A composition having synergistic anti-inflammatory, redness-reducing and soothing effects and uses thereof

Abstract

The present application provides a composition with anti-inflammatory, redness-relieving and soothing effects, which contains component a and component b, wherein component a is a compound gentian extract containing gentian, radix stephaniae tetrandrae and asiatic plant extract, and component b contains hydrolyzed ginsenosides. The present application also relates to the use of the composition in the preparation of a medicine or a cosmetic product, and further relates to a medicine or a cosmetic product containing the composition.

Description

TECHNICAL FIELD

[0001] The present application relates to a composition with anti-inflammatory, soothing and calming effects. The composition of the present application contains component a and component b, wherein component a is a compound gentian extract containing gentian, radix stephaniae tetrandrae and centella asiatica plant extracts, and component b contains hydrolyzed ginsenosides. The present application also relates to the use of the composition in the preparation of a medicine or a cosmetic product, and the present application further relates to a medicine or a cosmetic product containing the composition. BACKGROUND

[0002] With the development of economy and the improvement of living standards, environmental pollution, climate change, diversified raw materials of cosmetics and frequent medical beauty have become the main external factors for the formation of sensitive skin. In addition, the weak skin barrier caused by psychological stress, irregular work and rest and genetic factors is also the internal cause of the sensitive skin problem of contemporary people. Sensitive skin is prone to external stimuli due to damaged skin barrier and thin stratum corneum, and the tolerance is low. External stimuli can trigger non-specific immune mechanisms to produce skin inflammation, promote the release of inflammatory factors, and thus cause a vicious cycle of further damage to the barrier, resulting in subjective feelings such as stinging, itching, tightness, and possibly accompanied by symptoms such as dryness, desquamation, redness, etc. Therefore, detecting the modulation effect of the test substance on the release of pro-inflammatory factors by skin cells is one of the mainstream cell biology methods for evaluating the anti-inflammatory activity and soothing efficacy of cosmetic raw materials.

[0003] Interleukin-6 (IL-6) is a cytokine of the chemokine family, which can be produced by almost all immune system cells and stromal cells such as mononuclear macrophages, mast cells, keratinocytes, fibroblasts, etc. As a pro-inflammatory cytokine, IL-6 is in a central position in the inflammatory cascade, and after infection and inflammation, IL-6 is generated first and the level rises rapidly, which can reach a peak within two hours. Due to the early generation, high expression and important role in promoting the inflammatory cascade, IL-6 has become one of the most commonly detected inflammatory indicators. In commonly used mouse macrophage in vitro inflammation models, IL-6 can be induced by various immunogenic stimulants (such as lipopolysaccharide (LPS)) to generate immune cells.

[0004] In the current research on soothing cosmetics, the main active ingredients of gentian extract are widely used for their anti-inflammatory, soothing and antipruritic effects. For example, Han Ping et al. proposed a gentian extract with gentiopicroside as the main active ingredient, which can inhibit the secretion and release of cytokine IL-6 and itch regulatory factor TRPV1, inhibit the inflammatory response caused by dry skin pruritus, and thus improve the skin barrier function (Han Ping et al., Perfume & Flavor, 2021(6):27-31); CN112656731A discloses a composition containing gentian extract with soothing red effect. However, the current research also has some problems, such as (1) the bioavailability of gentian extract is low, and the anti-inflammatory activity is not obvious when added alone; (2) the plant extract composition containing gentian has the problems of large addition amount in the formula and high cost.

[0005] Therefore, there is an urgent need for a composition with strong synergistic anti-inflammatory, soothing red and / or soothing activity. Due to the synergistic effect, it can effectively reduce the addition amount of single component (such as gentian extract) in the composition, thereby achieving the purpose of reducing the addition amount of single component in cosmetics while maintaining high efficient activity (such as anti-inflammatory, soothing red and / or soothing activity), thereby reducing the cost of raw materials. SUMMARY

[0006] The present application relates to a composition comprising component a and component b, wherein component a is a compound gentian extract comprising gentian plant extract, radix Stephaniae tetrandrae plant extract and gynura plant extract, and component b comprises hydrolyzed ginsenosides. The composition of the present application has the following advantages:

[0007] (1) The composition of the present application has anti-inflammatory, soothing red and soothing activity;

[0008] (2) The composition of the present application comprising component a and component b has synergistic effect in anti-inflammatory activity, so it can reduce the addition amount of single component (such as component a or component b) in cosmetics while maintaining high efficient activity, thereby achieving the purpose of reducing the cost of raw materials.

[0009] The first aspect of the present application relates to a composition comprising component a and component b, wherein component a is a compound gentian extract comprising gentian plant extract, radix Stephaniae tetrandrae plant extract and gynura plant extract, and component b comprises hydrolyzed ginsenosides.

[0010] The second aspect of the present application relates to a composition comprising component a and component b, wherein component a is a compound gentian extract comprising gentian plant extract, radix stephaniae tetrandrae plant extract and gynura plant extract, and component b comprises hydrolyzed ginsenosides, wherein the content of each plant extract in component a is: gentian 30-70%, preferably 40-65%; radix stephaniae tetrandrae 10-40%, preferably 15-30%; gynura 10-40%, preferably 15-30%; wherein the plant extract is prepared by the following method: the pulverized plant material is extracted with water, the water extraction temperature is 60-80°C, then extracted with 20%-70%, preferably 20%-50% C2-6 alcohol aqueous solution, and further extracted with 50%-70% C2-6 alcohol aqueous solution, wherein the extraction temperature of the C2-6 alcohol aqueous solution is 40-85°C, preferably 50-75°C, and the C2-6 alcohol is propylene glycol or ethanol, and the three extracts are mixed to obtain the compound gentian extract.

[0011] The third aspect of the present application relates to the use of the composition according to the first aspect or the second aspect of the present application in the preparation of a medicament or a cosmetic having anti-inflammatory, redness-relieving and / or soothing effect.

[0012] The fourth aspect of the present application relates to a cosmetic comprising the composition according to the first aspect or the second aspect of the present application, wherein the composition accounts for about 0.1%-20% by weight of the weight of the cosmetic.

[0013] The fifth aspect of the present application relates to a medicament comprising the composition according to the first aspect or the second aspect of the present application, wherein the composition accounts for about 0.1%-20% by weight of the weight of the medicament. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 Effect of component a (compound gentian extract) in the composition according to the present application on the cell viability of RAW264.7 cells.

[0015] Figure 2 Effect of component b (GLF) in the composition according to the present application on the cell viability of RAW264.7 cells.

[0016] Figure 3 Effect of the composition according to the present application comprising component a (compound gentian extract) and component b (GLF) on the soothing redness degree of the skin of a subject with redness caused by occluded patch. DETAILED DESCRIPTION

[0018] DEFINITIONS

[0019] The terms "s", "min" and "h" as used herein represent "second", "minute" and "hour", respectively.

[0020] The term "comprising", as used herein, is synonymous with "including", "containing", or "characterized by", is inclusive or open, and does not exclude additional, unrecited elements or method steps. However, in the context of this document, each time "comprising" is recited, alternative embodiments in which "consisting essentially of" and "consisting of" are contemplated, in which "consisting of" excludes any element or step not specified and "consisting essentially of" permits the inclusion of additional elements or steps that do not materially affect the essential or novel characteristics of the composition or method at issue.

[0021] The term "about" as used herein means ± 10% of the numerical value of the term that it modifies, more preferably ± 5%, most preferably ± 2%, so that a person of ordinary skill in the art is clearly informed of the range intended for the term "about".

[0022] COMPOSITION

[0023] A first aspect of the present application relates to a composition comprising component a and component b, wherein component a is a compound gentian extract comprising gentian plant extract, Stephania tetrandra plant extract and Centella asiatica plant extract, and component b comprises hydrolyzed ginsenosides.

[0024] A second aspect of the present application relates to a composition comprising component a and component b, wherein component a is a compound gentian extract comprising gentian plant extract, Stephania tetrandra plant extract and Centella asiatica plant extract, and component b comprises hydrolyzed ginsenosides, wherein the content of each plant extract in component a is: gentian 30-70%, preferably 40-65%; Stephania tetrandra 10-40%, preferably 15-30%; and Centella asiatica 10-40%, preferably 15-30%; wherein the plant extracts are prepared by the following method: the pulverized plant material is extracted with water at a temperature of 60-80°C, then extracted with a 20%-70%, preferably 20%-50%, aqueous solution of C2-6 alcohol, and further extracted with a 50%-70% aqueous solution of C2-6 alcohol, wherein the extraction temperature of the aqueous solution of C2-6 alcohol is 40-85°C, preferably 50-75°C, and the C2-6 alcohol is propylene glycol or ethanol, and the three extracts are mixed to obtain the compound gentian extract.

[0025] In some embodiments, component a is a composition of plant extracts prepared according to the patent CN113041286B of Xiangyi Herbal. In some embodiments, component a is a composition of plant extracts prepared according to Example 3 of the patent CN113041286B of Xiangyi Herbal.

[0026] In some embodiments, component a is a compound gentian extract comprising gentian plant extract, Stephania tetrandra plant extract, and Gynura plant extract. In some embodiments, component a is a compound gentian extract consisting of gentian plant extract, Stephania tetrandra plant extract, and Gynura plant extract.

[0027] In some embodiments, the compound gentian extract is prepared by extracting gentian, Stephania tetrandra, and Gynura together. In some embodiments, the compound gentian extract is prepared by extracting gentian, Stephania tetrandra, and Gynura separately and then mixing them together.

[0028] In some embodiments, the content of the Gentianna extract, the Stephania tetrandra extract, and the Centella asiatica extract in component a is independently selected from the following weight percentages: 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 33.3%, 33.4%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, and 99%, and ranges between any of the aforementioned weight percentages. In some embodiments, the content of each plant extract in component a is: Gentianna 1-99%; Stephania tetrandra 1-99%; Centella asiatica 1-99%. In some embodiments, the content of each plant extract in component a is: Gentianna 30-70%; Stephania tetrandra 10-40%; Centella asiatica 10-40%. In some embodiments, the content of each plant extract in component a is: Gentianna 40-65%; Stephania tetrandra 15-30%; Centella asiatica 15-30%. In some embodiments, the content of each plant extract in component a is: Gentianna 45%; Stephania tetrandra 30%; Centella asiatica 25%.

[0029] In some embodiments, the temperature of the water extraction is 40-100°C. In some embodiments, the temperature of the water extraction is 40-45°C, 45-50°C, 50-55°C, 55-60°C, 60-65°C, 65-70°C, 70-75°C, 75-80°C, 80-85°C, 85-90°C, 90-95°C, or 95-100°C. In some embodiments, the temperature of the water extraction is 60-80°C. In some embodiments, the temperature of the water extraction is about 80°C.

[0030] In some embodiments, after the comminuted plant material is extracted with water, the comminuted plant material is then extracted with a 10%-70% aqueous C2-6alcohol solution, and further extracted with a 50%-90% aqueous C2-6alcohol solution. In some embodiments, after the comminuted plant material is extracted with water, the comminuted plant material is then extracted with a 10%-15%, 15%-20%, 20%-25%, 25%-30%, 30%-35%, 35%-40%, 40%-45%, 45%-50%, 50%-55%, 55%-60%, 60%-65%, or 65%-70% aqueous C2-6alcohol solution, and further extracted with a 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, or 85%-90% aqueous C2-6alcohol solution. In some embodiments, after the comminuted plant material is extracted with water, the comminuted plant material is then extracted with a 20%-50% aqueous C2-6alcohol solution, and further extracted with a 50%-70% aqueous C2-6alcohol solution. In some embodiments, after the comminuted plant material is extracted with water, the comminuted plant material is then extracted with a 40% aqueous C2-6alcohol solution, and further extracted with a 70% aqueous C2-6alcohol solution.

[0031] In some embodiments, the aqueous C2-6alcohol solution is extracted at a temperature of 30-100°C. In some embodiments, the aqueous C2-6alcohol solution is extracted at a temperature of 30-35°C, 35-40°C, 40-45°C, 45-50°C, 50-55°C, 55-60°C, 60-65°C, 65-70°C, 70-75°C, 75-80°C, 80-85°C, 85-90°C, 90-95°C, or 95-100°C. In some embodiments, the aqueous C2-6alcohol solution is extracted at a temperature of 40-85°C. In some embodiments, the aqueous C2-6alcohol solution is extracted at a temperature of 50-75°C. In some embodiments, the aqueous C2-6alcohol solution is extracted at a temperature of 60°C.

[0032] In some embodiments, the C2-6alcohol includes, but is not limited to, ethanol, propanol, butanol, pentanol, hexanol, ethylene glycol, propylene glycol, glycerol, butylene glycol, pentylene glycol, hexylene glycol. In some embodiments, the C2-6alcohol is propylene glycol or ethanol. In some embodiments, the C2-6alcohol is ethanol.

[0033] In some embodiments, in the method of preparing component a, the weight ratio between each plant material or each plant extract is: gentiana 3-7: Stephania 1-4: Centella asiatica 1-4. In some embodiments, in the method of preparing component a, the weight ratio between each plant material or each plant extract is: gentiana 4-6: Stephania 1-3: Centella asiatica 1-3. In some embodiments, in the method of preparing component a, the weight ratio between each plant material or each plant extract is: gentiana 4.5: Stephania 3: Centella asiatica 2.5.

[0034] In some embodiments, the method of preparing plant extracts further comprises filtering the obtained compound gentiana extract for purification, and obtaining a concentrated solution. In some embodiments, the filtering is membrane filtration. In some embodiments, first filtering with 5000-10000 Dalton ultrafiltration membrane, collecting the filtrate, then filtering with 100-1000 Dalton nanofiltration membrane, recovering the solvent, and obtaining the concentrated solution. In some embodiments, first filtering with 5000 Dalton ultrafiltration membrane, collecting the filtrate, then filtering with 1000 Dalton nanofiltration membrane, recovering the solvent, and obtaining the concentrated solution.

[0035] In some embodiments, the method of preparing plant extracts further comprises drying the obtained concentrated solution, optionally, adding water to the concentrated solution before drying.

[0036] In some embodiments, the drying is selected from freeze-drying, vacuum drying, spray drying, or a combination thereof.

[0037] In some embodiments, in the method of preparing plant extracts, an external circulation extraction method is used, an online spectrophotometer is used for monitoring, the end point is determined according to the absorbance, at which time the residue is removed, and the extract is obtained.

[0038] In some embodiments, component a comprises gentiopicroside, tetrandrine, and total centella asiatica glycosides. In some embodiments, the weight ratio between gentiopicroside, tetrandrine, and total centella asiatica glycosides in component a is gentiopicroside 3-7: tetrandrine 1-4: total centella asiatica glycosides 1-4. In some embodiments, the weight ratio between gentiopicroside, tetrandrine, and total centella asiatica glycosides in component a is gentiopicroside 4-6: tetrandrine 1-3: total centella asiatica glycosides 1-3. In some embodiments, the weight ratio between gentiopicroside, tetrandrine, and total centella asiatica glycosides in component a is gentiopicroside 6: tetrandrine 1: total centella asiatica glycosides 4. In some embodiments, the weight ratio between gentiopicroside, tetrandrine, and total centella asiatica glycosides in component a is gentiopicroside 4-7: tetrandrine 0-2: total centella asiatica glycosides 3-5.

[0039] In some embodiments, component b comprises hydrolyzed panax notoginseng saponins and butanediol.

[0040] In some embodiments, the content of hydrolyzed ginsenosides in component b ranges from 0.1-50%. In some embodiments, the content of hydrolyzed ginsenosides in component b ranges from 0.1-0.5%, 0.5-1%, 1-1.5%, 1.5-2%, 2-2.5%, 2.5-3%, 3-3.5%, 3.5-4%, 4-4.5%, 4.5-5%, 5-5.5%, 5.5-6%, 6-6.5%, 6.5-7%, 7-7.5%, 7.5-8%, 8-8.5%, 8.5-9%, 9-9.5%, 9.5-10%, 10-15%, 15-20%, 20-25%, 25-30%, 30-35%, 35-40%, 40-45%, or 45-50%. In some embodiments, the content of hydrolyzed ginsenosides in component b ranges from 1-20%. In some embodiments, the content of hydrolyzed ginsenosides in component b ranges from 5.5-6.5%. In some embodiments, the content of hydrolyzed ginsenosides in component b is 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, or 6.5%.

[0041] In some embodiments, the content of butanediol in component b ranges from 50-99%. In some embodiments, the content of butanediol in component b ranges from 50-55%, 55-60%, 60-65%, 65-70%, 70-75%, 75-80%, 80-85%, 85-90%, 90-95%, or 95-99%. In some embodiments, the content of butanediol in component b ranges from 80-99%. In some embodiments, the content of butanediol in component b ranges from 93.5-94.5%. In some embodiments, the content of butanediol in component b is 93.5%, 93.6%, 93.7%, 93.8%, 93.9%, 94.0%, 94.1%, 94.2%, 94.3%, 94.4%, or 94.5%.

[0042] In some embodiments, component b is a cosmetic raw material rare ginsenoside GLF with trade name Suni Jinsenoside GLF-HJ(BG). The component hydrolyzed ginsenosides in rare ginsenoside GLF are obtained by solvent extraction and hydrolysis of Panax ginseng C. A. Mey. The component hydrolyzed ginsenosides in rare ginsenoside GLF contain a certain amount of rare ginsenoside RG3.

[0043] In some embodiments, the ratio of the amount of component a to component b in the composition is from 20:1 to 1:20. In some embodiments, the ratio of the amount of component a to component b in the composition is from 5:1 to 1:20. In some embodiments, the ratio of the amount of component a to component b in the composition is from 1:1 to 1:8. In some embodiments, the ratio of the amount of component a to component b in the composition is 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.

[0044] In some embodiments, the composition of the present application has an anti-inflammatory, redness-reducing, and / or soothing effect.

[0045] In some embodiments, component a and component b in the composition of the present application have a synergistic effect in terms of anti-inflammation, wherein the anti-inflammatory effect of the composition comprising component a and component b is significantly better than the simple addition of the anti-inflammatory effect of component a alone and the anti-inflammatory effect of component b alone.

[0046] Use of the composition of the present application for the manufacture of a medicament or a cosmetic

[0047] A third aspect of the present application relates to the use of the composition according to the first aspect or the second aspect of the present application in the preparation of a medicament or a cosmetic product having an anti-inflammatory, redness-reducing, and / or soothing effect.

[0048] In some embodiments, the anti-inflammatory effect is an anti-inflammatory effect of inhibiting the secretion of pro-inflammatory factor IL-6.

[0049] In some embodiments, the soothing effect is a soothing effect on skin redness.

[0050] Medicaments and cosmetics

[0051] A fourth aspect of the present application relates to a medicament or a cosmetic product comprising the composition according to the first aspect or the second aspect of the present application, wherein the composition accounts for about 0.1% to 20% by weight of the medicament or the cosmetic product.

[0052] In some embodiments, the composition comprises about 0.1-1%, 1-2%, 2-3%, 3-4%, 4-5%, 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%, 13-14%, 14-15%, 15-16%, 16-17%, 17-18%, 18-19%, or 19-20% by weight of the pharmaceutical or cosmetic. In some embodiments, the composition comprises about 0.6% by weight of the pharmaceutical or cosmetic.

[0053] In some embodiments, the pharmaceutical and cosmetic are for skin care. In some embodiments, the pharmaceutical and cosmetic are for anti-inflammatory, redness reduction, and / or soothing. In some embodiments, the pharmaceutical and cosmetic comprise the composition comprising component a and component b of the present application as an active ingredient, and one or more pharmaceutically or cosmetically acceptable carriers, diluents, or excipients. In some embodiments, the pharmaceutical and cosmetic are creams, lotions, pastes, ointments, packs, gels, lotions, or serums. In some embodiments, the pharmaceutical and cosmetic of the present application can further contain one or more other ingredients, such as plant extracts, nutritional additives, surfactants, fragrances and flavors, pigments, preservatives, antioxidants, humectants, ultraviolet absorbers, astringents, penetration aids, pH adjusters, and the like. The selection can be made by those skilled in the art according to their common general knowledge and specific needs.

[0054] In some embodiments, the cosmetic comprises the composition according to the first or second aspect of the present application and one or more cosmetically acceptable carriers, diluents, or excipients. In some embodiments, the one or more cosmetically acceptable carriers, diluents, or excipients include thickening agents, humectants, emollients, emulsifiers, pH adjusters, chelating agents, water, or a combination of one or more thereof.

[0055] In some embodiments, the cosmetic comprises the composition according to the first or second aspect of the present application, thickening agents, humectants, emollients, emulsifiers, pH adjusters, chelating agents, and water.

[0056] In some embodiments, the thickening agent is Acrylates / C 10-30 Alkyl Acrylate Crosspolymer. In some embodiments, the humectant includes glycerin, glycereth-26, 1,2- hexanediol, D-panthenol, SPHINGOMONAS Ferment Extract, or a combination of one or more thereof. In some embodiments, the emollient is squalane. In some embodiments, the emulsifier is polyglyceryl-10 stearate. In some embodiments, the pH adjuster is tetrahydroxypropyl ethylenediamine. In some embodiments, the chelating agent is disodium EDTA.

[0057] In some embodiments, the cosmetic includes the composition according to the first or second aspect of the application, Acrylates / C 10-30 Alkyl Acrylate Crosspolymer, SPHINGOMONAS Ferment Extract, glycerin, disodium EDTA, glycereth-26, 1,2-hexanediol, p- hydroxyacetophenone, polyglyceryl-10 stearate, squalane, tetrahydroxypropyl ethylenediamine, D-panthenol, and water.

[0058] In some embodiments, the cosmetic includes about 0.1-1% of the composition according to the first or second aspect of the application, about 0.1-1% of Acrylates / C 10-30 Alkyl Acrylate Crosspolymer, about 0.01-1% of SPHINGOMONAS Ferment Extract, about 1-10% of glycerin, about 0.01-1% of disodium EDTA, about 1-10% of glycereth-26, about 0.1-1% of 1,2-hexanediol, about 0.1-1% of p-hydroxyacetophenone, about 0.1-1% of polyglyceryl-10 stearate, about 0.1-1% of squalane, about 0.01-1% of tetrahydroxypropyl ethylenediamine, about 1-10% of D-panthenol, and about 50-95% of water.

[0059] In some embodiments, the cosmetic includes about 0.6% of the composition according to the first or second aspect of the application, about 0.15% of Acrylates / C 10-30 Alkyl Acrylate Crosspolymer, about 0.05% of SPHINGOMONAS Ferment Extract, about 3% of glycerin, about 0.05% of disodium EDTA, about 4% of glycereth-26, about 0.4% of 1,2-hexanediol, about 0.4% of p-hydroxyacetophenone, about 0.2% of polyglyceryl-10 stearate, about 0.5% of squalane, about 0.07% of tetrahydroxypropyl ethylenediamine, about 2% of D-panthenol, and about 88.58% of water.

[0060] In some embodiments, the cosmetic product comprises the composition according to the first or second aspect of the application, wherein the content of component a is 0.01-1% and the content of component b is 0.1-1%. In some embodiments, the cosmetic product comprises the composition according to the first or second aspect of the application, wherein the content of component a is 0.1% and the content of component b is 0.5%. DETAILED DESCRIPTION

[0061] The present application can be implemented by the following embodiments, but the present application is not limited thereto.

[0062] The instruments and equipment used in the embodiments of the present application are all conventional instruments and equipment in the art and can be replaced by instruments and equipment meeting the corresponding standards.

[0063] The Chinese herbal medicine raw materials used in the embodiments of the present application are all commercially available if not specifically indicated, and the reagents used in the embodiments of the present application are all commercially available analytical pure chemical reagents if not specifically indicated.

[0064] The raw materials and instruments and equipment mentioned in the present application are all conventional raw materials and instruments and equipment in the art, which are only examples and are not intended to limit the protection scope of the present application. Those skilled in the art can select equivalent raw materials and related instruments and equipment based on the disclosure of the present application.

[0065] Materials, apparatus, and statistical analysis methods

[0066] Experimental reagents and apparatus: RAW264.7 cells (Fu Heng Biological), DMEM cell culture medium (Gibco), fetal bovine serum (Corning), DPBS (Gibco), LPS (Sigma), cell incubator (Thermo Scientific), fluorescence microplate reader (Molecular Devices), 96-well plate (Titan), T25 cell culture bottle (Corning), cell scraper (JET BIOFIL), 15 ml centrifuge tube (Titan), CCK-8 kit (Bi Yun Tian), mouse IL-6 ELISA kit (Invitrogen).

[0067] Statistical method: the experimental results are the mean ± standard deviation of multiple parallel experimental data, and the paired t-test is used to test whether the difference between groups has statistical significance. The statistical software is GraphPad 5.0.

[0068] Examples

[0069] Preparations of the composition

[0070] Component a: Compound gentian extract containing gentiopicroside, Hanfenside and total ginsenoside prepared by the extraction process of Example 3 disclosed in the patent CN113041286B of Xiangyi Herbal. The specific preparation process is as follows: take 1000g of plant material, of which gentian is 45%, Stephania tetrandra 30%, and Centella asiatica 25%, crush into 20 mesh coarse powder, add about 5 times water for external circulation extraction, the extraction temperature is about 80℃, and obtain extract A1; add about 3 times about 40% ethanol to the residue for external circulation extraction, the extraction temperature is about 60℃, and obtain extract A2; add about 2 times about 70% ethanol to the residue for external circulation extraction, the extraction temperature is about 60℃, and obtain extract A3. Combine A1, A2 and A3, mix and pass through a 5000 dalton ultrafiltration membrane, collect the filtrate; filter with a 1000 dalton nanofiltration membrane, recover the solvent, obtain a concentrated solution, adjust to 1000ml with water, and freeze-dry the solution to obtain the compound gentian extract.

[0071] Component b: commercially available rare ginsenoside from Sanmit Technology Co., Ltd., trade name Suni Jinsenoside GLF-HJ(BG) (hereinafter referred to as GLF), which contains 5.5-6.5% hydrolyzed ginsenosides and 93.5-94.5% butanediol as the main components.

[0072] Preparation of the composition: components a and b are prepared according to the ratio of 1:1, 1:4, 1:8 and 1:30, respectively, at a total composition concentration of 100 and 250μg / ml, to obtain Examples 1-8. When components a and b are applied alone at the same concentration, Comparative Examples 1-4 are obtained. The specific conditions of each prepared composition are shown in Table 1.

[0073] Table 1: Composition of each prepared composition, ratio of each component and composition concentration

[0074]

[0075]

[0076] Examples of effects

[0077] 1. Cytotoxicity evaluation of component a and component b

[0078] 1.1 Experimental method:

[0079] Dilute components a (compound gentian extract) and b to the concentration to be tested with DMEM cell culture medium containing 10% fetal bovine serum, and perform RAW264.7 cell toxicity test when the two components are applied alone.

[0080] RAW264.7 cells were inoculated into T25 culture flask and placed in a cell incubator at 37℃, 5% CO2, when the cells grew to 70-80% confluence, the cells were subcultured. The adherent RAW264.7 cells were scraped with a spatula, and the cell suspension was collected into a 15 mL centrifuge tube, centrifuged at 1000 rmp for 4 min, the supernatant was discarded, 4 mL of complete medium (DMEM cell culture medium containing 10% fetal bovine serum) was added to resuspend, and the cell density was adjusted to 6*10 5 Each well of 96-well plate was inoculated with 100 μL of the cell suspension, and the cells were cultured in the cell incubator overnight.

[0081] After overnight culture, the original culture medium in the wells was discarded, and 100 ul of component a (0, 25, 50, 100, 250, 500 μg / mL) or component b (0, 83.3, 166.7, 416.7, 833.3, 1666.7 μg / mL) at the concentration to be tested was added to each well, 6 replicate wells were set for each concentration of each sample to be tested, and the culture was continued in the cell incubator for 24 h.

[0082] After the culture was completed, 100 ul of CCK8 solution was added to the cells in the wells after the cell supernatant was collected, and the plate was placed in a 37℃ incubator for 40 min. The operation was carried out according to the requirements of the CCK8 kit instructions, and the absorbance was measured at 450 nm.

[0083] 1.2 Experimental results:

[0084] The results of the effect of component a (compound gentian extract) on the activity of RAW264.7 cells are shown in Figure 1 , and the results of the effect of component b (GLF) on the activity of RAW264.7 cells are shown in Figure 2 . As shown in the results, the survival rate of RAW264.7 cells in the group with component a (compound gentian extract) at a concentration of 250 μg / mL and below was more than 80%, and the survival rate of RAW264.7 cells in the group with component b (GLF) at a concentration of 1666.7 μg / mL and below was close to 100%, indicating that components a and b had no toxic effect on RAW264.7 cells within the above concentration range, and did not affect cell growth.

[0085] 2. Anti-inflammatory activity evaluation

[0086] The anti-inflammatory activity of the compositions of Examples 1-8 and the compositions of Comparative Examples 1-4 was evaluated by determining the effect of the compositions on LPS-induced IL-6 secretion from RAW264.7 cells. The interaction between component a and component b in the composition was calculated by the formula for the drug interaction index disclosed in the literature (Liu Jin et al., Foreign Medical Sciences, 2006(33):459-461) to determine whether the two components in the composition have a synergistic effect in terms of anti-inflammation.

[0087] 2.1 Experimental method

[0088] RAW264.7 cells were inoculated into T25 culture bottles and placed in a cell incubator at 37°C, 5% CO2, and when the cells grew to 70-80% confluence, the cells were subcultured. The adherent RAW264.7 cells were gently scraped off with a spatula, and the cell suspension was collected into a 15 mL centrifuge tube and centrifuged at 1000 rpm for 4 min. The supernatant was discarded, 4 mL of complete medium (DMEM cell culture medium containing 10% fetal bovine serum) was added to resuspend the cells, and the cell density was adjusted to 6*10 5 6 / mL, 100 μL per well was inoculated into a 96-well plate, and incubated in the incubator overnight.

[0089] The experiment was divided into a blank control group, a LPS model group, and a sample group. In the sample group, 100 μl of each concentration of the sample to be tested (Examples 1-8, Comparative Examples 1-6) was added to the well plate, and then 11 μL of LPS (10 μg / mL) was added to the well plate. In the blank control group, 100 μL of complete medium was added, and in the LPS model group, 50 μL of complete medium and 50 μL of LPS (4 μg / mL) were added. The culture was continued in the incubator for 24 h, and 3 replicate wells were set up for each sample to be tested.

[0090] After the culture was completed, the cell supernatant of each sample to be tested was collected, and the operation was performed according to the requirements of the mouse IL-6 ELISA kit instructions. The absorbance was measured at 450 nm, and the IL-6 concentration in the cell supernatant of each sample was calculated according to the standard curve to detect the effect of the sample to be tested on the secretion of the inflammatory factor IL-6 (n=2).

[0091] 2.2 Calculation of the inhibition rate of the sample to be tested on the IL-6 secretion of RAW264.7 cells

[0092] IL-6 secretion inhibition rate (%) = (LPS model group IL-6 concentration - sample to be tested group IL-6 concentration) / LPS model group IL-6 concentration * 100%

[0093] 2.3 Calculation of the interaction index between component a and component b

[0094] The interaction between component a and component b in the composition is calculated according to the calculation formula of the interaction index (γ) disclosed in the literature (Liu Jin et al., Foreign Medical Sciences Pharmacology, 2006 (33): 459-461). The specific calculation formula is γ = a / A + b / B. Wherein, a and b are the respective doses of the two components when used in combination, and A and B are the doses of the two components when used alone to produce the same effect as when used in combination. When γ = 1, the interaction of the two drugs is additive; when γ < 1, the interaction of the two drugs is synergistic, and the smaller the γ value, the stronger the synergistic effect; when γ > 1, the interaction of the two drugs is antagonistic.

[0095] 2.4 Experimental results

[0096] According to the above experimental method and the above calculation formula of IL-6 secretion inhibition rate and the calculation formula of interaction index, the determination results of each test sample on the amount of IL-6 secretion of LPS-induced RAW264.7 cells, IL6 secretion inhibition rate and the interaction index between component a and component b are shown in Table 2.

[0097] Table 2. The amount of IL-6 secretion of LPS-induced RAW264.7 cells, IL6 secretion inhibition rate and the interaction index of two components of each test sample

[0098]

[0099]

[0100] The data in Table 2 shows that the IL-6 concentration of the LPS model group is significantly higher than that of the blank control group. The IL-6 average inhibition rate data of Comparative Examples 1-2 shows that the inhibition rate of component a (compound gentian extract) alone on LPS-induced IL-6 secretion of RAW264.7 cells at concentrations of 100 and 250 μg / mL is 20.2% and 52.8%, respectively, indicating that component a has certain anti-inflammatory activity. The IL-6 average inhibition rate data of Comparative Examples 3-4 shows that the inhibition rate of component b (GLF) alone on LPS-induced IL-6 secretion of RAW264.7 cells at concentrations of 100 and 250 μg / mL is only 3.4% and 3.8%, respectively, indicating that the anti-inflammatory activity of component b is not obvious. The IL-6 average inhibition rate results of Examples 1-3 and Examples 6-8 show that compared with the use of component a or b alone, the inhibition of IL-6 secretion of the composition containing component a (compound gentian extract) and component b (GLF) at the same concentration is greater than that of the two components alone when the ratio of the two components is between 1:1 and 1:8. For example, in Example 1, the composition concentration is 100 μg / mL and the content ratio of component a and component b is 1:1, the average inhibition rate of the composition on IL-6 is 35.9%, which is greater than that of component a alone (i.e. Comparative Example 1, the average inhibition rate is 14.5%) and that of component b alone (Comparative Example 3, the average inhibition rate is 2.1%) at the same concentration. The IL-6 average inhibition rates of Example 4 and Example 8, in which the content ratio of component a and component b is 1:30, are 2.5% and 4.8%, respectively, which are close to those of Comparative Example 3 and Comparative Example 4 in which component b is used alone, indicating that when the concentration ratio of the two components is 1:30, the anti-inflammatory activity of the composition is close to that of component b alone.

[0101] According to the calculation results of the interaction index γ of Examples 1-8, when the content ratio of component a and component b in the composition is between 1:1 and 1:8, γ < 1, indicating that component a (compound gentian extract) and component b (GLF) have synergistic anti-inflammatory effect; and when the content ratio of component a and component b is 1:30, γ > 1, indicating that there is no synergistic anti-inflammatory effect between the two components.

[0102] 3. Evaluation of soothing efficacy in humans

[0103] The soothing redness efficacy of the composition containing component a (compound gentian extract) and component b (GLF) in humans was evaluated by the closed patch erythema model.

[0104] 3.1 Subjects and test samples

[0105] Subjects: healthy Chinese people aged 19-60 years old, in accordance with the selection of subjects in 5. Selection of subjects in Human Skin Patch Test in Cosmetic Safety Technology Specification 2015 Edition.

[0106] Erythema-inducing agent: 99% pure Sodium Dodecyl Sulfate (SDS) (from SIGMA-ALDRICH)

[0107] Patch tester: Finn Chamber 8mm

[0108] Test sample: Self-made water-in-oil emulsion cream containing 0.1% compound gentiana extract and 0.5% GLF

[0109] Blank control: Self-made water-in-oil emulsion cream

[0110] Medical tape: Low-allergenic tape (from 3M Transpore™)

[0111] 3.2 Test method

[0112] Day 1: Skin screening (BL) was performed on the test area on the back of the subjects by the staff. The skin area without influence evaluation factors was selected for patch test. 25 μL of 1% SDS aqueous solution was placed in the center of the 8mm patch adapter chamber with a pipette, and was attached to the back of the human body for 24h. In order to prevent the patch tester from falling off, medical tape was fixed around the four corners.

[0113] Day 2: 0.5h-1h (mainly based on the indentation disappearance time) after the patch tester was removed, the skin of the test area on the back was scored by the clinical dermatologist according to the judgment standard of the cosmetic safety technical specification (2015) (after erythema-T0.5). Then 25 μL of test sample and blank control material was placed in the center of the 8mm patch adapter chamber with a pipette, and was attached to the SDS stimulation site for 24h. In order to prevent the patch tester from falling off, medical tape was fixed around the four corners.

[0114] Day 3: 0.5h-1h (mainly based on the indentation disappearance time) after the patch tester was removed, the skin of the test area on the back was scored by the clinical dermatologist according to the judgment standard of the cosmetic safety technical specification (2015) (after erythema-T0.5).

[0115] 3.3 Test results

[0116] The clinical doctors scored the redness of the blank control group and the experimental group at two time points of 0.5h after erythema and 24h after erythema, and the average results of redness evaluation were as shown in Table 3. Figure 3

[0117] There were 23 subjects in the blank control group who were successfully activated redness at the test point. The test results of the blank control group showed that the redness value of the water-in-oil emulsion cream before and after the action did not change obviously, and there was no significant difference (p>0.05).​

[0118] The experimental group had 22 subjects successfully activated red at the test point. The test results of the experimental group showed that: after the action of 0.1% compound gentian extract + 0.5% GLF water-in-oil self-prepared cream, the redness evaluation value of the skin was significantly reduced, and there was a significant difference (p<0.01). It shows that the composition containing component a (compound gentian extract) and component b (GLF) of the present application can effectively relieve skin redness.

[0119] Cosmetic examples Figure 1

[0120] The soothing and relieving repair mask containing the composition of the present application can be prepared according to the following preparation process with the formula shown in Table 3.

[0121] Table 3. Soothing and relieving repair mask formula

[0122]

[0123]

[0124] The preparation process of the soothing and relieving repair mask is as follows:

[0125] 1. Add deionized water to the main pot, add Carbopol U-20 to the main pot, soak for 10 min, and then add the remaining A-phase raw materials under the condition of homogenization at 1500 rpm. Turn on the main pot stirring at 25 rpm, start heating, heat to 65℃, homogenize for 3000 min, 2 min, homogenization is completed, continue to warm up, heat to 80-85℃, keep warm for 10 min;

[0126] 2. Add the B-phase raw materials to the oil pot one by one, heat to 80-85℃, and stir to dissolve and disperse evenly;

[0127] 3. Under the condition of homogenization at 1500 rpm, add the B-phase raw materials to the main pot, homogenize at 3000 rpm for 2 min, and keep warm for 20 min, then start cooling;

[0128] 4. When the main pot is reduced to 65℃, turn off the cooling, add C-phase, stir for 5 min, and continue to cool;

[0129] 5. Reduce the temperature to 45℃, add the D-phase raw materials, and circulate at 1000 rpm for 2 min, then stir for 15 min;

[0130] 6. When the main pot is reduced to about 40℃, filter with a 200-mesh sieve, and then discharge after detection.

Claims

1. A composition comprising component a and component b, wherein component a is a compound gentian extract containing gentian plant extract, fangchi plant extract and centella asiatica plant extract, and component b is ginsenoside GLF, wherein the ginsenoside GLF consists of 5.5%-6.5% hydrolyzed ginsenosides and 93.5%-94.5% butylene glycol, wherein the content ratio of component a to component b in the composition is 1:1-1:8, and wherein component a is prepared by the following method: taking 1000g of plant material, including 45% gentian, 30% fangchi, and 25% centella asiatica. The powder was pulverized into a 20-mesh coarse powder, and extracted with 5 times its volume of water using an external circulation system at 80°C to obtain extract A1. The residue was then extracted with 3 times its volume of 40% ethanol using an external circulation system at 60°C to obtain extract A2. The residue was then extracted with 2 times its volume of 70% ethanol using an external circulation system at 60°C to obtain extract A3. A1, A2, and A3 were combined, mixed, and filtered through a 5000 Dalton ultrafiltration membrane. The filtrate was collected and filtered through a 1000 Dalton nanofiltration membrane to recover the solvent and obtain a concentrated solution. The solution was adjusted to 1000 ml with water and then freeze-dried to obtain the compound gentian extract.

2. Use of the composition according to claim 1 in the preparation of a medicament or cosmetic having anti-inflammatory, soothing, and / or calming effects.

3. A drug or cosmetic made from the composition according to claim 1, wherein the composition comprises 0.1%-20% by weight of the drug or cosmetic.

4. The drug or cosmetic according to claim 3, wherein the drug or cosmetic is a cream, lotion, paste, ointment, mask, gel, lotion, or serum.

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