Dendrobium officinale exosome-like vesicle composition, preparation method and application

Abstract

The invention relates to the technical field of cosmetics, and provides a dendrobium officinale exosome-like vesicle composition which comprises dendrobium officinale exosome-like vesicles, dendrobium officinale polysaccharide and hyaluronic acid, and the weight ratio of the dendrobium officinale exosome-like vesicles to the dendrobium officinale polysaccharide to the hyaluronic acid is (0.01-1): (0.5-10): (0.5-10). Or the exosome-like vesicle composition of the dendrobium officinale is prepared from the exosome-like vesicles of the dendrobium officinale, dendrobium officinale polysaccharide and sodium hyaluronate, and the weight ratio of the exosome-like vesicles of the dendrobium officinale to the dendrobium officinale polysaccharide to the sodium hyaluronate is (0.01-1): (0.5-10): (0.5-10); the three components are compounded and have a synergistic effect, so that the moisturizing and synergistic effects can be achieved, the skin quality can be improved, and the moisturizing and synergistic lotion is good in mildness, safe and non-irritant.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, and in particular to an exosome-like vesicle composition of Dendrobium officinale, a preparation method and application thereof. Background Art

[0002] Dendrobium candidum is a plant of the genus Dendrobium in the orchid family. It mainly contains active ingredients such as polysaccharides, flavonoids, phenols, lignin, stilbenes and alkaloids, and has anti-aging, anti-inflammatory and antibacterial, antioxidant, lipid-lowering, blood sugar-lowering and blood pressure-lowering effects. Plant exosome vesicles have a certain membrane structure and contain a variety of biological molecules, such as a variety of proteins, nucleic acids and other active small molecules derived from plants themselves. They have the biological properties of plants and are safe and non-toxic, and have great application potential in the field of cosmetics.

[0003] In the existing technology, Dendrobium officinale extracts are usually used as cosmetic raw materials, but there is a lack of research on the application of exosome-like vesicles of Dendrobium officinale as active ingredients in the cosmetic field. However, the extracts are generally single-category extracts, such as polysaccharides, flavonoids, phenols and other active ingredients, and their stability, safety and efficacy in cosmetic formulas still need to be further improved. Summary of the invention

[0004] One of the purposes of the present invention is to provide an exosome-like vesicle composition of Dendrobium officinale, aiming to provide a composition with moisturizing and synergistic effects, good stability, and safety and non-irritation.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] A Dendrobium officinale exosome-like vesicle composition, comprising:

[0007] Exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and hyaluronic acid, wherein the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the hyaluronic acid is (0.01-1): (0.5-10): (0.5-10);

[0008] Or, the exosome-like vesicle composition of Dendrobium officinale includes exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and sodium hyaluronate, and the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the sodium hyaluronate is (0.01-1): (0.5-10): (0.5-10).

[0009] Preferably, the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the hyaluronic acid is (0.1-0.3): (5-7): (4.9-6.5);

[0010] Or, the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the sodium hyaluronate is (0.1-0.3):(5-7):(4.9-6.5).

[0011] Preferably, the weight ratio of the exosome-like vesicles of the Dendrobium officinale, the Dendrobium officinale polysaccharide and the hyaluronic acid is (0.01-1): (3-5): (1-4);

[0012] Or, the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the sodium hyaluronate is (0.01-1):(3-5):(1-4).

[0013] The second object of the present invention is to provide a preparation method, aiming to prepare an exosome-like vesicle composition of Dendrobium officinale.

[0014] To achieve the above object, the present invention adopts the following technical solutions:

[0015] A method for preparing an exosome-like vesicle composition of Dendrobium officinale comprises the following steps:

[0016] (1) Preparing an exosome-like vesicle suspension of Dendrobium officinale: washing, pulping, filtering, and collecting the filtrate of fresh Dendrobium officinale stems, centrifuging the filtrate, collecting the precipitate after centrifugation, and dissolving it with an appropriate amount of phosphate buffer to obtain an exosome-like vesicle suspension of Dendrobium officinale;

[0017] (2) Preparation of Dendrobium officinale polysaccharide: fresh stems of Dendrobium officinale are washed, pulped, defatted, filtered, dried, water extracted, filtered, concentrated, precipitated with alcohol, allowed to stand, centrifuged, washed, and the precipitate is collected, and the precipitate is redissolved and freeze-dried to obtain Dendrobium officinale polysaccharide;

[0018] (3) Preparing a composition: dissolving Dendrobium officinale polysaccharide and hyaluronic acid in a suspension of exosome-like vesicles of Dendrobium officinale, or dissolving Dendrobium officinale polysaccharide and sodium hyaluronate in a suspension of exosome-like vesicles of Dendrobium officinale, adding water and stirring until completely dissolved, and then freeze-drying to obtain a Dendrobium officinale exosome-like vesicle composition.

[0019] Preferably, in step (1), the same mass of dilute phosphate buffer and crushed and juiced Dendrobium officinale stems are used for pulping;

[0020] The filtrate is centrifuged for the first time at 4°C and 1500-2500g for 20-30min to collect the first supernatant; the first supernatant is centrifuged for the second time at 4°C and 3000-12000g for 30-60min to collect the second supernatant; the second supernatant is centrifuged for the third time at 4°C and 100000-150000g for 60-90min to collect the first precipitate; the first precipitate is suspended with dilute phosphate buffer, and then centrifuged for the fourth time at 4°C and 100000-150000g for 60-120min to collect the second precipitate, and the second precipitate is dissolved with an appropriate amount of dilute phosphate buffer to obtain an exosome-like vesicle suspension of Dendrobium officinale.

[0021] Preferably, in step (2), 95% ethanol and the washed stems of Dendrobium officinale are crushed and juiced in a ratio of 2:1 for pulping; 80% to 95% ethanol is used for heating and degreasing twice, and the heating temperature is 60 to 80°C; the filter residue is filtered and collected, and then dried; 20 times the amount of distilled water is added to the dried filter residue, and water extraction is performed at 60 to 100°C, the number of extractions is 1 to 3, and the time of each extraction is 100 to 150 minutes, and the water extract is collected; the water extract is concentrated to 1 / 10 of the extract, and 4 times the amount of 75 to 95% ethanol is added to the concentrate for alcohol precipitation; the centrifugal treatment speed is 10000r / min, 15min; the precipitate is collected, and washed twice, with 1 to 2 times the volume of the precipitate and 75 to 95% ethanol.

[0022] The third object of the present invention is to provide a skin external preparation, aiming to achieve the effect of moisturizing and enhancing synergy.

[0023] To achieve the above object, the present invention adopts the following technical solutions:

[0024] A skin topical preparation having the function of moisturizing and enhancing efficacy, comprising the above-mentioned exosome-like vesicle composition of Dendrobium officinale;

[0025] The amount of the exosome-like vesicle composition of Dendrobium officinale added to the external skin preparation is 0.0001 to 20% by weight.

[0026] Preferably, the skin topical preparation is a face cream, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of cetearyl olive oil ester, and 10 parts of exosome-like vesicle composition of Dendrobium officinale;

[0027] Or, the facial cream comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of sorbitan olivate, and 10 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0028] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0029] Preferably, the skin topical preparation is an emulsion, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of cetearyl olivetate, and 6 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0030] Or, the emulsion comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of sorbitan olivate, and 6 parts of exosome-like vesicle composition of Dendrobium officinale;

[0031] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0032] Preferably, the skin topical preparation is an essence, which comprises, by weight, 3 parts of 1,3-butanediol, 5 parts of glycerol, 72 parts of deionized water, and 20 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0033] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0034] After adopting the above technical solution, the present invention has the following advantages compared with the background technology:

[0035] 1. The exosome-like vesicle composition of Dendrobium officinale of the present invention is a compound of exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharides and hyaluronic acid, or exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharides and sodium hyaluronate, which work synergistically to play a moisturizing and synergistic role, improve skin quality, and have good mildness, safety and non-irritation.

[0036] 2. The preparation method of the present invention can effectively protect the active ingredients from being destroyed by precisely controlling parameters such as temperature and time during the preparation process, and improve the efficiency and purity of the separation of exosome-like vesicles of Dendrobium officinale through multiple centrifugation to maximize its effect.

[0037] 3. The skin external preparation prepared by the present invention is safe and mild, and can provide long-lasting hydration to the skin after use, thereby maintaining the moisturizing property of the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The radar chart of skin feel test of the composition of Example 2, exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and hyaluronic acid described in Test Example 1 of the present invention;

[0039] Figure 2 It is a radar chart of skin feel test of the composition of Example 2, Comparative Example 1 and Comparative Example 2 described in Test Example 1 of the present invention;

[0040] Figure 3 This is the investigation result of the trial test described in Test Example 4 of the present invention. DETAILED DESCRIPTION

[0041] For ease of understanding of the present invention, the present invention will be described more fully below. Although the present application provides preferred embodiments of the present invention, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive. Where specific techniques or conditions are not indicated in the embodiments, the technical conditions described in the documents in this area or the product specifications are carried out.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0043] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0044] In the present invention, when it comes to numerical ranges, unless otherwise specified, the above numerical ranges are deemed to be continuous and include the minimum and maximum values ​​of the range, as well as each value between such minimum and maximum values. Further, when a range refers to an integer, each integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0045] Only some numerical ranges are specifically disclosed herein. However, any lower limit can be combined with any upper limit to form an unspecified range; and any lower limit can be combined with other lower limits to form an unspecified range, and likewise any upper limit can be combined with any other upper limit to form an unspecified range. In addition, each separately disclosed point or single value can itself be combined as a lower limit or upper limit with any other point or single value or with other lower limits or upper limits to form an unspecified range.

[0046] The temperature parameters in the present invention, if not specifically limited, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows the temperature to fluctuate within the precision range controlled by the instrument.

[0047] In the description of the invention, “plurality” means at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0048] If there is no special description, all embodiments and optional embodiments of the present invention can be combined with each other to form a new technical solution. If there is no special description, all technical features and optional technical features of the present invention can be combined with each other to form a new technical solution.

[0049] If not specifically stated, all steps of the present invention may be performed sequentially or randomly, but are preferably performed sequentially.

[0050] The reagents and instruments used without indicating the manufacturer are all conventional products obtained from the market.

[0051] Embodiment 1

[0052] This embodiment provides a Dendrobium officinale exosome-like vesicle composition, comprising:

[0053] The weight ratio of exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharides and hyaluronic acid is 0.1:5:4.9.

[0054] The preparation method comprises the following steps:

[0055] (1) Preparation of exosome-like vesicle suspension of Dendrobium officinale: fresh stems of Dendrobium officinale were washed, crushed and juiced with an equal mass of dilute phosphate buffer and the stems of Dendrobium officinale for pulping, filtered and the filtrate was collected, and then the filtrate was centrifuged for the first time at 4°C and 2500g for 30 min to collect the first supernatant; the first supernatant was centrifuged for the second time at 4°C and 12000g for 60 min to collect the second supernatant; the second supernatant was centrifuged for the second time at 4°C and 150000g for The first precipitate was collected by centrifugation for 90 minutes three times; the first precipitate was suspended with a dilute phosphate buffer, and then centrifuged for 120 minutes at 4°C and 150,000 g for a fourth time to collect the second precipitate, and the second precipitate was dissolved with an appropriate amount of dilute phosphate buffer (it should be noted that the appropriate amount means that the added dilute phosphate buffer can completely dissolve the second precipitate, and the dilute phosphate buffer can be gradually added until the second precipitate is observed to be completely dissolved), so as to obtain a suspension of exosome-like vesicles of Dendrobium officinale;

[0056] (2) Preparation of Dendrobium officinale polysaccharide: fresh stems of Dendrobium officinale are washed; 95% ethanol and the washed stems of Dendrobium officinale are crushed and juiced in a ratio of 2:1 for pulping; 80% ethanol is used for heating and degreasing twice at a heating temperature of 60°C; the residue is then filtered and collected; the residue is dried; 20 times the amount of distilled water is added to the dried residue, and water extraction is performed at 80°C for 2 times, each extraction time is 120 minutes, and the water extract is collected; the water extract is filtered and concentrated to 1 / 10 of the water extract to obtain a concentrated solution; 4 times the amount of 95% ethanol is added to the concentrated solution for alcohol precipitation; after standing, the concentrated solution is centrifuged at a speed of 10000 r / min for 15 minutes, and the precipitate is collected; then the concentrated solution is washed twice with 95% ethanol in an amount of 1 times the volume of the precipitate, the precipitate is collected, and the precipitate is redissolved and freeze-dried to obtain Dendrobium officinale polysaccharide;

[0057] (3) Preparing a composition: Dissolving Dendrobium officinale polysaccharide and hyaluronic acid in a suspension of exosome-like vesicles of Dendrobium officinale, adding water and stirring until completely dissolved, and then freeze-drying to obtain a Dendrobium officinale exosome-like vesicle composition.

[0058] Also provided is a facial cream comprising the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of cetearyl olivate, and 10 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0059] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0060] Also provided is an emulsion containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of cetearyl olivate, and 6 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0061] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0062] Also provided is an essence containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butanediol, 5 parts of glycerol, 72 parts of deionized water, and 20 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0063] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0064] Embodiment 2

[0065] This embodiment provides a Dendrobium officinale exosome-like vesicle composition, comprising:

[0066] The weight ratio of exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharides and hyaluronic acid is 1:5:4.

[0067] The preparation method comprises the following steps:

[0068] (1) Preparation of exosome-like vesicle suspension of Dendrobium officinale: fresh stems of Dendrobium officinale were washed, and the same mass of dilute phosphate buffer was used to crush and squeeze the stems of Dendrobium officinale for pulping, and the filtrate was filtered and collected. The filtrate was then centrifuged for the first time at 4°C and 1500g for 25 min to collect the first supernatant; the first supernatant was centrifuged for the second time at 4°C and 10000g for 45 min to collect the second supernatant; the second supernatant was centrifuged for the second time at 4°C and 120000g for The first precipitate was collected by centrifugation three times for 90 minutes; the first precipitate was suspended with a dilute phosphate buffer, and then centrifuged for a fourth time at 4°C and 120,000 g for 120 minutes to collect the second precipitate, and the second precipitate was dissolved with an appropriate amount of dilute phosphate buffer (it should be noted that the appropriate amount means that the added dilute phosphate buffer can completely dissolve the second precipitate, and the dilute phosphate buffer can be gradually added until the second precipitate is observed to be completely dissolved), so as to obtain a suspension of exosome-like vesicles of Dendrobium officinale;

[0069] (2) Preparation of Dendrobium officinale polysaccharide: fresh stems of Dendrobium officinale are washed; 95% ethanol and the washed stems of Dendrobium officinale are crushed and juiced in a ratio of 2:1 for pulping; 95% ethanol is used for heating and degreasing twice at a heating temperature of 70°C; then the residue is filtered and collected; the residue is dried; 20 times the amount of distilled water is added to the dried residue, and water extraction is performed at 100°C, the number of extractions is 1 time, and the time of each extraction is 100 minutes, and the water extract is collected; the water extract is filtered and concentrated to 1 / 10 of the water extract to obtain a concentrated solution; 4 times the amount of 90% ethanol is added to the concentrated solution for alcohol precipitation; after standing, the concentrated solution is centrifuged at a speed of 10000 r / min for 15 minutes; after centrifugation, the precipitate is collected and washed twice with 90% ethanol of 1 times the volume of the precipitate, the precipitate is collected, and the precipitate is redissolved and freeze-dried to obtain Dendrobium officinale polysaccharide;

[0070] (3) Preparing a composition: Dissolving Dendrobium officinale polysaccharide and hyaluronic acid in a suspension of exosome-like vesicles of Dendrobium officinale, adding water and stirring until completely dissolved, and then freeze-drying to obtain a Dendrobium officinale exosome-like vesicle composition.

[0071] Also provided is a facial cream comprising the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of cetearyl olivate, and 10 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0072] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0073] Also provided is an emulsion containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of cetearyl olivate, and 6 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0074] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0075] Also provided is an essence containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butanediol, 5 parts of glycerol, 72 parts of deionized water, and 20 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0076] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0077] Embodiment 3

[0078] This embodiment provides a Dendrobium officinale exosome-like vesicle composition, comprising:

[0079] The weight ratio of exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharides and sodium hyaluronate is 0.01:0.5:0.5.

[0080] The preparation method comprises the following steps:

[0081] (1) Preparation of exosome-like vesicle suspension of Dendrobium officinale: fresh stems of Dendrobium officinale were washed, crushed and juiced with an equal mass of dilute phosphate buffer and the stems of Dendrobium officinale for pulping, filtered and the filtrate was collected, and then the filtrate was centrifuged for the first time at 4°C and 2000g for 20 min to collect the first supernatant; the first supernatant was centrifuged for the second time at 4°C and 6000g for 40 min to collect the second supernatant; the second supernatant was centrifuged for the second time at 4°C and 100000g for 40 min to collect the second supernatant; The first precipitate was collected by centrifugation three times for 60 minutes; the first precipitate was suspended with a dilute phosphate buffer, and then centrifuged for a fourth time at 4°C and 100,000 g for 60 minutes to collect the second precipitate, and the second precipitate was dissolved with an appropriate amount of dilute phosphate buffer (it should be noted that the appropriate amount means that the added dilute phosphate buffer can completely dissolve the second precipitate, and the dilute phosphate buffer can be gradually added until the second precipitate is observed to be completely dissolved), so as to obtain a suspension of exosome-like vesicles of Dendrobium officinale;

[0082] (2) Preparation of Dendrobium officinale polysaccharide: fresh stems of Dendrobium officinale are washed; 95% ethanol and the washed stems of Dendrobium officinale are crushed and juiced in a ratio of 2:1 for pulping; 85% ethanol is used for heating and degreasing twice at a heating temperature of 75°C; the residue is then filtered and collected; the residue is dried; 20 times the amount of distilled water is added to the dried residue, and water extraction is performed at 60°C for 3 times, each extraction time is 150 minutes, and the water extract is collected; the water extract is filtered and concentrated to 1 / 10 of the water extract to obtain a concentrated solution; 4 times the amount of 75% ethanol is added to the concentrated solution for alcohol precipitation; after standing, the concentrated solution is centrifuged at a speed of 10000 r / min for 15 minutes; after centrifugation, the precipitate is collected and washed twice with 80% ethanol of 1.5 times the volume of the precipitate, the precipitate is collected, and the precipitate is redissolved and freeze-dried to obtain Dendrobium officinale polysaccharide;

[0083] (3) Preparing a composition: Dissolving Dendrobium officinale polysaccharide and sodium hyaluronate in a suspension of exosome-like vesicles of Dendrobium officinale, adding water and stirring until completely dissolved, and then freeze-drying to obtain a Dendrobium officinale exosome-like vesicle composition.

[0084] Also provided is a facial cream comprising the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of sorbitan olivate, and 10 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0085] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0086] Also provided is an emulsion containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of sorbitan olivate, and 6 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0087] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0088] Also provided is an essence containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butanediol, 5 parts of glycerol, 72 parts of deionized water, and 20 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0089] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0090] Embodiment 4

[0091] This embodiment provides a Dendrobium officinale exosome-like vesicle composition, comprising:

[0092] The weight ratio of exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharides and sodium hyaluronate is 1:10:10.

[0093] The preparation method comprises the following steps:

[0094] (1) Preparation of exosome-like vesicle suspension of Dendrobium officinale: fresh stems of Dendrobium officinale were washed, crushed and juiced with an equal mass of dilute phosphate buffer and the stems of Dendrobium officinale for pulping, filtered and the filtrate was collected, and then the filtrate was centrifuged for the first time at 4°C and 1700g for 28 min to collect the first supernatant; the first supernatant was centrifuged for the second time at 4°C and 3000g for 30 min to collect the second supernatant; the second supernatant was centrifuged for the second time at 4°C and 120000g for The first precipitate was collected by centrifugation for 70 minutes three times; the first precipitate was suspended with dilute phosphate buffer, and then centrifuged for a fourth time at 4°C and 110,000 g for 90 minutes to collect the second precipitate, and the second precipitate was dissolved with an appropriate amount of dilute phosphate buffer (it should be noted that the appropriate amount means that the added dilute phosphate buffer can completely dissolve the second precipitate, and the dilute phosphate buffer can be gradually added until the second precipitate is observed to be completely dissolved), so as to obtain an exosome-like vesicle suspension of Dendrobium officinale;

[0095] (2) Preparation of Dendrobium officinale polysaccharide: fresh stems of Dendrobium officinale are washed; 95% ethanol and the washed stems of Dendrobium officinale are crushed and juiced in a ratio of 2:1 for pulping; 90% ethanol is used for heating and degreasing twice at a heating temperature of 80°C; then the residue is filtered and collected; the residue is dried; 20 times the amount of distilled water is added to the dried residue, and water extraction is performed at 90°C for 2 times, each extraction time is 110 minutes, and the water extract is collected; the water extract is filtered and concentrated to 1 / 10 of the water extract to obtain a concentrated solution; 4 times the amount of 80% ethanol is added to the concentrated solution for alcohol precipitation; after standing, the concentrated solution is centrifuged at a speed of 10000 r / min for 15 minutes; after centrifugation, the precipitate is collected and washed twice with 75% ethanol in an amount of 2 times the volume of the precipitate, the precipitate is collected, the precipitate is redissolved, and freeze-dried to obtain Dendrobium officinale polysaccharide;

[0096] (3) Preparing a composition: Dissolving Dendrobium officinale polysaccharide and sodium hyaluronate in a suspension of exosome-like vesicles of Dendrobium officinale, adding water and stirring until completely dissolved, and then freeze-drying to obtain a Dendrobium officinale exosome-like vesicle composition.

[0097] Also provided is a facial cream comprising the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of sorbitan olivate, and 10 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0098] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0099] Also provided is an emulsion containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of sorbitan olivate, and 6 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0100] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0101] Also provided is an essence containing the above-mentioned exosome-like vesicle composition of Dendrobium officinale, which comprises, by weight, 3 parts of 1,3-butanediol, 5 parts of glycerol, 72 parts of deionized water, and 20 parts of the exosome-like vesicle composition of Dendrobium officinale;

[0102] The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

[0103] Comparative Example 1

[0104] This comparative example provides a composition, including Dendrobium officinale alcohol extract, Dendrobium officinale polysaccharide and hyaluronic acid, and the weight ratio of the Dendrobium officinale alcohol extract, Dendrobium officinale polysaccharide and hyaluronic acid is 1:5:4.

[0105] The preparation steps are: adding water to the dendrobium officinale alcohol extract, the dendrobium officinale polysaccharide and the hyaluronic acid, stirring until completely dissolved, and then freeze-drying to obtain the composition.

[0106] The preparation method of the alcohol extract of Dendrobium officinale is as follows: using the dried stems of Dendrobium officinale as raw materials, crushing and sieving; stirring and mixing 10 times the volume of 80% concentration ethanol with the sieved dried stems of Dendrobium officinale, heating at 60°C for 2 hours with ultrasonic-assisted extraction to obtain an extract; centrifuging and filtering the extract, collecting the filtrate, concentrating under reduced pressure, and freeze-drying to obtain the alcohol extract of Dendrobium officinale.

[0107] The preparation method of Dendrobium officinale polysaccharide is shown in Example 1.

[0108] Comparative Example 2

[0109] This comparative example provides a composition, including Dendrobium officinale polysaccharide and hyaluronic acid, and the weight ratio of Dendrobium officinale polysaccharide to hyaluronic acid is 6:4.

[0110] The preparation steps are as follows: adding water to the Dendrobium officinale polysaccharide and hyaluronic acid, stirring until complete, and then freeze-drying to obtain the product.

[0111] Wherein, the preparation method of Dendrobium officinale polysaccharide refers to Example 1.

[0112] Test Example 1

[0113] The present invention was subjected to a skin feel test, and the test samples were the exosome-like vesicle composition of Dendrobium officinale in Example 2, the composition of Comparative Example 1, the composition of Comparative Example 2, the exosome-like vesicle of Dendrobium officinale, Dendrobium officinale polysaccharide and hyaluronic acid. Among them, the exosome-like vesicle of Dendrobium officinale and Dendrobium officinale polysaccharide were prepared according to the preparation method of Example 2.

[0114] The above test samples are all prepared as aqueous solutions with a concentration of 1%.

[0115] The number of volunteers was 20, and 6 test areas of 3cm×3cm were randomly drawn on the arms of each volunteer, and each test area corresponded to a different sample to be tested. The skin feeling after the trial was scored, with a total of five items, each with a maximum score of 5 points. The test results are as follows Figure 1 and Figure 2 As shown, Figure 1 and Figure 2 The comparative composition 1 indicated in the table is the composition of comparative example 1, and the comparative composition 2 is the composition of comparative example 2.

[0116] According to the test results, compared with Comparative Example 1, Comparative Example 2 and the use of each component alone, the exosome-like vesicle composition of Dendrobium officinale of Example 2 of the present invention shows better effects in terms of viscosity, moisturizing degree, absorption speed and skin softness.

[0117] Test Example 2

[0118] The present invention was tested for moisturizing efficacy, and the composition of Example 1, the composition of Example 2, the composition of Example 3, the composition of Example 4, the composition of Comparative Example 1, the composition of Comparative Example 2, the exosome-like vesicles of Dendrobium officinale, the polysaccharide of Dendrobium officinale and hyaluronic acid were respectively prepared into an aqueous solution with a concentration of 1%. The exosome-like vesicles of Dendrobium officinale and the polysaccharide of Dendrobium officinale were prepared according to the preparation method of Example 2.

[0119] The samples to be tested were placed at a temperature of 25°C and a relative humidity of 43% (in a dryer filled with a saturated solution of NaCO3 (full name in Chinese: sodium carbonate)) for one set of tests; the samples to be tested were placed at a temperature of 25°C and a relative humidity of 81% (in a dryer filled with a saturated solution of (NH4)2SO4 (full name in Chinese: ammonium sulfate)) for two sets of tests.

[0120] The samples were taken out after 48 hours and weighed, and repeated three times. The moisture retention rate was calculated according to the following formula: moisture retention rate (%) = final weight of water in the sample / initial weight of water in the sample × 100%. The test results are shown in Table 1.

[0121] Table 1 Moisture retention test results

[0122]

[0123] According to the test results in Table 1, the moisturizing rates of Examples 1, 2, 3 and 4 of the present invention are all higher than those of other test samples, confirming that the exosome-like vesicle composition of Dendrobium officinale of the present invention has a better moisturizing effect. In addition, the moisturizing effects of Examples 1, 2, 3 and 4 are significantly better than those of the exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and hyaluronic acid, indicating that the formula of the exosome-like vesicle composition of Dendrobium officinale of the present invention has a synergistic effect on the moisturizing effect.

[0124] Test Example 3

[0125] The test was conducted in accordance with the "QB / T 4256-2011 Guidelines for Evaluation of Moisturizing Efficacy of Cosmetics", and the test samples were the composition of Example 1, the composition of Example 2, the composition of Example 3, the composition of Example 4, the composition of Comparative Example 1, the composition of Comparative Example 2, the exosome-like vesicles of Dendrobium officinale, the polysaccharide of Dendrobium officinale and hyaluronic acid, which were prepared into a 1% aqueous solution. The exosome-like vesicles of Dendrobium officinale and the polysaccharide of Dendrobium officinale were prepared according to the preparation method of Example 2.

[0126] The test environment temperature was 25±2°C, and the relative humidity was 40%-60%. The number of volunteers was 10. The Corneometer probe of the multifunctional skin tester of the German CK company was used to measure the skin moisture content.

[0127] Each volunteer washed his arms and dried them uniformly, and 10 test areas of 3cm×3cm were randomly drawn on the arms of each volunteer, and each test area corresponded to a different sample to be tested or a blank. After the volunteers sat quietly in a constant temperature and humidity environment for 30 minutes, each area was randomly marked with a serial number (1, 2, 3, 4, 5, 6, 7, 8, 9, 10), and the moisture content of each area was tested as the initial value (i.e., the measured value at 0h). Subsequently, each volunteer's No. 2 marked area was smeared with 18 μL of the sample to be tested in Example 1, the No. 3 marked area was smeared with 18 μL of the sample to be tested in Example 2, the No. 4 marked area was smeared with 18 μL of the sample to be tested in Example 3, the No. 5 marked area was smeared with 18 μL of the sample to be tested in Example 4, the No. 6 marked area was smeared with 18 μL of the sample to be tested in Comparative Example 1, the No. 7 marked area was smeared with 18 μL of the sample to be tested in Comparative Example 2, the No. 8 marked area was smeared with 18 μL of the sample to be tested of the exosome-like vesicles of Dendrobium officinale, the No. 9 marked area was smeared with 18 μL of the sample to be tested of Dendrobium officinale polysaccharide, and the No. 10 marked area was smeared with 18 μL of the sample to be tested of hyaluronic acid. The skin moisture content of 10 areas of each volunteer was measured at 1h, 2h and 4h after smearing. The test results are shown in Table 2.

[0128] Table 2 Moisture content test

[0129]

[0130]

[0131] According to the test results in Table 2, the exosome-like vesicle compositions of Dendrobium officinale of Examples 1, 2, 3 and 4 of the present invention all have good moisturizing effects, and the moisture content is significantly increased after 4 hours of use. In addition, the moisturizing effect of the exosome-like vesicle composition of Dendrobium officinale of the present invention is significantly better than that of the exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and hyaluronic acid used alone, which can effectively improve the moisturizing effect, significantly improve the skin barrier, and improve the skin's water-locking ability.

[0132] Test Example 4

[0133] The facial cream of Example 2 of the present invention was used as a test sample for a trial test. This test required the test subjects to use recent daily cosmetics normally, but stop using facial cream, and only use the facial cream test sample of Example 2 of the present invention, once in the morning and evening every day, for 28 consecutive days. The number of subjects was 35. Questionnaires were conducted on the 0th day, the 14th day and the 28th day, respectively, and data were collected and analyzed for efficacy based on the questionnaire.

[0134] The experimental results are as follows Figure 3 As shown, the testers' feedback approval is high, indicating that the facial cream prepared by the present invention can effectively achieve moisturizing effect, has good mildness, is safe and non-irritating.

[0135] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A composition of exosome-like vesicles of Dendrobium officinale, characterized in that: include: Exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and hyaluronic acid, wherein the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the hyaluronic acid is (0.01-1): (0.5-10): (0.5-10); Or, the exosome-like vesicle composition of Dendrobium officinale includes exosome-like vesicles of Dendrobium officinale, Dendrobium officinale polysaccharide and sodium hyaluronate, and the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the sodium hyaluronate is (0.01-1): (0.5-10): (0.5-10).

2. The exosome-like vesicle composition of Dendrobium officinale according to claim 1, characterized in that: The weight ratio of the exosome-like vesicles of the Dendrobium officinale, the Dendrobium officinale polysaccharide and the hyaluronic acid is (0.1-0.3): (5-7): (4.9-6.5); Or, the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the sodium hyaluronate is (0.1-0.3):(5-7):(4.9-6.5).

3. The exosome-like vesicle composition of Dendrobium officinale according to claim 1, characterized in that: The weight ratio of the exosome-like vesicles of the Dendrobium officinale, the Dendrobium officinale polysaccharide and the hyaluronic acid is (0.01-1): (3-5): (1-4); Or, the weight ratio of the exosome-like vesicles of Dendrobium officinale, the Dendrobium officinale polysaccharide and the sodium hyaluronate is (0.01-1):(3-5):(1-4).

4. A method for preparing an exosome-like vesicle composition of Dendrobium officinale, characterized in that: The following steps are involved: (1) Preparing an exosome-like vesicle suspension of Dendrobium officinale: washing, pulping, filtering, and collecting the filtrate of fresh Dendrobium officinale stems, centrifuging the filtrate, collecting the precipitate after centrifugation, and dissolving it with an appropriate amount of phosphate buffer to obtain an exosome-like vesicle suspension of Dendrobium officinale; (2) Preparation of Dendrobium officinale polysaccharide: fresh stems of Dendrobium officinale are washed, pulped, defatted, filtered, dried, water extracted, filtered, concentrated, precipitated with alcohol, allowed to stand, centrifuged, washed, and the precipitate is collected, and the precipitate is redissolved and freeze-dried to obtain Dendrobium officinale polysaccharide; (3) Preparing a composition: dissolving Dendrobium officinale polysaccharide and hyaluronic acid in a suspension of exosome-like vesicles of Dendrobium officinale, or dissolving Dendrobium officinale polysaccharide and sodium hyaluronate in a suspension of exosome-like vesicles of Dendrobium officinale, adding water and stirring until completely dissolved, and then freeze-drying to obtain a Dendrobium officinale exosome-like vesicle composition.

5. The method for preparing the exosome-like vesicle composition of Dendrobium officinale according to claim 4, characterized in that: In step (1), equal weight of dilute phosphate buffer and crushed and juiced Dendrobium officinale stems are used for pulping; The filtered juice was centrifuged for the first time at 4°C, 1500-2500g for 20-30 min, and the first supernatant was collected; Centrifuge the first supernatant for a second time at 4°C, 3000-12000g for 30-60 min, and collect the second supernatant; The second supernatant was centrifuged for a third time at 4°C, 100,000-150,000 g for 60-90 min to collect the first precipitate; After suspending the first precipitate with dilute phosphate buffer, centrifugation is performed for the fourth time at 4°C, 100,000-150,000g for 60-120 min, the second precipitate is collected, and the second precipitate is dissolved with an appropriate amount of dilute phosphate buffer to obtain an exosome-like vesicle suspension of Dendrobium officinale.

6. The method for preparing the exosome-like vesicle composition of Dendrobium officinale according to claim 4, characterized in that: In step (2), 95% ethanol and the washed stems of Dendrobium officinale are crushed and juiced in a ratio of 2:1 for pulping; 80% to 95% ethanol is used for heating and degreasing twice, and the heating temperature is 60 to 80° C.; the filter residue is filtered and collected, and then dried; 20 times the amount of distilled water is added to the dried filter residue, and water extraction is performed at 60 to 100° C., the number of extractions is 1 to 3, and the time of each extraction is 100 to 150 minutes, and the water extract is collected; the water extract is concentrated to 1 / 10 of the water extract, and 4 times the amount of 75 to 95% ethanol is added to the concentrate for alcohol precipitation; the speed of the centrifugal treatment is 10000 r / min, and the extraction time is 15 minutes; The precipitate was collected and washed twice with 75-95% ethanol in an amount of 1-2 times the volume of the precipitate.

7. A skin external preparation having the function of moisturizing and enhancing the effect, characterized in that: An exosome-like vesicle composition comprising the Dendrobium officinale according to any one of claims 1 to 3; The amount of the exosome-like vesicle composition of Dendrobium officinale added to the external skin preparation is 0.0001 to 20% by weight.

8. The skin external preparation according to claim 7, characterized in that: The skin topical preparation is a face cream, which includes, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of cetearyl olivate, and 10 parts of exosome-like vesicle composition of Dendrobium officinale; Or, the facial cream comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 67.5 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of sorbitan olivate, and 10 parts of the exosome-like vesicle composition of Dendrobium officinale; The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

9. The external skin preparation according to claim 7, characterized in that The skin topical preparation is an emulsion, which includes, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of cetearyl olivetate, and 6 parts of exosome-like vesicle composition of Dendrobium officinale; Or, the emulsion comprises, by weight, 3 parts of 1,3-butylene glycol, 6 parts of glycerol, 72.8 parts of deionized water, 0.2 parts of xanthan gum, 5 parts of caprylic / capric triglyceride, 4 parts of dicaprylyl carbonate, 3 parts of sorbitan olivate, and 6 parts of exosome-like vesicle composition of Dendrobium officinale; The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

10. The external skin preparation according to claim 7, characterized in that The skin topical preparation is an essence, which comprises, by weight, 3 parts of 1,3-butylene glycol, 5 parts of glycerol, 72 parts of deionized water, and 20 parts of the exosome-like vesicle composition of Dendrobium officinale; The exosome-like vesicle composition of Dendrobium officinale is an aqueous solution with a concentration of 10%.

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