A composition with anti-allergic soothing effect and application thereof

Abstract

The present application relates to a composition with anti-allergic soothing effect and application thereof, the composition comprising red mushroom extract, willow extract and hydrolyzed royal jelly protein. The present application creatively realizes the synergistic anti-allergic soothing effect of multi-target and multi-path by scientifically compounding three anti-stimulating active ingredients with different mechanisms and targets. The red mushroom extract mainly plays a role by regulating skin stress hormone level and inhibiting macrophage inflammatory polarization, the willow extract mainly plays a role by inhibiting inflammatory mediator release and antioxidant, and the hydrolyzed royal jelly protein mainly plays a role by regulating NLRP3 inflammasome-mediated inflammatory response and enhancing cell antioxidant capacity. The target points and paths of the three have complementarity, and the combination of the three has synergistic effect in anti-allergic soothing effect compared with single active component or scheme lacking any active component.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology and relates to a composition with anti-allergic and soothing effects and its application. Background Technology

[0002] As the largest organ in the human body, the skin is directly exposed to the external environment, constantly facing attacks from various internal and external stimuli such as ultraviolet rays, pollutants, irritating ingredients in cosmetics, and climate change. When the skin barrier function is impaired, external irritants can easily penetrate the skin, triggering the release of inflammatory factors by keratinocytes and immune cells, leading to discomfort such as redness, swelling, itching, burning, and stinging. With the increasing number of people with sensitive skin, cosmetics with anti-irritant and soothing effects have become a market hotspot.

[0003] Given the prevalence of sensitive skin among modern people, moisturizing, soothing, and repairing ingredients in skincare products are indispensable. These ingredients replenish the skin with moisture and nutrients, restoring its health and vitality, improving comfort and appearance, and fulfilling people's pursuit of healthy skin. However, current products with moisturizing, soothing, and repairing effects often have a relatively single pathway, resulting in limited improvement. Although some existing technologies have attempted to combine multiple active ingredients, these combinations are mostly based on simple, empirical pairings and have failed to achieve true multi-target synergistic effects. The components may have overlapping or interfering effects, making it difficult to achieve a synergistic effect. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a composition with anti-allergic and soothing effects and its application.

[0005] To achieve this objective, the present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides a composition having anti-allergic and soothing effects, the composition comprising Ganoderma lucidum extract, willowherb extract and hydrolyzed royal jelly protein.

[0007] This invention creatively combines three anti-irritant active ingredients with different mechanisms of action and targets to achieve a synergistic anti-allergic and soothing effect through multiple targets and pathways. Specifically, Ganoderma lucidum extract primarily works by regulating skin stress hormone levels and inhibiting macrophage inflammatory polarization; Willowherb extract primarily works by inhibiting the release of inflammatory mediators and providing antioxidant effects; and hydrolyzed royal jelly protein primarily works by regulating NLRP3 inflammasome-mediated inflammatory responses and enhancing cellular antioxidant capacity. The targets and pathways of these three ingredients are complementary, and their combined use provides a synergistic effect in anti-allergic and soothing efficacy compared to using a single active ingredient or lacking any one of the active ingredients.

[0008] Preferably, the composition comprises, by weight, 1-10 parts of Ganoderma lucidum extract, 10-20 parts of willowherb extract, and 0.5-5 parts of hydrolyzed royal jelly protein.

[0009] Based on the potential interaction between the three components in terms of anti-allergic and soothing effects, the effect is better when they meet the above-mentioned specific mass ratio.

[0010] The mass fractions of the Ganoderma lucidum extract can be selected from 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, etc.; the mass fractions of the Willowherb extract can be selected from 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, etc.; the mass fractions of the hydrolyzed royal jelly protein can be selected from 0.5 parts, 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, etc.

[0011] The Ganoderma lucidum extract involved in this invention is not limited to a specific preparation process. When it is prepared using the following process (i.e., compound enzymatic hydrolysis + segmented alcohol extraction process), its synergistic effect in anti-allergy and soothing is more obvious when used in combination with willowherb extract and hydrolyzed royal jelly protein.

[0012] Preferably, the Ganoderma lucidum extract is prepared by a method comprising the following steps:

[0013] (1) After the dried Ganoderma lucidum fruiting body is crushed, it is mixed with water and compound enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the enzyme is inactivated and the residue is collected by filtration.

[0014] (2) Mix the filter residue with a 50-60% (e.g., 50%, 54%, 55%, 57%, 60%, etc.) ethanol aqueous solution and reflux to separate the solid and liquid, and collect the supernatant and filter residue.

[0015] (3) The filter residue collected in step (2) is mixed with 75-90% (e.g., 75%, 80%, 84%, 85%, 90%, etc.) ethanol aqueous solution and refluxed to collect the supernatant;

[0016] (4) The supernatant collected in step (2) and the supernatant collected in step (3) are concentrated and dried separately and then mixed, or mixed and concentrated and dried to obtain the Ganoderma lucidum extract.

[0017] This invention combines active components obtained from alcohol extraction at different specific concentrations, resulting in superior anti-allergic and soothing effects compared to single alcohol extracts.

[0018] Preferably, the complex enzyme is a combination of cellulase, pectinase and papain.

[0019] Preferably, the mass ratio of cellulase, pectinase and papain is (1-2):(1-2):1, for example, 1:1:1, 1:2:1, 2:1:1, 2:2:1, etc.

[0020] Preferably, the amount of the compound enzyme added is 0.3%-0.8% of the mass of the dried Ganoderma lucidum fruiting body powder, such as 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, etc.

[0021] Preferably, the ratio of dried Ganoderma lucidum fruiting body to water is 1:(8-15) g / mL, for example, 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:11 g / mL, 1:12 g / mL, 1:13 g / mL, 1:14 g / mL, 1:15 g / mL, etc.

[0022] Preferably, the enzymatic hydrolysis treatment is carried out at pH=4.5-5.5 (e.g., pH=4.5, pH=4.7, pH=4.8, pH=5.0, pH=5.5, etc.) and 45-50℃ (e.g., 45℃, 46℃, 47℃, 48℃, 49℃, 50℃, etc.) for 2-4 h (e.g., 2 h, 2.5 h, 3 h, 3.5 h, 4 h, etc.).

[0023] Preferably, the ratio of the filter residue to the ethanol aqueous solution in step (2) is 1:(8-15) g / mL, for example, 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:11 g / mL, 1:12 g / mL, 1:13 g / mL, 1:14 g / mL, 1:15 g / mL, etc.

[0024] Preferably, the reflux process in step (2) is performed 1-3 times, for example, once, twice or three times; each time for 0.5-1.5 h, for example, 0.5 h, 0.7 h, 0.8 h, 0.9 h, 1.0 h, 1.2 h, 1.3 h, 1.4 h, 1.5 h, etc.

[0025] Preferably, the ratio of the filter residue to the ethanol aqueous solution in step (3) is 1:(8-15) g / mL, for example, 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:11 g / mL, 1:12 g / mL, 1:13 g / mL, 1:14 g / mL, 1:15 g / mL, etc.

[0026] Preferably, the reflux process in step (3) is performed 1-3 times, for example, once, twice or three times; each time for 0.5-1.5 h, for example, 0.5 h, 0.7 h, 0.8 h, 0.9 h, 1.0 h, 1.2 h, 1.3 h, 1.4 h, 1.5 h, etc.

[0027] The preparation process of the willowherb extract involved in this invention is not limited. When it is prepared by the following process (i.e., alcohol extraction + macroporous resin elution), its synergistic effect on anti-allergy and soothing is more obvious when it is used in combination with willowherb extract and hydrolyzed royal jelly protein.

[0028] Preferably, the willowherb extract is prepared by a method comprising the following steps:

[0029] (1) After the dried whole plant of Willowherb is crushed, it is mixed with 60-75% (e.g., 60%, 62%, 65%, 70%, 75% etc.) ethanol aqueous solution and refluxed. Solid-liquid separation is performed, the alcohol extract is collected, and the extract is shrunk under reduced pressure to obtain the extract.

[0030] (2) The extract was purified and eluted using macroporous resin. First, it was eluted with 20-40% (e.g., 20%, 25%, 30%, 35%, 40%) ethanol aqueous solution, and then eluted with 60-70% (e.g., 60%, 62%, 65%, 70%) ethanol aqueous solution. The eluent was collected, concentrated and dried to obtain the willowherb extract.

[0031] Preferably, the ratio of the dried willowherb whole herb to the ethanol aqueous solution is 1:(8-15) g / mL, for example, 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:12 g / mL, 1:13 g / mL, 1:14 g / mL, 1:15 g / mL, etc.

[0032] Preferably, the reflux treatment is performed 1-3 times, for example, once, twice or three times; each time for 0.5-1.5 hours, for example, 0.5 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1.0 hours, 1.2 hours, 1.3 hours, 1.4 hours, 1.5 hours, etc.

[0033] Preferably, the elution method is as follows: first, elute 3-8 BV with a 20-40% ethanol aqueous solution, then elute 3-8 BV with a 60-70% ethanol aqueous solution and collect the eluent.

[0034] Preferably, the macroporous resin is a mixture of AB-8 resin and HPD450 resin.

[0035] The present invention also creatively discovers that when a mixture of AB-8 resin and HPD450 resin is used to purify and elute willowherb extract, the eluted product has a superior anti-allergic and soothing effect compared to a single resin elution method.

[0036] Preferably, the mass ratio of AB-8 resin to HPD450 resin is (1-2):1.

[0037] Other specific point values ​​not listed in all the numerical ranges involved in this invention may be selected and are all within the protection scope of this invention, and will not be described in detail here.

[0038] In a second aspect, the present invention provides the use of the composition according to the first aspect in the preparation of products having anti-allergic and soothing effects.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] This invention creatively combines three anti-irritant active ingredients with different mechanisms of action and targets to achieve a synergistic anti-allergic and soothing effect through multiple targets and pathways. Specifically, Ganoderma lucidum extract primarily works by regulating skin stress hormone levels and inhibiting macrophage inflammatory polarization; Willowherb extract primarily works by inhibiting the release of inflammatory mediators and providing antioxidant effects; and hydrolyzed royal jelly protein primarily works by regulating NLRP3 inflammasome-mediated inflammatory responses and enhancing cellular antioxidant capacity. The targets and pathways of these three ingredients are complementary, and their combined use provides a synergistic effect in anti-allergic and soothing efficacy compared to using a single active ingredient or lacking any one of the active ingredients. Detailed Implementation

[0041] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0042] The sources of the active ingredients in the products involved in the following examples, comparative examples, application examples, and comparative application examples are as follows (only the active ingredients are shown; other necessary excipients contained in commercially available raw materials are not described).

[0043]

[0044] Preparation Example 1-1

[0045] This preparation example provides a Ganoderma lucidum extract, which is prepared by the following method:

[0046] (1) After the dried Ganoderma lucidum fruiting body is crushed to 80 mesh, it is mixed with water at a material-to-liquid ratio of 1:10 g / mL. Then, 0.5% of the mass of the crushed dried Ganoderma lucidum fruiting body is added with a compound enzyme (cellulase, pectinase and papain in a mass ratio of 2:2:1). The enzyme is hydrolyzed at pH=5.0 and 48℃ for 3 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 90℃ for 10 min. The residue is collected by filtration.

[0047] (2) Mix the filter residue with 60% ethanol aqueous solution at a material-to-liquid ratio of 1:12 g / mL and reflux extract twice, each time for 1 hour. Separate the solid and liquid, combine and collect the supernatant, and separate the filter residue.

[0048] (3) The filter residue collected in step (2) is mixed with 85% ethanol aqueous solution at a material-to-liquid ratio of 1:12 g / mL and refluxed twice, each time for 1 h, and the supernatant is collected.

[0049] (4) The supernatant collected in step (2) is mixed with the supernatant collected in step (3) and concentrated, then freeze-dried to obtain the Ganoderma lucidum extract.

[0050] Preparation Examples 1-2

[0051] This preparation example provides a Ganoderma lucidum extract, which is prepared by the following method:

[0052] (1) After the dried Ganoderma lucidum fruiting body is crushed to 80 mesh, it is mixed with water at a material-to-liquid ratio of 1:8 g / mL. Then, 0.8% of the mass of the crushed dried Ganoderma lucidum fruiting body is added with a compound enzyme (cellulase, pectinase and papain in a mass ratio of 1:1:1). The enzyme is hydrolyzed at pH=5.5 and 50℃ for 2 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 90℃ for 10 min. The residue is collected by filtration.

[0053] (2) Mix the filter residue with 50% ethanol aqueous solution at a material-to-liquid ratio of 1:10 g / mL and reflux extract twice, each time for 1.5 h. Separate the solid and liquid, combine and collect the supernatant, and separate the filter residue.

[0054] (3) The filter residue collected in step (2) is mixed with 80% ethanol aqueous solution at a material-to-liquid ratio of 1:10 g / mL and refluxed twice, each time for 1.5 h, and the supernatant is collected.

[0055] (4) The supernatant collected in step (2) is mixed with the supernatant collected in step (3) and concentrated, then freeze-dried to obtain the Ganoderma lucidum extract.

[0056] Preparation Examples 1-3

[0057] This preparation example provides a Ganoderma lucidum extract, which is prepared by the following method:

[0058] (1) After the dried Ganoderma lucidum fruiting body is crushed to 80 mesh, it is mixed with water at a material-to-liquid ratio of 1:15 g / mL. Then, 0.4% of the mass of the crushed dried Ganoderma lucidum fruiting body is added with a compound enzyme (cellulase, pectinase and papain in a mass ratio of 2:1:1). The enzyme is hydrolyzed at pH=4.5 and 45℃ for 4 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 90℃ for 10 min. The residue is collected by filtration.

[0059] (2) Mix the filter residue with 55% ethanol aqueous solution at a material-to-liquid ratio of 1:15 g / mL and reflux extract twice, each time for 1 hour. Separate the solid and liquid, combine and collect the supernatant, and separate the filter residue.

[0060] (3) The filter residue collected in step (2) is mixed with 90% ethanol aqueous solution at a material-to-liquid ratio of 1:15 g / mL and refluxed twice, each time for 1.5 h, and the supernatant is collected.

[0061] (4) The supernatant collected in step (2) and the supernatant collected in step (3) are concentrated and freeze-dried respectively, and then mixed evenly to obtain the Ganoderma lucidum extract.

[0062] Preparation Examples 1-4

[0063] This preparation example provides a Ganoderma lucidum extract, the only difference between which is the preparation method and that of Preparation Example 1-1: the complex enzyme is a 1:1 mass ratio of cellulase and pectinase, and the total amount of complex enzyme added remains unchanged. All other conditions remain unchanged.

[0064] Preparation Examples 1-5

[0065] This preparation example provides a Ganoderma lucidum extract, the only difference between which is the preparation method and that of Preparation Example 1-1 is the absence of the enzymatic hydrolysis treatment in step (1), that is, the dried Ganoderma lucidum fruiting body is pulverized to 80 mesh and then directly subjected to the alcohol extraction treatment in step (2). All other conditions remain unchanged.

[0066] Preparation Examples 1-6

[0067] This preparation example provides a Ganoderma lucidum extract, which is prepared by the following method:

[0068] (1) After the dried Ganoderma lucidum fruiting body is crushed to 80 mesh, it is mixed with water at a material-to-liquid ratio of 1:10 g / mL. Then, 0.5% of the mass of the crushed dried Ganoderma lucidum fruiting body is added with a compound enzyme (cellulase, pectinase and papain in a mass ratio of 2:2:1). The enzyme is hydrolyzed at pH=5.0 and 48℃ for 3 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 90℃ for 10 min. The residue is collected by filtration.

[0069] (2) The filter residue was mixed with 60% ethanol aqueous solution at a material-to-liquid ratio of 1:12 g / mL and refluxed twice for 1 hour each time. The solid and liquid were separated, the supernatant was collected, concentrated, and freeze-dried to obtain the Ganoderma lucidum extract.

[0070] Preparation Examples 1-7

[0071] This preparation example provides a Ganoderma lucidum extract, which is prepared by the following method:

[0072] (1) After the dried Ganoderma lucidum fruiting body is crushed to 80 mesh, it is mixed with water at a material-to-liquid ratio of 1:10 g / mL. Then, 0.5% of the mass of the crushed dried Ganoderma lucidum fruiting body is added with a compound enzyme (cellulase, pectinase and papain in a mass ratio of 2:2:1). The enzyme is hydrolyzed at pH=5.0 and 48℃ for 3 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 90℃ for 10 min. The residue is collected by filtration.

[0073] (2) The filter residue was mixed with 85% ethanol aqueous solution at a material-to-liquid ratio of 1:12 g / mL and refluxed twice for 1 hour each time. The solid and liquid were separated, the supernatant was collected, concentrated, and freeze-dried to obtain the Ganoderma lucidum extract.

[0074] Preparation Example 2-1

[0075] This preparation example provides a willowherb extract, which is prepared by the following method:

[0076] (1) After the dried whole plant of Willowherb was crushed, it was mixed with 70% ethanol aqueous solution at a material-to-liquid ratio of 1:10 g / mL and refluxed for 1.5 h. The solid and liquid were separated, the ethanol extract was collected, and the extract was shrunk under reduced pressure to obtain the extract.

[0077] (2) The extract was purified and eluted using macroporous resin (a mixture of AB-8 and HPD450 resins in a mass ratio of 3:2). First, 7 BV of the extract was eluted with 20% ethanol aqueous solution, and then 5 BV of the extract was eluted with 70% ethanol aqueous solution. The eluent was collected, concentrated, and freeze-dried to obtain the willowherb extract.

[0078] Preparation Example 2-2

[0079] This preparation example provides a willowherb extract, which is prepared by the following method:

[0080] (1) After pulverizing the dried whole plant of Willowherb, it was mixed with 65% ethanol aqueous solution at a material-to-liquid ratio of 1:15 g / mL and refluxed for 1.5 h. The solid and liquid were separated, the ethanol extract was collected, and the extract was shrunk under reduced pressure to obtain the extract.

[0081] (2) The extract was purified and eluted using macroporous resin (a mixture of AB-8 and HPD450 resins in a mass ratio of 1:1). First, 7 BV of the extract was eluted with 25% ethanol aqueous solution, and then 5 BV of the extract was eluted with 70% ethanol aqueous solution. The eluent was collected, concentrated, and freeze-dried to obtain the willowherb extract.

[0082] Preparation Examples 2-3

[0083] This preparation example provides a willowherb extract, which is prepared by the following method:

[0084] (1) After pulverizing the dried whole plant of Willowherb, it was mixed with 75% ethanol aqueous solution at a material-liquid ratio of 1:8 g / mL and refluxed for 1.5 h. The solid and liquid were separated, the ethanol extract was collected, and the extract was shrunk under reduced pressure to obtain the extract.

[0085] (2) The extract was purified and eluted using macroporous resin (a mixture of AB-8 and HPD450 resins in a mass ratio of 2:1). First, 7 BV of the extract was eluted with 25% ethanol aqueous solution, and then 5 BV of the extract was eluted with 70% ethanol aqueous solution. The eluent was collected, concentrated, and freeze-dried to obtain the willowherb extract.

[0086] Preparation Examples 2-4

[0087] This preparation example provides a willowherb extract, the preparation method of which differs from that of preparation example 2-1 only in that the volume fraction of the ethanol aqueous solution in step (1) is 85%, while other conditions remain unchanged.

[0088] Preparation Examples 2-5

[0089] This preparation example provides a willowherb extract, the preparation method of which differs from that of preparation example 2-1 only in that the volume fraction of the ethanol aqueous solution in step (1) is 50%, while all other conditions remain unchanged.

[0090] Preparation Examples 2-6

[0091] This preparation example provides a willowherb extract, the only difference between which is the preparation method and that of Preparation Example 2-1, except that the macroporous resin in step (2) is a single AB-8 resin. All other conditions remain unchanged.

[0092] Preparation Examples 2-7

[0093] This preparation example provides a willowherb extract, the only difference between which is the preparation method and that of Preparation Example 2-1, except that the macroporous resin in step (2) is a single HPD450 resin. All other conditions remain unchanged.

[0094] Example 1

[0095] This embodiment provides a composition, which, by mass parts, is obtained by mixing 5 parts of Ganoderma lucidum extract from Preparation Example 1-1, 15 parts of Willowherb extract from Preparation Example 2-1, and 2 parts of hydrolyzed royal jelly protein.

[0096] Example 2

[0097] This embodiment provides a composition, which, by mass parts, is obtained by mixing 8 parts of Ganoderma lucidum extract from Preparation Example 1-2, 13 parts of Willowherb extract from Preparation Example 2-2, and 1 part of hydrolyzed royal jelly protein.

[0098] Example 3

[0099] This embodiment provides a composition, which, by mass parts, is obtained by mixing 3 parts of Ganoderma lucidum extract from Preparation Examples 1-3, 18 parts of Willowherb extract from Preparation Examples 2-3, and 3 parts of hydrolyzed royal jelly protein.

[0100] Examples 4-7

[0101] This embodiment provides four compositions, which differ from Example 1 only in that the same mass fractions of Ganoderma lucidum extract in Preparation Example 1-1 are replaced with the Ganoderma lucidum extracts in Preparation Examples 1-4 to 1-7.

[0102] Examples 8-11

[0103] This embodiment provides four compositions, which differ from Example 1 only in that the same mass fraction of the willowherb extract in Example 2-1 is replaced with the willowherb extracts in Preparation Examples 2-4 to 2-7.

[0104] Example 12

[0105] This embodiment provides a composition, which is obtained by mixing 5 parts of Ganoderma lucidum extract (not self-made, but commercially available), 15 parts of Willowherb extract (not self-made, but commercially available) and 2 parts of hydrolyzed royal jelly protein by mass.

[0106] Comparative Example 1

[0107] This comparative example provides a composition that differs from Example 1 only in that it lacks Ganoderma lucidum extract, and the reduced mass fraction of it is proportionally allocated to the mass fractions of willowherb extract and hydrolyzed royal jelly protein.

[0108] Comparative Example 2

[0109] This comparative example provides a composition that differs from Example 1 only in that it lacks willowherb extract, and the reduced mass fraction of willowherb extract is proportionally allocated to the mass fractions of Ganoderma lucidum extract and hydrolyzed royal jelly protein.

[0110] Comparative Example 3

[0111] This comparative example provides a composition that differs from Example 1 only in that it lacks hydrolyzed royal jelly protein, and the reduced mass fraction of this protein is proportionally allocated to the mass fractions of Ganoderma lucidum extract and Willowherb extract.

[0112] Test Example 1

[0113] The capsaicin receptor inhibitory effects of the compositions prepared in Examples 1-12 and Comparative Examples 1-3 were tested.

[0114] The testing procedure was as follows: Each group of compositions was prepared into a 1% concentration sample for testing. A keratinocyte model was selected to determine the inhibitory effect of each composition on the capsaicin receptor (TRPV1). Keratinocytes were selected, revived, and seeded into 24-well plates, incubated overnight in CO2. When the cell deposition rate in the 24-well plates reached 40%-60%, 1 mL of culture medium containing 2 μM capsaicin was added to each well, along with each composition for stimulation and intervention (final concentration of the composition was 1%). The control group received no intervention. After intervention, the 24-well plates were cultured for 24 h, and the TRPV1 expression level was detected using a fluorescent probe. The TRPV1 receptor inhibition rate (%) was calculated, and the results are shown in Table 1.

[0115] Table 1

[0116]

[0117] As shown in Table 1, compared with the compositions of Comparative Examples 1-3, the present invention combines Ganoderma lucidum extract, willowherb extract, and hydrolyzed royal jelly protein, which synergistically enhance the effect of the three in inhibiting capsaicin receptors, thus exhibiting superior skin-soothing potential. A comparison of the composition data from Example 1 and Examples 4-12 reveals that the preparation processes of Ganoderma lucidum extract and willowherb extract also affect the skin-soothing efficacy of the compositions to some extent.

[0118] Test Example 2

[0119] Hyaluronidase activity inhibition tests were performed on the compositions obtained in Examples 1-12 and Comparative Examples 1-3:

[0120] The testing process is as follows: each group of compositions is prepared into a 0.5% concentration sample for later use.

[0121] Test tubes A and B contain 0.5 mL of sample solution and 0.5 mL of hyaluronidase (500 U / mL), and test tubes C and D contain 0.5 mL of distilled water and 0.5 mL of acetate buffer solution. Incubate at 37°C for 20 min. Add 0.1 mL (2.5 mmol / L) CaCl2 solution and incubate at 37°C for 20 min. Add 0.5 mL of sodium hyaluronate (0.5 mg / mL) to test tubes A and C, and 0.5 mL of acetate buffer solution to test tubes B and D. Incubate at 37°C for 40 min. Add 0.5 mL of acetylacetone solution, 0.1 mL of NaOH solution (5 mol / L), and 0.5 mL of distilled water. Incubate in a boiling water bath for 15 min, then immediately cool in ice water for 5 min. Add 1 mL of P-DAB colorimetric reagent, shake thoroughly, add 3.5 mL of anhydrous ethanol, and incubate for 30 min. Develop color and measure the OD values ​​of test tubes A, B, C, and D at 530 nm. The formula for calculating the hyaluronidase activity inhibition rate is:

[0122] Hyaluronidase inhibition rate (%) = [(CD) - (AB)] / (CD) × 100

[0123] The results are shown in Table 2.

[0124] Table 2

[0125]

[0126] As shown in Table 2, compared with the compositions of Comparative Examples 1-3, the present invention combines Ganoderma lucidum extract, willowherb extract, and hydrolyzed royal jelly protein, which synergistically enhance the effect of inhibiting hyaluronidase, thus exhibiting superior skin-soothing potential. A comparison of the composition data from Examples 1 and 4-12 reveals that the preparation processes of Ganoderma lucidum extract and willowherb extract also influence the skin-soothing efficacy of the compositions to some extent.

[0127] Test Example 3

[0128] The inflammatory factor inhibitory capacity of the compositions prepared in Examples 1-12 and Comparative Examples 1-3 was tested:

[0129] The testing process involved using an LPS-induced mouse macrophage (RAW264.7) inflammation model to determine the inhibitory effect of the sample on tumor necrosis factor-α (TNF-α).

[0130] (1) Preparation of test samples: Prepare each group of compositions into a 1% concentration test sample solution for later use.

[0131] (2) Adjust the RAW264.7 cell density to 5×10⁻⁶. 4Cells were seeded at a rate of 200 μL per well into a 96-well plate and incubated in an incubator (37°C, 5% CO2).

[0132] (3) Grouping: Blank control group (no LPS stimulation or sample intervention, 180 μL of basal culture medium was given), negative control group (1 μg / mL LPS stimulation but no sample intervention was given), experimental groups 1-15 (1 μg / mL LPS stimulation and 180 μL of 0.5% of the sample solutions of Examples 1-12 and Comparative Examples 1-3 were given as intervention), and 3 cell-free wells were set up as zeroing groups. Plates were arranged with 3 replicates per group.

[0133] (4) Add basic culture medium or sample solution to each group first, and then put it back into the incubator for incubation (37℃, 5%CO2); after 1 h, take out the 96-well plate, add 20 μL of basic culture medium to each well of the zeroing well and blank control group, add 20 μL of LPS (1 μg / mL) to each well of the negative control group and experimental groups 1-15, and then put it back into the incubator for incubation.

[0134] (5) After 24 hours, the 96-well plate was removed, and the cell supernatant of each sample group was collected into a centrifuge tube. The tube was centrifuged at 1000 rpm for 10 min, and the supernatant was collected and placed in a 1.5 mL centrifuge tube. The TNF-α concentration in the cell supernatant was measured using an ELISA kit, and the inhibition rate of TNF-α in each experimental group compared with the negative control group was calculated.

[0135] TNF-α inhibition rate (%) = TNF-α concentration in negative control group - TNF-α concentration in experimental group / TNF-α concentration in negative control group × 100

[0136] The results are shown in Table 3.

[0137] Table 3

[0138]

[0139] As shown in Table 3, compared with the compositions of Comparative Examples 1-3, the present invention combines Ganoderma lucidum extract, willowherb extract, and hydrolyzed royal jelly protein, which synergistically enhance the effect of the three in inhibiting inflammatory factors, thus exhibiting superior skin anti-inflammatory and soothing potential. A comparison of the composition data from Example 1 and Examples 4-12 reveals that the preparation process of Ganoderma lucidum extract and willowherb extract also affects the skin stress-relieving and soothing efficacy of the composition to some extent.

[0140] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the product of this invention, addition of auxiliary components, and selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0141] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0142] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A composition having anti-allergic and soothing effects, characterized in that, The composition includes Ganoderma lucidum extract, willowherb extract and hydrolyzed royal jelly protein.

2. The composition according to claim 1, characterized in that, The composition comprises, by weight, 1-10 parts of Ganoderma lucidum extract, 10-20 parts of willowherb extract, and 0.5-5 parts of hydrolyzed royal jelly protein.

3. The composition according to claim 1 or 2, characterized in that, The Ganoderma lucidum extract is prepared by a method comprising the following steps: (1) After the dried Ganoderma lucidum fruiting body is crushed, it is mixed with water and compound enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the enzyme is inactivated and the residue is collected by filtration. (2) Mix the filter residue with a 50-60% ethanol aqueous solution and reflux to separate the solid and liquid, and collect the supernatant and filter residue; (3) Mix the filter residue collected in step (2) with 75-90% ethanol aqueous solution and reflux, and collect the supernatant; (4) The supernatant collected in step (2) and the supernatant collected in step (3) are concentrated and dried separately and then mixed, or mixed and concentrated and dried to obtain the Ganoderma lucidum extract.

4. The composition according to claim 3, characterized in that, The complex enzyme is a combination of cellulase, pectinase and papain. Preferably, the mass ratio of cellulase, pectinase and papain is (1-2):(1-2):

1.

5. The composition according to claim 3, characterized in that, The amount of the compound enzyme added is 0.3%-0.8% of the weight of the dried Ganoderma lucidum fruiting body powder; Preferably, the ratio of the dried Ganoderma lucidum fruiting body to water is 1:(8-15) g / mL; Preferably, the enzymatic hydrolysis is carried out at pH 4.5-5.5 and 45-50°C for 2-4 hours.

6. The composition according to claim 3, characterized in that, The ratio of the filter residue to the ethanol aqueous solution in step (2) is 1:(8-15) g / mL; Preferably, the reflux treatment in step (2) is performed 1-3 times, each time for 0.5-1.5 hours; Preferably, the ratio of the filter residue to the ethanol aqueous solution in step (3) is 1:(8-15) g / mL; Preferably, the reflux treatment in step (3) is performed 1-3 times, each time for 0.5-1.5 hours.

7. The composition according to claim 1 or 2, characterized in that, The willowherb extract is prepared by a method comprising the following steps: (1) After the dried whole plant of Willowherb is crushed, it is mixed with 60-75% ethanol aqueous solution and refluxed. Solid-liquid separation is performed, the alcohol extract is collected, and the extract is shrunk under reduced pressure to obtain the extract. (2) The extract was purified and eluted using macroporous resin. First, it was eluted with 20-40% ethanol aqueous solution, then with 60-70% ethanol aqueous solution and the eluent was collected, concentrated and dried to obtain the willowherb extract.

8. The composition according to claim 7, characterized in that, The ratio of dried willowherb whole herb to ethanol aqueous solution is 1:(8-15) g / mL; Preferably, the reflux treatment is performed 1-3 times, each time for 0.5-1.5 hours; Preferably, the elution method is as follows: first, elute 3-8 BV with a 20-40% ethanol aqueous solution, then elute 3-8 BV with a 60-70% ethanol aqueous solution and collect the eluent.

9. The composition according to claim 7, characterized in that, The macroporous resin is a mixture of AB-8 resin and HPD450 resin; Preferably, the mass ratio of AB-8 resin to HPD450 resin is (1-2):

1.

10. The use of the composition according to any one of claims 1-9 in the preparation of a product having anti-allergic and soothing effects.

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