Application of a plant extract containing geniposide

Abstract

The present invention discloses the use of a plant extract containing geniposide in inhibiting and / or improving angiogenesis, as well as the use of a plant extract containing geniposide in preparing a cosmetic having the efficacy of inhibiting and / or improving angiogenesis, and provides a cosmetic containing a plant extract containing geniposide and a method for inhibiting and / or improving angiogenesis. By adopting the technical solution provided by the present invention, angiogenesis in the skin can be inhibited and / or improved, thereby improving problems such as telangiectasia, skin erythema, dark circles under the eyes, etc. caused by angiogenesis.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and particularly to the application of a plant extract containing geniposide. Background Art

[0002] Traditional Chinese medicine Gardenia jasminoides Ellis is the dried ripe fruit of the plant Gardenia jasminoides in the Rubiaceae family. It belongs to the first batch of dual-purpose resources of medicine and food promulgated by the Ministry of Health. It can clear heat and purge fire, cool blood and detoxify, activate blood circulation and disperse stasis, and has the effects of protecting the liver, promoting bile secretion, reducing blood pressure, sedation, hemostasis, and detumescence. It is commonly used in traditional Chinese medicine clinics to treat jaundice hepatitis, contusions and strains, hypertension, diabetes, etc.

[0003] The chemical components contained in traditional Chinese medicine Gardenia jasminoides Ellis mainly include three categories: iridoid glycosides, organic acids, and pigments, etc. Among them, iridoid glycosides include geniposide, genipin glycoside, genipinic acid, etc.; organic acids are esters formed by phenylpropionic acid derivatives and quinic acid, such as chlorogenic acid, etc.; pigments include flavonoid gardenin, diterpenoid crocin, etc.

[0004] At present, the effective components of traditional Chinese medicine Gardenia jasminoides Ellis recognized by domestic and foreign research are iridoid substances, and the one with the highest activity is geniposide. Geniposide is also an intermediate for the production of gardenia blue, gardenia red, and gardenia purple, and an effective monomer for pharmaceuticals. Therefore, it is of great significance to extract from traditional Chinese medicine Gardenia jasminoides Ellis and develop a gardenia fruit extract with geniposide as the main active ingredient and its efficacy applications.

[0005] Gardenia jasminoides Ellis as traditional Chinese medicine is widely used in traditional Chinese medicine compatibility, and its main efficacy application is for oral administration, such as Qinghuo Zhimai tablets, etc. However, the research on gardenia fruit extract as a skin external preparation is relatively less. From the literature found, it is known that gardenia used externally has the effect of detumescence and pain relief. More in-depth research mainly involves the anti-inflammatory effect of geniposide. There is no report on the efficacy research of geniposide or gardenia fruit extract for improving the neovascularization of cutaneous capillaries when used externally.

[0006] The neovascularization of cutaneous capillaries is the process of the human body generating new capillaries, which is affected by various physiological and pathological conditions of the skin. Research shows that the skin can induce the neovascularization of cutaneous capillaries under external pressure (such as ultraviolet irradiation, inflammation, hypoxia, etc.). Some research shows that the neovascularization of capillaries will lead to the generation of telangiectasia, resulting in skin erythema or dark circles. Summary of the Invention

[0007] The object of the present invention is to overcome the deficiencies of the prior art and provide the use of a plant extract containing geniposide in inhibiting and / or improving angiogenesis, as well as the use of a plant extract containing geniposide in preparing a cosmetic having the efficacy of inhibiting and / or improving angiogenesis. In addition, a cosmetic containing a plant extract containing geniposide and a method for inhibiting and / or improving angiogenesis are provided. By adopting the technical solution provided by the present invention, angiogenesis of the skin can be inhibited and / or improved, thereby improving problems such as telangiectasia, skin erythema, and dark circles caused by angiogenesis.

[0008] In order to achieve the above object, in the first aspect of the present invention, the use of a plant extract containing geniposide in inhibiting and / or improving angiogenesis is provided.

[0009] As a preferred embodiment of the present invention, the content of geniposide in the plant extract containing geniposide is ≥ 80 wt.%.

[0010] As a preferred embodiment of the present invention, the content of geniposide in the plant extract containing geniposide is ≥ 98 wt.%.

[0011] As a preferred embodiment of the present invention, the plant extract containing geniposide is a gardenia fruit extract.

[0012] As a preferred embodiment of the present invention, the preparation method of the gardenia fruit extract comprises the following steps:

[0013] S1. Crushing the gardenia fruit, performing water extraction after sieving, and after solid-liquid separation, the obtained liquid is the extract;

[0014] S2. Adsorbing the extract obtained in step S1 with a resin, and then eluting with a solvent to obtain an eluate;

[0015] S3. Concentrating the eluate obtained in step S2, and then drying to obtain the gardenia fruit extract.

[0016] As a preferred embodiment of the present invention, in step S2 of the preparation method of the gardenia fruit extract, the resin is a macroporous adsorption resin.

[0017] As a preferred embodiment of the present invention, in step S2 of the preparation method of the gardenia fruit extract, the macroporous adsorption resin is a non-polar resin or a weakly polar resin.

[0018] As a preferred embodiment of the present invention, in step S2 of the preparation method of the gardenia fruit extract, the solvent is water and / or an alcohol solvent.

[0019] As a preferred embodiment of the present invention, in step S2, the adsorption method is as follows:

[0020] Take a qualified resin column after cleaning, load it with pure water, and then load the extraction solution onto the column at room temperature, controlling the column flow rate at 1 - 3 BV / h; after the extraction solution loading is completed, keep the column flow rate unchanged, wash the resin column bed with pure water until the effluent from the column bed shows no amino acid color reaction, and discard the pure water washing solution after the pure water washing of the resin column is completed.

[0021] The elution method is as follows:

[0022] Elute the resin column with 1500 ml of 5 - 10% ethanol aqueous solution, control the elution flow rate at 1 - 3 BV / h for impurity removal; after recovering and removing alcohol from the 10% ethanol eluate, discard the residue; then elute the resin column with 2000 ml of 60 - 65% ethanol aqueous solution, control the elution flow rate at 1 - 3 BV / h, and collect the eluate to obtain an eluate containing Gardenia fruit extract.

[0023] In the second aspect of the present invention, there is provided the use of a plant extract containing geniposide in the preparation of a cosmetic having the efficacy of inhibiting and / or improving angiogenesis.

[0024] In the third aspect of the present invention, there is provided a cosmetic containing a plant extract containing geniposide.

[0025] As a preferred embodiment of the present invention, the dosage form of the cosmetic is aqueous solution, emulsion, cream, gel, mask or essence.

[0026] As a preferred embodiment of the present invention, the content of the plant extract containing geniposide in the cosmetic is 0.1 - 2 wt.%.

[0027] As a preferred embodiment of the present invention, the content of the plant extract containing geniposide in the cosmetic is 0.5 - 1 wt.%.

[0028] As a preferred embodiment of the present invention, the cosmetic further contains excipients acceptable in the cosmetic field.

[0029] As a preferred embodiment of the present invention, the excipients include at least one of solvent, thickener, humectant, emollient, emulsifier, preservative, antioxidant, pH regulator, penetration enhancer, liposome, chelating agent, surfactant, fragrance, pigment.

[0030] As a preferred embodiment of the present invention, the cosmetic comprises the following components in weight percentages:

[0031] 0.1 - 2% plant extract containing geniposide, 3 - 10% humectant, 0.1 - 20% thickener, and the balance water.

[0032] In a fourth aspect of the present invention, a method for inhibiting and / or improving angiogenesis is provided, comprising the following steps: contacting a cosmetic with the skin where angiogenesis occurs, and the cosmetic is the above-mentioned cosmetic.

[0033] The beneficial effects of the present invention are as follows:

[0034] (1) The present invention discovers that the plant extract containing geniposide has good efficacy in inhibiting and / or improving angiogenesis, and has a significant inhibitory effect on angiogenesis-related target genes.

[0035] (2) The plant extract containing geniposide can be used as an active ingredient in cosmetics to prepare cosmetics with the efficacy of inhibiting and / or improving angiogenesis. The cosmetics containing the plant extract containing geniposide provided by the present invention have good improvement and alleviation effects on telangiectasia, skin erythema, and dark circles caused by angiogenesis.

[0036] (3) By using the cosmetics of the present invention, skin telangiectasia can be soothed, the paroxysmal pain and fever symptoms of erythematous skin can be alleviated, the redness can be rapidly subsided, and the dark circles caused by insufficient skin microcirculation can also be lightened. Description of the Drawings

[0037] Figure 1 It is the HPLC chromatogram of the gardenia fruit extract in Example 1;

[0038] Figure 2 It is the test result of the expression level of angiogenesis-related genes in Example 2;

[0039] Figure 3 It is the test result of the left arm of the erythematous skin for the cosmetic aqueous solution A and deionized water in Example 3;

[0040] Figure 4 It is the test result of the right arm of the erythematous skin for the cosmetic aqueous solution A and deionized water in Example 3;

[0041] Figure 5 It is the test result of the effect of the cosmetic gel A on dark circles in Example 4;

[0042] Figure 6 It is the test result of the effect of the cosmetic emulsion A and the control emulsion on telangiectasia in Example 5. Detailed Embodiments

[0043] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0044] In the present invention, among the technically described features in an open-ended manner, it includes a closed technical solution composed of the listed features, and also includes an open technical solution containing the listed features.

[0045] In the present invention, regarding the numerical range, unless otherwise specified, the above numerical range is considered continuous and includes the minimum and maximum values of this range, as well as each value between such minimum and maximum values. Further, when the range refers to an integer, it includes each integer between the minimum and maximum values of this range. In addition, when providing multiple range descriptions of features or characteristics, these ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all sub-ranges subsumed therein.

[0046] In the present invention, there is no particular limitation on the specific dispersion and stirring treatment methods.

[0047] The reagents or instruments used in the present invention that are not indicated by the manufacturer can all be conventional products obtained through commercial purchase.

[0048] To solve the problem of capillary angiogenesis in the prior art, the embodiments of the present invention provide the application of a plant extract containing geniposide in inhibiting capillary angiogenesis and / or improving capillary angiogenesis.

[0049] Capillary angiogenesis is closely related to the expression of various factors in the skin. Platelet endothelial cell adhesion molecule (PECAM1 / CD31) is a marker antigen expressed by capillary endothelial cells and is also a marker of capillary angiogenesis. Smooth muscle actin (α-SMA) is an important functional protein in vascular smooth muscle cells, and the change in its expression is of great significance for indicating the changes in the structure and function of vascular smooth muscle cells. α-SMA can promote the generation of extracellular matrix.

[0050] Cyclooxygenase 2 (COX-2), also known as prostaglandin endoperoxide reductase 2, is a key enzyme that catalyzes the conversion of arachidonic acid into prostaglandins. Endothelial nitric oxide synthase (eNOS) is secreted and expressed by vascular endothelial cells and is an important isoenzyme that regulates vascular function. eNOS catalyzes the production of nitric oxide (NO) from L-arginine. Vascular endothelial cells participate in regulating vascular tone, hemostasis, blood pressure, and vascular remodeling by synthesizing and secreting NO.

[0051] Through creative experiments, the inventors found that plant extracts containing geniposide can significantly inhibit the expression of target genes related to angiogenesis (α-SMA gene, PECAM1 gene, eNOS gene, COX-2 gene), and this inhibitory effect shows an obvious dose-dependent relationship.

[0052] Among them, COX-2 is a key enzyme for prostaglandin synthesis. Plant extracts containing geniposide can inhibit prostaglandin synthesis by inhibiting COX-2, thereby alleviating the symptoms of erythema skin paroxysmal pain and fever caused by skin redness agents (such as methyl nicotinate) based on vasodilation effects, achieving the effect of rapid redness reduction, and thus improving capillary dilation, that is, improving capillary neogenesis.

[0053] In one embodiment, the content of geniposide in the plant extract containing geniposide is ≥80 wt.%. Optionally, the content of geniposide in the plant extract containing geniposide is ≥90 wt.%; optionally, the content of geniposide in the plant extract containing geniposide is ≥98 wt.%.

[0054] The inventors found that the inhibitory or improvement effect of plant extracts containing geniposide on capillary neogenesis is not only related to the content of geniposide, but also related to other trace components in the plant extract. Geniposide and other trace components in the plant extract achieve the inhibition or improvement of capillary neogenesis through synergistic effects.

[0055] In one embodiment, the plant extract containing geniposide is gardenia fruit extract.

[0056] The chemical components contained in gardenia fruit include iridoid glycosides 、 organic acids, pigments, etc. Among them, iridoid glycosides include geniposide. In the gardenia fruit extract obtained by a suitable extraction method, in addition to containing a high content of geniposide, it also contains different contents of cellobiose, ferulic acid, genipin-1-gentiobioside, geniposidic acid, sinapic acid, methyl ester of scandoside methyl ester, shanzhiside, genipin gentiobioside, isopropyl 4-hydroxybenzoate, dimethoxy-4-hydroxycinnamaldehyde, physcion, etc. trace components.

[0057] In one embodiment, the preparation method of the gardenia fruit extract includes the following steps:

[0058] S1. Crush the gardenia fruit, sieve it, then perform water extraction, and after solid-liquid separation, the obtained liquid is the extract;

[0059] S2. Adsorb the extract obtained in step S1 with resin, and then elute it with a solvent to obtain the eluate;

[0060] S3. Concentrate the eluate obtained in step S2, and then dry it to obtain the Gardenia fruit extract.

[0061] Through the steps of water extraction, resin adsorption, elution, concentration, and drying, a Gardenia fruit extract rich in geniposide can be prepared.

[0062] In one embodiment, the particle size of the crushed Gardenia fruit is 40-60 mesh.

[0063] In one embodiment, in step S1 of the method for preparing the Gardenia fruit extract, the extraction is carried out by stirring at 50-60 °C for at least 2 times.

[0064] In one embodiment, step S1 in the method for preparing the Gardenia fruit extract is as follows:

[0065] The crushed Gardenia fruit is dispersed in water, stirred at 50-60 °C for 1-3 hours, filtered, and the liquid obtained is the primary extract, and the solid is the filter residue, where the weight ratio of Gardenia fruit to water is 1:(4-6);

[0066] The filter residue is redispersed in water, stirred at 50-60 °C for 1-3 hours, filtered, and the liquid obtained is the secondary extract, where the weight ratio of the filter residue to water is 1:(2-4);

[0067] The primary extract and the secondary extract are mixed to obtain the extract.

[0068] By performing water extraction multiple times, the active ingredients in the Gardenia fruit can be greatly extracted.

[0069] In one embodiment, in step S2 of the method for preparing the Gardenia fruit extract, the resin is a macroporous adsorption resin.

[0070] In one embodiment, in step S2 of the method for preparing the Gardenia fruit extract, the macroporous adsorption resin is a non-polar resin or a weakly polar resin. The non-polar resin or weakly polar resin can be a conventional non-polar resin or weakly polar resin in the art. Optionally, the non-polar resin is D101 resin, D4020 resin; optionally, the weakly polar resin is AB-8 resin.

[0071] In one embodiment, in step S2 of the method for preparing the Gardenia fruit extract, the macroporous adsorption resin is AB-8 resin.

[0072] A suitable macroporous adsorption resin can effectively adsorb the Gardenia fruit extract in the extract, which helps to improve the yield and purity of the Gardenia fruit extract.

[0073] In one of the embodiments, in step S2 of the preparation method of the cape jasmine fruit extract, the method for adsorbing the extract with resin is as follows:

[0074] Take a qualified resin column, load it with pure water, and then load the extract onto the column at room temperature, with the column flow rate controlled at 1 - 3 BV / h;

[0075] After the extract loading is completed, keep the column flow rate unchanged, wash the resin column bed with pure water until the effluent from the column bed shows no amino acid color reaction, and the pure water washing of the resin column is completed. Discard the pure water washing solution.

[0076] In one of the embodiments, in step S2 of the preparation method of the cape jasmine fruit extract, the solvent is water and / or alcohol solvent.

[0077] In one of the embodiments, in step S2 of the preparation method of the cape jasmine fruit extract, the solvent is a mixed solution of water and ethanol.

[0078] In one of the embodiments, in step S2 of the preparation method of the cape jasmine fruit extract, the solvent is an ethanol aqueous solution with a concentration of 60 - 65%.

[0079] In one of the embodiments, in step S2 of the preparation method of the cape jasmine fruit extract, the elution method is as follows:

[0080] Elute the resin column with 1500 ml of ethanol aqueous solution with a concentration of 5 - 10%, control the elution flow rate at 1 - 3 BV / h, and elute to remove impurities; after the 10% ethanol eluate is recovered and de - alcoholized, discard the residual liquid;

[0081] Then elute the resin column with 2000 ml of ethanol aqueous solution with a concentration of 60 - 65%, control the elution flow rate at 1 - 3 BV / h, collect the eluate, and obtain the eluate containing the cape jasmine fruit extract.

[0082] On the one hand, the ethanol aqueous solution can elute the cape jasmine fruit extract well, and on the other hand, it will not destroy the activity of the cape jasmine fruit extract and does not affect the subsequent application of the cape jasmine fruit extract in inhibiting and / or improving capillary neovascularization.

[0083] In one of the embodiments, in step S3 of the preparation method of the cape jasmine fruit extract, the concentration method is vacuum distillation.

[0084] In one of the embodiments, in step S3 of the preparation method of the cape jasmine fruit extract, the drying method is vacuum drying.

[0085] By removing the solvent and drying, the water content of the prepared cape jasmine fruit extract is ≤10%, and the content of geniposide in the cape jasmine fruit extract is ≥80 wt.%.

[0086] An embodiment of the present invention provides an application of a plant extract containing geniposide in the preparation of a cosmetic having the efficacy of inhibiting angiogenesis and / or improving angiogenesis.

[0087] The plant extract containing geniposide is the above-mentioned plant extract containing geniposide.

[0088] In one embodiment, the content of geniposide in the plant extract containing geniposide is ≥80 wt.%. Optionally, the content of geniposide in the plant extract containing geniposide is ≥90 wt.%; Optionally, the content of geniposide in the plant extract containing geniposide is ≥98 wt.%.

[0089] An embodiment of the present invention provides a cosmetic, which contains a plant extract containing geniposide.

[0090] The plant extract containing geniposide is the above-mentioned plant extract containing geniposide.

[0091] In one embodiment, the content of geniposide in the plant extract containing geniposide is ≥80 wt.%. Optionally, the content of geniposide in the plant extract containing geniposide is ≥90 wt.%; Optionally, the content of geniposide in the plant extract containing geniposide is ≥98 wt.%.

[0092] In one embodiment, the dosage form of the cosmetic is aqueous solution, emulsion, cream, gel, mask or essence.

[0093] In one embodiment, the content of the plant extract containing geniposide in the cosmetic is 0.1-2 wt.%.

[0094] In one embodiment, the content of the plant extract containing geniposide in the cosmetic is 0.5-1 wt.%.

[0095] The inventors have found through research that the content of the plant extract containing geniposide in the cosmetic shows a dependence relationship with the efficacy of inhibiting angiogenesis and / or improving angiogenesis. When the content of the plant extract containing geniposide is too low, the efficacy of the cosmetic in inhibiting angiogenesis and / or improving angiogenesis is less significant or less efficient, and longer time or larger amount of use is required to achieve the corresponding efficacy; when the content of the plant extract containing geniposide is too low, the improvement of the efficacy of the cosmetic in inhibiting angiogenesis and / or improving angiogenesis is limited.

[0096] In one embodiment, the cosmetic further contains excipients acceptable in the cosmetic field.

[0097] In one embodiment, the adjuvants include at least one of a solvent, a thickening agent, a humectant, an emollient, an emulsifier, a preservative, an antioxidant, a pH regulator, a penetration enhancer, a liposome, a chelating agent, a surfactant, a fragrance, and a pigment.

[0098] It should be noted that those skilled in the art can select the types of adjuvants according to actual needs as long as the corresponding effects are achieved.

[0099] In one embodiment, the cosmetic includes the following components in weight percentages:

[0100] 0.1 - 2% of a plant extract containing geniposide, 3 - 10% of a humectant, 0.1 - 20% of a thickening agent, and the balance being water.

[0101] In one embodiment, the cosmetic further includes the following components in weight percentages: 5 - 15% of an emollient, 1 - 10% of an emulsifier.

[0102] In one embodiment, the cosmetic includes the following components in weight percentages:

[0103] 0.5 - 1% of a plant extract containing geniposide, 5 - 8% of a humectant, 0.3 - 0.6% of a thickening agent, 10 - 13% of an emollient, 5 - 7% of an emulsifier, and the balance being water.

[0104] In one embodiment, the humectant includes at least one of 1,3 - butanediol, propylene glycol, hexylene glycol, dipropylene glycol, glycerol, polyethylene glycol, xylitol, diglycerol, maltitol, maltose, D - mannitol, pentylene glycol, dipropylene glycol, polypropylene glycol, ethylene glycol, neopentyl glycol, erythritol, sorbitol, mannitol, xylitol, glucose, lactose, mannose, maltose, fructose, inositol, sorbitol, sorbitan, trehalose, and hydrolyzed starch sugar.

[0105] It should be noted that those skilled in the art can select the specific type of humectant according to actual needs as long as the corresponding moisturizing effect is achieved.

[0106] In one embodiment, the thickening agent includes at least one of carbomer, hydroxyethyl cellulose, xanthan gum, sodium polyacrylate, acrylic acid (ester) cross - linked polymer, and xanthan gum.

[0107] It should be noted that those skilled in the art can select the specific type of thickening agent according to actual needs as long as the corresponding thickening effect is achieved.

[0108] In one embodiment, the emollient includes at least one of triglyceride caprylate / caprate, isocetane, squalane, polydimethylsiloxane, cyclopentasiloxane, diisostearyl malate, dicetostearyl dimer dilinoleate, hexyl laurate, polyglyceryl-2 diisostearate, hydrogenated polyisobutene, pentaerythritol tetra(isostearate), triglyceride caprylate / caprate, octyldodecanol, dicaprylyl carbonate, macadamia nut oil, shea butter, jojoba oil, lanolin, white oil, almond oil, olive oil, jojoba oil, castor oil, C12-15 alkyl benzoate, isononyl isononanoate, dioctyl adipate, octyl stearate, hexyl laurate, coco-caprylate, and caprate.

[0109] It should be noted that those skilled in the art can select the specific type of emollient according to actual needs as long as the corresponding emollient effect is achieved.

[0110] In one embodiment, the emulsifier includes at least one of polyglyceryl-3 diisostearate, polyglyceryl-10 stearate, polyglyceryl-10 laurate, polyglyceryl-10 myristate, polyglyceryl-5 trioleate, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-30 soy sterol, PPG-13-decyltetradeceth-24, nonoxynol-12, PEG-35 castor oil / PEG-50 hydrogenated castor oil / noxynol-12 / propylene glycol, steareth-2, steareth-21, and hydrogenated lecithin.

[0111] It should be noted that those skilled in the art can select the specific type of emulsifier according to actual needs as long as the corresponding emulsifying effect is achieved.

[0112] One embodiment of the present invention provides a method for inhibiting and / or improving angiogenesis, comprising the following steps: contacting the skin surface where angiogenesis occurs with a cosmetic, and the cosmetic is the above-mentioned cosmetic.

[0113] It should be noted that, according to the dosage form of the cosmetic, the cosmetic can be applied, applied by dressing, or sprayed onto the skin surface where angiogenesis occurs.

[0114] The present invention will be further illustrated by specific examples as follows:

[0115] Example 1

[0116] This example provides a gardenia fruit extract, which is prepared by the following method:

[0117] (1) Pre-crushing:

[0118] Take the dried gardenia fruits and pre-crush them with a traditional Chinese medicine coarse crusher into a coarse powder of 40-60 meshes.

[0119] (2) Water extraction:

[0120] After 600 g of gardenia fruits are crushed, 3000 mL of water is added, and stirring and extraction are carried out at 55°C ± 2°C for 2 hours, followed by precise filtration to obtain the primary extraction liquor;

[0121] Another 2400 mL of pure water is added to the filter residue, and stirring and extraction are carried out at 55°C ± 2°C for 1 hour, followed by precise filtration to obtain the secondary extraction liquor;

[0122] The primary extraction liquor and the secondary extraction liquor are combined to obtain 5100 mL of water extraction liquor containing geniposide for standby.

[0123] (3) Resin adsorption and elution of geniposide:

[0124] Take 500 g of qualified AB-8 resin and load it onto the column with pure water; load the above 5100 mL of extraction solution onto the column at room temperature, and control the column flow rate at 1 - 3 BV / h;

[0125] After all the extraction solution is loaded onto the column, while keeping the flow rate unchanged, rinse the resin column bed with pure water until there is no amino acid color reaction in the effluent from the column bed. The pure water washing of the resin column is completed, and the pure water washing solution is discarded;

[0126] Then, elute the resin column with 1500 ml of 5 - 10% ethanol aqueous solution, control the elution flow rate at 1 - 3 BV / h, and elute to remove impurities; after the eluate is recovered and de-alcoholized, the residue is discarded;

[0127] Finally, elute the resin column with 2000 ml of 60 - 65% ethanol aqueous solution, control the elution flow rate at 1 - 3 BV / h, collect the eluate, and obtain the eluate containing gardenia fruit extract.

[0128] (4) Solvent removal and drying:

[0129] The eluate is concentrated and de-alcoholized by vacuum distillation to remove the solvent, obtaining a fluid extract;

[0130] The above fluid extract is vacuum dried until the water content is not more than 10%; the dried product is pulverized to obtain a solid powder, which is the gardenia fruit extract.

[0131] The yield of the gardenia fruit extract is 3.67%. After HPLC detection, the geniposide content in the gardenia fruit extract is 88.21%. The HPLC chromatogram of the gardenia fruit extract is shown in Figure 1 .

[0132] Specifically, the conditions for HPLC detection are:

[0133] (1) Preparation of geniposide standard curve:

[0134] Weigh approximately 0.1 g of geniposide standard (mass fraction ≥ 98%, accurate to 0.0001 g), dissolve it with the mobile phase and make up the volume to 100 mL to obtain the stock solution A of the standard sample; Pipette 0.5 mL, 1.0 mL, 2.0 mL, 4.0 mL, 5.0 mL, 8.0 mL, 10.0 mL of the stock solution A, dilute each with the mobile phase and make up the volume to 10 mL respectively to obtain 7 standard samples; Under the reference chromatographic conditions, measure the standard samples with gradient concentrations, repeat the experiment twice, and obtain the average peak area value of the standard samples; Use the peak area of the standard sample as the ordinate and the geniposide mass concentration (g / mL) of the standard sample as the abscissa to plot the standard curve;

[0135] (2) Preparation of the test solution:

[0136] Weigh an appropriate amount of the gardenia fruit extract sample (accurate to 0.0001 g), dissolve it with the mobile phase and make up the volume to 100 mL, filter the resulting solution through a 0.45 μm filter membrane, and reserve the filtrate;

[0137] (3) Determination:

[0138] Under the reference chromatographic conditions, measure the test solution, and identify it qualitatively according to the retention time of the geniposide standard; Inject the sample again, and obtain the average peak area value of geniposide; According to the linear relationship between the peak area of the standard sample and the geniposide mass concentration of the standard sample, obtain the geniposide mass concentration (g / mL) in the test solution; If the geniposide concentration (g / mL) in the test solution is not within the range of the standard curve, the concentration of the test solution should be adjusted or the standard curve should be redesigned;

[0139] (4) Result calculation:

[0140] The content X1 of geniposide in the sample is calculated according to the following formula:

[0141] In the formula:

[0142] X1 —— the content of geniposide in the sample, %;

[0143] c1 —— the geniposide concentration obtained from the standard curve, in grams per milliliter (g / mL);

[0144] c2 —— the concentration of the test solution, in grams per milliliter (g / mL).

[0145] The experimental results are based on the arithmetic mean of the parallel determination results. The absolute difference between the two independent determination results obtained under repeatability conditions is not greater than 0.1%;

[0146] In the conditions of this HPLC detection, the reference chromatographic conditions are as follows:

[0147] Chromatographic column: ODS C18, 4.6 mm × 15 cm, particle size 5 μm; mobile phase: acetonitrile∶water = 15∶85; mix 150 mL of chromatographically pure acetonitrile with 850 mL of water evenly, filter through a 0.45 μm filter membrane, and use it after ultrasonic degassing; column temperature: 40 °C; flow rate: 0.7 mL / min; injection volume: 10 μL.

[0148] In addition, HPLC-MS analysis was also performed on the Gardenia fruit extract to detect other trace component components besides geniposide. Through HPLC-MS analysis, in addition to a relatively high purity of geniposide, the Gardenia fruit extract also contains different contents of cellobiose, ferulic acid, genipin-1-glucoside, gardenic acid, sinapic acid, methyl ester of scandoside methyl ester, shanzhiside, genipin gentiobioside, isopropyl 4-hydroxybenzoate, dimethoxy-4-hydroxycinnamaldehyde, and physcion.

[0149] Example 2

[0150] This example uses a human primary dermal vascular endothelial cell model to detect the effect of the geniposide-containing extract (specifically the Gardenia fruit extract prepared in Example 1) on capillary angiogenesis-related target genes.

[0151] Human primary dermal vascular endothelial cells were cultured in an incubator at 37 °C, containing 5% CO2, and saturated humidity. After the cells grew to more than 80% confluence, they were digested and seeded in a 12-well plate and continued to be cultured for 48 h for testing. On the day of the experiment, appropriate sample concentrations were selected to treat the keratinocytes for 24 h, the supernatant was discarded, and each well was washed 2 times with 1 mL of pre-cooled phosphate buffer (PBS), and then cell lysates were prepared using RNA extraction reagents. After collecting the lysate samples, total cellular RNA was uniformly extracted, reverse transcribed into cDNA, and then the expression levels of angiogenesis-related genes were detected by real-time fluorescence quantitative PCR. The obtained results were analyzed and quantified by the 2 -ΔΔCt method to obtain the gene expression levels of each group of cells.

[0152] The test results are as follows:

[0153] After treating the cells with aqueous solutions of Gardenia fruit extract at three concentrations of 1 wt.%, 0.5 wt.%, and 0.1 wt.% for 24 h, the cells were collected to extract total RNA, and after reverse transcription, the expression levels of each gene were detected. As Figure 2 shown in Table 2, the aqueous solutions of Gardenia fruit extract at the three concentrations in this test all had an inhibitory effect on capillary angiogenesis-related target genes, and this effect showed an obvious dose-dependent relationship.

[0154] Table 2

[0155]

[0156]

[0157]

[0158]

[0159]

[0160] α-SMA gene: The gene expression levels after treatment with 1 wt.% and 0.5 wt.% Gardenia fruit extract samples were 18.46% and 34.25% respectively, and the inhibition rates were 81.54% (P<0.001) and 65.75% (P<0.001) respectively. The 0.1% Gardenia fruit extract had no significant effect on the gene expression.

[0161] PECAM1 gene: The gene expression levels after treatment with 1 wt.% and 0.5 wt.% Gardenia fruit extract samples were 39.41% and 54.95% respectively, and the inhibition rates were 60.59% (P<0.001) and 45.05% (P<0.01) respectively. The 0.1% Gardenia fruit extract had no significant effect on the gene expression.

[0162] eNOS gene: The gene expression level after treatment with 1 wt.% Gardenia fruit extract sample was 34.09%, and the inhibition rate was 65.91% (P<0.001). The 0.5 wt.% and 0.1 wt.% Gardenia fruit extracts had no significant effect on the gene expression.

[0163] COX-2 gene: The gene expression levels after treatment with 1 wt.% and 0.5 wt.% Gardenia fruit extract samples were 53.07% and 75.57% respectively, and the inhibition rates were 46.93% (P<0.01) and 24.43% (P<0.05) respectively. The 0.1% Gardenia fruit extract had no significant effect on the gene expression.

[0164] Through Figure 2 and the results in Table 2 show that Gardenia fruit extract can significantly inhibit the expression of angiogenesis-related genes in human primary dermal vascular endothelial cells.

[0165] Example 3

[0166] This example provides a cosmetic aqueous solution. The component contents of the cosmetic aqueous solution are shown in Table 3, and the preparation method of the cosmetic aqueous solution is as follows:

[0167] Disperse the Gardenia fruit extract in water and mix evenly to obtain the cosmetic aqueous solution.

[0168] Table 3 Component contents (wt.%) of the cosmetic aqueous solution

[0169]

[0170] Taking cosmetic aqueous solution A as an example, the effect of this cosmetic on erythematous skin was detected.

[0171] Test method:

[0172] There were 13 subjects, including 3 males and 10 females, with no smoking history and no coffee-drinking habit. They had not used any oral medications in the week before the test, were in good health, and had normal diets.

[0173] A total of two tests were conducted on the left arm and the right arm. The specific test steps were as follows:

[0174] The skin area for testing was the left forearm:

[0175] (1) First, apply cosmetic aqueous solution A to the skin test area, with deionized water as the blank control;

[0176] (2) After 30 minutes, stimulate the skin test area with 3% methyl nicotinate solution for 5 minutes to make the skin turn red;

[0177] (3) Observe the degree and duration of skin redness and take photos for recording.

[0178] The skin area for testing was the right forearm:

[0179] (1) First, stimulate the skin with 1.0% methyl nicotinate solution to make the skin turn red;

[0180] (2) After 10 minutes, apply cosmetic aqueous solution A to the skin redness area respectively, with deionized water as the blank control; apply it once every 15 minutes.

[0181] (3) Observe the time of skin redness and redness subsidence and take photos for recording.

[0182] The schematic diagram of the results of the left arm test is shown in Figure 3 ; the schematic diagram of the results of the right arm test is shown in Figure 4 .

[0183] The test results of the 13 subjects were similar. In the left arm test, after stimulating the skin test area with 3% methyl nicotinate solution, the skin at the corresponding positions of the subjects all turned red. After 60 minutes, the redness of the skin area treated with cosmetic aqueous solution A was significantly lighter than that of the skin area treated with deionized water, indicating that using cosmetics containing Gardenia fruit extract helps prevent skin redness; in the right arm test, after first stimulating the skin with 1.0% methyl nicotinate solution and then treating the corresponding skin area with cosmetic aqueous solution A or deionized water, it can be seen that after 120 minutes, the redness of the skin area treated with cosmetic aqueous solution A was significantly lighter than that of the skin area treated with deionized water, indicating that using cosmetics containing Gardenia fruit extract helps improve skin redness.

[0184] Methyl nicotinate's main function in cosmetics and skin care products is as a skin reddening agent and is commonly used in lip products. Methyl nicotinate's vasodilatory effect in the skin is mainly mediated through the prostaglandin pathway. Cyclooxygenase (COX-2) is a key enzyme in prostaglandin synthesis. Therefore, extracts containing Gardenia fruit may be similar to non-steroidal anti-inflammatory drugs (NSAIDs) and inhibit prostaglandin synthesis by inhibiting COX-2, reducing the paroxysmal pain and fever symptoms of erythematous skin caused by methyl nicotinate and achieving the effect of rapid redness reduction.

[0185] After testing, compared with deionized water, cosmetic aqueous solutions B, C, and D all have similar effects to cosmetic aqueous solution A in preventing and improving skin redness, but it takes longer for cosmetic aqueous solutions B and C to achieve the same efficacy as cosmetic aqueous solution A.

[0186] Example 4

[0187] This example provides a cosmetic gel. The component contents of the cosmetic gel are shown in Table 4. The preparation method of the cosmetic gel is as follows:

[0188] (1) Weigh deionized water, preservative HTsoft_5688, humectant isopentylene glycol, and thickener carbomer into a beaker in sequence, heat to above 80 °C, and keep warm for 20 minutes;

[0189] (2) Homogenize at a speed of 5000 rpm for 5 minutes. After fully dissolving arginine with a small amount of deionized water, add it to the homogenizing material and continue to homogenize for 5 minutes;

[0190] (3) Start stirring. After cooling to below 40 °C, add the dissolved Gardenia fruit extract and stir evenly to complete the preparation.

[0191] Table 4 Component contents of the cosmetic gel (wt.%)

[0192]

[0193]

[0194] Taking cosmetic gel A as an example, detect the effect of this cosmetic on dark circles.

[0195] Test method:

[0196] There were 16 subjects, including 11 females and 5 males, with obvious dark circles under the eyes. After simple tests, they were found to have vascular dark circles and a mixed type of vascular / pigment deposition dark circles. The method for judging the type of dark circles is as follows: Use a transparent glass slide and gently press on the area with dark circles. If the black or bluish-purple color of the dark circles fades away and reveals the original skin color, it is a blood stasis type of dark circles; if it fades by half and a little brown remains, it belongs to the mixed type of vascular dark circles and pigment deposition dark circles.

[0197] The skin area tested was around the eyes. The test sample was cosmetic gel A, and no other eye care products were used during the test period.

[0198] The specific method is as follows:

[0199] (1) Twice a day, one pump each time, evenly apply it to the skin around the eyes;

[0200] (2) Continuously use it for 28 days;

[0201] (3) Observe the skin condition around the eyes and take photos for record;

[0202] (4) There were no situations such as business trips or staying up late during the test period. Ensure to observe and take photos in the same room, with the same light and the same angle.

[0203] The test results of the 16 subjects were similar. The schematic diagram of the results of this example is shown in Figure 5 .

[0204] It can be seen that with the use of cosmetic gel A, the dark circles of the subjects were significantly alleviated.

[0205] Vascular dark circles, also known as blue circles under the eyes, are caused by insufficient microcirculation in the skin around the eyes. The skin around the eyes is relatively thin, only about 1 / 4 of the skin thickness of other parts of the face, and there are very rich capillary and venous blood vessels around the eyes. When there is neovascularization of capillaries in the area around the eyes, it will cause the oxygen content of hemoglobin in red blood cells accumulated in these capillaries to be relatively low, so the color is bluish or purple, forming the color of dark circles. Cosmetic gel A containing gardenia fruit extract can effectively improve vascular dark circles caused by neovascularization of capillaries.

[0206] After testing, cosmetic gel B, cosmetic gel C, and cosmetic gel D all have similar effects of improving dark circles as cosmetic gel A, but it takes longer for cosmetic gel B and cosmetic gel C to achieve the same efficacy as cosmetic gel A.

[0207] Example 5

[0208] This example provides a cosmetic lotion. The component contents of the cosmetic lotion are shown in Table 5. The preparation method of the cosmetic lotion is as follows:

[0209] (1) Weigh deionized water, stabilizer (chelating agent EDTA-2Na), preservative HTsoft_5688, humectant and thickener into a beaker in sequence as the aqueous phase, and weigh emulsifier and emollient into another beaker as the oil phase. Heat the materials in both beakers to above 80 °C and keep warm for 20 minutes.

[0210] (2) Take out the aqueous phase and homogenize it at a speed of 5000 rpm for 5 minutes. Add the heated and well-mixed oil phase, continue to homogenize and emulsify for 5 minutes, and then add the dissolved antioxidant sodium metabisulfite and homogenize for another 2 minutes.

[0211] (3) Start stirring. After cooling to below 40 °C, add the dissolved Gardenia fruit extract and stir evenly to complete the preparation.

[0212] Table 5 Component contents (wt.%) of cosmetic emulsion

[0213]

[0214] Take cosmetic emulsion A as an example to detect the effect of this cosmetic on telangiectasia.

[0215] Use the Coloemeter probe of CK Company in Germany to detect the reflected light on the skin surface structure. In the L*a*b* colorimetric system, L* represents black and white degree, a* represents red and green degree, and b* represents blue and yellow degree. The comprehensive evaluation of the three is used to reflect the change of skin color. Express the color change in the form of a quantitative value, which can accurately reflect the change of skin erythema, so as to intuitively reflect the effect of the product in soothing telangiectasia.

[0216] Test method:

[0217] The subjects are 5 females with obvious facial flushing. They did not use other skin-soothing and redness-reducing skin care products and oral medications within one week before the test, are in good health, and have normal diet.

[0218] The specific test method is as follows:

[0219] (1) The subjects clean their faces with the designated cleansing product and sit still in the laboratory at room temperature of 21 ± 1 °C and 50 ± 10% RH for 30 min.

[0220] (2) Test the basic a value and basic skin temperature of the left and right sides of the subjects' faces respectively, and take photos for record. The test sites on both sides of the face are kept consistent.

[0221] (3) Adopt the double-blind method. Apply cosmetic emulsion A on one side of the face and the control group emulsion on the other half of the face.

[0222] (4) After sitting still for 30 min, test the a value and skin temperature and take photos for record.

[0223] The average results of the a values and skin temperatures of 5 subjects are shown in Table 6. The schematic diagram of the results of this example is shown in Figure 6 .

[0224] Table 6

[0225]

[0226] According to Table 6 and Figure 6 the test results, it can be seen that the skin of the experimental group using cosmetic lotion A has a lower a value, a lower skin temperature, less skin redness, and a better effect of soothing telangiectasia after 30 minutes. The cosmetic lotion A containing Gardenia fruit extract can effectively improve telangiectasia caused by neovascularization of capillaries.

[0227] After testing, cosmetic lotion B, cosmetic lotion C, and cosmetic lotion D all have a similar effect of soothing telangiectasia to cosmetic lotion A, but it takes longer for cosmetic lotion B and cosmetic lotion C to achieve the same efficacy as cosmetic lotion A.

[0228] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. Use of a Gardenia fruit extract containing geniposide in the preparation of a product for non-therapeutic improvement of skin redness caused by telangiectasia, capillary dilation or dark circles under the eyes, wherein the content of geniposide in the Gardenia fruit extract containing geniposide is ≥ 80 wt.%.

2. The application according to claim 1, wherein The content of geniposide in the Gardenia fruit extract containing geniposide is ≥ 98 wt.%.

3. The application according to claim 1, wherein The preparation method of the Gardenia fruit extract comprises the following steps: S1. Crush the Gardenia fruit, perform water extraction after sieving, and after solid-liquid separation, the obtained liquid is the extract; S2. Adsorb the extract obtained in step S1 using resin, and then elute with a solvent to obtain an eluate; S3. Concentrate the eluate obtained in step S2, and then dry it to obtain the Gardenia fruit extract.

4. The application according to claim 3, characterized in that, In step S2, the solvent is water and / or an alcohol solvent; the resin is a macroporous adsorption resin.

5. The application according to claim 4, wherein The macroporous adsorption resin is a non-polar resin or a weakly polar resin.

6. The application according to claim 3, characterized in that, In step S2, the adsorption method is as follows: Take a qualified resin column washed with pure water, load the column with pure water, and then load the extract onto the column at room temperature, controlling the column flow rate at 1 - 3 BV / h; after the extract loading is completed, keep the column flow rate unchanged, wash the resin column bed with pure water until the amino acid color reaction is not detected in the column effluent, and discard the pure water washing solution after the pure water washing of the resin column is completed; The elution method is as follows: Elute the resin column with 1500 ml of an ethanol aqueous solution with a concentration of 5 - 10%, control the elution flow rate at 1 - 3 BV / h, and elute to remove impurities; after the 10% ethanol eluate is recovered and de-alcoholized, discard the residual liquid; then elute the resin column with 2000 ml of an ethanol aqueous solution with a concentration of 60 - 65%, control the elution flow rate at 1 - 3 BV / h, collect the eluate, and obtain the eluate containing the Gardenia fruit extract.

7. Use of a Gardenia fruit extract containing geniposide in the preparation of a cosmetic for non-therapeutic improvement of skin redness caused by telangiectasia, capillary dilation or dark circles under the eyes, wherein the content of geniposide in the Gardenia fruit extract containing geniposide is ≥ 80 wt.%.

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