Tissue repair components

Abstract

The present invention relates to a tissue repair composition comprising nucleic acid and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan.

Description

Technical Field

[0001] The present invention relates to a tissue repair composition comprising a nucleic acid and at least one substance selected from the group consisting of glycol chitosan (GC), chitosan oligosaccharide lactate (COL), trimethyl chitosan (TMC), methyl glycol chitosan (MGC), carboxymethyl chitosan (CMC), N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan.

Background Art

[0002] As the social structure changes and the population increases, the number of patients with burns, pressure ulcers, trauma, plastic surgery, refractory ulcers, diabetic skin necrosis, etc. is gradually increasing, and accordingly, the treatment methods for damaged skin are also developing. For example, when more than 60% of the body surface area is damaged by burns, it was usually fatal due to sepsis, but recently, with the development of artificial skin, water loss and infection can be prevented, and the mortality rate has decreased significantly. In addition, as the interest in beauty increases, fillers are widely used for cosmetic purposes.

[0003] Currently, the hyaluronic acid filler, which is the most widely used filler, occupies more than 90% of the global filler market. In addition, products in which hyaluronic acid and a cross-linking substance are cross-linked to extend the resorption period are on the market (Patent Document 1). However, even in such cross-linked products, problems due to the toxicity of the cross-linking substance have been reported. Therefore, the development of a tissue repair composition that has excellent physical properties and safety with biocompatibility and no risk of toxicity generation is still required.

Prior Art Documents

[0004] [Patent Document 1] Korean Published Patent Publication No. 10-2014-0072008 [Overview of the Initiative] [Problems that the invention aims to solve]

[0005] The problem that the present invention aims to solve is to provide a tissue repair composition comprising nucleic acid and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. [Means for solving the problem]

[0006] The present invention aims to provide a tissue repair composition comprising nucleic acid and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. [Effects of the Invention]

[0007] The composition of the present invention does not require chemical crosslinking, has excellent stability, does not require a separate sterilization process, and possesses functionality, physical properties, and safety suitable for biomaterials for tissue repair, making it useful as a composition for repairing various tissues. [Brief explanation of the drawing]

[0008] [Figure 1] This figure shows the physical properties of the tissue repair composition of the present invention (experimental example) and the conventional products Rejuran and Conjuran (comparative example). [Figure 2] This is a schematic diagram of an experimental design for evaluating the tissue repair ability and biodegradability of tissue repair compositions. [Figure 3] This figure shows a volume measurement image using a 3D imaging system (PRIMOSCR). [Figure 4] This figure shows the volume change analysis results for the untreated group (Saline) using a 3D imaging system (PRIMOSCR). [Figure 5] This figure shows the volume change analysis results graph for test group 1 (CJR) using a 3D imaging system (PRIMOSCR). [Figure 6] This figure shows the volume change analysis results graph for test group 2 (RJR) using a 3D imaging system (PRIMOSCR). [Figure 7] This figure shows the volume change analysis results for test group 3 (GC) using a 3D imaging system (PRIMOSCR). [Figure 8] This figure shows the volume change analysis results graph for test group 4 (TMC) using a 3D imaging system (PRIMOSCR). [Figure 9] This figure shows the volume change analysis results graph for test group 5 (CMC) using a 3D imaging system (PRIMOSCR). [Figure 10] This figure shows an image of mouse volume change analysis using a 3D imaging system (PRIMOSCR). [Figure 11] This figure shows graphs of group-specific half-life analysis results using a 3D imaging system (PRIMOSCR). [Modes for carrying out the invention]

[0009] These will be explained in detail below. Note that each description and embodiment disclosed in this invention applies to other descriptions and embodiments. That is, any combination of the various elements disclosed in this invention is included. Furthermore, this invention is not limited to the following specific descriptions.

[0010] Furthermore, a person with ordinary skill in the art would be able to recognize and confirm many equivalents of the specific embodiments of the present invention described herein using only ordinary experiments. Moreover, these equivalents are also intended to be included in the present invention.

[0011] One aspect of the present invention provides a tissue repair composition comprising nucleic acid and at least one substance selected from the group consisting of glycol chitosan (GC), chitosan oligosaccharide lactate (COL), trimethyl chitosan (TMC), methyl glycol chitosan (MGC), carboxymethyl chitosan (CMC), N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan.

[0012] In any of the above-described examples, the nucleic acid contained in the composition of the present invention may be present in an amount of 0.1% to 5% by weight relative to the total weight of the composition.

[0013] In any of the foregoing embodiments, any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan contained in the composition of the present invention may be contained in a content of 0.01% to 2% by weight based on the total weight of the composition.

[0014] In any of the foregoing embodiments, the weight ratio of the nucleic acid contained in the composition of the present invention to any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan may be 500:1 to 1:20.

[0015] The nucleic acid of the present invention may be deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or a mixture thereof. Specifically, it is deoxyribonucleic acid, but is not limited thereto.

[0016] In any of the foregoing embodiments, the nucleic acid of the present invention may be selected from oligonucleotide, polynucleotide, and polydeoxyribonucleotide.

[0017] In this invention, "polynucleotide" is also called "PN" and refers to a DNA or RNA chain, which is a polymer of nucleotides in which nucleotide monomers are covalently linked together in a chain. In this invention, "polydeoxyribonucleotide" is also called "PDRN" and refers to a type of low molecular weight DNA complex having a specific molecular weight, but is not limited to these.

[0018] For example, the aforementioned polynucleotides have a relatively longer nucleic acid length or a larger molecular weight compared to polydeoxyribonucleotides, and they act as physical supports, providing cell adhesion, lubrication, and buffering effects, and are therefore used as raw materials for medical devices. Polydeoxyribonucleotides are used as raw materials for pharmaceuticals for cell proliferation and tissue regeneration, but are not limited to these uses.

[0019] In any of the above-described examples, the nucleic acid of the present invention may have a molecular weight of approximately 1 to 100,000 kDa, 5 to 50,000 kDa, 50 to 10,000 kDa, or 50 to 1,500 kDa.

[0020] In any of the embodiments described above, the nucleic acid of the present invention may be obtained by extraction from the testes or semen of a fish. Specifically, the fish may be a salmonid fish. More specifically, it may be salmon or trout, but is not limited to these.

[0021] In the present invention, "glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, or N-hydroxypropyl ether chitosan," which are components of the tissue repair composition together with nucleic acids, are substances that can be mixed with nucleic acids to provide a tissue repair composition.

[0022] Specifically, the substance of the present invention is selected from, but is not limited to, glycol chitosan (GC), trimethyl chitosan (TMC), or carboxymethyl chitosan (CMC).

[0023] In the present invention, "glycol chitosan (GC)" may be a compound represented by the following formula (1).

[0024] [ka]

[0025] In the present invention, "trimethyl chitosan (TMC)" is a multifunctional polymer used in various nanoparticle forms in the fields of pharmaceuticals, functional foods, and biopharmaceuticals. The trimethyl chitosan may also be a compound represented by the following formula (2).

[0026] [ka]

[0027] The "carboxymethyl chitosan (CMC)" in this invention is prepared by bonding carboxymethyl groups to the C-2 and C-6 positions of chitosan, and possesses characteristics such as biodegradability, biocompatibility, and antibacterial properties, and is water-soluble. The carboxymethyl chitosan may also be a compound represented by the following formula (3).

[0028] [ka]

[0029] In any of the embodiments described above, the substances of the present invention, glycol chitosan, chitosan oligosaccharide lactic acid, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan, may be water-soluble.

[0030] In any of the above-described examples, any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactic acid, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan of the present invention may be soluble in a neutral solvent with a pH of about 5.0 to 9.0, and more specifically, may be soluble in a neutral solvent with a pH of about 6.0 to 8.0.

[0031] The composition of the present invention may contain nucleic acids and any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan in specific amounts, and may be used as a tissue repair composition.

[0032] In any of the embodiments described above, the nucleic acid contained in the composition of the present invention may be present in an amount of 0.1% to 5% by weight relative to the total weight of the composition.

[0033] Specifically, the nucleic acids contained in the composition of the present invention are approximately 0.3-5% by weight, 0.5-4% by weight, 0.7-3% by weight, 1-2.8% by weight, 1.2-2.7% by weight, or 1.5-2.5% by weight, based on the total weight of the composition, but are not limited to these amounts.

[0034] In any of the embodiments described above, any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactic acid, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan may be included in the composition of the present invention in an amount of 0.01% to 2% by weight relative to the total weight of the composition.

[0035] Specifically, the composition of the present invention contains any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactic acid, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan in an amount of about 0.01 to 1.8% by weight, 0.05 to 1.6% by weight, 0.07 to 1.4% by weight, 0.08 to 1.2% by weight, 0.09 to 1.0% by weight, or 0.1 to 0.5% by weight, relative to the total weight of the composition, but is not limited to these amounts.

[0036] In any of the embodiments described above, the weight ratio of nucleic acids contained in the composition of the present invention to at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan may be 500:1 to 1:20.

[0037] Specifically, the weight ratio of nucleic acids contained in the composition of the present invention to at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan is about 500:1, 400:1, and 300 :1, 200:1, 150:1, 100:1, 50:1, 25:1, 20:1, 15:1, 10:1, 5:1, 4:1, 3:1, 2:1, 5:4, 4:3, 3:2, 1:1, 2:3, 3:4, 4:5, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:14, 1:16, 1:18, or 1:20, but not limited to these.

[0038] The term "about" as used above includes not only the exact number that follows the term "about," but also a range that is close to or near that number. Considering the context in which the number is used, it is possible to determine whether it is close to or near the specific number mentioned. For example, "about" indicates a range of -10% to +10% of a given number. Another example is that "about" indicates a range of -5% to +5% of a given number. Yet another example is a range that includes, but is not limited to, ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc.

[0039] The compositions of the present invention may dissolve in a neutral solvent without precipitation.

[0040] In this invention, "neutral" means conditions with a pH of approximately 5.0 to 9.0. More specifically, it means conditions with a pH of approximately 5.5 to 8.5, and more specifically, conditions with a pH of approximately 6.0 to 8.0, but is not limited to these conditions.

[0041] The solvent used in the present invention may be any solvent that can dissolve nucleic acids and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. For example, it may be a polar solvent or a hydrophilic solvent, and water is a specific example, but it is not limited to these.

[0042] As one example, the composition of the present invention dissolves without precipitation in a polar solvent with a pH of 5.0 to 9.0, and more specifically, dissolves without precipitation in water with a pH of 6.0 to 8.0, but is not limited to these.

[0043] In any of the embodiments described above, sterile water for injection is used as the solvent for the composition of the present invention, and unless otherwise specified, physiological saline injection, Ringer's solution, or other suitable aqueous solvent may be used instead of sterile water for injection, but is not limited to these.

[0044] The composition of the present invention may further contain a water-soluble polymer.

[0045] Specifically, the water-soluble polymer may be at least one selected from the group consisting of hyaluronic acid, chondroitin sulfate, glycogen, dextrin, dextran, dextran sulfate, hydroxypropyl methylcellulose, alginic acid, chitin, pullulan, collagen, gelatin and its hydrolysates, polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid, and carboxylvinyl polymer.

[0046] The water-soluble polymer of the present invention may be soluble or degradable in living organisms. The water-soluble polymer is used, for example, to control the degradation rate or lubricate the tissue repair composition of the present invention, but is not limited to these uses.

[0047] The term "for tissue repair" is used to encompass all bio-derived materials used for the replacement, repair, and reconstruction of human tissues and organs such as blood vessels, heart, diaphragm, fascia, and skin (dermal, cutaneous, etc.). It is also used in a broad sense to include products containing human-derived skin that has been secondarily processed by mixing in additives to improve its function. It is also used interchangeably with "biomaterials for tissue repair."

[0048] Specifically, it refers to a substance with components similar to human tissue that is inserted into a specific area to expand soft tissue, thereby filling in sunken or missing areas, and is used for wrinkle improvement, contour correction, synovial fluid replenishment, etc. It is used temporarily or semi-permanently for the improvement or repair of wrinkles in the skin of the body, improvement of contour, creation of tissue volume, healing of scars, and other tissue regeneration. The skin and tissues mentioned above include the face, chest, buttocks, genitals, and other body parts.

[0049] The tissue repair composition of the present invention is used in cosmetic fillers, prostheses, joint cavity repair, or combinations thereof, but is not limited to these uses, and may be any composition that is applicable to biomaterials.

[0050] The compositions of the present invention may have physical properties suitable for use in tissue repair. For example, the compositions of the present invention may have elasticity and / or viscosity at a level equivalent to or better than conventional products that can be injected into target tissue.

[0051] In this invention, "elasticity" refers to the solid properties of an object when force is applied, that is, the property of changing shape when force is applied but returning to its original shape when the force is removed. Elasticity refers to the degree to which a desired shape can be maintained under skin tissue, and the higher the elasticity, the longer the initial shape can be maintained. Such elasticity is expressed by the storage modulus (G'), and its unit is Pascal (Pa).

[0052] In one embodiment, the composition of the present invention may have an elasticity of about 80 to 250 Pa under conditions of a temperature of 25°C. For example, the composition exhibits an elastic modulus of about 80 Pa to about 250 Pa, about 90 Pa to about 225 Pa, about 100 Pa to about 200 Pa, about 110 Pa to about 190 Pa, about 120 Pa to about 180 Pa, or about 130 Pa to about 160 Pa. In another embodiment, the tissue repair composition of the present invention exhibits an elastic modulus of about 80 Pa or more, about 90 Pa or more, about 100 Pa or more, about 110 Pa or more, about 120 Pa or more, about 130 Pa or more, about 140 Pa or more, about 150 Pa or more, about 160 Pa or more, about 170 Pa or more, about 180 Pa or more, about 190 Pa or more, about 200 Pa or more, about 225 Pa or more, or about 250 Pa or more.

[0053] Within the elastic range described above, when injected into the body, the tissue repair effect will be uniform, and the higher the elastic modulus, the longer the effect will last in the body.

[0054] In this invention, "viscosity" refers to a quantity that indicates the magnitude of viscosity, which is the resistance to fluid flow. Higher viscosity makes the material easier to work with and is useful for creating delicate shapes.

[0055] In one embodiment, the composition of the present invention may have a viscosity of about 100 to 400 Pa·s under conditions of a temperature of 25°C and a shear rate of 0.1·s-1. For example, the composition exhibits viscosity coefficients of approximately 100 Pa·s to approximately 400 Pa·s, approximately 120 Pa·s to approximately 380 Pa·s, approximately 130 Pa·s to approximately 360 Pa·s, approximately 140 Pa·s to approximately 340 Pa·s, approximately 150 Pa·s to approximately 320 Pa·s, approximately 160 Pa·s to approximately 300 Pa·s, approximately 170 Pa·s to approximately 280 Pa·s, approximately 180 Pa·s to approximately 260 Pa·s, approximately 190 Pa·s to approximately 240 Pa·s, approximately 200 Pa·s to approximately 260 Pa·s, approximately 220 Pa·s to approximately 280 Pa·s, approximately 240 Pa·s to approximately 300 Pa·s, or approximately 260 Pa·s to approximately 320 Pa·s. In other embodiments, the tissue repair composition of the present invention exhibits a viscosity coefficient of approximately 100 Pa·s or more, approximately 120 Pa·s or more, approximately 150 Pa·s or more, approximately 180 Pa·s or more, approximately 200 Pa·s or more, approximately 220 Pa·s or more, approximately 240 Pa·s or more, approximately 260 Pa·s or more, approximately 280 Pa·s or more, approximately 300 Pa·s or more, approximately 320 Pa·s or more, approximately 340 Pa·s or more, approximately 360 Pa·s or more, approximately 380 Pa·s or more, or approximately 400 Pa·s or more.

[0056] It is important to adjust the viscosity and elasticity ranges within the above ranges to inject a tissue repair composition suitable for the target site.

[0057] In any of the embodiments described above, the composition of the present invention may be characterized by volume retention.

[0058] In any of the embodiments described above, the composition of the present invention may be characterized by its biodegradability.

[0059] For use in tissue repair, a material must possess biocompatibility and appropriate volume retention to create a sense of volume in human tissue. It is also preferable that it possesses biodegradability, meaning it breaks down after a predetermined time and is naturally removed through the body's metabolic processes. Without biocompatibility and appropriate volume retention, tissue regeneration, such as improving or repairing wrinkles in the skin, improving facial contours, creating a sense of volume, and healing scars, becomes difficult. Furthermore, a lack of biodegradability in the body may cause harmful side effects such as allergic reactions.

[0060] The composition of the present invention has been confirmed to be a tissue repair composition that does not pose a risk of toxicity, possesses excellent biocompatibility, safety, appropriate tissue repair ability, and / or biodegradability.

[0061] For example, animal experiments evaluated the tissue repair ability and / or biodegradability of the composition of the present invention and confirmed that it exhibited tissue repair ability and biodegradability at a level equivalent to that of the comparative examples, Conjuran and Rejuran. Therefore, it was found that the composition of the present invention can repair tissue for a predetermined time due to its volume persistence, and can be stably used as a biomaterial for tissue repair due to its appropriate biodegradability.

[0062] Another aspect of the present invention provides a method for producing a tissue repair composition, comprising the step of dissolving nucleic acid and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan in a solvent.

[0063] The solvent used in the manufacturing method of the present invention may be any solvent that can dissolve nucleic acids and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. Specifically, a neutral solvent can be used. Specifically, the solvent is water, but is not limited to water.

[0064] The solvent of the present invention is characterized by being a mixture of nucleic acids and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. Therefore, it does not require the addition of a separate acidic solution for dissolution, and does not require the addition of raw materials and solutions for neutralizing the effects of the composition itself or its in vivo administration, making it suitable for use as an injectable preparation.

[0065] In the dissolution step, the order in which nucleic acids and at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan are mixed with a solvent is not particularly defined (in any order), and they may be dissolved simultaneously.

[0066] The method for producing the tissue repair composition of the present invention may further include processing steps such as dust removal, disinfection, and sterilization.

[0067] In one embodiment, the manufacturing method of the present invention may further include a filtration step before and / or after dissolution. The type of filter used here can be appropriately changed depending on the purpose, and for example, a 0.20um to 0.50um filter can be used, but is not limited thereto.

[0068] In one embodiment, the manufacturing method of the present invention may include a sterilization step.

[0069] The aforementioned sterilization method may be any known method, such as autoclaving, high-temperature sterilization, filtration, chemical treatment, and / or radiation treatment, but is not limited to these.

[0070] The sterilization step of the present invention is not limited to this, but may be performed after the dissolution step. For example, the sterilization step after the dissolution step is high-temperature sterilization, but is not limited to this.

[0071] In any of the embodiments described above, the sterilization process may be performed as the final step of the manufacturing process. A sterilization process performed as the final step of the manufacturing process is called a "post-sterilization process."

[0072] Specifically, the post-sterilization process of the present invention includes a high-temperature sterilization process.

[0073] The manufacturing method of the present invention is characterized in that no precipitate is formed in the sterilization step after mixing nucleic acids, which are characteristic raw materials, with at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. Therefore, the manufacturing method of the present invention may not require the step of sterilizing each raw material before mixing nucleic acids with at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan, and mixing them in a sterile facility.

[0074] The tissue repair composition produced by the manufacturing method of the present invention may be filled into a container suitable for use. For example, the composition may be provided in the form of a syringe, with single or multiple doses of the composition filled into a syringe barrel. Here, the syringe may be equipped with a drug extrusion plunger or the like, and capable of extruding the composition of the present invention.

[0075] A further aspect of the present invention provides a tissue repair method comprising the step of administering the tissue repair composition of the present invention to an individual.

[0076] In this invention, "tissue repair" means the replacement, repair, or reconstruction of human tissues and organs such as blood vessels, heart, diaphragm, fascia, and skin (dermal, cutaneous, etc.), and may also refer to physical repair.

[0077] In this invention, "individual" refers to any animal, including humans, that requires tissue repair. This animal may be a mammal, but is not limited to humans.

[0078] In this invention, "administration" means introducing the tissue repair composition of the present invention into an individual by an appropriate method, specifically by injection, but is not limited thereto. The administration route of the composition of the present invention can be various routes, such as subcutaneous, dermal, blood vessels, biological membranes, tissue fibers, and synovial fluid, as long as it is capable of performing tissue repair. The dosage forms are topical preparations, subcutaneous injections, dermal injections, intravascular injections, intramuscular injections, intraarticular injections, tendon sheath (tendon) injections, and ligament injections, but are not limited thereto.

[0079] Yet another aspect of the present invention provides a tissue repair application for the tissue repair composition of the present invention.

[0080] The tissue repair composition and tissue repair described above are as previously stated. [Examples]

[0081] The present invention will be described in more detail below with reference to examples and experimental cases. However, these examples and experimental cases are merely illustrative of the present invention, and the present invention is not limited to these examples and experimental cases.

[0082] (Example 1) Preparation of tissue repair compositions containing nucleic acids and GC, TMC, or CMC. A tissue repair composition containing nucleic acid and glycol chitosan (GC), trimethyl chitosan (TMC), or carboxymethyl chitosan (CMC) was prepared. As comparative examples, Conjuran (PN 2 wt, PharmaResearch Co., Ltd.), which is used for joint cavity repair, and Rejuran (PN 2 wt, PharmaResearch Co., Ltd.), which is used for facial repair, were used.

[0083] Table 1 shows the constituent components used in each composition and the content of each component relative to the total composition.

[0084] [Table 1]

[0085] (Example 2) Physical property evaluation of tissue repair compositions The physical properties of test groups 3-5, prepared in Example 1, were compared and evaluated with those of comparative examples, test groups 1 and 2. The evaluated items and results are shown in Table 2.

[0086] [Table 2]

[0087] The experimental results showed that test groups 3-5, newly discovered in this invention, exhibited physical properties equivalent to, or even higher than, those of the comparative examples Conjuran (test group 1) and Rejuran (test group 2), which have been used as conventional tissue repair compositions. Therefore, it was found that they can be injected into target tissues that require volume and shape retention after treatment.

[0088] (Example 3) Evaluation of tissue repair ability and biodegradability of tissue repair compositions 3-1: Evaluation of tissue repair ability of tissue repair compositions To evaluate the tissue repair ability of the composition of the present invention, a 3D imaging system (PRIMOS) was used. CR The presence or absence of volume retention was measured by evaluation.

[0089] Specifically, 18 six-week-old male SKH-1 hairless mice were purchased from Orient Bio Co., Ltd. and reared under the following conditions: temperature: 20-24°C, humidity: 40-60%, differential pressure: negative pressure, noise: less than 60 dB, 12-hour light-dark cycle. After purchasing the experimental animals, a one-week acclimatization period was followed before use in the experiment. Anesthesia was administered by inhalation, maintaining an isoflurane concentration of 3.0%. Once anesthesia was confirmed, the experimental animals were moved to the operating table, and then 100 μl of the substance was administered subcutaneously to three mice per group using a 1 mL syringe for the untreated group (Saline), test group 1 (CJR), test group 2 (RJR), test group 3 (GC), test group 4 (TMC), and test group 5 (CMC). The administration times of the test substance are shown in Table 3, and a schematic diagram of the experiment is shown in Figure 2.

[0090] [Table 3]

[0091] As shown in Table 3, experimental animals were anesthetized by inhalation with isoflurane 3.0% at 0, 4, 8, 12, 24, 36, 48, and 60 hours after administration of the test substance, a camera was fixed in place, and then a dimensional imaging system (PRIMOS) using a high-resolution sensor was used. CR Images were captured using Canfield (USA), and volume measurement and analysis were performed using PRIMOS 5.0 software. Furthermore, tissue repair capacity was judged to be present if the volume was sustained, with 70% being the baseline.

[0092] All calculated data were validated for statistical significance using SPSS Package Program version 20 (IBM, USA). The evaluation results were compared with the control group using the ANOVA method (*p<0.05, **p<0.01, ***p<0.001).

[0093]

number

[0094] 3D imaging system (PRIMOS CRAnalysis of volume changes using ) revealed that in the untreated group (Saline), tissue repair ability was maintained for more than 4 hours and degraded within 12 hours; in test group 1 (CJR), tissue repair ability was maintained for more than 24 hours and degraded within 48 hours; in test group 2 (RJR), tissue repair ability was maintained for more than 12 hours and degraded within 48 hours; in test group 3 (GC), tissue repair ability was maintained for more than 24 hours and degraded within 48 hours; in test group 4 (TMC), tissue repair ability was maintained for more than 24 hours and degraded within 48 hours; and in test group 5 (CMC), tissue repair ability was maintained for 8 to 12 hours or more and degraded within 48 hours (Table 4 and Figures 4-10).

[0095] [Table 4] JPEG2026512089000010.jpg117170

[0096] 3-2: Evaluation of biodegradability of tissue repair compositions To evaluate the biodegradability of the tissue repair composition of the present invention, half-life analysis was performed.

[0097] Specifically, the test substance of the present invention was administered, and the trend of change between the time of maximum volume and the time of minimum volume at 0, 4, 8, 12, 24, 48, and 60 hours was expressed as an exponential function. Then, the X (time) value at which the Y (volume) value became half of the maximum volume was determined and analyzed.

[0098] All calculated data were validated for statistical significance using SPSS Package Program version 20 (IBM, USA). The evaluation results were compared with the control group using the ANOVA method (*p<0.05, **p<0.01, ***p<0.001).

[0099] Analysis of the half-lives revealed that the half-lives were 5.75 hours for the untreated group (Saline), 21.45 hours for test group 1 (CJR), 17.15 hours for test group 2 (RJR), 18.40 hours for test group 3 (GC), 20.59 hours for test group 4 (TMC), and 16.69 hours for test group 5 (CMC) (Table 5 and Figure 11).

[0100] [Table 5]

[0101] In summary, test groups 1 (CJR), 3 (GC), and 4 (TMC) of the present invention exhibited tissue repair ability for approximately 24 hours, test groups 2 (RJR) and 5 (CMC) exhibited tissue repair ability for 8 to 12 hours or more, and all groups were confirmed to decompose within 48 hours.

[0102] These results show that the newly discovered test groups 3 to 5 in the present invention also exhibit tissue repair ability and biodegradability at a level equivalent to that of Conjuran (test group 1) and Rejuran (test group 2), which are comparative examples used as conventional tissue repair compositions.

[0103] This suggests that the test material of the present invention, due to its volume-sustaining properties, can repair tissue for a predetermined period of time, and, due to its appropriate biodegradability, can be stably used as a biomaterial for tissue repair.

[0104] From the above description, those skilled in the art in the field to which the present invention pertains will understand that the present invention can be implemented in other specific forms without altering its technical idea or essential features. It should be understood that the above embodiments are merely illustrative and not limiting. The present invention should be interpreted as encompassing all modified or altered forms derived from the meaning and scope of the claims and their equivalent concepts, rather than the specification.

Claims

1. nucleic acids and, It comprises at least one substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan. Composition for tissue repair.

2. The composition according to claim 1, wherein the nucleic acid is contained in an amount of 0.1% to 5% by weight relative to the total weight of the composition.

3. The composition according to claim 1, wherein any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan is included in an amount of 0.01% to 2% by weight of the total weight of the composition.

4. The composition according to claim 1, wherein the weight ratio of the nucleic acid to any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan is 500:1 to 1:

20.

5. The composition according to claim 1, wherein the nucleic acid has a molecular weight of 50 kDa to 10,000 kDa.

6. The composition according to claim 1, wherein the nucleic acid is isolated from the testes or semen of a fish.

7. The composition according to claim 6, characterized in that the fish is of the Salmonidae family.

8. The composition according to claim 1, wherein the substance is one or more selected from glycol chitosan, trimethyl chitosan, or carboxymethyl chitosan.

9. The composition according to claim 1, wherein any substance selected from the group consisting of glycol chitosan, chitosan oligosaccharide lactate, trimethyl chitosan, methyl glycol chitosan, carboxymethyl chitosan, N-hydroxymethyl chitosan, N-hydroxypropyl chitosan, and N-hydroxypropyl ether chitosan is soluble in a neutral solvent with a pH of 5 to 9.

10. The composition according to claim 1, further comprising a water-soluble polymer.

11. The composition according to claim 10, wherein the water-soluble polymer is at least one selected from the group consisting of hyaluronic acid, chondroitin sulfate, glycogen, dextrin, dextran, dextran sulfate, hydroxypropyl methylcellulose, alginic acid, chitin, pullulan, collagen, gelatin and its hydrolysates, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylic acid, and carboxylvinyl polymer.

12. The composition according to claim 1, wherein the elasticity of the composition is 90 to 250 Pa.

13. The composition according to claim 1, wherein the viscosity of the composition is 100 to 400 Pa·s.

14. The composition according to claim 1, characterized by its volume retention properties.

15. The composition according to claim 1, characterized by its biodegradability.

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