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CN-121971356-A - Multi-source stem cell composite extract freeze-dried powder and composition, preparation method and application thereof, and repair essence

CN121971356ACN 121971356 ACN121971356 ACN 121971356ACN-121971356-A

Abstract

The invention relates to the field of biotechnology and skin health products, and discloses multi-source stem cell composite extract freeze-dried powder, a composition, a preparation method and application thereof, and repair essence containing the multi-source stem cell composite extract freeze-dried powder. The composition comprises a compound extract, a lyoprotectant and a buffer solution, wherein the content of the lyoprotectant is 2-4 mg and the content of the buffer solution is 0.1-1 mL relative to the compound extract of 1 mg based on the total content of protein. The freeze-dried powder of the multi-source stem cell composite extract provided by the invention is prepared by compounding the pilose antler extract, the ginseng extract and the centella asiatica extract, and obviously improving the effects of skin repair, anti-inflammatory, anti-aging and barrier reconstruction through the cooperation of multiple paths of regeneration promotion, anti-inflammatory, anti-oxidation and anti-fibrosis.

Inventors

  • XU ZHONGHUA
  • DU WENJING

Assignees

  • 广州森野生物医药科技有限公司

Dates

Publication Date
20260505
Application Date
20260409

Claims (10)

  1. 1. A composition of freeze-dried powder of a multi-source stem cell composite extract is characterized by comprising the composite extract, a freeze-drying protective agent and a buffer solution, wherein the content of the freeze-drying protective agent is 2-4 mg and the content of the buffer solution is 0.1-1 mL relative to the composite extract of 1 mg based on the total protein content; The compound extract contains velvet antler extract, ginseng extract and centella asiatica extract, and the content mass ratio of the velvet antler extract, the ginseng extract and the centella asiatica extract is 1-5:0.5-3:1 based on the protein content; Based on the total mass of the lyoprotectant, the lyoprotectant comprises 37-72wt% of trehalose, 20-35wt% of mannitol, 5-15wt% of human serum albumin, 1-5wt% of polyvinylpyrrolidone, 1-3wt% of butylhydroxytoluene, 1-5wt% of reduced glutathione and/or sodium ascorbate.
  2. 2. The composition according to claim 1, wherein the content mass ratio of the velvet antler extract, the ginseng extract, and the centella asiatica extract is 1-3:0.5-2:1, based on the protein content; And/or the buffer solution is Tris-HCl buffer solution, and the pH value of the buffer solution is 7-7.5; and/or, the buffer solution contains 3-8wt% of trehalose based on the total mass of the buffer solution.
  3. 3. The composition of claim 1, wherein each of the deer antler extract, the ginseng extract, and the centella asiatica extract is independently prepared by a method comprising the steps of: (a) Inoculating stem cells or centella asiatica callus cells into a culture medium, and culturing in a constant temperature incubator to obtain a conditional culture medium, wherein the stem cells are deer antler mesenchymal stem cells or ginseng cambium stem cells; (b) Filtering the conditioned medium to obtain filtrate; (c) Carrying out tangential flow filtration and concentration on the filtrate to obtain concentrated solution, wherein an ultrafiltration membrane with the molecular weight cut-off of 100-500 kDa is adopted for the tangential flow filtration and concentration; (d) Subjecting the concentrate to size exclusion chromatography to obtain the extract, wherein the size exclusion chromatography is performed by agarose gel chromatography column.
  4. 4. A method for preparing a multi-source stem cell composite extract lyophilized powder, wherein the method is performed using the components of the composition of any one of claims 1-3, the method comprising: (1) Firstly mixing the composite extract with a buffer solution to obtain a material I; (2) Carrying out second mixing on the material I and a freeze-drying protective agent to obtain a material II; (3) And performing co-freeze-drying treatment on the material II to obtain the freeze-dried powder of the multi-source stem cell composite extract.
  5. 5. The method according to claim 4, wherein in the step (3), the specific operation of the co-lyophilization treatment comprises subjecting the material II to prefreezing, primary drying and secondary drying in this order to obtain the multi-source stem cell composite extract lyophilized powder; The pre-freezing condition comprises the steps of quick freezing by liquid nitrogen, cooling to the temperature of less than or equal to-80 ℃ at the cooling rate of more than 50 ℃ per min, and keeping the temperature at the temperature of less than or equal to-80 ℃ to be 1-3 h; The primary drying condition comprises that the temperature is between minus 35 ℃ and minus 25 ℃, the vacuum degree is between 80 and 120 mTorr, and the time is between 15 and 20 h; The secondary drying condition comprises heating to 20-30deg.C at a heating rate of 4-6deg.C/h, vacuum degree of <50 mTorr, and time of 8-12 h.
  6. 6. A multi-source stem cell composite extract lyophilized powder prepared by the method of claim 4 or 5.
  7. 7. The multi-source stem cell composite extract freeze-dried powder of claim 6, wherein the particle size of the multi-source stem cell composite extract freeze-dried powder after reconstitution is 50-120 nm, and the polydispersity index PDI is less than or equal to 0.25.
  8. 8. Use of the multi-source stem cell composite extract freeze-dried powder according to claim 6 or 7 for preparing skin repair, anti-inflammatory, anti-aging and skin barrier reconstruction products.
  9. 9. The repairing essence for repairing skin is characterized by comprising 0.1-5wt% of active core component, 5-20wt% of moisturizing matrix, 0.1-1wt% of preservative and 74-94.8wt% of water based on the total mass of the repairing essence; The active core component contains a redissolution stabilizer and the multi-source stem cell composite extract freeze-dried powder of claim 6, wherein the content of the multi-source stem cell composite extract freeze-dried powder is 0.5-1.5 mg relative to the redissolution stabilizer of 1 mL; the redissolution stabilizer comprises PBS buffer solution, L-glutamic acid and polysorbate 20, wherein the content of the L-glutamic acid is 7-22 g and the content of the polysorbate 20 is 1-10 g relative to the PBS buffer solution of 1L.
  10. 10. The repair essence according to claim 9, wherein the moisturizing matrix comprises sodium hyaluronate, glycerol and butanediol in a mass ratio of 1:4-8:8-12; and/or the preservative is methyl parahydroxybenzoate.

Description

Multi-source stem cell composite extract freeze-dried powder and composition, preparation method and application thereof, and repair essence Technical Field The invention relates to the field of biotechnology and skin health products, in particular to multi-source stem cell composite extract freeze-dried powder and a composition, a preparation method and application thereof, and repair essence. Background The active ingredient is the core of cosmetic and dermatological health products, the efficacy of which depends on the activity, stability and bioavailability of the ingredient. Traditional plant extracts (e.g. ginseng, centella asiatica) or animal derived components (e.g. deer antler extract) are active, but the following challenges are often faced in practical applications: (1) The efficacy singleness and mechanism are limited, namely, single active ingredients often aim at specific skin problems (such as moisturizing and antioxidation), face complex networks of multi-factor (inflammation, oxidative stress and fibrosis) interweaving in skin aging and injury repair, have limited single component action paths, and are difficult to realize systematic and multi-target cooperative regulation. (2) The component utilization efficiency is low, the traditional extract is mostly small molecules or crude extracts, and the problems of low transdermal absorption rate, poor targeting property, insufficient stability and the like exist, so that the efficacy performance of the final product is affected. (3) Simple compounding lacks a synergistic effect in that the active ingredients from different sources are physically mixed, possibly simply as a pile-up of ingredients. The components have differences in physical and chemical properties, and are easy to generate interaction, sedimentation or functional mutual interference after being mixed, so that a stable composite system with synergistic interaction capability can not be formed. (4) The preparation has the difficult stability that a plurality of active ingredients have fragile structures, and are easy to degrade and lose activity after long-term storage under liquid state. Repeated freeze thawing can further compromise its integrity. There is a lack of a formulation suitable for industrial production and commercialization that can maintain the stability and bioactivity of multi-source composite systems for a long period of time. Lyophilization is a common option, but conventional lyoprotectants have non-uniform protective effects on extracts of different sources and different properties. Therefore, there is an urgent need to develop a novel composition and a preparation method capable of integrating advantages of extracts from different sources, achieving stable coexistence and functional synergy through a nano preparation technology, and having excellent long-term stability. Disclosure of Invention The invention aims to solve the problems that in the prior art, when a multi-source extract compound is applied to repairing essence, the efficacy is single, the synergistic effect is poor, and multi-channel synergistic repairing is difficult to realize, and provides multi-source synergistic stem cell compound extract freeze-dried powder with the efficacy exceeding that of single or simple mixed extracts. In order to achieve the above object, a first aspect of the present invention provides a composition for freeze-dried powder of a multi-source stem cell complex extract, which comprises a complex extract, a lyoprotectant and a buffer solution, wherein the content of the lyoprotectant is 2-4 mg and the content of the buffer solution is 0.1-1 mL relative to the complex extract of 1 mg based on total protein content; The compound extract contains velvet antler extract, ginseng extract and centella asiatica extract, and the content mass ratio of the velvet antler extract, the ginseng extract and the centella asiatica extract is 1-5:0.5-3:1 based on the protein content; Based on the total mass of the lyoprotectant, the lyoprotectant comprises 37-72wt% of trehalose, 20-35wt% of mannitol, 5-15wt% of human serum albumin, 1-5wt% of polyvinylpyrrolidone, 1-3wt% of butylhydroxytoluene, 1-5wt% of reduced glutathione and/or sodium ascorbate. In a second aspect, the present invention provides a method for preparing a multi-source stem cell composite extract lyophilized powder, the method being carried out using the components of the composition of the first aspect, the method comprising: (1) Firstly mixing the composite extract with a buffer solution to obtain a material I; (2) Carrying out second mixing on the material I and a freeze-drying protective agent to obtain a material II; (3) And performing co-freeze-drying treatment on the material II to obtain the freeze-dried powder of the multi-source stem cell composite extract. The third aspect of the invention provides multi-source stem cell composite extract freeze-dried powder prepared by the method of the second aspect. The fourt