It is rich in amino acids, proteins, and polysaccharides. The research shows that the extract and the chemical components of the plant extract mainly have the biological activities of anti-inflammation, anti-diabetes, anti-oxidation, anti-cancer, antibiosis and the like, can reduce the redness on the aspect of skin and is beneficial to the improvement of the skin barrier.

Based on this, the present invention provides a composition for inhibiting the TRPV1 pathway to reduce skin irritation, capable of inhibiting stinging, itching and burning of the skin due to TRPV1 channel activation.

Disclosure of Invention

The invention aims to provide a composition for inhibiting a TRPV1 pathway to reduce skin irritation, and solves the technical problem that the existing cosmetics are less deficient in reducing skin irritation by inhibiting a TRPV1 pathway.

The second purpose of the invention is to provide a skin care product which inhibits TRPV1 pathway and reduces skin irritation.

The purpose of the invention is realized by the following technical scheme:

a composition for inhibiting a TRPV1 pathway to reduce skin irritation comprises a camellia extract and a purslane extract, wherein the mass ratio of the camellia extract to the purslane extract is 0.01-10: 0.01-10.

Further, the mass ratio of the camellia extract to the purslane extract is 1-10: 1-10.

The camellia sinensis flower extract in the composition for inhibiting the TRPV1 pathway and reducing the skin irritation contains rich polyphenol and flavonoid compounds, and has good antioxidant activity. The purslane extract contains abundant flavones, alkaloids and polysaccharides, and has good antibacterial, antioxidant and anti-inflammatory effects. The camellia extract and the purslane extract selected by the invention have an inhibiting effect on TRPV1 channels, can improve the threshold value of the stimulation response, and can reduce the skin sensitivity response. The two extracts, when combined, have a synergistic effect in inhibiting skin stinging, itching and burning due to activation of TRPV1 channel.

In the invention, the protocatechuic acid content of the camellia extract is not lower than 1%.

The invention can be improved as follows, and the camellia extract is prepared by the following steps:

(1) drying flos Camelliae Japonicae, removing impurities, and pulverizing to 20-100 mesh;

(2) adding 10-30 times of water and ethanol mixture, decocting under reflux at normal pressure, filtering, and concentrating the filtrate by vacuum distillation to 80% -200% of the weight of flos Camelliae Japonicae to obtain crude extract;

(3) adding adsorbent 10-20% of the crude extract, microwave shaking to colorless, filtering, and vacuum distilling the filtrate to dry to obtain flos Camelliae Japonicae extract.

Further, the volume ratio of water to ethanol in the mixture of water and ethanol is 20:80-80: 20.

Preferably, the volume ratio of water to ethanol is 40:60 to 60: 40.

The temperature of reflux decoction is 80-95 deg.C.

In the invention, the polysaccharide content of the purslane extract is not less than 10%.

The invention can also be improved as follows, and the purslane extractive is prepared by the following steps:

(1) drying herba Portulacae, removing impurities, and pulverizing to 20-100 mesh;

(2) adding a mixture of water and ethanol with the weight of 10-60 times of that of the purslane, carrying out reflux decoction under normal pressure, filtering, and carrying out reduced pressure distillation and concentration on the filtrate until the weight of the purslane is 80-200% of that of the purslane to obtain a crude extract;

(3) adding adsorbent 10-20% of the weight of the crude extract, performing microwave oscillation until the crude extract is colorless, filtering, and distilling the filtrate under reduced pressure until the filtrate is dried to obtain the purslane extract.

Further, the weight ratio of the added mixture of water and ethanol to the purslane is 40-60 times.

Further, the volume ratio of the water to the ethanol is 10:90-90: 20.

Preferably, the volume ratio of water to ethanol is 70:30 to 90: 20.

Further, the temperature of reflux decoction is 80-95 ℃.

Preferably, the reflux decoction temperature is 90-95 ℃.

A skin care product for inhibiting TRPV1 pathway to reduce skin irritation comprises the composition for inhibiting TRPV1 pathway to reduce skin irritation.

Further, the skin care product for inhibiting the TRPV1 pathway to reduce skin irritation also comprises the following components in percentage by mass: 1 to 5 percent of glycerin, 1 to 5 percent of butanediol, 0.1 to 1 percent of allantoin, 0.1 to 1 percent of thickening agent, 0.01 to 0.5 percent of sodium hyaluronate, 0.1 to 5 percent of squalane, 0.5 to 1.5 percent of emulsifier and the balance of water.

In the invention, the thickening agent is one or more of carbomer and a neutralizing agent thereof, xanthan gum and acrylate/C10-30 alkyl acrylate cross-linked polymer (copolymer).

The emulsifier is one or more of olem 1000 and Montanov L.

Compared with the prior art, the invention has the following beneficial effects:

(1) the composition for inhibiting the TRPV1 channel and reducing skin irritation comprises a camellia extract and a purslane extract which can inhibit the TRPV1 channel, and the camellia extract and the purslane extract have the combined action, so that the threshold value of a irritation response can be increased, the skin sensitivity response can be reduced, and the skin irritation, pruritus and burning sensation caused by the activation of the TRPV1 channel can be reduced.

(2) The composition for inhibiting the TRPV1 pathway to reduce skin irritation, combined with the application of the composition in cosmetics, can be widely applied to cosmetics and medicines, and solves various skin problems.

Drawings

FIG. 1 is a fluorescent detection map of TRPV1 protein from Blank Control (BC);

fig. 2 is a fluorescent detection map of TRPV1 protein for Negative Control (NC);

FIG. 3 is a fluorescent detection map of TRPV1 protein of negative control (4-tert-butylcyclohexanol);

FIG. 4 is a fluorescent detection map of TRPV1 protein of comparative example 1;

FIG. 5 is a fluorescent detection map of TRPV1 protein of comparative example 2;

fig. 6 is a fluorescent detection map of TRPV1 protein of the composition of example 1 that inhibits the TRPV1 pathway to reduce skin irritation;

fig. 7 is a fluorescent detection map of TRPV1 protein from the composition of example 2 that inhibits the TRPV1 pathway to reduce skin irritation;

fig. 8 is a fluorescent detection map of TRPV1 protein from the composition of example 3 that inhibits the TRPV1 pathway to reduce skin irritation;

FIG. 9 is a graph showing the results of the TRPV1 protein content assay of different working solutions tested;

FIG. 10 is a graph of the glow-on-capsaicin results for the skin care product of comparative example 1;

FIG. 11 is a graph of the hot pepper results of comparative example 2;

FIG. 12 is a graph of the burning results of capsaicin in the skin care product of example 1;

FIG. 13 is a graph of the burning results of capsaicin in the skin care product of example 2;

fig. 14 is a graph of the burning results of capsaicin for the skin care product of example 3.

Detailed Description

The present invention is further described below in conjunction with specific examples to better understand and implement the technical solutions of the present invention for those skilled in the art. In the following examples, the content of protocatechuic acid in the camellia extract is not less than 1%, and the content of polysaccharide in the purslane extract is not less than 10%.

The camellia extract is prepared by the following steps:

(2) adding 10-30 times of water and ethanol mixture at a volume ratio of 50:50, decocting under reflux at 85 deg.C under normal pressure, filtering, and concentrating the filtrate by vacuum distillation to 150% of the weight of flos Camelliae Japonicae to obtain crude extract;

(3) adding 15% adsorbent activated carbon particles, microwave shaking to colorless, filtering, and vacuum distilling the filtrate to dry to obtain flos Camelliae Japonicae extract.

The purslane extract is prepared by the following steps:

(2) adding 40-60 times of water and ethanol mixture at a volume ratio of 80:20, decocting under reflux at 90 deg.C under normal pressure, filtering, and concentrating the filtrate by vacuum distillation to 150% of herba Portulacae weight to obtain crude extract;

(3) adding 20% adsorbent activated carbon particles based on the weight of the crude extract, performing microwave oscillation until the mixture is colorless, filtering, and continuously distilling the filtrate under reduced pressure until the filtrate is dried to obtain the purslane extract.

Example 1

A composition (Redsnow Relief) for inhibiting TRPV1 pathway and reducing skin irritation comprises flos Camelliae Japonicae extract and herba Portulacae extract, wherein the mass percent of flos Camelliae Japonicae extract is 0.0125%, and the mass percent of herba Portulacae extract is 0.004375%.

The preparation method of the composition for inhibiting the TRPV1 pathway to reduce skin irritation comprises the following steps: dissolving camellia extract powder in water to prepare a solution containing 1% of camellia extract, which is called as camellia extract rednow, dissolving purslane extract powder in water to prepare a solution containing 3.5% of purslane extract, which is called as purslane extract protulaca, diluting the camellia extract rednow to the mass fraction of 0.025%, diluting the purslane extract protulaca to the mass fraction of 0.00875%, and then mixing in equal amount to obtain the composition (rednow) for inhibiting TRPV1 pathway and reducing skin irritation.

A skin care product for inhibiting a TRPV1 pathway and reducing skin irritation comprises the following components in percentage by mass:

example 2

A composition (Redsnow Relief) for inhibiting TRPV1 pathway and reducing skin irritation comprises flos Camelliae Japonicae extract and herba Portulacae extract, wherein the mass percent of flos Camelliae Japonicae extract is 0.006% and the mass percent of herba Portulacae extract is 0.021%.

The preparation method of the composition for inhibiting the TRPV1 pathway to reduce skin irritation comprises the following steps: dissolving camellia extract powder in water to prepare a solution containing 1% of camellia extract, which is called as camellia extract rednow, dissolving purslane extract powder in water to prepare a solution containing 3.5% of purslane extract, which is called as purslane extract protulaca, diluting the camellia extract rednow to the mass fraction of 0.012%, diluting the purslane extract protulaca to the mass fraction of 0.042%, and then mixing in equal amount to obtain the composition (rednow) for inhibiting TRPV1 pathway and reducing skin irritation.

example 3

A composition (Redsnow Relief) for inhibiting TRPV1 pathway and reducing skin irritation comprises flos Camelliae Japonicae extract and herba Portulacae extract, wherein the mass percent of flos Camelliae Japonicae extract is 0.0006%, and the mass percent of herba Portulacae extract is 0.021%.

The preparation method of the composition for inhibiting the TRPV1 pathway to reduce skin irritation comprises the following steps: dissolving camellia extract powder in water to prepare a solution containing 1% of camellia extract, which is called as camellia extract rednow, dissolving purslane extract powder in water to prepare a solution containing 3.5% of purslane extract, which is called as purslane protulaca, diluting the camellia extract rednow to the mass fraction of 0.0012%, diluting the purslane extract protulaca to the mass fraction of 0.042%, and then mixing in equal amount to obtain the composition (rednow) for inhibiting TRPV1 pathway and reducing skin irritation.

comparative example 1

A composition comprises flos Camelliae Japonicae extract 0.0125 wt%. The preparation method comprises the following steps: dissolving the camellia extract powder in water to obtain a solution containing 1% of camellia extract, which is marked as camellia extract rednow, and diluting the camellia extract rednow to the mass fraction of 0.0125%.

A skin care product comprising the above composition, which is different from example 1 in that there is no purslane extract, and the contents of camellia extract and water are different.

Comparative example 2

A composition comprises herba Portulacae extract 0.004375%. The preparation method comprises the following steps: dissolving herba Portulacae extract powder in water to obtain a solution containing 3.5% of herba Portulacae extract, which is called herba Portulacae protulacae, and diluting herba Portulacae extract protulaca to 0.004375% by mass.

A skin care product containing the composition is different from the skin care product in example 1 in that the camellia extract is not contained, and the purslane extract and water are different in content.

Performance testing

1. Assay for TRPV1 receptor protein of keratinocytes

Keratinocytes are the most prominent cells found in the epidermis of the skin, making up 90% of the epidermal cells of the skin. By inhibiting the expression of epidermal cells, the activation of TRPV1 pathway caused by stimulation to skin is reduced. For inhibition verification of TRPV1 pathway, inhibition of TRPV1 pathway was detected by testing the expression of TRPV1 receptor protein using a keratinocyte assay for assaying the content of TRPV1 receptor (protein).

The cytotoxic concentrations of the camellia extract rednow and the purslane extract protulaca were first tested, and the specific results are shown in tables 1 and 2.

TABLE 1 cytotoxic concentration profile of Camellia extract redsinow

TABLE 2 cytotoxic concentration profile of Portulaca oleracea extract protulaca

As can be seen from the data in tables 1 and 2, the redmouth of the camellia extract and the protulaca extract at the test concentrations are not higher than 1.25% and not higher than 0.625%.

By 1.8X 10 ⁵ The density of the cells/well was inoculated with keratinocytes in 6-well plates, incubators (37 ℃, 5% CO) ₂ ) And incubated overnight. The working fluid usage of the test is shown in table 3 below.

TABLE 3 table of the working solution usage

According to the test scheme of the table, when the cell plating rate in the 6-well plate reaches 40% -50%, the administration amount of each well is 2mL, and each group is provided with 3 multiple wells. After completion of the administration, the 6-well plate was placed in an incubator (37 ℃ C., 5% CO) ₂ ) And (5) performing medium incubation and culture for 24 h.

TRPV1 detection: fixing with 4% paraformaldehyde for 30min, performing immunofluorescence detection of capsaicin receptor (TRPV1), taking pictures under microscope, and analyzing.

The results are expressed as Mean ± SD using GraphPad Prism mapping. The comparison between groups was performed by statistical analysis of t-test. Statistical analysis was double-tailed. P <0.05 was considered to have significant differences, and P <0.01 was considered to have very significant differences.

Based on the test scheme, TRPV1 detection is carried out according to the immunofluorescence operation procedure, the fluorescence detection graphs of the TRPV1 protein are shown in figures 1-8, and the change trend is shown in figure 9. Wherein (# representing significant differences between NC groups and BC groups, <0.05, <0.01, <0.001) it can be seen that both camellia extract and purslane extract alone had inhibitory effects on TRPV 1. When the two are compounded in a certain proportion, the synergistic effect is achieved, the inhibition effect is more obvious, and the effect of the embodiment 2 is the best. This indicates that the combination of the two groups of plant extracts can effectively inhibit TRPV1 channel, thereby achieving the effect of reducing skin irritation.

2. Human capsaicin test

Capsaicin is a classical stimulus that activates TRPV1 and produces stinging, burning and redness when a person comes into contact with capsaicin. In the test process, 0.01% of capsaicin is used as a stimulus source, and after the nasolabial sulcus parts on two sides are stimulated, a test sample is used on one side, and the improvement condition of a blank part is compared. Wherein the skin care products prepared in examples 1-3 and comparative examples 1-2 were tested.

In the test, a piece of filter paper (about 0.8cm in diameter) soaked with 50. mu.L of a 0.01% capsaicin aqueous solution was applied simultaneously to the nasolabial folds on both sides of the subject, and the filter paper was removed when the subject felt that the sensory values in the nasolabial folds were both equal to or greater than 3 points (moderate sensory values and above). The test sample was applied by the tester to the test area on one side in an amount of 50. mu.L, and left untreated on the other side (blank). The burning pain at the nasolabial sulcus on both sides was subjectively evaluated by the subject before use of the sample (after capsaicin challenge, before use of the sample), 0.5min, 5min, 10min, 15min, 20min and 30min after use of the sample, and was scored by 5 points, with the scoring criteria: a score of 0 is no sensation, a score of 1 is barely detectable, a score of 2 is mildly detectable, a score of 3 is moderately detectable, a score of 4 is severely detectable, and a score of 5 is pain. The specific results are shown in FIGS. 10-14.

As can be seen from FIGS. 10 to 14, the comparative examples and the examples have certain improvements on the burning sensation caused by capsaicin, wherein the examples have better irritation response reduction effects than the comparative examples, wherein the example 2 has the best effect, and the burning sensation is obviously reduced relative to the blank at 5 min.

The above embodiments are described in detail for different implementation procedures of the present invention, but the implementation manner of the present invention is not limited thereto, and those skilled in the art can achieve the object of the present invention based on the disclosure of the present invention, and any modifications and variations based on the concept of the present invention fall within the protection scope of the present invention, which is defined by the claims.

Claims

1. A composition for inhibiting a TRPV1 pathway to reduce skin irritation is characterized by comprising a camellia extract and a purslane extract, wherein the mass ratio of the camellia extract to the purslane extract is 0.01-10: 0.01-10.

2. The composition for reducing skin irritation through inhibition of the TRPV1 pathway according to claim 1, wherein the ratio of the mass of the camellia extract to the mass of the purslane extract is 1-10: 1-10.

3. The composition for inhibiting the TRPV1 pathway to reduce skin irritation according to claim 2, wherein the camellia extract has a protocatechuic acid content of not less than 1%.

4. The composition for reducing skin irritation that inhibits the TRPV1 pathway according to claim 3, wherein the camellia extract is prepared by:

5. The composition for inhibiting the TRPV1 pathway for reducing skin irritation according to claim 4, wherein the volume ratio of water to ethanol in the mixture of water and ethanol is from 20:80 to 80: 20; the temperature of reflux decoction is 80-95 deg.C.

6. The composition for inhibiting the TRPV1 pathway to reduce skin irritation according to any one of claims 3-5, wherein the purslane extract contains no less than 10% polysaccharides.

7. The composition for inhibiting the TRPV1 pathway to reduce skin irritation according to claim 6, wherein the purslane extract is prepared by:

8. The composition for inhibiting the TRPV1 pathway for reducing skin irritation according to claim 7, wherein the volume ratio of water to ethanol in the mixture of water and ethanol is from 20:80 to 80: 20; the temperature of reflux decoction is 80-95 deg.C.

9. A skin care product for reducing skin irritation which inhibits the TRPV1 pathway comprising the composition for reducing skin irritation which inhibits the TRPV1 pathway of any one of claims 1 to 8.

10. The skin care product for inhibiting the TRPV1 pathway to reduce skin irritation according to claim 9, wherein the skin care product for inhibiting the TRPV1 pathway to reduce skin irritation further comprises the following components by mass percent: 1 to 5 percent of glycerin, 1 to 5 percent of butanediol, 0.1 to 1 percent of allantoin, 0.1 to 1 percent of thickening agent, 0.01 to 0.5 percent of sodium hyaluronate, 0.1 to 5 percent of squalane, 0.5 to 1.5 percent of emulsifier and the balance of water.