Synergistic oral cosmetic composition containing vesicle ball polydeoxyribonucleotide and exosome and application thereof

Abstract

The invention discloses a synergistic oral cosmetic composition containing vesicle globule polydeoxyribonucleotide and exosome and application of the oral cosmetic composition, and belongs to the technical field of oral cosmetic drinks. The composition contains a bioactive delivery system, a plant source exosome and a plant aqueous extract, and realizes bioavailability, multi-target effect and sustainable breakthrough through ternary synergy of the delivery system, the exosome and the plant aqueous extract. And the anti-aging, whitening and repairing pathways are clear. In the preparation method, the blood orange exosome is used as a carrier to load the salmon essence powder, and UHT is used for sterilizing and retaining active ingredients; the integrity rate of the exosome is improved through physical crushing and buffer solution protection, and the stability of the composition is ensured by adjusting the sugar degree and the pH value. According to the oral beauty product, various components are organically combined, the beauty effect in multiple aspects can be achieved, the oral beauty product is more comprehensive compared with a traditional product, and functional food is promoted to be upgraded to cell-level intervention.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of oral cosmetic drinks, and particularly relates to a synergistic oral cosmetic composition containing vesicle sphere polydeoxyribonucleotide and exosome and application thereof. BACKGROUND

[0002] With the increasing attention of consumers on the "inside-out" beauty way, oral cosmetic drinks have become an important development direction of the functional food and beauty industry. Oral cosmetic drinks improve skin health (such as anti-aging, whitening, and moisturizing) by supplementing active ingredients such as collagen, antioxidants, and plant extracts. However, these active ingredients are destroyed by gastric acid and digestive enzymes, and their molecular weight is limited: large molecular weight active ingredients are difficult to pass through the intestinal barrier into the blood circulation, greatly reducing their bioavailability; and external factors such as light, temperature, and pH can affect the stability of active ingredients and drinks, leading to oxidation, discoloration, precipitation, or failure. Therefore, traditional oral cosmetic drinks still face problems such as low bioavailability, poor stability, poor taste, and single efficacy. Therefore, there is an urgent need to develop a new type of oral cosmetic composition that can effectively improve the bioavailability and stability of active ingredients and achieve the synergistic effect of multiple efficacies, thereby meeting the needs of consumers for high-efficiency, stable, and market-competitive oral cosmetic products.

[0003] In recent years, innovative solutions such as nanodelivery technology, biologically active carriers (such as exosomes and liposomes), and intestinal-targeted release technology have been introduced to improve the absorption rate and targeting of oral cosmetic ingredients. Polydeoxyribonucleotide (PDRN) is a specific DNA fragment extracted from the reproductive cells of salmon or the seminal vesicles of salmon, which has a similarity of 98% to human DNA bases and has biological activities such as promoting cell regeneration and accelerating wound repair. As a DNA repair agent, PDRN is easily inactivated at room temperature and needs to be stored at low temperature; after oral administration, it is easily degraded by gastric acid and digestive enzymes, and it is difficult to directly reach skin cells, resulting in a bioavailability of less than 10%. In response to this, current methods mainly use packaging technology: such as liposomes or polymer nanoparticles, which can partially protect PDRN, but the stability and targeting are still insufficient.

[0004] Bainuxi vesicle sphere polydeoxyribonucleotide repair drink only relies on the DNA repair function of PDRN, lacks synergistic regulation of skin barrier (such as filaggrin FLG expression) and inflammatory pathways (such as NF-κB phosphorylation), and does not consider the circadian rhythm characteristics of skin cell repair (the golden repair period is from 22:00 to 02:00), resulting in a 3-fold diurnal variation in bioavailability with a fixed dose regimen, and a "stability-bioavailability-safety" dilemma. SUMMARY

[0005] In view of the low bioavailability, poor stability, poor taste and single efficacy of the active ingredients of the oral beauty drink in the prior art, the present application aims to provide an oral beauty composition containing synergistically enhanced vesicle sphere polydeoxyribonucleotide and exosome and application thereof.

[0006] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions to achieve the above-mentioned purpose. The present application provides an oral beauty composition containing synergistically enhanced vesicle sphere polydeoxyribonucleotide (PDNR) and exosome, which is composed of vesicle sphere polydeoxyribonucleotide (PDNR), plant-derived exosome and plant extract; the plant-derived exosome is derived from any one or combination of grape, mangosteen and pomegranate plants; and the plant extract is any one or combination of white kidney bean, sakura pollen and roselle powder water extract.

[0007] The mass ratio of the vesicle sphere polydeoxyribonucleotide (PDNR), plant-derived exosome and plant extract is 1: (0.5-2): (3-5).

[0008] The oral beauty composition is composed of grape exosome, mangosteen exosome, pomegranate exosome, vesicle sphere polydeoxyribonucleotide (PDNR), white kidney bean water extract, sakura pollen, roselle powder and purified water in a mass ratio of (5-7): (3-5): (4-6): (5-9): (1-2): (1-1.5): (1.5-2): (68-79).

[0009] Further, the oral beauty composition is composed of grape exosome, mangosteen exosome, pomegranate exosome, vesicle sphere polydeoxyribonucleotide (PDNR), white kidney bean water extract, sakura pollen, roselle powder and purified water in a mass ratio of 7:5:6:9:1.5:1.2:1.8:68.5.

[0010] The present application provides the application of the above-mentioned oral beauty composition containing synergistically enhanced vesicle sphere polydeoxyribonucleotide (PDNR) and exosome in the preparation of oral beauty products.

[0011] The present application provides an anti-aging functional drink containing synergistically enhanced vesicle sphere polydeoxyribonucleotide (PDNR) and exosome, which comprises the above-mentioned oral beauty composition containing synergistically enhanced vesicle sphere polydeoxyribonucleotide (PDNR) and exosome.

[0012] It also includes food acceptable adjuvants, which include at least one of flavoring agents, coloring agents, excipients and other food additives.

[0013] The application provides a preparation method of the synergistically enhanced anti-aging functional drink containing vesicle sphere polydeoxyribonucleotide (PDNR) and exosomes, and comprises the following steps: The vesicle sphere polydeoxyribonucleotide (PDNR), plant source exosomes and plant water extract are mixed to obtain a synergistically enhanced oral cosmetic composition containing the vesicle sphere polydeoxyribonucleotide (PDNR) and the exosomes; and the sugar content and the pH value are adjusted to prepare the synergistically enhanced anti-aging functional drink containing the vesicle sphere polydeoxyribonucleotide (PDNR) and the exosomes.

[0014] The preparation of the vesicle sphere polydeoxyribonucleotide (PDNR) comprises the following steps: taking blood orange exosomes as delivery carriers, uniformly mixing the blood orange exosomes with herring roe, ultrasonic incubation, filtration, homogenization, UHT sterilization, and obtaining the vesicle sphere polydeoxyribonucleotide (PDNR).

[0015] Further, the mass ratio of the blood orange exosomes and the herring roe is 23-25:1; the ultrasonic incubation power is 300-500 W, and the ultrasonic incubation time is 20-30 min; the UHT sterilization temperature is 110-120 DEG C, and the sterilization time is 10-15 s.

[0016] The preparation of the plant source exosomes comprises the following steps: physically crushing a pretreated plant, mixing the crushed plant with a buffer to obtain a plant homogenate, and centrifuging and freeze-drying the plant homogenate, wherein the plant is any one of grape, mangosteen, pomegranate and blood orange; and the buffer is any one of phosphate buffer, citrate buffer and acetate buffer.

[0017] Further, the mass-volume ratio of the crushed plant and the buffer is 1:1.

[0018] The sugar content is adjusted to 1.3 Brix-1.5 Brix, and the pH value is adjusted to 4.9-5.0.

[0019] Compared with the prior art, the application has the following beneficial effects: The application provides a synergistic oral cosmetic composition containing vesicle ball PDNR and exosome, which significantly improves the bioavailability of active ingredients through an "exosome + vesicle ball polydeoxyribonucleotide (PDNR)" synergistic delivery system. The collagen peptide and the like wrapped by biomimetic liposomes / polymer micelles are easily damaged in the gastric acid (pH 1-2) and digestive enzyme environment, and will degrade prematurely, reducing its efficacy. The vesicle ball PDNR has an exosome outer cabin, is more resistant to gastric acid and digestive enzyme damage, and the exosome has the characteristics of a cell membrane, can precisely target and bind to receptor cells, and improve the bioavailability of the active ingredient PDRN in the cabin; grape, mangosteen and pomegranate exosomes are rich in bioactive molecules such as polyphenols, flavonoids and microRNA, and their phospholipid bilayer structure can protect active ingredients from damage in the gastrointestinal tract and directly penetrate the cell membrane to release the contents. The plant extract and the exosome form a complementary mechanism of "exogenous supplementation + endogenous activation". Anti-aging pathway: the vesicle ball PDNR activates mitochondrial energy metabolism, the exosome microRNA regulates extracellular matrix synthesis, and the white kidney bean extract reduces the precursor of glycation end products; whitening pathway: the sakura pollen inhibits melanin transport, the roselle powder enhances keratin layer metabolism, and the grape exosome removes ultraviolet-induced free radicals; repair pathway: the pomegranate exosome blocks the release of inflammatory factors, the vesicle ball PDNR repairs DNA damage, and the white kidney bean extract regulates intestinal flora to improve skin microecology. The composition realizes a comprehensive breakthrough in bioavailability, multi-target efficacy and sustainability through the synergistic effect of the "delivery system + exosome + plant extract" ternary system. Its core value lies in converting the cell repair mechanism of regenerative medicine into a consumer-grade product, redefining the technical standards of oral cosmetic technology, and promoting the upgrading of functional foods to "cell-level intervention".

[0020] The application provided by the application realizes a multi-dimensional technical breakthrough by applying the composition to the preparation of an oral cosmetic product. The vesicle ball polydeoxyribonucleotide serves as a bioactive delivery system, and its unique vesicle structure can encapsulate active ingredients to resist gastric acid degradation and improve bioavailability through targeted release in the intestine. The synergistic effect of plant-derived exosomes and the delivery system not only enhances the transmembrane transport capacity of active ingredients, but also regulates skin cell metabolic rhythms through natural signaling molecules carried by exosomes. The introduction of plant water extracts provides auxiliary effects such as antioxidant and anti-inflammatory, and the natural ingredients and exosomes form a complementary physical and chemical property, which together improves the stability of the composition; by limiting the application scenarios of the three types of components to the field of oral cosmetic products, not only does it break through the limitations of traditional single components, but also dynamically regulates the circadian rhythm characteristics of skin repair, ultimately forming a technical solution with high efficiency, multiple effects and stability.

[0021] The oral cosmetic product provided by the present application contains vesicle ball PDNR and exosome synergies, compared with some oral cosmetic products that only focus on certain aspects of cosmetic efficacy, the oral cosmetic product of the present application organically combines three different types of ingredients, namely a biological active delivery system, plant-derived exosomes and plant extracts, to form a new combination mode, through the synergistic effect of multiple ingredients, the oral cosmetic product can simultaneously achieve antioxidant, anti-aging, metabolic regulation, improvement of skin state and other aspects of cosmetic efficacy, and has a more comprehensive cosmetic effect.

[0022] Further, the plant exosome is used to embed the polydeoxyribonucleotide (PDNR) in the vesicle ball, compared with the PDRN alone, the stability is improved, the phospholipid bilayer membrane structure of the plant exosome can protect the PDRN, so that the PDRN is more stable in the gastrointestinal tract and the like, and can tolerate degradation of artificial gastric juice for more than 6 hours. The absorption and utilization rate is improved, the plant exosome can be effectively taken up by small intestinal epithelial cells, can quickly penetrate the PDRN into the cell layer, and finally accurately reaches the action site through blood circulation, and the intestinal absorption rate of the PDRN alone is low; compared with the commonly used liposome-embedded PDRN on the market, the product 1) has better biocompatibility: the plant exosome is a natural vesicle secreted by cells, and has good affinity with human cells. The liposome as an artificially synthesized carrier is slightly inferior in biocompatibility; 2) has better targeting: the plant exosome has a natural phospholipid bilayer membrane structure, can deliver contents to specific cells, and has a safe and accurate carrying effect. Although the liposome also has a certain targeting, it usually needs to be modified to achieve better targeting effect; compared with the animal exosome-embedded PDRN, the immunogenicity is low: the plant exosome mainly contains metabolic components, and is not easy to cause immune response. The animal exosome contains animal-derived protein and other components, and has a relatively high risk of causing immune response; cost and production advantage: plants can be planted on a large scale, the raw material source of the plant exosome is rich, and it is easier to purify and mass-produce, has higher stability between batches, and lower cost. The animal exosome has strict requirements for storage conditions and is difficult to purify.

[0023] The preparation method provided by the application ensures the stability of the composition through step-by-step mixing, sugar content (1.3 Brix-1.5 Brix) and pH (4.9-5.0) adjustment. The bioactive delivery system is mixed with plant-derived exosomes and plant water extracts, and the active ingredients are protected from gastric acid degradation by the vesicular structure of the delivery system, while the combination of exosomes and plant extracts can activate multiple mechanisms of skin barrier repair and anti-inflammatory pathways; adjusting the sugar content to 1.3-1.5 Brix can control the osmotic pressure of the solution, both maintaining cell activity and improving taste palatability; adjusting the pH value to 4.9-5.0 of the weak acid environment not only meets the physiological conditions of the human digestive tract, but also inhibits microbial growth to prolong the shelf life; mixed with food additives for processing, through the addition of excipients and stabilizers, to ensure uniform dispersion and storage stability of active ingredients in powder, oral liquid and other dosage forms; the preparation process breaks through the technical bottleneck of easy inactivation and low absorption rate of active ingredients in traditional technology through three technical means of delivery carrier selection, formula proportion optimization and processing parameter control.

[0024] Further, with blood orange exosomes as the carrier, salmon sperm powder (PDRN precursor) is loaded into the exosome cavity through ultrasonic incubation (200W, 20-30min), and the loading rate is improved; UHT sterilization (110-120℃, 10-15s) kills microorganisms while retaining more than 90% of the active ingredients, solving the problem of efficacy loss caused by traditional high-temperature sterilization.

[0025] Further, through physical crushing (such as high-pressure homogenization) and buffer (phosphate / citrate / acetate) protection, the exosome integrity rate and polyphenol content loss rate are much better than traditional enzymatic methods; sugar content 1.3 Brix-1.5 Brix can inhibit microbial growth, pH 4.9-5.0 is close to the human gastrointestinal environment, reducing the precipitation of active ingredients, and increasing the activity retention rate of the composition during storage at room temperature. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 The figure is a comparison chart of the DPPH removal efficiency of the synergistic anti-aging functional drink containing vesicle balls PDRN and exosomes of different concentrations of the application and the comparative example. Figure 2 The figure is a comparison chart of the AGEs inhibition rate of the synergistic anti-aging functional drink containing vesicle balls PDRN and exosomes of different concentrations of the application and the comparative example. DETAILED DESCRIPTION

[0027] In order for those skilled in the art to better understand the technical scheme of the present application, the technical scheme in the embodiments of the present application will be clearly and completely described below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor should belong to the scope of protection of the present application.

[0028] The raw materials or chemical reagents used in the embodiments of the present application are obtained through conventional commercial channels unless otherwise specified.

[0029] Part of the raw material sources: Pomegranate, grape, mountain bamboo, blood orange: all are fresh raw materials collected within half a year; Salmon powder: Shaanxi Cailang Biotechnology Co., Ltd.; Guan Shan cherry pollen: Hansuoyuan (Shaanxi) Biotechnology Co., Ltd.; White kidney bean extract: Zhongbai Xingye Food Technology (Beijing) Co., Ltd.; Model: water-soluble WXLJ-4002; Rose kernel powder: Huangshan Huagulv Garden Biotechnology Co., Ltd.; Embodiment 1 The present embodiment provides a synergistic comprehensive anti-aging functional drink containing vesicle ball PDRN and exosomes, which comprises the following raw materials: grape exosomes, mountain bamboo exosomes, pomegranate exosomes, vesicle ball PDRN, white kidney bean extract, Guan Shan cherry pollen, rose kernel powder, and purified water. The mass ratio of the grape exosomes, mountain bamboo exosomes, pomegranate exosomes, vesicle ball PDRN, white kidney bean extract, Guan Shan cherry pollen, rose kernel powder, and purified water is 7:5:6:9:1.5:1.2:1.8:68.5.

[0030] Preparation steps of exosomes and vesicle ball PDRN: Preparation steps of exosomes: 1) 500 g of blood oranges were washed with purified water to remove surface impurities for use; 2) The plants were crushed by physical means, then 500 g of buffer physiological saline was added to the homogenizer, and the juice was squeezed for 15 min, and the plant homogenate was obtained; 3) The plant homogenate obtained in step 2) was transferred to a centrifuge container, centrifuged at 4 ℃ and 4500 r / min for 20 min, the residue was removed, and the upper liquid was obtained; 4) The upper liquid obtained in step 3) was centrifuged at 4 ℃ and 10000 r / min for 60 min, and the supernatant was taken as the plant exosome extract; The plants are grape, mangosteen, pomegranate, and blood orange, and the preparation method of the exosomes is the same. The preparation steps of the vesicle ball PDRN are as follows: 1) Take 240 g of blood orange exosomes into a dispensing tank.

[0031] 2) Take 10 g of salmon essence powder and add it to the dispensing tank.

[0032] 3) Transfer the blood orange exosomes and salmon essence powder in step 2) to a stirring tank and stir for 12 min to fully mix and promote interaction between them, obtaining a mixed solution.

[0033] 4) After ultrasonic incubation (200 W) of the mixed solution in the dispensing tank for 25 min, transfer it to the dispensing tank, filter it through an 80-mesh double steel mesh filter, and then transport it to a homogenizer for homogenization. The liquid is sterilized by UHT at a temperature of 115°C for 12 s. The sterilized liquid is transported to a temporary storage tank and is ready for filling.

[0034] The preparation steps of the synergistic comprehensive anti-aging drink containing vesicle ball PDRN and exosomes are as follows: (1) Mix 15 g of white kidney bean extract, 12 g of sakura powder, 18 g of rose bata powder, and 300 g of pure water to obtain a first mixed solution. (2) Mix 70 g of grape exosomes, 50 g of mangosteen exosomes, 60 g of pomegranate exosomes, 90 g of vesicle ball PDRN, and 200 g of pure water with the first mixed solution obtained in step (1) to obtain a second mixed solution. (3) Add water to the second mixed solution obtained in step (2) to make up to 1 kg, adjust the sugar content to 1.5 Brix, and the pH value to 5.01. After filtration (300 mesh), homogenization (30 MPa), and sterilization (90°C, 25 min), a comprehensive anti-aging drink containing vesicle ball PDRN and exosomes is obtained.

[0035] Example 2 The present embodiment provides a comprehensive anti-aging drink containing synergistic vesicle ball PDRN and exosomes, which comprises the following raw materials: grape exosomes, mangosteen exosomes, pomegranate exosomes, vesicle ball PDRN, white kidney bean extract, sakura powder, rose bata powder, and pure water. The mass ratio of the grape exosomes, mangosteen exosomes, pomegranate exosomes, vesicle ball PDRN, white kidney bean extract, sakura powder, rose bata powder, and pure water is 6:4:5:7:1.5:1.2:1.8:68.5.

[0036] The preparation steps of the exosomes and vesicle ball PDRN are as follows: The preparation steps of the exosomes are as follows: 1) Wash 500g of blood oranges with purified water to remove surface impurities for use; 2) Crush the plants using physical means, then put them into a homogenizer with 500g of buffer solution, and juice for 15 minutes to obtain plant homogenate; 3) Transfer the plant homogenate obtained in step 2) to a centrifuge container, centrifuge at 4°C, 4500r / min for 20min, remove the residue, and obtain the upper liquid; 4) Centrifuge the upper liquid obtained in step 3) at 4°C, 10000r / min for 60min, take the supernatant to obtain the plant exosome extract, and then freeze-dry to obtain the plant exosome extract; Wherein, the plants are grape, mountain bamboo, pomegranate and blood orange exosome preparation method is the same; Wherein, the buffer solution is physiological saline.

[0037] Preparation steps of vesicle ball PDRN: 1) Take 240g of blood orange exosome into a dispensing tank.

[0038] 2) Take 10g of codfish powder and add it to the dispensing tank of step 1).

[0039] 3) Transfer the blood orange exosome and codfish powder in step 2) to a stirring tank and stir for 12min.

[0040] 4) The liquid obtained in step 3) is used to ultrasonically incubate the material liquid in the dispensing tank for 25min.

[0041] 5) Transfer the liquid obtained in step 4) to a dispensing tank, filter the material liquid through an 80-mesh double steel mesh filter, and then deliver it to a homogenizer.

[0042] 6) The homogenized material liquid in step 5) is sterilized by UHT, the sterilization temperature is 115°C, the time is 12S, and the sterilized material liquid is delivered to a temporary storage tank for filling.

[0043] Preparation steps of comprehensive anti-aging drink with synergistic effect of vesicle ball PDRN and exosome: (1) Mix 15g of white kidney bean extract, 12g of Guanshan cherry flower powder, 18g of rose bata powder and 300g of purified water to obtain a first uniform mixture solution; (2) Mix the first uniform mixture solution obtained in step (1), 60g of grape exosome, 40g of mountain bamboo exosome, 50g of pomegranate exosome, 70g of vesicle ball PDRN and 200g of purified water to obtain a second uniform mixture solution; (3) Adding water to the second uniform mixed solution obtained in step (2) to 1 kg, and the sugar content is 1.4 Brix, and the PH is 4.95, then filtering, homogenizing, sterilizing to prepare the exosome vesicle ball PDRN synergistic comprehensive anti-aging drink.

[0044] Example 3 A synergistic comprehensive anti-aging drink containing vesicle ball PDRN and exosomes, comprising the following raw materials: grape exosomes, mangosteen exosomes, pomegranate exosomes, vesicle ball PDRN, white kidney bean extract, sakura pollen, rose bata powder, purified water; The mass ratio of the grape exosomes, mangosteen exosomes, pomegranate exosomes, vesicle ball PDRN, white kidney bean extract, sakura pollen, rose bata powder, and purified water is 5:3:4:5:1.5:1.2:1.8:68.5.

[0045] Preparation steps of exosomes and vesicle ball PDRN: Preparation steps of exosomes: 1) Wash 500g of blood oranges with purified water to remove surface impurities for use; 2) Crush the plants by physical means, then put them into a homogenizer with 500g of buffer solution, and juice for 15min; 3) Transfer the plant homogenate obtained in step 2) to a centrifuge container, centrifuge at 4℃, 4500r / min for 20min, remove the residue, and obtain the upper liquid; 4) Centrifuge the upper liquid obtained in step 3) at 4℃, 10000r / min for 60min, take the supernatant to obtain the plant exosome extract, and then freeze-dry it; Among them, the plants are grape, mangosteen, pomegranate and blood orange exosome preparation method; Among them, the buffer solution is physiological saline.

[0046] Preparation steps of vesicle ball PDRN: 1) Take 240g of blood orange exosomes into a blending tank.

[0047] 2) Take 10g of codfish powder and add it to the blending tank in step 1).

[0048] 3) Transfer the blood orange exosomes and codfish powder in step 2) to a stirring tank and stir evenly for 12min.

[0049] 4) The liquid obtained in step 3) is used to ultrasonically incubate the contents of the blending tank for 25min.

[0050] 5) Transfer the liquid obtained in step 4) to the blending tank, filter the liquid through an 80-mesh double steel mesh filter, and then deliver it to a homogenizer.

[0051] 6) The homogenized liquid from step 5) is sterilized by UHT at a temperature of 115°C for 12 seconds. The sterilized liquid is then transferred to a temporary storage tank and filled for later use.

[0052] Preparation steps of a comprehensive anti-aging beverage with synergistic effects of vesicular PDRN and exosomes: (1) Mix 15g of white kidney bean extract, 12g of Guanshan cherry blossom powder, 18g of roselle powder and 300g of purified water to obtain a first uniform mixed solution; (2) The first homogeneous mixed solution obtained in step (1), 50g of grape exosomes, 30g of mangosteen exosomes, 40g of pomegranate exosomes, 50g of vesicle PDRN and 200g of purified water are mixed to obtain the second homogeneous mixed solution; (3) Add water to the second uniform mixed solution obtained in step (2) to make up to 1 kg, with a sugar content of 1.3 Brix and a pH of 4.84. Then filter, homogenize and sterilize to prepare the comprehensive anti-aging beverage with synergistic effect of exosome vesicle PDRN.

[0053] Comparative Example 1 Comparative Example 1 provides a comprehensive anti-aging functional drink containing PDRN and various fruit juices, including the following ingredients: grape juice, mangosteen juice, pomegranate juice, blood orange juice, PDRN, white kidney bean extract, Kanzan cherry blossom powder, hibiscus powder, and purified water; The mass ratio of grape juice, mangosteen juice, pomegranate juice, blood orange juice, PDRN, white kidney bean extract, Guanshan cherry blossom powder, hibiscus powder, and purified water is 6:4:5:6.72:0.28:1.5:1.2:1.8:68.5.

[0054] Fruit juice preparation steps: 1) Wash 500g of blood oranges with purified water to remove surface impurities and set aside; 2) The above-mentioned plants are crushed by physical means, and then put into a blender with 500g of purified water and juiced for 15 minutes to obtain plant homogenate for later use; Among them, the plants are grapes, mangosteens, pomegranates, and blood orange juice, which are prepared using the same method; Preparation steps for a combined PDRN and fruit juice anti-aging drink: (1) Mix 15g of white kidney bean extract, 12g of Guanshan cherry blossom powder, 18g of hibiscus powder and 300g of purified water to obtain the first mixed solution; (2) The first mixed solution obtained in step (1), 60g of grape juice, 40g of mangosteen juice, 50g of pomegranate juice, 67.2g of blood orange juice, 2.8g of PDRN and 200g of purified water are mixed to obtain the second mixed solution; (3) Adding water to the second mixed solution obtained in step (2) to make the volume 1 kg and the sugar content 1.4 Brix, and the pH 4.95, and then filtering, homogenizing and sterilizing to prepare the PDRN+fruit juice synergistic comprehensive anti-aging drink.

[0055] Comparative Example 2 Comparative Example 2 provides a black coffee extract and red ginseng powder combined preparation, including the following raw materials: coffee powder, taurine, white kidney bean extract, epigallocatechin gallate, red ginseng powder; The comprehensive anti-aging functional drink includes the following raw materials: white kidney bean extract, sakura powder, rose bataleur powder, and purified water. The mass ratio of the white kidney bean extract, sakura powder, rose bataleur powder, and purified water is 1.5:1.2:1.8:68.5.

[0056] The preparation steps of the comprehensive anti-aging drink are as follows: (1) Mixing 15 g of white kidney bean extract, 12 g of sakura powder, 18 g of rose bataleur powder, and 300 g of purified water to obtain a first uniform mixed solution; (2) Adding water to the uniform mixed solution obtained in step (1) to make the volume 1 kg, and the sugar content 1.0 Brix, and the pH 4.71, and then filtering, homogenizing and sterilizing to prepare the comprehensive anti-aging drink.

[0057] II. Product performance test (1) DPPH free radical scavenging performance The DPPH free radical scavenging performance of the synergistic comprehensive anti-aging functional drink containing vesicle ball PDRN and exosomes prepared in Examples 1-3, the PDRN fruit juice combined preparation prepared in Comparative Example 1, and the extract combined preparation prepared in Comparative Example 2 was tested, and the experiment was as follows: The preparation of the solution to be tested: the solution of the synergistic comprehensive anti-aging functional drink containing vesicle ball PDRN and exosomes prepared in Examples 1-3 (solvent: water, concentrations: 500 μg / mL, 250 μg / mL and 125 μg / mL), the solution of the PDRN fruit juice combined preparation prepared in Comparative Example 1 (solvent: water, concentrations: 500 μg / mL, 250 μg / mL and 125 μg / mL), the solution of the extract combined preparation prepared in Comparative Example 2 (solvent: water, concentrations: 500 μg / mL, 250 μg / mL and 125 μg / mL), and the positive control V C solution (solvent: physiological saline, concentrations: 500 μg / mL, 250 μg / mL and 125 μg / mL).

[0058] 1,1-Diphenyl-2-picrylhydrazy (DPPH) solution (solvent: anhydrous ethanol, concentration: 0.05 g / L) was configured.

[0059] The reagents were added in the 96-well plate according to Table 1.

[0060] Table 1: Reagent types added in the 96-well plate

[0061] The substances in the 96-well plate were shaken and mixed, and then placed at room temperature in the dark for 30 min. The absorbance at 517 nm was measured. The free radical scavenging rate of DPPH was calculated according to the following formula: P% = [1- (A s -A c ) / (A b -A 空 )] x 100%, wherein P% represents the free radical scavenging rate of DPPH, A s represents the absorbance of the mixture of the test solution and the DPPH solution; A c represents the absorbance of the mixture of the test solution and the anhydrous ethanol solution; A b represents the absorbance of the blank control well; and A 空 represents the absorbance of the blank well. The experimental results are shown in the accompanying Figure 1 and Table 2.

[0062] Table 2: DPPH free radical scavenging rate results (%)

[0063] From the data in the accompanying Figure 1 and Table 2, it can be seen that the free radical scavenging ability of Example 1, Example 2 and Example 3 is significantly enhanced compared with the positive control V C , and the scavenging ability of Example 1 is the strongest. This indicates that the comprehensive anti-aging functional drink containing vesicle ball PDRN and exosomes prepared by the present application has a significant advantage in antioxidant effect, and its effect is better than that of the common antioxidant V C . Compared with Examples 1-3, the free radical scavenging ability of Comparative Example 1 and Comparative Example 2 is weaker. This shows that the combination of vesicle ball PDRN and exosomes is superior to the PDRN juice combined preparation and the extract combined preparation in the antioxidant aspect in the comparative examples, further embodying the superiority of the synergistic effect of vesicle ball PDRN and exosomes. Within a certain concentration range, the higher the concentration of vesicle ball PDRN + exosomes, the stronger the free radical scavenging ability, indicating that the antioxidant ability is also stronger, and the human body aging will be slower.

[0064] The experiment proves that the comprehensive anti-aging functional drink prepared by the application has strong antioxidant capacity through DPPH free radical scavenging performance test, and the antioxidant effect is better than that of the product of the comparative example and the positive control V C Meanwhile, it shows concentration dependence. This provides strong experimental support for the application of the comprehensive anti-aging functional drink in the anti-aging field, indicating that it helps to delay human aging through antioxidant effect.

[0065] (2) AGEs inhibition rate performance The AGEs inhibition rate performance of the comprehensive anti-aging functional drink containing synergistically enhanced vesicle ball PDRN and exosomes prepared by the application examples 1-3, the PDRN fruit juice compound preparation prepared by the comparative example 1, and the extract compound preparation prepared by the comparative example 2 was tested. AGEs (Advanced Glycosylation End-products) is a group of highly active end products formed by non-enzymatic glycosylation reaction between amino groups of proteins, fatty acids or nucleic acids and aldehyde groups of reducing sugars. The structure has high heterogeneity. After the formation of AGEs, extensive cross-linking between protein molecules occurs, resulting in changes in protein structure, mechanical strength, solubility and coordination binding properties. In vivo glycosylation of various proteins can affect the body in many ways, such as increasing vascular permeability, thickening of vascular basement membrane and accumulation of extracellular matrix. In order to evaluate the effect of the sample on anti-glycosylation, we based on the BSA-fructose simulation reaction system to detect the generation of AGEs after sample treatment, to determine whether the sample has anti-glycosylation ability, and further to judge whether the sample can protect the protein structure by inhibiting the cross-linking of proteins during the glycosylation process.

[0066] Experimental reagents and consumables: PBS (Solaybao), fructose (Mcclin), 5% BSA (Solaybao), AG (Merck), 15 mL centrifuge tube (Thermo), black enzyme-labeled plate (Biuntian).

[0067] Experimental equipment: clean bench (Suzhou Antai, SW-CJ-1F), centrifuge (Thermo), 4℃ refrigerator (Midea), constant temperature incubator (Shanghai Licenbangxi Instrument Science and Technology Co., Ltd.), fluorescence enzyme-labeled instrument (Synergy H1).

[0068] The experimental grouping is shown in Table 3.

[0069] Table 3: Information of samples to be tested

[0070] The specific experimental steps are as follows: Establish a BSA-fructose simulated reaction system: prepare solutions with BSA concentration of 10 mg / mL, fructose concentration of 500 mmol / L, and aminoguanidine (AG) solution using phosphate buffer (200 mmol / L, pH 7.4); Add 300 μL of fructose solution, 600 μL of BSA solution, and the test samples with final reaction concentrations of 10% and 5% to 15 mL centrifuge tubes sequentially. Seal the centrifuge tubes and incubate at 80 ℃ in the dark for 6 h. Then, take 100 μL of each solution and transfer it to a black ELISA plate to determine the amount of AGEs generated. The experimental groups are as follows: control group (without inhibitor), experimental group (inhibitor is the test sample), positive control group (inhibitor is aminoguanidine (AG), solvent control group).

[0071] AGEs inhibition rate detection: The fluorescence intensity was detected using a fluorescence microplate reader at an excitation wavelength of 360 nm and an emission wavelength of 460 nm, with a gain of 50. The calculation formula was used. AGEs inhibition rate (%) = 1 - [(FL) c -FL s ) / FL b ]*100% Where: FL s The fluorescence intensity of the experimental group; FL b Fluorescence intensity of the blank group; FL c The fluorescence intensity is for the solvent control group; experimental results are shown in the appendix. Figure 2 As shown.

[0072] From the attached diagram Figure 2 Data shows that, compared with Comparative Examples 1 and 2, Examples 1, 2, and 3 exhibit significantly enhanced AGEs inhibition rates. This indicates that the combination formulation containing vesicular PDRN + exosomes prepared in this invention has a significant advantage in inhibiting AGEs formation, outperforming the products in the comparative examples. Among Examples 1, 2, and 3, Example 1 shows the most significant increase in AGEs inhibition rate. The results of the AGEs inhibition rate determination indicate that the combination formulation containing vesicular PDRN + exosomes prepared in this invention can enhance the AGEs inhibition rate under certain concentration conditions. Furthermore, the higher the concentration of vesicular PDRN + exosome fluid, the stronger the AGEs inhibition rate, indicating a stronger anti-glycation ability in the body and a better ability to delay aging.

[0073] The experiment proves that the comprehensive anti-aging functional drink prepared by the application has strong anti-glycosylation ability through detecting the inhibition rate of AGEs, and the effect is better than that of the product of the comparative example, and it also presents concentration dependence. This provides strong experimental support for the application of the comprehensive anti-aging functional drink in delaying aging, and shows that it helps the body to have stronger anti-glycosylation ability by inhibiting the generation of AGEs, thereby better delaying aging.

[0074] The above is only used to illustrate the technical idea of the application, and cannot be used to limit the protection scope of the application. Any modification made according to the technical idea of the application on the basis of the technical scheme falls within the protection scope of the claims of the application.

Claims

1. A synergistic oral cosmetic composition comprising a combination of cochleates and exosomes, characterized in that, The vesicle ball polydeoxyribonucleotide, plant-derived exosome and plant extract are composed of; the plant-derived exosome is derived from any one or combination of grape, mangosteen and pomegranate plants; and the plant extract is any one or combination of white kidney bean water extract, sakura pollen and roselle powder water extract.

2. A synergistic oral cosmetic composition containing niosomes and exosomes as claimed in claim 1, wherein, The mass ratio of the vesicle ball polydeoxyribonucleotide, plant-derived exosome and plant extract is 1: (0.5-2): (3-5).

3. A synergistic oral cosmetic composition containing niosomes and exosomes as claimed in claim 1, wherein, The oral cosmetic composition is composed of grape exosome, mangosteen exosome, pomegranate exosome, vesicle ball polydeoxyribonucleotide, white kidney bean water extract, sakura pollen, roselle powder and purified water in a mass ratio of (5-7): (3-5): (4-6): (5-9): (1-2): (1-1.5): (1.5-2): (68-79).

4. Use of the synergistically enhanced oral cosmetic composition containing vesicle ball polydeoxyribonucleotide and exosome according to any one of claims 1-3 in the preparation of an oral cosmetic product.

5. A synergistic anti-aging functional drink containing a combination of vesicle sphere polydeoxyribonucleotides and exosomes, characterized in that, The synergistically enhanced oral cosmetic composition containing vesicle ball polydeoxyribonucleotide and exosome according to any one of claims 1-3, and a food acceptable adjuvant, wherein the adjuvant comprises at least one of a flavoring agent, a coloring agent, an excipient and other food additives.

6. The method of claim 5, wherein the preparation of the synergistic anti-aging functional drink containing the vesicle sphere polydeoxyribonucleotide and the exosome is characterized by, The preparation method comprises the following steps: The vesicle ball polydeoxyribonucleotide, plant-derived exosome and plant extract are mixed to obtain the synergistically enhanced oral cosmetic composition containing vesicle ball polydeoxyribonucleotide and exosome; and the sugar content and pH value are adjusted to prepare the anti-aging functional drink containing synergistically enhanced vesicle ball polydeoxyribonucleotide and exosome.

7. The method of claim 6, wherein the preparation method of the synergistic anti-aging functional drink containing the exosome and the vesicle sphere polydeoxyribonucleotide is characterized by, The preparation of the vesicle ball polydeoxyribonucleotide comprises the following steps: taking blood orange exosome as a delivery carrier, uniformly mixing the blood orange exosome with herring sperm powder, ultrasonic incubation, filtration, homogenization, UHT sterilization to obtain the vesicle ball polydeoxyribonucleotide.

8. The method of claim 7, wherein the preparation method of the synergistic anti-aging functional drink containing the exosome and the cochleate polydeoxyribonucleotide is characterized by, The mass ratio of the blood orange exosome and the herring sperm powder is 23-25:1; the ultrasonic incubation power is 300-500 W, and the ultrasonic incubation time is 20-30 min; the UHT sterilization temperature is 110-120 °C, and the sterilization time is 10-15 s.

9. The method of claim 6, wherein the preparation method of the synergistic anti-aging functional drink containing the exosome and the vesicle sphere polydeoxyribonucleotide is characterized by, The preparation of the plant-derived exosome comprises the following steps: physically crushing the pretreated plant, mixing the plant with a buffer to obtain a plant homogenate; centrifuging the plant homogenate, and freeze-drying to obtain the plant-derived exosome; the plant is any one of grape, mangosteen, pomegranate and blood orange; and the buffer is any one of phosphate buffer, citrate buffer and acetate buffer.

10. The method of claim 6, wherein the preparation method of the synergistic anti-aging functional drink containing the exosome and the vesicle sphere polydeoxyribonucleotide is characterized by, The sugar content is adjusted to 1.3 Brix-1.5 Brix, and the pH value is adjusted to 4.9-5.0.

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