Biomaterial composition for tissue repair comprising DNA fragment mixture and polyhydric alcohol

Abstract

The present invention relates to: a composition for tissue repair comprising a DNA fragment mixture and a C3 or C4 polyhydric alcohol; a filler composition; a method for tissue repair using same; and a use for tissue repair.

Description

A biomaterial composition for tissue repair comprising a mixture of DNA fragments and a polyhydric alcohol

[0001] The present invention relates to a composition for tissue repair comprising a mixture of DNA fragments and a C3 or C4 polyhydric alcohol; a filler composition; a method for tissue repair using the same; and a use for tissue repair.

[0002]

[0003] As interest in anti-aging grows, the number of procedures aimed at improving deficiencies in areas of the body, such as joints, cardiovascular system, and skin, is increasing. For example, cosmetic procedures aimed at improving wrinkles and other physical appearances fall into this category. Recently, methods that inject highly biocompatible materials have been increasingly used to replace damaged tissue and restore volume to desired areas of the body.

[0004] For example, filler compositions containing hyaluronic acid as the main ingredient exist. However, some hyaluronic acid fillers have high viscosity and low elasticity, making them difficult to inject into the skin. Furthermore, even if injected, they do not maintain their injected shape for long and have a short shape retention period. Various compounds are being added to address this issue (see U.S. Patent No. 11,154,481), but it remains challenging to address these issues while minimizing side effects.

[0005]

[0006] The development of improved biomaterials for tissue repair is still needed.

[0007]

[0008] One object of the present invention is to provide a tissue repair composition comprising a DNA fragment mixture and a C3 or C4 polyhydric alcohol, wherein the DNA fragment mixture is contained in an amount of 2 to 5 wt% based on the total composition, and the polyhydric alcohol is contained in an amount of 0.5 to 4 wt% based on the total composition.

[0009] Another object of the present invention is to provide a filler composition comprising a DNA fragment mixture and a C3 or C4 polyhydric alcohol, wherein the DNA fragment mixture is contained in an amount of 2 to 5 wt% based on the total composition, and the polyhydric alcohol is contained in an amount of 0.5 to 4 wt% based on the total composition.

[0010] Another object of the present invention is to provide a tissue repair method comprising a step of administering the tissue repair composition or filler composition to a subject.

[0011] Another object of the present invention is to provide a composition comprising a mixture of DNA fragments; and a polyhydric alcohol of C3 or C4, wherein the mixture of DNA fragments is contained in an amount of 2 to 5 wt% based on the total composition, and the polyhydric alcohol is contained in an amount of 0.5 to 4 wt% based on the total composition, for use in tissue repair.

[0012]

[0013] The biomaterial for tissue repair of the present invention has an excellent effect of not only having tissue repair ability but also having a moisturizing effect, thereby preventing the tissue from feeling dry.

[0014]

[0015] Figure 1 is a diagram showing the results of confirming the effect of increasing moisture content through animal testing.

[0016] Figure 2 is a diagram showing the standardized injection force evaluation results for manufactured comparative examples and examples.

[0017] Figures 3 to 6 are diagrams showing the phase angle evaluation results for manufactured comparative examples and examples.

[0018] Figure 7 is a diagram showing the results of biodegradability evaluation for manufactured comparative examples and examples.

[0019] Figure 8 is a diagram showing the results of comparing the height and width of the injection site after injection of the liquid composition according to the manufactured comparative example and example.

[0020] Figure 9 is a diagram showing the results of comparing the properties of each composition when preparing a liquid composition.

[0021]

[0022] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below. In addition, numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated into this specification in their entirety by reference to more clearly explain the level of the technical field to which the present invention belongs and the contents of the present invention.

[0023]

[0024] One aspect of the present invention provides a tissue repair composition comprising a mixture of DNA fragments and an alcohol. Specifically, the alcohol may be a polyhydric alcohol, and more specifically, a C3 or C4 polyhydric alcohol.

[0025] As one specific example, the tissue repair composition of the present invention is characterized by a tissue repair composition comprising an active ingredient comprising a mixture of DNA fragments and a C3 or C4 polyhydric alcohol.

[0026] In the present invention, the term "DNA fragment mixture" refers to a mixture of nucleic acid fragments having a molecular weight range that includes DNA, which is a biopolymer composed of phosphate, four types of bases, and deoxyribose, and exists as a polymer of nucleotides. The DNA fragment mixture may exist in the form of fragments with reduced molecular weight, but is not limited thereto, and may be used interchangeably with terms such as 'DNA fragment', 'DNA fraction', 'nucleic acid fragment', and 'nucleic acid fragment mixture'.

[0027] The DNA fragment mixture according to any one of the preceding specific examples may be a polynucleotide (PN), a polydeoxyribonucleotide (PDRN), or a mixture thereof.

[0028] In the present invention, the term "polynucleotide" is referred to as "PN" and may mean a DNA or RNA strand, which is a polymer of nucleotides in which nucleotide units are covalently bonded to form a chain. In addition, the term "polydeoxyribonucleotide" in the present invention is also referred to as "PDRN" and may be a type of low molecular weight DNA complex having a specific molecular weight, but is not limited thereto. For example, the polynucleotide may have a relatively longer nucleic acid length or a larger molecular weight than polydeoxyribonucleotide, and may be used as a raw material for medical devices due to its physical support role, cell fixation, lubrication, and buffering effects, and polydeoxyribonucleotide may be used as a raw material for pharmaceuticals for cell proliferation and tissue regeneration, but is not limited thereto.

[0029] The solution of the DNA fragment mixture according to any one of the preceding specific examples may include a buffer, and the buffer may be at least one selected from the group consisting of sodium phosphate monobasic dihydrate, sodium phosphate dibasic dodecahydrate, sodium chloride, magnesium chloride, potassium chloride, and phosphate buffer saline or HEPES (N-(2-hydroxyethyl)-piperazine-N'-2-ethanesulfonic acid), but is not limited thereto.

[0030] The DNA fragment mixture according to any of the preceding specific examples may be obtained by extraction from the testis or semen of a fish. Specifically, the fish may be a salmonid. More specifically, it may be salmon or trout, but is not limited thereto.

[0031] The DNA fragment mixture according to any one of the preceding specific examples may have a molecular weight of about 1 to 100,000 kDa, 5 to 50,000 kDa, 50 to 10,000 kDa, or 50 to 1,500 kDa.

[0032] As an embodiment according to any one of the preceding specific examples, the DNA fragment mixture of the present invention may be, but is not limited to, a polynucleotide (PN).

[0033] The DNA fragment mixture according to any one of the preceding specific examples may be included in an amount of 2 to 7 wt% relative to the total tissue repair composition, and specifically, may be included in an amount of 2 to 7 wt%, 2 to 6 wt%, 2 to 5 wt%, 2 to 4 wt%, 2 to 3 wt%, 3 to 7 wt%, 3 to 6 wt%, 3 to 5 wt%, 3 to 4 wt%, 4 to 7 wt%, 4 to 6 wt%, 4 to 5 wt%, 5 to 7 wt%, 5 to 6 wt%, 6 to 7 wt%, or 6 to 7 wt%.

[0034] The C3 or C4 polyhydric alcohol of the present invention refers to an alcohol having 3 or 4 carbon atoms and two or more hydroxyl groups (-OH).

[0035] The polyhydric alcohol according to any one of the preceding specific examples may be included in an amount of 0.5 to 8 wt% based on the total composition, specifically 0.5 to 8 wt%, 0.5 to 7 wt%, 0.5 to 6 wt%, 0.5 to 5 wt%, 0.5 to 4 wt%, 0.5 to 3 wt%, 0.5 to 2 wt%, 0.5 to 1 wt%, 1 to 8 wt%, 1 to 7 wt%, 1 to 6 wt%, 1 to 5 wt%, 1 to 4 wt%, 1 to 3 wt%, 1 to 2 wt%, 2 to 8 wt%, 2 to 7 wt%, 2 to 6 wt%, 2 to 5 wt%, 2 to 4 wt%, 2 to 3 wt%, 3 to 8 wt%, 3 to 7 wt%, 3 to It may be comprised in an amount of 6 wt%, 3 to 5 wt%, 3 to 4 wt%, 4 to 7 wt%, 4 to 6 wt%, 4 to 5 wt%, 5 to 7 wt%, or 5 to 6 wt%.

[0036] The polyhydric alcohol according to any one of the preceding specific examples may be, but is not limited to, one or more selected from the group consisting of glycerin, propylene glycol, and butylene glycol.

[0037] In the present invention, it was confirmed that polyhydric alcohol alone cannot be used for tissue repair purposes, whereas when a mixture of DNA fragments and polyhydric alcohols of C3 or C4 are combined, viscosity is reduced, thereby increasing ease of injection.

[0038] In addition, in the case of a combination of a DNA fragment mixture and polyhydric alcohols such as polyethylene glycol (PEG) and triethylene glycol, a large amount of foam is generated during the preparation of the liquid composition, which increases the probability of defective products during subsequent product production, or the viscosity approaches 0, making it impossible to use for tissue repair. On the other hand, it was confirmed that the combination of a DNA fragment mixture of the present invention and a C3 or C4 polyhydric alcohol has a significantly lower probability of this.

[0039]

[0040] Meanwhile, the tissue repair composition of the present invention may have tissue moisturizing properties.

[0041] The term "tissue moisturizing ability" in the present invention refers to increasing the moisture content of tissue. The tissue moisturizing ability of the present invention may be an increase in the moisture content of tissue upon injection of the composition of the present invention compared to when the composition of the present invention is not injected into the tissue. Such moisturizing ability may contribute to an increase in the moisture content within the tissue, tissue repair with a high survival rate and increased moisture content while not causing a dry feeling in the surrounding tissue but rather alleviating the dry feeling.

[0042] As another specific example, the tissue repair composition may increase the moisture content of the tissue when injected into the tissue.

[0043] Examples of the above tissues may include, but are not limited to, skin.

[0044] In one embodiment of the present invention, it was confirmed that a tissue repair composition further comprising a C3 or C4 polyhydric alcohol (particularly at a specific content) compared to a DNA fragment mixture alone not only has an excellent tissue repair effect when injected into a tissue, but also increases moisture content, which suggests that the composition of the present invention provides moisture to the adjacent tissue without excessively absorbing moisture from the adjacent tissue when injected, so that it does not cause a dry feeling in the surrounding tissue but rather alleviates the dry feeling, and has a tissue repair effect with a high engraftment rate and good moisture content.

[0045] In particular, it was confirmed that a composition comprising a DNA fragment mixture; and a C3 or C4 polyhydric alcohol, wherein the DNA fragment mixture is included in an amount of 2 to 5 wt% based on the total composition, and the polyhydric alcohol is included in an amount of 0.5 to 4 wt% based on the total composition, has a critical significance for a moisturizing effect, and it was confirmed that a tissue repair effect is excellent in the same range, suggesting that in the range of the above-mentioned content combination, it is possible to relieve a dry feeling in the surrounding tissue without giving a dry feeling, and rather has a tissue repair effect with good moisturizing feeling.

[0046]

[0047] In the tissue repair composition according to any one of the preceding specific examples, the DNA fragment mixture and the polyhydric alcohol may be included in a weight ratio of 10:1 to 1:2, but is not limited thereto.

[0048] In one embodiment according to any one of the preceding specific examples, in the tissue repair composition of the present invention, the DNA fragment mixture and the polyhydric alcohol may be included in a weight ratio of about 10:1 to 1:2, 10:1 to 1:1, 8:1 to 1:2, 8:1 to 1:1, 6:1 to 1:2, 6:1 to 1:1, 5:1 to 1:2, 5:1 to 1:1, 4:1 to 1:2, or 4:1 to 1:1, and specifically, may be included in a weight ratio of about 10:1 to 1:2, 10:1 to 1:1, or 6:1 to 1:2.

[0049] The term "about" above includes not only the exact number stated after the term, but also a range that is or is nearly that number. Whether the number is or is nearly the specific number stated can be determined based on the context in which it is presented. For example, the term "about" may refer to a range of -10% to +10% of a given numerical value. As another example, the term "about" may refer to a range of -5% to +5% of a given numerical value. Other examples include, but are not limited to, a range that includes ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc.

[0050] Even if the word "about" is omitted before a number in the present invention, it is self-evident that it is included in the present invention to the extent that it is not omitted.

[0051]

[0052] Meanwhile, in the present invention, the term "tissue repair" means something that can be used for replacing, repairing, and / or reconstructing human tissues and organs such as blood vessels, heart, septum, fascia, and / or skin.

[0053] The tissue repair composition of the present invention can be used as a tissue repair biomaterial, and the tissue repair biomaterial refers to a bio-derived material used for replacement, repair, and reconstruction of human tissues and organs such as blood vessels, heart, septum, fascia, and skin, and is not limited to a cosmetic filler, and can refer to a tissue repair biomaterial specified by the Ministry of Food and Drug Safety.

[0054]

[0055] The tissue repair composition of the present invention may be a liquid composition or an injectable composition, but is not limited thereto.

[0056] In the present invention, the term "injectable" means a material having the properties required for administering a composition to a subject using an injection device having a needle.

[0057] In the present invention, the term "administration" means introducing the composition of the present invention into a subject by any appropriate method, and the administration route may be various routes such as application, subcutaneous injection, dermal injection, blood vessel, biological membrane, tissue fiber, synovial fluid, etc., as long as it can reach the target tissue. The formulation may be, but is not limited to, topical application, subcutaneous injection, dermal injection, blood vessel injection, intramuscular injection, joint injection, tendon injection, and ligament injection formulation. The preferred dosage of the composition of the present invention may vary depending on the condition of the subject.

[0058]

[0059] The tissue repair composition according to the present invention may exhibit an appropriate viscosity range that allows injection into a desired tissue.

[0060] In addition, the term 'viscosity' used in the present invention refers to the property of a fluid, that is, the appearance of a flow with viscosity, which is resistance to flow. This viscosity can be expressed as viscosity, and particularly complex shear viscosity (η*, Pa·s).

[0061] As another specific example, the tissue repair composition may exhibit a complex shear viscosity of 1 to 2,000 Pa·s, specifically 5 to 2,000 Pa·s, 8 to 2,000 Pa·s, 1 to 1,500 Pa·s, 5 to 1,500 Pa·s, 8 to 1,500 Pa·s, 1 to 1,300 Pa·s, 5 to 1,300 Pa·s, or 8 to 1,300 Pa·s.

[0062] The viscosity range of tissue repair compositions used as general tissue repair biomaterials is from about 1 Pa·s to about 8,000 Pa·s. When the viscosity range is included, the composition can be naturally settled in the tissue in the body after administration. In general, when the viscosity range of the tissue repair composition is about 1 Pa·s or less, although it is settled in the body, its ability to take its own shape is low, so it cannot function as a tissue repair biomaterial for creating volume, and there may be difficulties in controlling the injection amount, such as a large amount of injection solution being injected even with a small amount of force. In addition, when the viscosity range is high, such as about 8,000 Pa·s or more, it is difficult to extrude the syringe when injecting it into the body, and excessive force may be required during injection, making it difficult to inject an accurate amount. In addition, it is difficult to finely control the injection amount, which may cause side effects.

[0063] The tissue repair composition of the present invention can be injected at a rate of 50 mm / min with an injection force of 0 or more; and an extrusion force of about 60 N, about 55 N, about 50 N, about 45 N, about 40 N, about 35 N, about 30 N, or about 25 N or less. For example, the injection force may be obtained by injection through a 33 gauge needle, but is not limited thereto. Specifically, according to the guideline for approval review of fillers for plastic surgery, it may be appropriate for the injection force to be set to 40 N or less.

[0064]

[0065] The tissue repair composition of the present invention may additionally include, but is not limited to, one or more compounds selected from the group consisting of anesthetics, vitamins, amino acids, metals, antioxidants, and mineral salts.

[0066] The tissue repair composition of the present invention may additionally include any suitable excipient commonly used in the art, and such excipient may be, for example, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, or an isotonic agent, but is not limited thereto.

[0067]

[0068] Another aspect of the present invention provides a biomaterial for tissue repair comprising the composition for tissue repair of the present invention.

[0069] The above tissue repair composition and tissue repair biomaterial are as described in other aspects.

[0070]

[0071] Another aspect of the present invention provides a filler composition comprising a mixture of DNA fragments and an alcohol. Specifically, the alcohol may be a polyhydric alcohol, and more specifically, a C3 or C4 polyhydric alcohol.

[0072] Additionally, the DNA fragment mixture may be included in an amount of 2 to 5 wt% relative to the total composition, and the polyhydric alcohol may be included in an amount of 0.5 to 4 wt% relative to the total composition.

[0073] In the present invention, the term "filler" refers to a medical device that is injected into the skin as a material for tissue repair to restore volume, etc., and has an operating principle of maintaining skin volume through physical repair.

[0074] The above DNA fragment mixture, polyhydric alcohol, etc. are as described in other aspects, and the filler composition may be a specific example of a tissue repair composition, and all of the above-described contents of the tissue repair composition of the present invention (DNA fragment mixture, polyhydric alcohol, content thereof, weight ratio, tissue moisturizing ability, liquid composition, injectable composition, administration, viscosity in an appropriate range for injection, etc.) may also be applied to the filler composition.

[0075] The filler composition of the present invention may additionally include one or more compounds selected from the group consisting of anesthetics, vitamins, amino acids, metals, antioxidants, and mineral salts, but is not limited thereto.

[0076] The filler composition of the present invention may additionally include any suitable excipient commonly used in the art, and such excipient may be, for example, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, or an isotonic agent, but is not limited thereto.

[0077]

[0078] Another aspect of the present invention provides a method for tissue repair comprising administering to a subject a composition for tissue repair or a filler composition of the present invention.

[0079] The above administration, tissue repair, tissue repair composition, filler composition, etc. are as described in other aspects.

[0080] The term "subject" in the present invention refers to any animal, including rats, mice, and livestock, including humans, that requires or is likely to require tissue repair. Specifically, it may be a mammal, including humans.

[0081]

[0082] Another object of the present invention is to provide a composition for tissue repair comprising a mixture of DNA fragments and a polyhydric alcohol of C3 or C4, wherein the mixture of DNA fragments is contained in an amount of 2 to 5 wt% based on the total composition, and the polyhydric alcohol is contained in an amount of 0.5 to 4 wt% based on the total composition.

[0083] The above DNA fragment mixture, polyhydric alcohol, and tissue repair are as described in other aspects.

[0084]

[0085] The present invention is described in more detail below through examples. However, the following examples are merely preferred embodiments intended to illustrate the present invention and are therefore not intended to limit the scope of the present invention. Furthermore, technical details not described herein can be readily understood and implemented by those skilled in the technical field of the present invention or similar fields.

[0086]

[0087] Manufacturing example: Manufacturing of a biomaterial for tissue repair comprising a mixture of DNA fragments and a polyhydric alcohol.

[0088] A DNA fragment mixture solution was prepared by dissolving the DNA fragment mixture in a buffer solution at a high temperature of 60-80°C using a heat stirrer. Polyhydric alcohol was added to the DNA fragment mixture solution prepared in a heat stirrer at 60-80°C and mixed, and then the temperature of the mixed solution was lowered to room temperature to prepare a liquid formulation.

[0089] At this time, PN (polynucleotide; manufacturer: Pharma Research) was used as a representative example of a DNA fragment mixture, and the sample name and concentration were prepared as shown in Tables 1 and 2 below.

[0090]

[0091] Sample name DNA fragment mixture (wt%) Polyvalent alcohol type; and content (wt%) Example 12 Glycerin; 2 Example 22 Propylene glycol; 2 Example 32 Butylene glycol; 2 Example 4 PEG400; 2 Example 5 PEG4000; 2 Example 6 PEG6000; 2 Example 7 2 Triethylene glycol; 2

[0092] Sample name DNA fragment mixture (wt%) Glycerin content (wt%) Comparative Example 120 Example 8 20.5 Example 9 21.0 Example 10*22.0 Example 11 23.0 Example 12 24.0 Example 13 28.0 Comparative Example 230 Example 14 30.5 Example 15 31.0 Example 16 32.0 Example 17 33.0 Example 18 34.0 Example 19 38.0 Comparative Example 340 Example 20 40.5 Example 21 41.0 Example 22 42.0 Example 23 43.0 Example 24 44.0 Example 25 48.0 Comparative Example 450 Example 26 50.5 Example 27 51.0 Example 28 52.0 Example 2953.0 Example 3054.0 Example 3158.0 Comparative Example 570 Example 3274.0 Example 3378.0 Comparative Example 602.0

[0093] * Example 10 has the same composition and content as Example 1.

[0094]

[0095] Experimental Example 1: Confirmation of moisturizing effect through animal testing

[0096] A skin moisture loss model was created using mice as a representative example of tissue. This was created by referencing a previously published method (Park, No-June, et al., "Compound K improves skin barrier function by increasing SPINK5 expression." Journal of Ginseng Research 44.6 (2020): 799-807).

[0097] After the liquid composition prepared in the manufacturing example was injected into the body of the manufactured animal model, the moisture content of the mouse skin was measured under conditions of 25±5℃ and 50%±5% RH using a moisture measuring device (Corneometer® CM 825 (Courage + Khazaka electronic GmbH, Germany).

[0098] As a result, as shown in Fig. 1, it was confirmed that the DNA fragment mixture and the example including glycerin in a specific content or ratio imparted a higher moisture content (moisturizing effect) to the tissue.

[0099] In particular, in the case of a composition in which the DNA fragment mixture is included in an amount of 2 to 5 wt% based on the total composition and the polyhydric alcohol is included in an amount of 0.5 to 4 wt% based on the total composition, it was confirmed that there was a critical significance for a moisturizing effect, and as described below, it was confirmed that the tissue repair effect was excellent in terms of viscosity, injection force, injection feeling, biodegradability, etc. in the same content range, suggesting that the tissue repair effect with a higher survival rate and better moisturizing feeling was present in the range of the above-mentioned content combination.

[0100]

[0101] Experimental Example 2: Viscosity Evaluation

[0102] The liquid compositions prepared in Comparative Examples 1 to 6 and Examples 8 to 33, which were prepared using only the DNA fragment mixture, only glycerin, and a combination of the DNA fragment mixture and glycerin, were subjected to measurement of complex viscosity (η*, Pa·s; hereinafter, viscosity) using a rheometer (NETZSCH (Germany), Kinexus Ultra+ Rheometer) 48 hours after preparation. Specifically, the viscosity was measured under the following conditions: measurement temperature: 25°C, geometry used: PU20, gap: 1.0 mm, measured Hz: 0.1 Hz, shear strain: 0.5%, and analysis program: rSpace for Kinexus. The results are shown in Table 3 below.

[0103]

[0104] Sample viscosity (η*, Pa·s) Comparative example 1195.2 Comparative example 2482.0 Comparative example 3777.9 Comparative example 4985.4 Comparative example 51924.0 Comparative example 60.03 Embodiment 8 1 22.2 Embodiment 9 43.12 Embodiment 108.176 Embodiment 115.697 Embodiment 123.266 Embodiment 132.651 Embodiment 14 276.1 Embodiment 15 88.98 Embodiment 16 40.08 Embodiment 17 38.46 Embodiment 18 35.21 Embodiment 19 28.83 Embodiment 20 456.5 Embodiment 21 312.6 Embodiment 22 185.5 Embodiment 23 160.6 Embodiment 24 147.1 Embodiment 25 125.9 Embodiment 26 636.7 Embodiment 27 464.0 Embodiment 28 361.6 Embodiment 29 329.3 Embodiment 30 276.7 Embodiment 31227.1 Example 321237.0 Example 331147.0

[0105] As a result, as shown in Table 3 above, it was confirmed that glycerin alone (Comparative Example 6) had a viscosity similar to purified water and therefore could not be used for tissue repair. On the other hand, it was confirmed that the viscosity of the DNA fragment mixture and glycerin combination (Examples 8 to 33) was reduced compared to the DNA fragment mixture alone (Comparative Examples 1 to 5). This suggests that the ease of injection of the DNA fragment mixture and glycerin combination (Examples 8 to 33) can be increased.

[0106]

[0107] Experimental Example 3: Injection Power Evaluation

[0108] Except for glycerin alone (Comparative Example 6), which was confirmed not to be usable for tissue repair in Experimental Example 2, the injection force was measured for Comparative Examples 1 to 5 and Examples 8 to 33.

[0109] The injection force represents the force (N) required to inject the injection solution, and the injection speed was measured at 50 mm / min. The liquid compositions prepared in all examples and comparative examples for which the injection force was confirmed were measured using an injection force measuring device (tensile and compression tester) 48 hours after preparation. Specifically, the injection force was measured using a tensile and compression tester (Universal Testing Machine (UTM), Dahwa Tester (Republic of Korea)) under the following conditions: measurement temperature: 25 ± 2 ℃, injection needle: JBP Korea (Republic of Korea), 33 G nano needle, injection speed: 50 mm / min. The results are shown in Table 4 and Fig. 2 below.

[0110]

[0111] Sample silk input (N) Comparative example 117.2 Comparative example 227.17 Comparative example 332.37 Comparative example 449.33 Comparative example 569.39 Example 8 15.22 Example 9 14.45 Example 10 12.47 Example 11 12.34 Example 12 11.88 Example 13 10.49 Example 14 21.3 Example 15 19.81 Example 16 18.15 Example 17 17.4 Example 18 16.21 Example 19 14.66 Example 20 28.09 Example 21 25.69 Example 22 24.79 Example 23 21.3 Example 24 20.68 Example 25 18.28 Example 26 36.46 Example 2735.87 Example 2833.66 Example 2931.13 Example 3029.4 Example 3126.88 Example 3252.43 Example 3348.05

[0112] The injection power values ​​of the glycerin-free compositions (Comparative Examples 1 to 5) were replaced with 100%, and the values ​​of the examples based on the same content of DNA fragment mixtures were standardized to the values ​​of the comparative examples. The results are shown as % values ​​in Figure 2.

[0113] As a result, as shown in Table 4 and Fig. 2, it was confirmed that the injection force was weakened in the case of the combination with glycerin compared to the DNA fragment mixture alone, and it was confirmed that even though the injection force of the comparative example was high, the injection force control effect of the example in which the DNA fragment mixture and glycerin were combined was large.

[0114]

[0115] Experimental Example 4: Evaluation of Injection Sensitivity

[0116] Except for glycerin alone (Comparative Example 6), which was confirmed not to be usable for tissue repair in Experimental Example 2, the injection feeling of representative comparative examples and examples was evaluated, excluding Examples 32 and 33, which did not satisfy the criteria for the guideline for approval of plastic fillers, which stipulates that the injection force should be set to 40 N or less.

[0117] A blind test was conducted on independently recruited experimenters (4 people), and the experimenters evaluated the injection sensation by dispensing the compositions of each comparative example and experimental example into a petri dish using a syringe needle (33 gauge nano needle) used in the actual product. The related conditions were all the same as the injection force test except that the device (equipment) was not used. The easier the injection (the more convenient), the more difficult the injection (the more uncomfortable), and the 5 points were indicated. The results are shown in Tables 5 and 6 below. Table 5 shows the number of people who indicated each score.

[0118]

[0119] Score comparison example 1 Example 8 Example 10 Example 12 Comparison example 2 Example 14 12 3 4 2 3 2 1 1 3 12 2 4 2 1 5 Total score 9 6 5 4 1 4 1 2 Main input (N) 17.2 15.2 2 12.4 7 11.8 8 27.1 7 2 1.3

[0120] Score Example 16 Example 18 Comparative Example 4 Example 26 Example 28 Example 3 0 1 1 2 3 3 1 1 2 4 2 3 2 5 4 2 Total Score 9 7 2 0 1 8 1 5 1 4 Main Input (N) 18.1 5 16.2 1 4 9.3 3 6.4 6 33.6 6 2 9.4

[0121] As shown in Tables 5 and 6 above, the actual injection sensation evaluation score was lower in the case of the combination with glycerin (Example 8, etc.) compared to the DNA fragment mixture alone (Comparative Examples 1, 2, and 4), confirming that the injection sensation was superior.

[0122]

[0123] Experimental Example 5: Phase Angle Evaluation

[0124] The liquid compositions prepared in each example and comparative example were measured for phase angle (δ, °) using a rheometer (NETZSCH (Germany), Kinexus Ultra+ Rheometer) 48 hours after preparation. Specifically, the viscosity was measured under the following conditions: measurement temperature: 25 ℃, geometry used: PU20, gap: 1.0 mm, measured Hz: 0.1 Hz, shear strain: 0.5%, and analysis program: rSpace for Kinexus. The results are shown in FIGS. 3 to 6.

[0125] As a result, as shown in FIGS. 3 to 6, the phase angle increases in the case of the combination with glycerin (Examples 8 to 31) compared to the DNA fragment mixture alone (Comparative Examples 1 to 4), suggesting that the ease of injection can be increased by further increasing fluidity.

[0126]

[0127] Experimental Example 6: Comparison of biodegradability of biorepair materials with and without DNA fragment mixtures and polyhydric alcohols.

[0128] A liquid composition containing a mixture of DNA fragments and / or polyhydric alcohols (glycerin, propylene glycol, butylene glycol) in the ranges of Comparative Example 6 and Example 28 was injected into a mouse. The mouse was a SKH-1 hairless mouse, 6 weeks old, female, and was housed in a cage with a temperature of 22 ± 2 °C and a relative humidity of 50 ± 10%, with free access to food and water. The experimental period was one week after the acclimatization period, after which the sample was injected, and sacrificed 60 hours later. Measurements were made by injecting 100 μL intradermally into the dorsal area of ​​the mouse.

[0129] After sample injection, the volume change at the injection site was observed, and the results confirmed by 3D photo simulation (Primos, Canfield (USA)) are shown in Figure 7.

[0130] As a result, as shown in Fig. 7, it was confirmed again that polyhydric alcohols (glycerin, propylene glycol, butylene glycol) alone were all decomposed immediately after injection and could not be used for tissue repair purposes. However, in the case of a combination of a DNA fragment mixture and polyhydric alcohols (glycerin, propylene glycol, butylene glycol), it was confirmed that they existed even after 60 hours, confirming their suitability as a material for biorepair.

[0131]

[0132] Experimental Example 7: Comparison of height and width after injection of liquid composition

[0133] If a biomaterial for tissue repair is injected into a living body and spreads too widely, the repair effect may be minimal. If the width is narrow and the height is high, the problem of it appearing like a protrusion or hive may occur because it is too prominent visually and aesthetically. Therefore, the width and height were evaluated after injection.

[0134] The mice used in this experiment were SKH-1 hairless mice, 6 weeks old, female. They were housed at a temperature of 22 ± 2 °C and a relative humidity of 50 ± 10%, with free access to food and water. The experimental period was one week after the acclimatization period, after which samples were injected, and measurements were taken immediately after injection. The mice were sacrificed after 7 days. Measurements were taken by injecting 100 μL intradermally into the dorsal area of ​​the mouse.

[0135] To this end, the liquid compositions of the comparative examples and examples prepared in the aforementioned manufacturing examples were injected into mice, and the height and width of the injected sample were measured using a vernier caliper (MITUTOYO, Japan) and analyzed using ImageJ software. The results are shown in Fig. 8 and Table 7.

[0136]

[0137] Area (mm) 2) Height (mm) RJR 7 3.0 2 2.63 Comparative Example 2 48.2 5 3.35 Exemplary Example 1 46 8.3 7 2.53 Exemplary Example 1 57 4.7 0 2.81 Exemplary Example 1 67 5.5 4 2.56 Exemplary Example 1 87 2.0 2 2.47 Exemplary Example 1 9 1 49.7 7 1.65

[0138] As shown in Table 7 and Figure 8 above, when compared with a commercialized product (Rejuran; RJR; Pharma Research Co., Ltd.), it was confirmed that the evaluated examples 14, 15, 16 and 18 had a similar level of repair range to the commercialized product, indicating that the composition of the present invention has an excellent tissue repair effect.

[0139]

[0140] Experimental Example 8: Comparison of Efficacy with Other Polyhydric Alcohols

[0141] In order to compare the efficacy of Example 1 manufactured using glycerin and Examples 2 to 7 manufactured using other polyhydric alcohols instead of glycerin, their properties were compared and shown in Figure 9.

[0142] As a result, as shown in Fig. 9, in the case of Examples 4 to 6 using different polyhydric alcohols, a large amount of foam was generated during the production of the liquid composition, which may increase the probability of defective products being produced in the future, and thus it was confirmed that these cannot be used as materials for tissue repair.

[0143]

[0144] In addition, the results of evaluating the complex shear viscosity in the same manner as in Experimental Example 2 for Examples 1 to 3, Example 7, and purified water, in which no foam was generated, are shown in Table 8 below.

[0145]

[0146] Test ItemsExample 1Example 2Example 3Example 7Purified Water Viscosity (η*, Pa·s)8.17610.1711.870.037710.02692

[0147] As a result, as shown in Table 8 above, it was confirmed that Example 7 could not be used as a biomaterial for tissue repair because its viscosity was so low that it was close to the level of purified water.

[0148]

[0149] According to the results described above, it was confirmed that the tissue repair composition comprising the DNA fragment mixture of the present invention and the polyhydric alcohol not only has an excellent tissue repair effect, but also has a moisturizing effect, especially in a specific content range combination. These results suggest that the composition does not excessively absorb moisture from adjacent tissues when injected, but provides moisture to adjacent tissues, so that it does not cause dryness in the surrounding tissues but rather alleviates the dryness, and has a high survival rate and good moisturizing effect.

[0150]

[0151] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A composition for tissue repair comprising a mixture of DNA fragments; and a polyhydric alcohol of C3 or C4, The above DNA fragment mixture is included in an amount of 2 to 5 wt% relative to the total composition, The above polyhydric alcohol is included in an amount of 0.5 to 4 wt% relative to the total composition, A composition for tissue repair, wherein the polyhydric alcohol is at least one selected from the group consisting of glycerin, propylene glycol, and butylene glycol.

2. A composition for tissue repair according to claim 1, wherein the composition has tissue moisturizing properties.

3. A composition for tissue repair in claim 1, which increases the moisture content of a tissue when injected into the tissue.

4. A composition for tissue repair, wherein the DNA fragment mixture and polyhydric alcohol are included in a weight ratio of 10:1 to 1:2 in paragraph 1.

5. A composition for tissue repair, wherein the DNA fragment mixture and polyhydric alcohol are included in a weight ratio of 6:1 to 1:1 in paragraph 1.

6. A tissue repair composition according to claim 1, wherein the DNA fragment mixture is a polynucleotide (PN), a polydeoxyribonucleotide (PDRN), or a mixture thereof.

7. A composition for tissue repair according to claim 1, wherein the DNA fragment mixture has a molecular weight of 50 to 10,000 kDa.

8. A biomaterial for tissue repair, comprising a composition according to any one of claims 1 to 7.

9. A filler composition comprising a mixture of DNA fragments; and a polyhydric alcohol of C3 or C4, The above DNA fragment mixture is included in an amount of 2 to 5 wt% relative to the total composition, The above polyhydric alcohol is included in an amount of 0.5 to 4 wt% relative to the total composition, A filler composition, wherein the above polyhydric alcohol is at least one selected from the group consisting of glycerin, propylene glycol, and butylene glycol.

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