KR102249241B1 - Method for extracting polydeoxyribonucleotides and polynucleotides derived from algae with the effect of neovascularization and cell regeneration - Google Patents

Abstract

The present invention provides a method, which extracts polydeoxyribonucleotides (PDRN) from seaweed, and through a process of making a small molecule with a size of less than 50 kDa with excellent skin and tissue penetration rate, produces low-molecular PDRN. More specifically, without using substances such as phenol that are harmful to the human body, by using eco-friendly and safe materials, the method of the present invention can extract the low-molecular PDRN economically and safely by minimizing the use of organic solvents and processing processes.

Description

Method for extracting polydeoxyribonucleotides and polynucleotides derived from algae with the effect of neovascularization and cell regeneration}

The present invention extracts polydeoxyribonucleotides (PDRN) from marine ecological disturbing species and year-round, economical and eco-friendly seaweeds, and has a small molecular weight process with a size of less than 50 kDa with excellent skin and tissue penetration. This is a method of preparing PDRN. More specifically, it relates to a method of extracting low-molecular weight PDRN from seaweeds economically and safely by minimizing the use of organic solvents and processing processes by using environmentally friendly and safe materials without using substances such as phenol that are harmful to the human body.

Polydeoxyribonucleotide (PDRN) and polynucleotide (PN) have viscoelastic properties, which are polymeric properties, and exhibit tissue repair effects when injected intradermally. In other words, it becomes the main component of a new concept of tissue repair medical devices that can induce the regeneration of skin tissues rather than simple skin fillers by promoting the secretion of extracellular matrix such as collagen by proliferating and increasing the activity of fibroblasts that make up the skin tissue.

It is a cell growth activator. As a tissue regenerating material, it has a special effect on the regeneration of tissues in our body such as ligaments, tendons, and skin, and in alleviating inflammation. This is due to the fact that substances contained in very small amounts in human placenta, ethical issues, and pharmaceutical productivity are difficult, so PDRN in fish came out as an alternative.

PDRN and PN contain phosphoric acid and four bases adenine, guanine, thymine, and cytosine. The structure in which four bases, deoxyribose, and phosphoric acid are combined constitute a repeating unit to form a high molecular weight double-stranded polymer. In addition, four types of bases, adenine (A), guanine (G), cytosine (C), and uracil (U), as well as sugars and phosphoric acid become repeat units to form a single-stranded polymer (hereinafter, abbreviated as a gene). )to be.

These genetic materials such as DNA or RNA can be hydrolyzed and bound to proteins or distributed freely in all living organisms. In the free single-stranded RNA, the complementary portions form a hydrogen bond with each other to form a hairpin structure and partially form a double-stranded RNA.

The growth point sites of animal sperm, eggs, or plants are relatively higher than those of other tissues, and the contents of PDRN and PN are relatively higher than those of other tissues, and these genetic materials are divided into small molecules, polymers, and extraction methods. It is known that PDRN and PN form a complex with adenosine A2 Receptor (receptor) with safety secured low-molecular process, promote VEGF secretion, induce angiogenesis, and activate fibroblasts, osteoblasts, and cartilage cells.

Korean Patent Publication No. 10-2019-0139633 and Korean Patent No. 10-2031152 disclose a method for extracting PDRN, but the prior document discloses a method for extracting animal-derived pdialene. It is different from the present invention.

Currently, in the case of animal-derived PDRN, there is a possibility of incorporation of viruses derived from animal tissues and it is difficult to obtain continuous raw materials. Therefore, by using marine ecological disturbance species and seaweeds that are economical and eco-friendly that can be produced year-round, we aim to produce functional low-molecular PDRNs that can secure both safety and stability by solving the above.

The present invention relates to a method for extracting PDRN from laver (inclusively algae) and a low molecular weight process, and in detail, such as phenol from laver (inclusively algae) or marine ecological disturbance species It is intended to provide a method of extracting PDRN, which is economically cost-saving, by extracting PDRN in a minimal process without using toxic substances.

In addition, it provides an eco-friendly method and a method of extracting and purifying genetic material such as PDRN (polydeoxyribonucleotide) or PN (polynucleotide) having a cellular activity effect from an environmentally friendly raw material called algae. In particular, the present invention separates genetic material such as PDRN and PN from all tissues other than specific tissues of seaweed (inclusively seaweed), and a mixture of small molecule PDRN and PN that activates adenosine A2 receptor from the secured seaweed-derived gene mixture. Provides a method of manufacturing.

Korean Patent Publication No. 10-2019-0139633 discloses a method for extracting high-efficiency PDRN from semen and testis of salmon family fish without the use of organic solvents such as phenol in a process and cost-effective manner. . In Korean Patent No. 10-2031152, a salmon-derived polydeoxyribonucleotide (PDRN, polydeoxyribonucleotide)-polymer complex, and salmon cartilage extract are hydrolyzed to treat salmon cartilage enzyme decomposition products and PDRN core-shell nanocapsules containing salmon oil It relates to a method for producing a functional cosmetic composition for improving skin and a cosmetic composition prepared according thereto. Accordingly, skin containing core-shell nanocapsules containing polydioxyribonucleotides capable of improving whitening, wrinkles, and pore improvement effects by protecting the ingredients of salmon's skin improvement ingredients to prevent degeneration and to penetrate into the dermis of the skin. Disclosed is an improved functional cosmetic composition and a method of manufacturing the same. Korean Patent Publication No. 10-2018-0060693 relates to a cosmetic composition with improved skin wrinkle removal and elasticity improvement effects, and includes a culture medium obtained by culturing perivascular stem cells derived from humans and polydeoxyribonucleotides. Disclosed is a cosmetic composition having improved skin wrinkle removal and elasticity improvement effects, which have the advantage of inhibiting the creation of skin wrinkles, removing generated wrinkles, and improving skin elasticity.

Jong-Guk Yoon and 4 others, Effect of Low Molecularized Polydeoxynucleotide (PDRN) on Surgical Wound Healing in Rats, J. Soc. Cosmet. Sci. Korea, Vol. 41, No. 4, December 2015, 401-411

In order to solve the risk of virus mixing in the existing salmon and trout (animal raw materials) and restrictions on the supply and demand of raw materials, etc., a comprehensive range of seaweeds and marine ecological disturbances such as Maesengi, maizean, kelp, seaweed, etc. A task is to provide a method for extracting genetic material having a molecular weight of between 50kDa and 20000kDa by adding adlay, green tea extract, soybean fatty acid, tocopherol cocamidopropyl betain, and Olive oil carboxylate.

As mentioned above, it does not use harmful substances such as phenol and sodium hydroxide, which are previously used to separate genetic material derived from tissues containing the growth point of natural seaweed that is safe from the risk of viral infection, not of animal origin, and at the same time, the manufacturing process and low molecular weight. We intend to provide a method of manufacturing PDRN and PN with secured price competitiveness by simplifying the process.

The method of extracting polydeoxyribonucleotides and polynucleotides derived from seaweeds having angiogenesis and cell regeneration effects according to an embodiment of the present invention includes the steps of securing raw materials for drying and pulverizing seaweeds (a); To prepare a mixed solution consisting of 10% by weight of soybean extract, 10% by weight of adlay extract, 20% by weight of green tea extract, 20% by weight of soybean fatty acid, 20% by weight of tocopherol cocamidopropyl betain, and 20% by weight of Olive oil carboxylate And, after dissolving the protein by adding it to the dried and pulverized seaweed raw material through step (a), adding a ribonuclease to separate a mixture of polynucleotides and polydeoxyribonucleotides. And decomposition step (b); An impurity removal and pure separation step of purely separating polydioxyribonucleotides and polynucleotides by centrifuging the mixture after step (b) to remove impurities, filtering, and drying (c); A washing and drying step (d) of washing and drying the polydeoxyribonucleotide and polynucleotide purely separated through the (c) step; The resultant dried in step (D) was diluted with 1 ml of 1% sodium purified water, treated with fermented acetic acid for 10 minutes, followed by sonication treatment for 10 minutes, and centrifuged to perform low-molecular polydeoxyribonucleotides and polynucleotides. The low-molecular process step (e) to secure the; It may consist of a quality inspection step (f) of measuring the DNA absorbance of the resultant obtained through the step (E), confirming and pulverizing 99% low-molecular polydeoxyribonucleotides and polynucleotides having a purity of 1.7 to 2.0.

The mixed solution prepared in step (b) may be added by 40 ml per 1 g of the weight of the seaweed raw material in step (a), and ribonuclease may be added by 0.1 ml per 1 g of the weight of the seaweed raw material in step (a). . In addition, the seaweed may be one or more selected from Maesengi, Hatban, kelp, seaweed, and laver.

In the present invention, safety is secured because eco-friendly seaweed or laver with smooth raw material supply and demand is used, and eco-friendly raw materials are used instead of harmful substances in the process. In addition, since physical methods are used in the extraction and low molecular weight processes, the conventional method More economical and effective PDRN can be used.

1 shows a schematic diagram of a method for extracting low-molecular polydeoxyribonucleotides and polynucleotides using seaweed.
2 shows the electrophoresis results according to Experimental Example 1 of the present invention.
Figure 3 shows the results of in vitro new angiogenesis experiments for the low-molecular-processed PDRN derived from seaweed (Maesengi, hatban, kelp, seaweed, seaweed, etc.), the existing salmon-derived polymer PDRN, and the control.
Figure 4 shows the results of in vitro cell regeneration experiments for the low-molecular-processed PDRN and the existing salmon-derived polymer PDRN, and a control derived from seaweed (meal saengi, hatban, kelp, seaweed, seaweed, etc.)

The terms referred to in the present invention, polydeoxyribonucleotide (PDRN) and polynucleotide (PN), include phosphoric acid and four bases adenine, guanine, thymine, and cytosine. The bonded structure constitutes a repeating unit to form a high molecular weight double-stranded polymer or a low molecular weight double-stranded polymer, and additionally, four types of adenine (A), guanine (G), cytosine (C), and uracil (U) It refers to a genetic material (hereinafter, abbreviated as a gene) in which a single-stranded polymer is formed by forming a single-stranded polymer by forming a repeating unit of the base of the sugar and phosphoric acid.

In addition, the molecular weight of the low-molecular polydeoxyribonucleotide and polynucleotide prepared in the present invention may be 5kDa-20000kDa, and more specifically, the low-molecular polydeoxyribonucleotide may be 5kDa or less.

The present invention minimizes the use of non-phenols and organic solvents by using seaweeds, including mascarpone, mascarpone, kelp, seaweed, laver, etc., in order to improve the problem of the supply and demand of raw materials other than animal raw materials and the risk of contamination of viruses, and the conventional centrifugation method The process and cost were minimized by using the vacuum extraction method. Additionally, the low molecular weight process was performed through non-toxic substances and physical processes without using poisons such as sodium hydroxide and hydrochloric acid.

Hereinafter, the accompanying drawings will be described in detail with reference to the method of extracting polydeoxyribonucleotides derived from seaweeds with the effect of neovascularization and cell regeneration of the present invention by the biphenol method.

1 shows a schematic diagram of a method for extracting polydeoxyribonucleotides and polynucleotides. The method for extracting polydeoxyribonucleotides derived from seaweeds of the present invention includes the step of securing raw materials for drying and pulverizing seaweeds, such as maesengyi, hatban, kelp, seaweed, etc. of the present invention; To prepare a mixed solution consisting of 10% by weight of soybean extract, 10% by weight of adlay extract, 20% by weight of green tea extract, 20% by weight of soybean fatty acid, 20% by weight of tocopherol cocamidopropyl betain, and 20% by weight of Olive oil carboxylate And, after dissolving the protein by adding it to the dried and pulverized seaweed raw material through step (a), adding a ribonuclease to separate a mixture of polynucleotides and polydeoxyribonucleotides. And decomposition step (b); An impurity removal and pure separation step of purely separating polydioxyribonucleotides and polynucleotides by centrifuging the mixture after step (b) to remove impurities, filtering, and drying (c); A washing and drying step (d) of washing and drying the polydeoxyribonucleotide and polynucleotide purely separated through the (c) step; The resultant dried in step (D) was diluted with 1 ml of 1% sodium purified water, treated with fermented acetic acid for 10 minutes, followed by sonication treatment for 10 minutes, and centrifuged to perform low-molecular polydeoxyribonucleotides and polynucleotides. The low-molecular process step (e) to secure the; By measuring the DNA absorbance of the resultant (E) step (e), the purity of 99% low-molecular polydeoxyribonucleotides and polynucleotides having a value of 1.7 to 2.0 can be identified and powdered.

(A) Raw material acquisition stage

The raw material securing step (a) of the present invention includes drying seaweed and removing and decomposing proteins. The seaweed referred to in the present invention may be one or more selected from Maesengi, Mochi, kelp, seaweed, and laver. The prepared seaweed is dried and pulverized.

(B) Protein removal and decomposition step

The protein removal and decomposition step (b) of the present invention includes 10% by weight of soybean extract, 10% by weight of adlay extract, 20% by weight of green tea extract, 20% by weight of soybean fatty acid, 20% by weight of tocopherol cocamidopropyl betain, To prepare a mixed solution consisting of 20% by weight of olive oil carboxylate. To dissolve the protein by adding the prepared mixed solution to the dried and pulverized seaweed raw material through the step (a), and to separate the polynucleotide and polydeoxyribonucleotide mixture by adding ribonuclease after dissolving the protein. It may include steps.

Each of the soybean extract, adlay extract, and green tea extract constituting the mixed solution is heated at 100 degrees for 2 to 3 hours by adding each ingredient and distilled water in a ratio of 1:5 to freeze-dry the solution extracted with hot water to obtain each powder. Each powder is mixed at each mixing ratio constituting the mixed solution, and the mixed powder and distilled water of the same weight are mixed to prepare a solution. Here, soybean fatty acid, tocopherol cocamidopropyl betain, and olive oil carboxylate are mixed in each ratio to complete a mixed solution. The sloybean) fatty acid, tocopherol cocamidopropyl betain and Olive oil carboxylate were commercially available products.

The mixed solution prepared for removal of the polysaccharide, cell lysis, and protein decomposition may dissolve the protein by adding 40 ml each based on 1 g of the seaweed raw material in step (a).

After the protein dissolution step, 0.1 ml of ribonuclease relative to the weight of 1 g of the seaweed raw material in step (a) may be added to separate a mixture of PN (Polynucleotide) / PDRN (Polydeoxyribonucleotide). In step (b) according to the present invention, the substance added to the protein removal and decomposition is an eco-friendly degrading enzyme, and stability can be secured compared to conventional methods.

(C) Removal of impurities and separation of pure water

The step (c) of removing impurities and separating pure water includes the step of centrifuging the mixture after step (b) to remove impurities, filtering, and drying to purely separate polydeoxyribonucleotides and polynucleotides.

The mixture after step (b) is centrifuged at 2000 rpm for 1 hour to obtain a supernatant. Subsequently, the obtained supernatant is diluted in a volume ratio of 1:1 with purified water and centrifuged at 5000 rpm for 30 minutes to separate the supernatant and remove the residue.

Thereafter, purified water is added twice the volume of the separated supernatant, and filtering is performed for 30 minutes using a vacuum pressure pump of 500 mbar using a micro filter. After filtering is completed, the filter paper is dried at room temperature for 1 hour.

The separation step is generally performed by centrifugation to recover the supernatant, but in the present invention, high purity polydeoxyribonucleotides and polynucleotides could be separated by using a vacuum pump pressure and a micro filter in parallel.

(D) washing and drying steps

The washing and drying step (D) of the present invention includes washing and drying the polydeoxyribonucleotide and polynucleotide purely separated through the step (c). Put 10 ml of purified water on the dried filter paper and dry the product filtered through step (C) for 1 hour at 30-60 degrees Celsius.

(E) Small molecule process step

In the low-molecular process step (e) of the present invention, the resultant dried in step (D) is diluted with 1 ml of purified sodium water, treated with fermented acetic acid for 10 minutes, and then sonicated for 10 minutes, and centrifuged. And securing a low molecular weight polydeoxyribonucleotide and a polynucleotide.

The resultant dried in step (D) is diluted by adding 10 ml of 1% sodium purified water, and 10 ml of fermented acetic acid obtained by mixing apples and rice in a ratio of 1:1 based on the volume ratio is added and treated at 30-60° C. for 30 minutes. .

After that, sonication is performed for 10 minutes, transferred to a micro-filter column, and centrifuged at 8000 rpm for 10 minutes. Thereafter, 5 ml of purified water is added and centrifuged at 12000 rpm for 2 minutes to obtain a final low-molecular polydeoxyribonucleotide and polynucleotide.

(F) Quality inspection stage

In the quality inspection step (f) of the present invention, a step of measuring the DNA absorbance of the resultant obtained through the step (e), confirming and pulverizing low molecular weight polydeoxyribonucleotides and polynucleotides having a purity of 99% having a value of 1.7 to 2.0. Includes. The DNA absorbance according to the present invention is measured using a Thermo's nanodrop-1000 device, and a purity of 99% is determined when the value of 260 (DNA)/280 nm (protein) is between 1.7 and 2.0.

For the concentration of the DNA stock solution, measure the absorbance (A) at 260 nm using a spectrophotometer for the DNA stock solution, and measure the absorbance at 260/280 nm to confirm the purity of the extracted PDRN, and the ratio is 1.7 to 2.0. It is judged to have a purity of 99% or higher. In the case of general purity measurement of PDRN, the content is measured using DNA absorbance, and when the value is 1.7 to 2.0, it can be considered to be an average of 99% or more high purity.

The method according to the present invention uses seaweed as a raw material and requires a process to remove it due to the generation of external mucous and polysaccharides, and because it decomposes without using harmful substances such as biphenols, sodium hydroxide, and hydrochloric acid, it is more environmentally friendly and stable. It is effective, and in the case of raw materials, it is effective in the possibility of introducing viruses and economical cost by using marine plants other than animals.

In addition, the present invention is capable of smooth supply of raw materials and economical and eco-friendly seaweeds such as seaweed, seaweed, kelp, seaweed, and seaweed in order to solve the risk of virus mixing in the existing salmon, trout, etc. (animal raw materials) and restrictions on the supply and demand of raw materials. And it is possible to extract a genetic material having a molecular weight of 5kDa-20000kDa or less than 5kDa by reducing environmentally friendly materials and process processes from the tissues of marine ecological disturbance species.

As described above, it is extracted by simplifying the manufacturing process without using harmful substances such as phenol and sodium hydroxide, which are used to separate genetic material derived from tissues containing the growth point of natural seaweed that is safe from the risk of viral infection, not of animal origin. It is possible to manufacture PDRN and PN with low molecular weight processing time and secured price competitiveness. Hereinafter, an experimental example of the present invention will be described in detail as follows.

Experimental Example 1. Low molecular weight electrophoresis experiment

According to Experimental Example 1 of the present invention, an electrophoresis experiment was performed on a polymer PDRN used for medical use and a low molecular weight PDRN derived from seaweed prepared by the method according to the present invention.

2 shows the electrophoresis results according to Experimental Example 1 of the present invention. In general, a low-molecularization process is necessary in order to effectively absorb skin, tissue penetration, and absorption. Therefore, as can be seen from the results of this experiment, it is judged that the genetic material produced by the low-molecular process of 5 kDa or less can increase the penetration and absorption rate in the tissues and skin in the future.

Experimental Example 2. In vitro new angiogenesis experiment

Experimental Example 2 of the present invention is a photograph of the results of in vitro neovascularization experiments for seaweed (meal saengi, hatban, kelp, seaweed, seaweed, etc.)-derived low-molecular-processed PDRN, the existing salmon-derived polymer PDRN, and a control. Silver was an essential process for supplying nutrients and oxygen in the healing process of wounds, and was analyzed microscopically through tube formation using human umbilical vein endothelial cells.

After adding 20% FBS, Growth factor, 1% penicillin/streptomycin to EGM2 medium, culture the cells, coat 50 µl of 1X Matrigel on the plate, and dispense HUVEC (3x10 ⁵ cells/ml) and dispense reagents (control, experimental group, VEH (experimental group) After 4 hours after treatment with only the solvent used in the control), the product of other company), the tube formation was confirmed.

Figure 3 shows the results of in vitro new angiogenesis experiments for the low-molecular-processed PDRN derived from seaweed (Maesengi, hatban, kelp, seaweed, seaweed, etc.), the existing salmon-derived polymer PDRN, and the control. As a result, the formation of new blood vessels is an essential process for the supply of nutrients and oxygen in the wound healing process. As a result of microscopic analysis through tube formation using human umbilical vein endothelial cells, the algae of the present invention It was confirmed that the low-molecular PDRN material increased by 30% or more, compared to the control, by more than 30% compared to other companies' products.

In the case of tube formation, it is a neovascularization experiment, which is essential for the supply of nutrients and oxygen and removal of waste products in the wound healing process.The experiment is a tube formation experiment using human umbilical vein endothelial cells (HUVEC). Through the verification of the migration of HUVEC cells by PDRN, the effect of PDRN inducing tube formation of human umbilical vein endothelial cells on Matrigel was confirmed.

Experimental Example 3. In vitro cell regeneration experiment

In Experimental Example 3 of the present invention, in vitro cell regeneration experiments were conducted for low-molecular-processed PDRN derived from algae (meal saengi, hatban, kelp, seaweed, seaweed, etc.) and the existing salmon-derived polymer PDRN, and a control.

In the case of wound healing, the continuity of the wound tissue means its original continuity due to external action, and the tissue mainly refers to the skin. The skin is the largest organ in our body, and from the risk of external irritation and infection. It plays a primary defense role. The skin consists of the epidermis and the dermis, and the epidermis is layered in the order of the basal layer, the play layer, the granule layer, and the stratum corneum from the inside, and the keratinocytes that make up the epidermis are the basal cells generated by cell division. , Granular cells and keratinocytes are differentiated in this order, and then metabolism (turnover) is repeated to separate from the surface as individual pieces. The process is as follows.

In the stage of wound healing, when a person's skin tissue is damaged for any reason, the reaction to heal the wound begins immediately. This is a process that occurs in all wounds regardless of the type of wound healing. As follows.

1) Inflammation stage: When a wound occurs, cells in damaged blood vessels around the wound (hemofarm, white blood cells, macrophages, etc.) are activated to remove bacteria, foreign substances, and necrotic tissue, and various active substances are secreted from these cells. To stimulate the skin cells around the wound

2) Epithelialization step: The step in which the skin cells (epithelial cells) on the wound surface differentiate and move through cell division, and the dividing epithelial cells are filled throughout the wound.

3) Proliferation stage: A stage in which the cytoplasm and the substrates of cells proliferate, mainly for collagen synthesis

4) Maturation Stage: A stage in which the balance between collagen production and decomposition in the scar tissue formed through the epithelial stage and proliferation stage is achieved.

The reactions are superimposed without distinct distinction and go through a series of steps.Wounds are destroyed by physical or chemical wounds or bacterial infection, resulting in various cellular and molecular sequelae, and wound healing is performed in a special way. As a systematic biochemical and cellular process that induces the growth and regeneration of wounded tissues, it is a complex process that induces the restoration of damaged skin or tissue to a normal state by accommodating the interaction between blood cells, cytokines, and growth factors.

In addition, it is important to heal the wound early, and if the wound is open for a long time, secondary complications such as infection may occur, and pain and discomfort in daily life are accompanied during the treatment period, so the wound should be closed early. do.

However, in some cases, the emergence of pathogenic microorganisms with immunity to antibiotics has increased due to the indiscriminate use of synthetic drugs, and the high price and side effects of synthetic drugs are emerging as major problems in the treatment of diseases, and better bioactivity potential and side effects. Continuous development of these small drugs is required.

Thus, Experimental Example 4 of the present invention was carried out by adding 10% FBS and 1% penicillin/streptomycin to DMEM medium, and then dispensing HaCaT to the plate after cell culture (2x10 ⁵ cells/ml), incubating for 4 hours, and starvation with HaCaT in a serum-free medium. After 16 hours of incubation, after scratching the plate, 24 hours after dispensing reagents (control group, experimental group, VEH (control group treated with only the solvent used in the experiment group), products from other companies), wound healing was confirmed, and human epithelial cell line (HaCaT)-wound Healing model: As a result of analyzing the extent of cell migration by scratching confluent cells, the narrower the scratch range is, the more effective it is to regenerate.The graph shows that the scratch area is quantified, and the lower the value, the more effective it is. can see.

Figure 4 shows the results of in vitro cell regeneration experiments for the low-molecular-processed PDRN and the existing salmon-derived polymer PDRN, and a control derived from seaweed (meal saengi, hatban, kelp, seaweed, seaweed, etc.). As a result of analyzing the degree of cell migration by scratching the human epithelial cell line (HaCaT) using the Wound healing model, the marine algae of the present invention (meal saengyi, mascarpone, kelp, seaweed, seaweed, etc.) It was confirmed that the wound healing effect was increased by more than 5% and by more than 25% compared to the control.

The present invention provides a method for preparing a low-molecular PDRN, PN mixture that activates adenosine A2 receptor from the obtained seaweed-derived gene mixture by isolating genetic material such as PDRN and PN from all tissues other than the specific tissues of Kim. As it has angiogenesis and cell regeneration effects, it meets the needs of related consumers and contributes to improving the profits of related industries, so it has industrial applicability.

Claims (4)

Securing raw materials for drying and pulverizing seaweed (a);
In the seaweed raw material dried and pulverized through the above step (a), 10% by weight of beans, 10% by weight of adlay, 20% by weight of green tea extract, 20% by weight of soybean fatty acid, 20% by weight of tocopherol cocamidopropyl betain, Protein removal comprising the step of separating and reacting a mixture of polynucleotides and polydeoxyribonucleotides by preparing and adding a protein dissolution mixed solution consisting of 20% by weight of olive oil carboxylate to dissolve the protein, and then adding ribonuclease to separate a mixture of polynucleotides and polydeoxyribonucleotides, and Decomposition step (b);
An impurity removal and pure separation step of purely separating polydioxyribonucleotides and polynucleotides by centrifuging the mixture after step (b) to remove impurities, filtering, and drying (c);
A washing and drying step (d) of washing and drying the polydeoxyribonucleotide and polynucleotide purely separated through the (c) step;
A low-molecular process step (e) of securing low-molecular polydeoxyribonucleotides and polynucleotides by centrifuging the resultant dried in step (d);
Characterized in that consisting of a quality inspection step (f) of measuring the DNA absorbance of the resultant obtained through the step (E), confirming and pulverizing 99% low-molecular polydeoxyribonucleotides and polynucleotides having a purity of 1.7 to 2.0. Seaweed-derived polydeoxyribonucleotide and polynucleotide extraction method having new angiogenesis and cell regeneration effects delete The method according to claim 1, wherein the seaweed is one or more selected from meesengyi, masaengi, kelp, seaweed, and seaweed.The method of extracting polydeoxyribonucleotides and polynucleotides derived from seaweeds having a new angiogenesis and cell regeneration effect. The method of claim 1, wherein the protein dissolution mixed solution prepared in step (b) is added by 40 ml per 1 g of the weight of the seaweed raw material in step (a), and ribonuclease is added relative to the weight of 1 g of the seaweed raw material in step (a). Seaweed-derived polydeoxyribonucleotide and polynucleotide extraction method having new angiogenesis and cell regeneration effects, characterized by adding 0.1 ml each

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