Medical Composition

Abstract

The present invention relates to a medical composition containing polyhydroxyalkanoate (PHA), which has excellent biodegradability, biocompatibility, adhesive strength to biological tissue, wound healing efficacy, and hemostatic efficacy. Therefore, the medical composition of the present invention can be used for hemostasis, wound healing, tissue adhesion, or bacterial infection prevention.

Description

[Technical Field]

[0001] The present invention relates to a medical composition that can be applied to the living tissue of humans or animals, and a method for producing the same. [Background technology]

[0002] When bleeding occurs due to an injury in daily life or in an industrial setting, surgical procedures are performed to quickly and safely stop the bleeding and prevent excessive bleeding from the wound site. Effective hemostasis and suturing of the wound site during such surgical procedures can reduce the amount of bleeding and the amount of blood transfusion required by the patient, thereby facilitating the patient's recovery, and therefore, these procedures must be performed appropriately.

[0003] Conventionally, medical devices such as gauze, bandages, surgical sutures, staplers, or electric / laser instruments have been used for the hemostasis and suturing treatments, but these have had the problem of causing side effects such as damage or infection at the wound site when the patient's biological tissue (e.g., skin) is weak.

[0004] To solve these problems, hemostatic agents or medical adhesives have been developed. When applied to a wound site where bleeding occurs, the hemostatic agents or medical adhesives exhibit physical, chemical, or physiologically active properties to temporarily prevent bleeding and help the wound site heal. Furthermore, because they are applied by coating to the wound site, they can minimize damage or infection to the wound site even if the patient's biological tissue is delicate.

[0005] However, currently developed hemostatic agents or medical adhesives have limitations such as not being able to exhibit the adhesive strength required for suturing wound sites, or causing immune reactions and poor biocompatibility. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Republic of Korea Patent Publication No. 2013-0055847 Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention aims to provide a medical composition that exhibits high adhesive strength to biological tissues while minimizing the occurrence of immune reactions, and has excellent hemostatic efficacy, wound healing efficacy, bacterial infection suppression efficacy, antibacterial efficacy, etc.

[0008] Another object of the present invention is to provide a method for producing the medical composition. [Means for solving the problem]

[0009] In order to solve the above problems, one aspect of the present invention provides a medical composition containing polyhydroxyalkanoate (PHA) having a particle size of 10,000 nm or less and containing 40 wt% or more of repeating units derived from 3-hydroxybutyrate (3HB) based on the total weight.

[0010] In one embodiment, the medical composition may have a tissue adhesive strength of 15 kPa or more measured at 100% relative humidity and room temperature.

[0011] In another embodiment, the polyhydroxyalkanoate (PHA) may further include repeating units derived from 4-hydroxybutyrate (4HB).

[0012] In yet another embodiment, the content of the repeating unit derived from 4-hydroxybutyrate (4HB) may be 0.1 to 60% by weight based on the total weight of the polyhydroxyalkanoate (PHA).

[0013] In yet another embodiment, the polyhydroxyalkanoate (PHA) may be poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer.

[0014] In yet another embodiment, the polyhydroxyalkanoate (PHA) may have a molecular weight of 10,000 to 1,200,000 g / mol.

[0015] In yet another embodiment, the concentration of the polyhydroxyalkanoate (PHA) contained in the medical composition may be 0.1 to 75% (w / v).

[0016] In yet another embodiment, the medical composition may be used for hemostasis.

[0017] In yet another embodiment, the medical composition may be used for wound healing.

[0018] In yet another embodiment, the medical composition may be used for tissue adhesion.

[0019] In yet another embodiment, the medical composition may be used to inhibit bacterial infection.

[0020] Meanwhile, another aspect of the present invention provides a method for preparing a medical composition, including the steps of: dissolving polyhydroxyalkanoate (PHA) in a solvent to prepare a dispersed phase solution; preparing a continuous phase solution containing a surfactant; passing the dispersed phase solution through a membrane having a pore size of 10,000 nm or less to form an emulsion in which dispersed phase particles are dispersed in the continuous phase solution; and solidifying the emulsion to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less.

[0021] In one embodiment, the content of the polyhydroxyalkanoate (PHA) contained in the dispersed phase solution may be 0.01 to 5 wt % based on the total weight of the dispersed phase solution.

[0022] In another embodiment, the dispersed phase solution may be passed through the membrane at a pressure of 2.5 to 320 kPa.

[0023] In accordance with yet another aspect of the present invention, there is provided a method for preparing a medical composition, the method comprising the steps of: dissolving polyhydroxyalkanoate (PHA) in a solvent to prepare a dispersed phase solution; preparing a continuous phase solution containing a surfactant; mixing the dispersed phase solution and the continuous phase solution to form a premixed emulsion; feeding the premixed emulsion into a high-pressure dispersing device (microfluidizer) to form an emulsion in which dispersed phase particles having a particle size of 10,000 nm or less are dispersed; and solidifying the emulsion to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less.

[0024] In one embodiment, the pressure applied to the premix emulsion by the high-pressure dispersion device may be 1 to 30 kpsi. [Effects of the Invention]

[0025] The medical composition of the present invention contains polyhydroxyalkanoate (PHA) having a particle size controlled within a specific range, and thus exhibits excellent biodegradability and biocompatibility (low immunogenicity) while also exhibiting strong adhesive strength to biological tissue.

[0026] Therefore, the medical composition of the present invention can be effectively used for hemostasis, healing, or suturing of human or animal biological tissues. In particular, the medical composition of the present invention exhibits strong adhesive strength even to biological tissues in the presence of water (e.g., tissues with wet surfaces or tissues submerged in water), and can exhibit excellent hemostatic efficacy when used to stop bleeding at a wound site in water.

[0027] Furthermore, the medical composition according to the present invention can be formulated into various forms depending on the characteristics of the application site of biological tissue. In particular, the medical composition according to the present invention contains polyhydroxyalkanoate (PHA), which can be mass-produced using a microbial system, making it possible to economically provide a formulated hemostatic agent or medical adhesive. [Brief explanation of the drawings]

[0028] [Figure 1] 1 is an image (SEM) showing polyhydroxyalkanoate (PHA) particles produced in Synthesis Examples 1 to 5 and 10 to 15 according to the present invention. [Figure 2] 1 is an image (SEM) showing polyhydroxyalkanoate (PHA) particles produced in Synthesis Examples 1 to 5 and 10 to 15 according to the present invention. [Figure 3] 1 shows the results of analyzing the changes in size and particle size distribution of polyhydroxyalkanoate (PHA) particles using a light scattering analysis method in Test Example 2 according to the present invention. [Figure 4] 1 shows the results of analyzing the changes in size and particle size distribution of polyhydroxyalkanoate (PHA) particles using a light scattering analysis method in Test Example 2 according to the present invention. [Figure 5] 1 shows the results of measuring the expression levels of inflammatory cytokine factors (TNF-α and IL-6) in Test Example 3 according to the present invention. [Figure 6] 1 shows the results of measuring the expression levels of inflammatory cytokine factors (TNF-α and IL-6) in Test Example 3 according to the present invention. [Figure 7] 1 shows the results of evaluation of the adhesive performance of a medical composition to pig skin in a 100% humid environment in Test Example 4 according to the present invention. [Figure 8] 1 shows the results of evaluating the adhesive performance of a medical composition to a hydrogel in Test Example 5 according to the present invention. [Figure 9] 1 shows images of the wound site over time in Test Example 6 according to the present invention. [Figure 10] 1 shows the results of calculations using the Image J program for changes in the wound site in Test Example 6 according to the present invention. [Figure 11] 1 is an image showing the skin tissue and blood vessels at the wound site in Test Example 6 according to the present invention. [Figure 12] 1 shows images of the wound site over time in Test Example 7 according to the present invention. [Figure 13] 1 shows images of the wound site over time in Test Example 7 according to the present invention. [Figure 14] 10 is an image showing the results of gene analysis in Test Example 8 according to the present invention. [Figure 15] 10 is an image showing the results of gene analysis in Test Example 8 according to the present invention. [Figure 16] 10 is an image showing the results of gene analysis in Test Example 8 according to the present invention. [Figure 17] 10 is an image showing the results of gene analysis in Test Example 8 according to the present invention. [Figure 18] 10 is an image showing the results of gene analysis in Test Example 8 according to the present invention. [Figure 19] 1 shows the results of evaluating the hemostatic efficacy of a medical composition in Test Example 9 according to the present invention. [Figure 20] 1 shows the results of evaluation of the hemostatic efficacy of the medical composition in Test Example 10 according to the present invention. [Figure 21] 1 shows the results of evaluating the bacterial infection suppression efficacy of a medical composition in Test Example 11 according to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0029] The present invention will be described below with reference to the following embodiments. However, the present invention is not limited to the following disclosed contents, and can be modified in various forms as long as the gist of the invention is not changed.

[0030] In this specification, when a component is described as being formed above / below another component, or as being connected or joined to each other, this includes all components that are formed, connected, or joined between these components directly or indirectly via other components. It should be understood that the reference to above / below each component may change depending on the direction in which the object is observed.

[0031] In this specification, the term "comprising" is intended to embody certain features, regions, steps, processes, elements and / or components, and does not exclude the presence or addition of other features, regions, steps, processes, elements and / or components, unless specifically stated to the contrary.

[0032] All numbers and expressions indicating amounts of components, reaction conditions, and the like described herein may be understood to be modified in all instances by the word "about," unless otherwise specified.

[0033] The present invention relates to a biocompatible composition containing biocompatible polymer particles. Specifically, the present invention relates to a medical composition that can bond biological tissues and exhibit hemostatic effects, wound healing effects, bacterial infection suppression effects, antibacterial effects, etc., on biological tissues, which will be described in detail below.

[0034] Composition of the medical composition A medical composition according to one embodiment of the present invention includes polyhydroxyalkanoate (PHA) having a particle size controlled within a specific range. Specifically, the medical composition according to one embodiment of the present invention may be a composition in which polyhydroxyalkanoate (PHA) particles having a particle size within a specific range are dispersed in a common solvent or solution.

[0035] The polyhydroxyalkanoate (PHA) is a particle having a particle size (average particle size) of 10,000 nm or less. Specifically, the particle size of the polyhydroxyalkanoate (PHA) may be 9,000 nm or less, 8,000 nm or less, 7,000 nm or less, 6,000 nm or less, 5,000 nm or less, 3,000 nm or less, 1,000 nm or less, 700 nm or less, 500 nm or less, 300 nm or less, or 200 nm or less. For example, the polyhydroxyalkanoate (PHA) may have a particle size (average particle size) of 10 to 10,000 nm, 10 to 9,000 nm, 13 to 8,500 nm, 13 to 6,500 nm, 15 to 5,000 nm, 15 to 4,000 nm, 18 to 3,500 nm, 18 to 2,000 nm, 20 to 1,500 nm, 23 to 1,000 nm, 23 to 950 nm, 25 to The particle size of the polyhydroxyalkanoate (PHA) may be 800 nm, 25-700 nm, 28-600 nm, 28-500 nm, 30-400 nm, 30-300 nm, 50-290 nm, 60-285 nm, 70-280 nm, 90-275 nm, 100-270 nm, 110-260 nm, 120-250 nm, 130-240 nm, 140-230 nm, or 150-260 nm. By having the particle size of the polyhydroxyalkanoate (PHA) within this range, the medical composition of the present invention can be applied uniformly (high density) to biological tissue, maximizing the adhesive area and adhesive strength. In particular, controlling the particle size of the polyhydroxyalkanoate (PHA) increases the surface area of the polyhydroxyalkanoate (PHA), which increases its interaction with biological tissue. By satisfying this requirement, the present invention can optimize the hemostatic and / or adhesive functions of the medical composition. Furthermore, since the particle size of the polyhydroxyalkanoate (PHA) is within the above range, the medical composition of the present invention can efficiently penetrate into injured biological tissue (biological tissue cells) and act as a nutrient, thereby allowing the medical composition of the present invention to have excellent wound healing efficacy.

[0036] The polyhydroxyalkanoate (PHA) may have a particle size variation of ±0.1 μm or less. Specifically, the particle size variation may be ±0.09 μm or less, ±0.07 μm or less, ±0.05 μm or less, or ±0.03 μm or less.

[0037] The particle size and particle size variation of the polyhydroxyalkanoate (PHA) may refer to values measured using a nanoparticle size analyzer. Specifically, the average particle size and particle size variation of the polyhydroxyalkanoate (PHA) can be measured using a Zetasizer Nano ZS (manufactured by Marven) nanoparticle size analyzer at a temperature of 25°C and a measurement angle of 175° using dynamic light scattering (DLS). In this case, the peak value derived from the polydispersity index (PDI) at a confidence interval of 0.5 can be defined as the particle size of the polyhydroxyalkanoate (PHA).

[0038] The polyhydroxyalkanoate (PHA) may have a polydispersity index (PDI) of 1 or less, specifically 0.001 to 1, 0.003 to 0.9, 0.005 to 0.8, or 0.005 to 0.6.

[0039] By ensuring that the particle size variation and polydispersity index of the polyhydroxyalkanoate (PHA) are within the above ranges, the dispersibility of the polyhydroxyalkanoate (PHA) particles and the processability of the medical composition can be improved.

[0040] The polyhydroxyalkanoate (PHA) contains repeating units (3HB repeating units) derived from 3-hydroxybutyrate (3HB). The content of the 3HB repeating units may be 40% by weight or more, specifically 40 to 99.9% by weight, 42 to 99.5% by weight, 45 to 99% by weight, 46 to 98.5% by weight, 47 to 98% by weight, 48 to 97% by weight, 50 to 96% by weight, 51 to 95% by weight, 52 to 94.5% by weight, 55 to 94% by weight, 60 to 93.5% by weight, 65 to 93% by weight, 70 to 93% by weight, 75 to 92.5% by weight, 80 to 92% by weight, or 82 to 91.5% by weight, based on the total weight of the polyhydroxyalkanoate (PHA).

[0041] The polyhydroxyalkanoate (PHA) may further contain repeating units derived from one or more selected from the group consisting of 4-hydroxybutyrate (4HB), 3-hydroxypropionate (3HP), 3-hydroxyhexanoate (3HH), 3-hydroxyvalerate (3HV), 4-hydroxyvalerate (4HV), 5-hydroxyvalerate (5HV), and 6-hydroxyhexanoate (6HH).

[0042] Specifically, the polyhydroxyalkanoate (PHA) may further contain a repeating unit (4HB repeating unit) derived from 4-hydroxybutyrate (4HB). By including the 4HB repeating unit in the polyhydroxyalkanoate (PHA), the biodegradability, biocompatibility, hemostatic efficacy, wound healing efficacy, and adhesive performance of the medical composition can be improved.

[0043] The content of the 4HB repeating unit may be 0.1 to 60 wt % based on the total weight of the polyhydroxyalkanoate (PHA), specifically 0.5 to 58 wt %, 1 to 55 wt %, 1.5 to 54 wt %, 2 to 53 wt %, 3 to 52 wt %, 4 to 50 wt %, 5 to 49 wt %, 6 to 48 wt %, 6 to 45 wt %, 6.5 to 40 wt %, 7 to 35 wt %, 7 to 30 wt %, 7.5 to 25 wt %, 8 to 20 wt %, or 8.5 to 18 wt %.

[0044] Specifically, the polyhydroxyalkanoate (PHA) may be poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer, which can improve the biodegradability, biocompatibility, hemostatic efficacy, wound healing efficacy, and adhesive performance of the medical composition.

[0045] Meanwhile, the crystallinity of the polyhydroxyalkanoate (PHA) is controlled depending on the content of the 4HB repeating unit, and it can be classified as semi-crystalline PHA (scPHA) or amorphous PHA (aPHA).

[0046] Specifically, the scPHA may have a 4HB repeat unit content of 0.1 to 30 wt%, 1 to 28 wt%, 3 to 26 wt%, 5 to 25 wt%, 6 to 23 wt%, 8 to 21 wt%, or 10 to 20 wt%, and the aPHA may have a 4HB repeat unit content of 15 to 60 wt%, 20 to 58 wt%, 25 to 55 wt%, 35 to 53 wt%, 40 to 50 wt%, 43 to 49 wt%, or 45 to 48 wt%.

[0047] The polyhydroxyalkanoate (PHA) may consist of the scPHA alone, the aPHA alone, or a mixture of these.

[0048] The polyhydroxyalkanoate (PHA) may have a crystallinity of 90% or less, 85% or less, 80% or less, 75% or less, or 70% or less, as measured by differential scanning calorimeter (DSC).

[0049] The molecular weight (weight average molecular weight) of the polyhydroxyalkanoate (PHA) may be 10,000 to 1,200,000 g / mol, specifically 20,000 to 1,100,000 g / mol, 30,000 to 1,000,000 g / mol, 40,000 to 900,000 g / mol, 45,000 to 850,000 g / mol, 50,000 to 80 The molecular weight of the polyhydroxyalkanoate (PHA) may be 0,000 g / mol, 60,000 to 750,000 g / mol, 70,000 to 700,000 g / mol, 80,000 to 600,000 g / mol, 90,000 to 500,000 g / mol, 100,000 to 400,000 g / mol, 200,000 to 800,000 g / mol, or 300,000 to 700,000 g / mol. By having the molecular weight of the polyhydroxyalkanoate (PHA) within the above range, the adhesive performance of the medical composition to biological tissue can be improved.

[0050] The polyhydroxyalkanoate (PHA) has a glass transition temperature (T g ) may be −45 to 80° C., −35 to 50° C., −20 to 20° C., or −15 to 0° C. Furthermore, the polyhydroxyalkanoate (PHA) may have a crystallization temperature (T c ) may not be measured, or may be 60 to 120°C, 70 to 115°C, 75 to 110°C, or 80 to 105°C. In addition, the polyhydroxyalkanoate (PHA) may have a melting temperature (T m ) may not be measured, or may be 100 to 170°C, 105 to 160°C, 110 to 150°C, or 115 to 140°C.

[0051] The concentration of the polyhydroxyalkanoate (PHA) may be 0.1 to 75% (w / v) (0.1 to 75 wt% based on the total weight of the medical composition). Specifically, the concentration of the polyhydroxyalkanoate (PHA) contained in the medical composition according to one embodiment of the present invention may be 0.2 to 70% (w / v), 0.2 to 60% (w / v), 0.2 to 50% (w / v), 0.3 to 40% (w / v), 0.3 to 30% (w / v), 0.3 to 25% (w / v), 0.3 to 16% (w / v), 0.35 to 15.6% (w / v), 0.7 to 13% (w / v), 3 to 11% (w / v), or 6 to 9% (w / v). When the concentration of the polyhydroxyalkanoate (PHA) is within the above range, the hemostatic effect, wound healing effect and adhesive performance of the medical composition can be significantly improved.

[0052] The medical composition according to one embodiment of the present invention, which includes the polyhydroxyalkanoate (PHA), may have a tissue adhesive strength of 15 kPa or more, measured at 100% relative humidity and room temperature (e.g., 20±5°C, specifically 25°C). Specifically, the medical composition according to one embodiment of the present invention may have an adhesive strength to biological tissue of 15 to 300 kPa, 18 to 200 kPa, 20 to 100 kPa, or 30 to 50 kPa. When the adhesive strength to biological tissue is within the above range, the medical composition can exhibit high adhesive performance not only to biological tissue in the absence of water, but also to biological tissue in the presence of water (e.g., tissue with a wet surface or tissue in water).

[0053] The medical composition according to one embodiment of the present invention may further include a biocompatible polymer and / or adhesive substance other than the polyhydroxyalkanoate (PHA). The biocompatible polymer may be, for example, catechol, caffeic acid, gallic acid, tannin; a polysaccharide-based polymer such as chitosan, hyaluronic acid, alginic acid, dextran, or a protein-based polymer such as collagen or gelatin. The adhesive substance may be, for example, protamine, mussel adhesive protein, or squid sucker ring tooth protein.

[0054] The medical composition according to one embodiment of the present invention may further contain a known pharmacologically active substance, which may be specifically an analgesic, anti-inflammatory, antibacterial, or antifungal agent.

[0055] The medical composition according to one embodiment of the present invention may be formulated into various forms, such as a powder form, a liquid form, a gel form, or a sheet form (film form).

[0056] Uses of the medical composition The medical composition according to one embodiment of the present invention may be used for various purposes, specifically for hemostasis, wound healing, bedsore healing, tissue adhesion, or bacterial infection suppression, which will be described in detail below.

[0057] Composition for hemostasis A medical composition according to one embodiment of the present invention may be a hemostatic composition containing polyhydroxyalkanoate (PHA). Polyhydroxyalkanoate (PHA) has low immunogenicity, excellent biocompatibility, and can effectively stop bleeding. By containing such polyhydroxyalkanoate (PHA), the hemostatic composition according to one embodiment of the present invention can efficiently stop bleeding in injured biological tissue.

[0058] The polyhydroxyalkanoate (PHA) contained in the hemostatic composition according to one embodiment of the present invention has substantially the same structure and characteristics as the polyhydroxyalkanoate (PHA) described above in the "Composition of the medical composition" section.

[0059] Preferably, the polyhydroxyalkanoate (PHA) contained in the hemostatic composition may contain 0.1 to 30 wt%, 1 to 28 wt%, 3 to 26 wt%, 5 to 25 wt%, 6 to 23 wt%, or 10 to 20 wt% of repeating units derived from 4-hydroxybutyrate (4HB), based on the total weight of the polyhydroxyalkanoate (PHA), and may have a molecular weight of 30,000 to 1,000,000 g / mol, 40,000 to 900,000 g / mol, 45,000 to 850,000 g / mol, or 50,000 to 800,000 g / mol. Furthermore, the polyhydroxyalkanoate (PHA) contained in the hemostatic composition may be particles having a particle size (average particle size) of 25 to 700 nm, 25 to 600 nm, 25 to 500 nm, 30 to 400 nm, 30 to 300 nm, 50 to 250 nm, 50 to 230 nm, 70 to 220 nm, 100 to 210 nm, 120 to 200 nm, 130 to 190 nm, 140 to 180 nm, or 150 to 180 nm.

[0060] The concentration of the polyhydroxyalkanoate (PHA) contained in the hemostatic composition according to one embodiment of the present invention may be 0.1 to 25% (w / v) (0.1 to 25 wt% based on the total weight of the hemostatic composition), specifically 0.3 to 16% (w / v), 0.5 to 15% (w / v), 0.7 to 13% (w / v), 1 to 10% (w / v), 2 to 9% (w / v), 3 to 9% (w / v), or 6 to 8% (w / v). When the concentration of the polyhydroxyalkanoate (PHA) is within the above range, the hemostatic efficacy of the hemostatic composition can be significantly improved.

[0061] The hemostatic composition according to one embodiment of the present invention may contain a common solvent or solution that is harmless to the living body (human body) and capable of uniformly dispersing polyhydroxyalkanoate (PHA). The solvent or solution is not particularly limited, but examples thereof include purified water, saline, and PBS buffer solution.

[0062] Furthermore, the hemostatic composition according to one embodiment of the present invention may further optionally contain the above-mentioned biocompatible polymer, the above-mentioned adhesive substance, and the above-mentioned pharmacologically active substance.

[0063] Wound healing composition A medical composition according to one embodiment of the present invention may be a wound healing composition containing the polyhydroxyalkanoate (PHA) described above. The polyhydroxyalkanoate (PHA) serves as a nutrient that enables injured biological tissue (biological tissue cells) to rapidly recover, and can act to accelerate the recovery rate of injured biological tissue. By containing such polyhydroxyalkanoate (PHA), the wound healing composition according to one embodiment of the present invention can efficiently heal (treat) injured biological tissue.

[0064] The polyhydroxyalkanoate (PHA) contained in the wound healing composition of one embodiment of the present invention has substantially the same structure and characteristics as the polyhydroxyalkanoate (PHA) described above in the "Composition of the medical composition" section.

[0065] Preferably, the polyhydroxyalkanoate (PHA) contained in the wound healing composition contains 0.1 to 30 wt%, 1 to 28 wt%, 3 to 26 wt%, 5 to 25 wt%, 6 to 23 wt%, or 10 to 20 wt% of repeating units derived from 4-hydroxybutyrate (4HB), based on the total weight of the polyhydroxyalkanoate (PHA), and may have a molecular weight of 30,000 to 1,000,000 g / mol, 40,000 to 900,000 g / mol, 45,000 to 850,000 g / mol, or 50,000 to 800,000 g / mol. Furthermore, the polyhydroxyalkanoate (PHA) contained in the wound healing composition may be particles having a particle size (average particle size) of 25 to 700 nm, 25 to 600 nm, 25 to 500 nm, 30 to 400 nm, 30 to 300 nm, 50 to 250 nm, 50 to 230 nm, 70 to 220 nm, 100 to 210 nm, 120 to 200 nm, 130 to 190 nm, 140 to 180 nm, or 150 to 180 nm.

[0066] The concentration of the polyhydroxyalkanoate (PHA) contained in the wound healing composition according to one embodiment of the present invention may be 0.1 to 25% (w / v) (0.1 to 25 wt% based on the total weight of the wound healing composition), specifically 0.3 to 16% (w / v), 0.5 to 15% (w / v), 0.7 to 13% (w / v), 1 to 12% (w / v), 2 to 11% (w / v), 3 to 10% (w / v), 5 to 9% (w / v), or 6 to 8% (w / v). By having the polyhydroxyalkanoate (PHA) concentration within the above range, the wound healing efficacy of the wound healing composition can be significantly enhanced.

[0067] The wound healing composition according to one embodiment of the present invention may contain a conventional solvent or solution that is harmless to the living body (human body) and capable of uniformly dispersing polyhydroxyalkanoate (PHA). The solvent or solution is not particularly limited, but examples thereof include purified water, saline, and PBS buffer solution.

[0068] Furthermore, the wound healing composition according to one embodiment of the present invention may further optionally contain the above-mentioned biocompatible polymer, the above-mentioned adhesive substance, and the above-mentioned pharmacologically active substance.

[0069] Composition for tissue adhesion A medical composition according to one embodiment of the present invention may be a tissue adhesive composition containing the polyhydroxyalkanoate (PHA). The polyhydroxyalkanoate (PHA) contains various functional groups (e.g., OH groups) and therefore can be applied not only to dry biological tissues (e.g., skin tissues) but also to wet biological tissues, whereby it can strongly bond (adhere) the tissue to other biological tissues. The tissue adhesive composition according to one embodiment of the present invention, containing such a polyhydroxyalkanoate (PHA), can efficiently suture (join) injured biological tissues.

[0070] The polyhydroxyalkanoate (PHA) contained in the tissue adhesive composition according to one embodiment of the present invention has substantially the same structure and characteristics as the polyhydroxyalkanoate (PHA) described above in the "Composition of the medical composition" section.

[0071] Preferably, the polyhydroxyalkanoate (PHA) contained in the tissue adhesive composition may contain 0.1 to 30 wt%, 1 to 28 wt%, 3 to 26 wt%, 4 to 25 wt%, 5 to 23 wt%, 6 to 20 wt%, 7 to 18 wt%, or 8 to 19 wt% of repeating units derived from 4-hydroxybutyrate (4HB) based on the total weight of the polyhydroxyalkanoate (PHA), and may have a molecular weight of 100,000 to 1,000,000 g / mol, 150,000 to 900,000 g / mol, 200,000 to 800,000 g / mol, or 300,000 to 700,000 g / mol. Furthermore, the polyhydroxyalkanoate (PHA) contained in the tissue adhesive composition may be particles having a particle size (average particle size) of 25 to 700 nm, 25 to 650 nm, 25 to 600 nm, 30 to 550 nm, 30 to 500 nm, 50 to 450 nm, 50 to 400 nm, 70 to 350 nm, 100 to 300 nm, 110 to 290 nm, 120 to 280 nm, 130 to 270 nm, or 140 to 260 nm.

[0072] The concentration of the polyhydroxyalkanoate (PHA) contained in the tissue adhesive composition according to one embodiment of the present invention may be 0.1 to 25% (w / v) (0.1 to 25 wt% based on the total weight of the tissue adhesive composition), and more specifically, may be 0.3 to 16% (w / v), 0.5 to 15% (w / v), 0.7 to 14% (w / v), 1 to 13% (w / v), 2 to 12% (w / v), 3 to 11% (w / v), 4 to 10% (w / v), or 5 to 10% (w / v). When the concentration of the polyhydroxyalkanoate (PHA) is within the above range, the biological tissue adhesive performance of the tissue adhesive composition can be significantly improved.

[0073] The tissue adhesive composition according to one embodiment of the present invention may have an adhesive strength to tissue (biological tissue) of 15 kPa or more, specifically 15 to 300 kPa, 16 to 270 kPa, 17 to 250 kPa, 18 to 200 kPa, 19 to 150 kPa, 20 to 100 kPa, 25 to 80 kPa, 28 to 60 kPa, or 30 to 50 kPa, measured at 100% relative humidity and room temperature (for example, 20±5°C, specifically 25°C). Furthermore, the tissue adhesive composition according to one embodiment of the present invention may have an adhesive strength to a hydrogel of 1 to 300 J / m 2 Specifically, it may be 2 to 280 J / m 2 , 3~250J / m 2 , 5~230J / m 2 , 6~200J / m 2 , 7~180J / m 2 , 8~150J / m 2 , 9~100J / m 2 , 10~80J / m 2 , 13~70J / m 2 , 15~60J / m 2 , 17~50J / m 2 , or 20-45J / m 2 When the adhesive strength is within the above range, the tissue adhesive composition can exhibit high adhesive performance not only to biological tissues that do not contain water, but also to biological tissues that contain water (for example, tissues with a wet surface or tissues in water).

[0074] The tissue adhesive composition according to one embodiment of the present invention may contain a common solvent or solution that is harmless to living organisms and capable of uniformly dispersing polyhydroxyalkanoate (PHA). The solvent or solution is not particularly limited, but specific examples thereof include purified water, saline, and PBS buffer solution.

[0075] Furthermore, the tissue adhesive composition according to one embodiment of the present invention may further optionally contain the above-mentioned biocompatible polymer, the above-mentioned adhesive substance, and the above-mentioned pharmacologically active substance.

[0076] Compositions for inhibiting bacterial infection (antibacterial or sterilizing compositions) A medical composition according to one embodiment of the present invention may be a composition for inhibiting bacterial infection, which contains polyhydroxyalkanoate (PHA). Polyhydroxyalkanoate (PHA) can act to inhibit the proliferation and metabolic function of bacteria (e.g., germs, bacteria, etc.). By including such polyhydroxyalkanoate (PHA), the composition for inhibiting bacterial infection according to one embodiment of the present invention can efficiently inhibit bacterial infection in injured or surgically treated biological tissue.

[0077] The polyhydroxyalkanoate (PHA) contained in the composition for inhibiting bacterial infection in one embodiment of the present invention has substantially the same structure and characteristics as the polyhydroxyalkanoate (PHA) described above in the "Composition of the medical composition" section.

[0078] Preferably, the polyhydroxyalkanoate (PHA) contained in the composition for inhibiting bacterial infection may contain 0.1 to 30 wt%, 1 to 28 wt%, 3 to 26 wt%, 5 to 25 wt%, 6 to 23 wt%, or 10 to 20 wt% of repeating units derived from 4-hydroxybutyrate (4HB), based on the total weight of the polyhydroxyalkanoate (PHA), and may have a molecular weight of 30,000 to 1,000,000 g / mol, 40,000 to 900,000 g / mol, 45,000 to 850,000 g / mol, or 50,000 to 800,000 g / mol. Furthermore, the polyhydroxyalkanoate (PHA) contained in the composition for inhibiting bacterial infection may be particles having a particle size (average particle size) of 25 to 700 nm, 25 to 600 nm, 25 to 500 nm, 30 to 400 nm, 30 to 300 nm, 50 to 250 nm, 50 to 230 nm, 70 to 220 nm, 100 to 210 nm, 120 to 200 nm, 130 to 190 nm, 140 to 180 nm, or 150 to 180 nm.

[0079] The concentration of the polyhydroxyalkanoate (PHA) contained in the bacterial infection-suppressing composition according to one embodiment of the present invention may be 0.1 to 75% (w / v) (0.1 to 75 wt% based on the total weight of the bacterial infection-suppressing composition), specifically 5 to 75% (w / v), 10 to 75% (w / v), 15 to 70% (w / v), 20 to 70% (w / v), 25 to 65% (w / v), 30 to 65% (w / v), or 35 to 55% (w / v). By ensuring that the concentration of the polyhydroxyalkanoate (PHA) is within the above range, the bacterial infection-suppressing efficacy of the bacterial infection-suppressing composition can be significantly enhanced.

[0080] The composition for inhibiting bacterial infection according to one embodiment of the present invention may contain a common solvent or solution that is harmless to the living body (human body) and can uniformly disperse polyhydroxyalkanoate (PHA). The solvent or solution is not particularly limited, but specific examples include purified water, saline, PBS buffer solution, etc.

[0081] Furthermore, the composition for inhibiting bacterial infection according to one embodiment of the present invention may further optionally contain the above-mentioned biocompatible polymer, the above-mentioned adhesive substance, and the above-mentioned pharmacologically active substance.

[0082] Method for producing a medical composition The present invention makes it possible to produce a medical composition through a process of granulating polyhydroxyalkanoate (PHA) to have a particle size that can exhibit optimal biocompatibility.

[0083] Specifically, a method for preparing a medical composition according to one embodiment of the present invention includes the steps of: (S-1) dissolving polyhydroxyalkanoate (PHA) in a solvent to prepare a dispersed phase solution; (S-2) preparing a continuous phase solution containing a surfactant; (S-3) passing the dispersed phase solution through a membrane having a pore size of 10,000 nm or less to form an emulsion in which dispersed phase particles are dispersed in the continuous phase solution; and (S-4) solidifying the emulsion to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less.

[0084] Step S-1 is a step of dissolving (dispersing) the initial raw material polyhydroxyalkanoate (PHA) in a solvent to prepare a dispersed phase solution in which polyhydroxyalkanoate (PHA) is dispersed.

[0085] The polyhydroxyalkanoate (PHA) used as the initial raw material may be obtained by cell disruption of microorganisms using mechanical or physical methods, or may be obtained by cell disruption of microorganisms using non-mechanical or chemical methods.

[0086] The solvent is not particularly limited as long as it can dissolve the initial raw material polyhydroxyalkanoate (PHA), but specifically may be one or more selected from the group consisting of chloroform, chloroethane, dichloromethane, dichloroethane, butyl acetate, and trichloroethane.

[0087] On the other hand, in order to make it easier for the polyhydroxyalkanoate (PHA) to be uniformly dissolved in the solvent, the polyhydroxyalkanoate (PHA) may be dissolved by stirring at 40 to 50° C. for 45 to 55 hours.

[0088] The dispersed phase solution prepared through this process may contain 0.01 to 5 wt% of the initial raw material, polyhydroxyalkanoate (PHA). Specifically, the dispersed phase solution may contain 0.02 to 4.5 wt%, 0.05 to 4 wt%, 0.08 to 3.5 wt%, 0.1 to 3 wt%, 0.2 to 2.5 wt%, 0.35 to 2 wt%, 0.4 to 1.5 wt%, 0.5 to 1.3 wt%, 0.6 to 1.2 wt%, 0.7 to 1 wt%, or 0.75 to 0.9 wt%, based on the total weight of the dispersed phase solution. By ensuring that the polyhydroxyalkanoate (PHA) content in the dispersed phase solution is within this range, polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less and a uniform (spherical) morphology can be obtained.

[0089] Step S-2 is a step of preparing a continuous phase solution containing a surfactant (emulsifier). Specifically, the continuous phase solution may contain a surfactant and an aqueous solvent.

[0090] Specifically, the surfactant may be at least one selected from the group consisting of sodium dodecyl sulfate, sodium lauryl sulfate, sodium laureth sulfate, sodium stearate, sodium cocoyl glycinate, and perfluorooctane sulfonate.

[0091] The aqueous solvent may be water, distilled water, deionized water, pure water, ultrapure water, or the like.

[0092] The stirring speed during the preparation of the continuous phase solution may be 1 to 800 rpm, 50 to 600 rpm, or 200 to 500 rpm. By keeping the stirring speed of the continuous phase solution within this range, polyhydroxyalkanoate (PHA) particles having a uniform nano-size can be obtained.

[0093] The step S-2 of preparing the continuous phase solution may be performed simultaneously with the step S-1, or may be performed before or after step S-1.

[0094] Step S-3 is a step in which the dispersed phase solution prepared in step S-1 is passed through a membrane having a pore size of 10,000 nm or less to form an emulsion in which dispersed phase particles (PHA dispersed phase particles) are dispersed in the continuous phase solution prepared in step S-2.

[0095] Specifically, the membrane may be an inorganic porous membrane (e.g., SPG (Shirasu Porous Glass)) having a pore size of 10,000 nm or less, so that the resulting polyhydroxyalkanoate (PHA) particles have a desired particle size of 10,000 nm or less.

[0096] The pressure at which the dispersed phase solution passes through the membrane may be 2.5 to 320 kPa, specifically 50 to 320 kPa, 100 to 320 kPa, or 200 to 320 kPa. More specifically, the pressure at which the dispersed phase solution passes through the membrane may be adjusted depending on the pore size; the larger the pores, the lower the pressure required. By keeping the pressure within this range, polyhydroxyalkanoate (PHA) particles with a uniform morphology (spherical shape) can be obtained.

[0097] Step S-4 is a step of solidifying the emulsion containing the dispersed phase particles to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less. Solidifying the emulsion may mean removing the solvent used to dissolve the polyhydroxyalkanoate (PHA) in step S-1. Solidifying the emulsion can be performed by air drying, vacuum drying using an oven, or drying using a rotary evaporator. By solidifying the emulsion, a product containing the polyhydroxyalkanoate (PHA) particles (e.g., an aqueous dispersion in which PHA particles having a particle size of 10,000 nm or less are dispersed) can be obtained.

[0098] A method for preparing a medical composition according to another embodiment of the present invention includes the steps of: dissolving polyhydroxyalkanoate (PHA) in a solvent to prepare a dispersed phase solution (S-1'); preparing a continuous phase solution containing a surfactant (S-2'); mixing the dispersed phase solution with the continuous phase solution to form a premix emulsion (S-3'); feeding the premix emulsion into a high-pressure dispersing device to form an emulsion in which dispersed phase particles having a particle size of 10,000 nm or less are dispersed (S-4'); and solidifying the emulsion to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less (S-5').

[0099] The steps S-1' and S-2' have substantially the same configurations and features as the steps S-1 and S-2, respectively, and a detailed description thereof will be omitted.

[0100] The step S-3' is a step of mixing the dispersed phase solution prepared in the step S-1' and the continuous phase solution prepared in the step S-2' to form a premix emulsion (A).

[0101] The mixing for forming the premix emulsion (A) may be performed using a conventional homogenizer at a stirring speed of 5,000 to 15,000 rpm for 1 to 5 minutes.

[0102] Step S-4' is a step of feeding the premix emulsion (A) into a high-pressure dispersion device to form an emulsion (B) in which dispersed phase particles having a particle size of 10,000 nm or less are dispersed.

[0103] The high-pressure dispersing device (high-pressure dispersing and emulsifying device) is a device that applies pressure to the premix emulsion (A) to make the droplets of the premix emulsion (A) finer, and a typical high-pressure dispersing device can be used.

[0104] The pressure applied to the premix emulsion (A) by the high-pressure dispersing device may be 1 to 30 kpsi, specifically 3 to 25 kpsi, 5 to 20 kpsi, or 9 to 15 kpsi. More specifically, the operating pressure of the high-pressure dispersing device may be adjusted according to the content of polyhydroxyalkanoate (PHA) contained in the dispersed phase solution, and the higher the content of polyhydroxyalkanoate (PHA), the higher the operating pressure. When the pressure is within this range, polyhydroxyalkanoate (PHA) particles having a uniform morphology (spherical shape) can be obtained.

[0105] The step S-5’ is a step of solidifying the emulsion B containing the dispersed phase particles to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less. The solidification of the emulsion may mean removing the solvent used to dissolve polyhydroxyalkanoate (PHA) in the step S-1’. The solidification of such an emulsion (B) can be carried out by natural drying, vacuum drying using an oven, drying using a rotary evaporation concentrator, or the like. By undergoing the solidification of the emulsion (B), a product containing the polyhydroxyalkanoate (PHA) particles (for example, an aqueous dispersion in which PHA particles having a particle size of 10,000 nm or less are dispersed) can be obtained.

[0106] By producing polyhydroxyalkanoate (PHA) particles through the membrane emulsification method using a membrane or the emulsification method using a high-pressure dispersion device in this way, the present invention can easily produce polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less and a uniform shape.

[0107] The polyhydroxyalkanoate (PHA) particles (products containing polyhydroxyalkanoate (PHA) particles) respectively obtained by the step S-4 and the step S-5’ can further undergo processes such as sterilization. Thereafter, the obtained polyhydroxyalkanoate (PHA) particles (products containing polyhydroxyalkanoate (PHA) particles) can be applied as a medical composition according to an embodiment of the present invention by themselves. Furthermore, the obtained polyhydroxyalkanoate (PHA) particles can be applied as a medical composition according to an embodiment of the present invention after undergoing dosage form formation such as powderization, liquefaction, gelation, sheet formation (film formation), and the like.

Example

[0108] Hereinafter, the present invention will be described more specifically through examples. However, the scope of the present invention is not limited by these examples.

[0109] <Production of PHA particles> Synthesis Example 1

[0110] PHA particles were prepared using membrane emulsification. Specifically, poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer ([P(3HB-co-4HB)], 4HB repeat unit: 17 wt%, molecular weight: 687,000 g / mol) was dissolved in chloroform to prepare 10 ml of dispersed phase solution (PHA content: 0.4 wt% or less). Furthermore, 100 ml of continuous phase aqueous solution containing 0.3 wt% of the surfactant sodium dodecyl sulfate was prepared.

[0111] The emulsion was induced by applying the prepared dispersed and continuous phase solutions to an IMK-40 (MCTech) emulsifier using an SPG membrane. The membrane pore size was 300 nm, and the membrane pressure was controlled at 150-320 kPa and the stirring speed at 100-300 rpm.

[0112] 200 ml of the formed emulsion was poured into a container capable of maintaining an interfacial height of less than 10 mm, and the chloroform was gradually evaporated at room temperature for 48 hours to induce solidification of the emulsion, thereby obtaining PHA particles (specifically, an aqueous dispersion containing PHA particles) with a particle size of 300 nm or less.

[0113] Synthesis Examples 2-5 PHA particles (specifically, aqueous dispersions containing PHA particles) were obtained by the same process as in Synthesis Example 1, except that membranes with pore sizes of 200 nm (Synthesis Example 2), 1 μm (Synthesis Example 3), 5 μm (Synthesis Example 4), and 10 μm (Synthesis Example 5), respectively, were used.

[0114] Synthesis Examples 6-9 PHA particles (specifically, PHA particle-containing aqueous dispersions) were obtained in the same manner as in Synthesis Example 1, except that the composition and molecular weight of the poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer dissolved in chloroform were adjusted as shown in Table 1 below.

[0115] Synthesis Example 10 PHA particles were produced using a membrane emulsification method. Specifically, poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer ([P(3HB-co-4HB)], 4HB repeat unit: 8.7 wt%, molecular weight: 390,000 g / mol) was dissolved in chloroform to prepare 10 ml of dispersed phase solution (PHA content: 1.0 wt% or less). Furthermore, 100 ml of continuous phase aqueous solution containing 0.3 wt% of the surfactant sodium dodecyl sulfate was prepared.

[0116] The emulsion was induced by applying the prepared dispersed and continuous phase solutions to an IMK-40 (MCTech) emulsifier using an SPG membrane. The membrane pore size was 300 nm, and the membrane pressure was controlled at 150–320 kPa and the stirring speed at 100–300 rpm.

[0117] 200 ml of the resulting emulsion was poured into a 1 L round-bottom flask, and the chloroform was evaporated at 57°C and 150 mbar for 4 hours to induce solidification of the emulsion, thereby obtaining PHA particles (specifically, an aqueous dispersion containing PHA particles) with a particle size of 300 nm or less.

[0118] Synthesis Examples 11-14 PHA particles (specifically, aqueous dispersions containing PHA particles) were obtained by the same process as in Synthesis Example 10, except that membranes with pore sizes of 1 μm (Synthesis Example 11), 2 μm (Synthesis Example 12), 5 μm (Synthesis Example 13), and 10 μm (Synthesis Example 14) were used.

[0119] Synthesis Examples 15-17 PHA particles (specifically, PHA particle-containing aqueous dispersions) were obtained by the same process as in Synthesis Example 10, except that the content (concentration) of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer dissolved in chloroform was adjusted as shown in Table 1 below.

[0120] Synthesis Examples 18-20 PHA particles (specifically, PHA particle-containing aqueous dispersions) were obtained by the same process as in Synthesis Example 11, except that the content (concentration) of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer dissolved in chloroform was adjusted as shown in Table 1 below.

[0121] Synthesis Examples 21-25 PHA particles (specifically, PHA particle-containing aqueous dispersions) were obtained by the same process as in Synthesis Example 10, except that the composition and molecular weight of the poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer dissolved in chloroform were adjusted as shown in Table 1 below.

[0122] Synthesis Example 26 PHA particles were produced by an emulsification method using a high-pressure dispersion device. Specifically, poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer ([P(3HB-co-4HB)], 4HB repeat unit: 8.7 wt%, molecular weight: 390,000 g / mol) was dissolved in chloroform to prepare 25 ml of dispersed phase solution (PHA content: 5 wt%). Furthermore, 500 ml of continuous phase aqueous solution containing 0.3 wt% of the surfactant sodium dodecyl sulfate was prepared.

[0123] The dispersed phase solution was added to the continuous phase aqueous solution and mixed for 2 minutes using a homogenizer (IKA ULTRA-TURRAX (registered trademark) T 25 digital, IKA) set at 13,600 rpm to form a premix emulsion.

[0124] Next, the premix emulsion was introduced into a high-pressure dispersion device (LM20, Microfluidics) and pressure was applied to induce the formation of a homogenized emulsion, which was controlled at 10 kpsi.

[0125] 200 ml of the resulting emulsion was poured into a 1 L round-bottom flask, and the chloroform was evaporated at 57°C and 150 mbar for 4 hours to induce solidification of the emulsion, thereby obtaining PHA particles (specifically, an aqueous dispersion containing PHA particles) with a particle size of 300 nm or less.

[0126] [Table 1]

[0127] Test Example 1 - Confirmation of PHA particles The PHA particles obtained in Synthesis Examples 1 to 5 and 10 to 15 were examined using a scanning electron microscope (SEM), and the results are shown in FIGS.

[0128] 1 and 2, it can be seen that PHA particles having a uniform size and a spherical shape can be appropriately produced by preparing PHA particles by the membrane emulsification method using a membrane.

[0129] Test Example 2 - Analysis of PHA particle distribution PHA particles were obtained by the same process as in Synthesis Example 1, except that the PHA content (concentration) was adjusted to 0.042-0.334 wt% when preparing the dispersed phase solution. The size and distribution of the obtained PHA particles were analyzed by light scattering analysis (Zetasizer Nano ZS, Malvern Instruments, UK), and the results are shown in Figure 3.

[0130] PHA particles were obtained through the same process as in Synthesis Examples 10 and 11, except that the PHA content (concentration) was adjusted to 0.1-0.8 wt% when preparing the dispersed phase solution. The size and distribution of the obtained PHA particles were analyzed using a light scattering analysis method (Zetasizer Nano ZS, Malvern Instruments, UK), and the results are shown in Figure 4.

[0131] 3, it can be seen that PHA particles having a particle size of 300 nm or less can be appropriately produced when the content of PHA (initial raw material) in the dispersed phase solution is within the range of 0.01 to 0.5 wt%. Also, FIG. 4, it can be seen that PHA particles having a particle size of 300 nm or less can be appropriately produced when the content of PHA (initial raw material) in the dispersed phase solution is within the range of 0.1 to 1 wt% and the membrane pore size is 0.3 μm.

[0132] Test Example 3 - Evaluation of PHA immunogenicity The immunogenicity of semi-crystalline PHA (scPHA, [P(3HB-co-4HB)] form, 17 wt% repeating units derived from 4HB, molecular weight: 687,000 g / mol) from Synthesis Example 1 and amorphous PHA (aPHA, [P(3HB-co-4HB)] form, 48 wt% repeating units derived from 4HB, molecular weight: 800,000 g / mol) from Synthesis Example 9 was evaluated, and the results are shown in Figures 5 and 6. Specifically, Raw 264.7 cells were suspended in DMEM (Dulbecco's Modified Eagle Medium) containing 10% FBS (Fetal Bovine Serum), and then 1 x 10 cells were plated in a 48-well plate. 5The cells were aliquoted to a cell count of 1000 cells / ml and cultured for 12 hours in a CO2 incubator at 37°C. Subsequently, scPHA or aPHA, dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4), was treated at concentrations of 10 μg / ml, 100 μg / ml, 500 μg / ml, and 1000 μg / ml, respectively, and cultured for 24 hours. After incubation, the cell culture medium was sampled and the contents of the inflammatory cytokines TNF-α and IL-6 were measured using an ELISA kit. The positive control was LPS (lipopolysaccharides from Escherichia coli O55:B5 (Cat# L2880, Sigma-Aldrich, St. Louis, MO, USA)), and the negative control was 0.1 M PBS buffer (pH 7.4) (NT).

[0133] 5 and 6, it can be seen that the PHAs according to the present invention, aPHA 10 μg / ml, aPHA 100 μg / ml, aPHA 500 μg / ml, aPHA 1000 μg / ml, scPHA 10 μg / ml, scPHA 100 μg / ml, and scPHA 1000 μg / ml, suppress the expression of inflammatory cytokine factors TNF-α and IL-6. In particular, when compared with aPHA 1000 μg / ml and scPHA 1000 μg / ml and LPS (positive control group) treated at the same concentration, it can be seen that the PHAs according to the present invention significantly reduced the expression of inflammatory cytokine factors compared to LPS.

[0134] These results support the fact that the PHA of the present invention has low immunogenicity and excellent biocompatibility.

[0135] <Production of medical composition> Example 1 PHA particles ([P(3HB-co-4HB)] with a particle size of 178 nm, 4HB repeating unit: 17 wt%, molecular weight: 687,000 g / mol) were obtained through the same process as in Synthesis Example 1, except that the pore size of the membrane was controlled. The obtained PHA particles were dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4) at concentrations ranging from 0.3 to 16% (w / v) to prepare a composition.

[0136] Example 2 PHA particles ([P(3HB-co-4HB)] with a particle size of 259 nm, 4HB repeating unit: 17 wt%, molecular weight: 687,000 g / mol) were obtained through the same process as in Synthesis Example 1, except that the pore size of the membrane was controlled. The obtained PHA particles were dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4) at concentrations ranging from 0.3 to 16% (w / v) to prepare a composition.

[0137] Example 3 PHA particles ([P(3HB-co-4HB)] with a particle size of 180 nm, 4HB repeating units: 8.7 wt%, molecular weight: 390,000 g / mol) were obtained through the same process as in Synthesis Example 11, except that the pore size of the membrane was controlled. The concentration of the obtained PHA particles was adjusted to a range of 0.1-10% (w / v) to prepare a composition. The concentration of the PHA particles was adjusted by concentrating them using a rotary evaporator or by diluting them with purified water.

[0138] Example 4 A composition was prepared in the same manner as in Example 3, except that PHA particles ([P(3HB-co-4HB)] with a particle size of 580 nm, 4HB repeating unit: 8.7 wt%, molecular weight: 390,000 g / mol) were obtained.

[0139] Example 5 A composition was prepared in the same manner as in Example 3, except that PHA particles ([P(3HB-co-4HB)] with a particle size of 850 nm, 4HB repeating unit: 8.7 wt%, molecular weight: 390,000 g / mol) were obtained.

[0140] Example 6 A composition was prepared in the same manner as in Example 3, except that PHA particles ([P(3HB-co-4HB)] with a particle size of 4740 nm, 4HB repeating unit: 8.7 wt%, molecular weight: 390,000 g / mol) were obtained.

[0141] Example 7 A composition was prepared in the same manner as in Example 3, except that PHA particles ([P(3HB-co-4HB)] with a particle size of 8190 nm, 4HB repeating unit: 8.7 wt%, molecular weight: 390,000 g / mol) were obtained.

[0142] Comparative Example 1 Fibrin glue (Green Cross, Korea) (FG) was applied.

[0143] Test Example 4 - Adhesion Performance Evaluation 1 The adhesive strength of the composition to biological tissue was evaluated through shear stress measurement using a universal testing machine (Instron 5544, Norwood, MA, US), and the results are shown in Figure 7. Specifically, pig skin (Stellen Medical, USA) was cut into 10 mm x 10 mm pieces and placed in 0.1 M PBS buffer (pH 7.4) at 37°C for 1 hour. After the aging process, the pig skin was attached to a 10 mm x 100 mm aluminum bar using instant adhesive (3M). The samples prepared in Examples 1 and 2 and the fibrin glue prepared in Comparative Example 1 were applied to the surface of the pig skin. Next, the aluminum bars with the pig skin without the sample attached were placed on top of each other, and the two aluminum bars were secured together using clips. The two fixed aluminum bars were incubated at 100% relative humidity and room temperature (RT) for 2 hours, and then the shear stress until the two aluminum bars were completely separated was measured using a 10 kN load cell at a crosshead speed of 10 mm / min.

[0144] 7, it can be seen that Examples 1 and 2, which are compositions according to the present invention, exhibit adhesive strength equal to or greater than that of the fibrin glue of Comparative Example 1. In particular, it can be seen that when the particle size of the PHA particles is 259 nm and the concentration of the PHA particles is 3 to 16% (w / v), a significantly high adhesive strength is exhibited.

[0145] These results support the need to control the particle size of PHA particles in order to increase their adhesive strength to biological tissue, and also support the fact that the medical composition (tissue adhesive composition) of the present invention exhibits strong adhesive strength even in environments where moisture is present.

[0146] Test Example 5 - Adhesion Performance Evaluation 2 The adhesive strength of the composition to biological tissue was evaluated through shear stress measurement using a universal testing machine (QC-508E, Cometech, Taiwan). The results are shown in Figure 8. Specifically, 27.6 mg of N,N'-methylenebis(acrylamide) and 4,920 mg of potassium peroxodisulfate were dissolved in 30 g of deionized water at 4°C, followed by the addition of 18.528 ml of N,N-dimethylacrylamide and 270 μl of N,N,N',N'-tetramethylethylenediamine. The resulting solution was poured into a 1.5 mm wide mold and crosslinked at 30°C for 16 hours to produce a hydrogel. The resulting hydrogel was cut into 5 mm x 30 mm pieces, and 10 μl of the compositions prepared in Examples 3 to 7 was applied to each 5 mm x 10 mm area on the surface of the hydrogel. Next, two hydrogel pieces with the applied samples were stacked and left at room temperature (RT) for 4 hours. After 4 hours, the shear stress was measured using a 50 N load cell at a cross head speed of 10 mm / min until the two hydrogel pieces were completely separated.

[0147] 8, it can be seen that the smaller the particle size of the PHA particles, the higher the adhesive strength. In particular, when the particle size of the PHA particles is 300 nm or less and the concentration of the PHA particles is 3 to 10% (w / v), a significantly high adhesive strength is exhibited.

[0148] These results also support the need to control the particle size of PHA particles in order to increase their adhesive strength to biological tissue, and that the medical composition (tissue adhesive composition) of the present invention exhibits strong adhesive strength even in environments where moisture is present.

[0149] Test Example 6 - Wound recovery evaluation 1 A wound healing test was conducted on a composition prepared by dissolving (dispersing) the 178 nm diameter PHA particles ([P(3HB-co-4HB)] obtained in Example 1 (4HB repeating unit: 17 wt%, molecular weight: 687,000 g / mol) in 0.1 M PBS buffer (pH 7.4) at a concentration of 7.8% (w / v). The results are shown in Figures 9 to 11. Specifically, Sprague-Dawley (SD) rats of matched sex and age, aged 7 weeks or less, and sterilized surgical instruments were prepared. The composition containing the PHA particles dissolved (dispersed) at a concentration of 7.8% (w / v) was sterilized with ultraviolet light. Next, the prepared rats were anesthetized using a respiratory anesthesia machine containing isoflurane solution. After shaving the backs of anesthetized rats, a circular wound was created using an 8mm diameter biopsy trephine (Kai Medical, Japan). The UV-sterilized composition was applied, followed by a sterile waterproof film to prevent scratching. Wound healing was monitored over 21 days by simultaneous photography and the wound area was calculated using the Image J program. On days 7 and 14, rats were euthanized with carbon dioxide. Skin tissue surrounding the wound was cut into 10mm x 10mm pieces, fixed in formalin, and stained with hematoxylin and eosin (H&E) and Masson's trichrome (MT). Images of the stained areas were confirmed using a Leica microscope. A negative control group (NT) was left untreated, and a positive control group (FG) was treated with fibrin glue (Green Cross, Korea).

[0150] 9 and 10, it can be seen that the medical composition (PHA) according to the present invention reduces the area of the wound site in a short time compared to the control group.

[0151] Referring to Figure 11(c) (hematoxylin-eosin (H&E) staining results), it can be seen that when treated with the medical composition (PHA) of the present invention, wound repair occurred rapidly, and numerous new blood vessels were generated. These newly generated blood vessels support the formation of temporary granulation tissue, promoting new tissue growth and delivering nutrients and oxygen to the expanded tissue necessary for the healing process. Furthermore, the angiogenesis process helps remove waste and dead cells from the wound site, speeding up the healing process. Meanwhile, when treated with the medical composition (PHA) of the present invention, faster re-epithelialization (epidermal restoration) occurred compared to the control group. In particular, newly formed epithelial tongues were only observed in the group treated with the medical composition (PHA) of the present invention from day 3 onwards.

[0152] Referring to Figure 11(d) (results of Masson's Trichrome (MT) staining), it can be seen that when the medical composition (PHA) according to the present invention is treated, the granulation tissue is remodeled more rapidly with a more mature collagen content structure.

[0153] These results support the fact that the medical composition (wound healing composition) of the present invention has wound healing efficacy.

[0154] Test Example 7 - Wound recovery evaluation 2 PHA particles having a particle size of 178 nm obtained in Example 1 ([P(3HB-co-4HB)], 4HB repeating unit: 17 wt%, molecular weight: 687,000 g / mol) and PHA particles having a particle size of 180 nm obtained in Example 3 ([P(3HB-co-4HB)], 4HB repeating unit: 8.7 wt%, molecular weight: 390,000 g / mol) were dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4) at a concentration of 25% (w / v). Compositions (PHA1 (4HB repeating unit: 17 wt%), PHA2 (4HB repeating unit: 8.7 wt%)) were used to conduct a wound healing test (evaluation of in vivo wound healing ability). The results are shown in Figures 12 and 13. Specifically, Sprague-Dawley (SD) rats were anesthetized with isoflurane, and their backs were shaved. A 1.5 cm long and 3.0 mm deep incision was made in the skin to create a thin wound. Next, 10 μl of PHA1 and PHA2 compositions were applied to the wound site, and the wound was allowed to set for 3 minutes. Seven days later, the rats were euthanized, and a 1 cm x 2 cm incision was made in the skin to encompass the wound site. The incised skin was then fixed in p-formaldehyde solution (3.7 wt%) and embedded in paraffin. Cross-sectional sections of the skin were stained with hematoxylin and eosin (H&E). Images of the stained areas were then confirmed using a Leica microscope.

[0155] 12 and 13, it can be seen that the medical compositions according to the present invention (PHA1 and PHA2) allowed the wound site to heal more cleanly than the control group.

[0156] Test Example 8 - Wound healing evaluation 3 (Analysis of differentially expressed genes (DEGs) in vitro cell lines using PHA particles) PHA particles having a particle size of 178 nm obtained in Example 1 ([P(3HB-co-4HB)], 4HB repeat unit: 17 wt%, molecular weight: 687,000 g / mol) and PHA particles having a particle size of 180 nm obtained in Example 3 ([P(3HB-co-4HB)], 4HB repeat unit: 8.7 wt%, molecular weight: 390,000 g / mol) were dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4) at a concentration of 25% (w / v). Compositions (PHA1 (4HB repeat unit: 17 wt%), PHA2 (4HB repeat unit: 8.7 wt%)) were analyzed to determine which genes the PHA particles promote in human wound healing. The results are shown in Figures 14 to 18. Specifically, human vascular cell line HMEC01 and human dermal fibroblast cell line HDfn were cultured in PRMI 1640 containing 10.0 vol% FBS and 1 vol% penicillin-streptomycin (in a humidified incubator at 37°C with 5% CO2, in the presence of LC nanoclusters). 5 10 mL of cell suspension with a density of 1000 cells / mL was added to each 10 cm diameter circular cell culture dish, and the outer region was filled with cell culture medium and cultured for 24 hours at 37°C and 5% CO2. After culture, PHA1 composition and PHA2 composition were added to cell culture medium and further cultured for 24 hours at 37°C and 5% CO2. After further culture, the medium was discarded, 1 mL of trizol was added, and the cells were flash-frozen at -70°C. ROKIT Genomics Inc. conducted a DEGs analysis, and the results were obtained.

[0157] 14 to 18, it can be seen that genes related to angiogenesis and wound healing were mainly overexpressed in the PHA particle treatment group.

[0158] Test Example 9 - Hemostatic Efficacy Evaluation 1 A hemostatic performance test was conducted on a composition prepared by dissolving (dispersing) the 178 nm diameter PHA particles ([P(3HB-co-4HB)] obtained in Example 1 (4HB repeating unit: 17 wt%, molecular weight: 687,000 g / mol) in 0.1 M PBS buffer (pH 7.4) at a concentration of 7.8% (w / v). The results are shown in Figure 19. Specifically, rabbit whole blood containing a 3.8% sodium citrate solution was centrifuged to separate red blood cells (RBCs) and platelet-rich plasma (PRP) for hemolysis and plasma coagulation analyses, respectively. Next, the red blood cells (RBCs) were washed four times with saline, and 500 μL of diluted red blood cells (1 mL of RBCs and 9 mL of saline) were added to a microtube containing 100 mg of the composition (PHA composition). 100 μL of 0.1% Triton and 100 μL of PBS were used as positive controls (PC) and negative controls (NC), respectively, and 100 μL of Fbrin glue was used as a comparison group. The microtubes were incubated at 37°C for 1 hour and then centrifuged at 3,500 rpm for 10 minutes. The optical density (OD) of the resulting supernatant was analyzed at 540 nm using a microplate reader, and the hemolysis rate (%) was calculated according to the following formula (N=3).

[0159] [Formula 1] Hemostasis rate (%) = {(OD sample - OD negative control group) / (OD positive control group - OD (negative control group)} × 100

[0160] Referring to FIG. 19, it can be seen that the medical composition (hemostatic composition) according to the present invention has excellent hemostatic efficacy.

[0161] Test Example 10 - Hemostatic Efficacy Evaluation 2 The 178 nm diameter PHA particles ([P(3HB-co-4HB)] obtained in Example 1, 4HB repeating units: 17 wt%, molecular weight: 687,000 g / mol) were dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4) at concentrations of 1% (w / v), 3% (w / v), 7% (w / v), and 10% (w / v). A hemostatic performance test was conducted on these compositions, and the results are shown in Figure 20. Specifically, rabbit whole blood containing a 3.8% sodium citrate solution was centrifuged to separate red blood cells (RBCs) and platelet-rich plasma (PRP) for hemolysis and plasma coagulation analyses, respectively. Next, 180 μL of plasma was added to microtubes containing the compositions (PHA compositions) prepared for each concentration and 20 μL of silica nanoparticle suspension. The microtubes were then incubated at 37°C for 30 minutes and centrifuged at 6,000 rpm for 5 minutes. Next, 100 μL of each supernatant was transferred to a well of a 96-well flat-bottom plate and recalcified with 65 μL of 50 mM CaCl2 solution. The degree of fibrin polymerization was assessed by measuring the absorbance at 405 nm every 50 seconds over 1 hour using a plate reader.

[0162] Referring to FIG. 20, it can be seen that the medical composition (hemostatic composition) according to the present invention has a high degree of fibrin polymerization and is excellent in hemostatic efficacy.

[0163] Test Example 11 - Evaluation of bacterial infection inhibition The PHA particles having a particle size of 178 nm obtained in Example 1 ([P(3HB-co-4HB)], 4HB repeat unit: 17 wt%, molecular weight: 687,000 g / mol) and the PHA particles having a particle size of 180 nm obtained in Example 3 ([P(3HB-co-4HB)], 4HB repeat unit: 8.7 wt%, molecular weight: 390,000 g / mol) were dissolved (dispersed) in 0.1 M PBS buffer (pH 7.4) at concentrations of 30% (w / v), 50% (w / v), and 70% (w / v), respectively, and the antibacterial activity of these compositions (PHA3 (4HB repeat unit: 17 wt%), PHA4 (4HB repeat unit: 8.7 wt%)) was evaluated. The results are shown in Figure 21. Specifically, when the OD600 value reached 0.5, each PHA composition was added to 0.2 mL of E. coli solution, which was then plated on LB solid medium. After culturing at 37°C for 12 hours, the colony formation value was analyzed to determine the colony formation rate of E. coli.

[0164] 21, it can be seen that the number of colonies decreases as the PHA concentration increases in the medical compositions according to the present invention (PHA3, PHA4). This result supports the efficacy of the medical composition according to the present invention (composition for inhibiting bacterial infection) in inhibiting bacterial infection.

Claims

1. A medical composition comprising a polyhydroxyalkanoate (PHA) having a particle size of 10,000 nm or less and containing 40% by weight or more of repeating units derived from 3-hydroxybutyrate (3HB) based on the total weight.

2. 2. The medical composition according to claim 1, which has a tissue adhesive strength of 15 kPa or more measured at 100% relative humidity and room temperature.

3. 2. The medical composition according to claim 1, wherein the polyhydroxyalkanoate (PHA) further comprises a repeating unit derived from 4-hydroxybutyrate (4HB).

4. 4. The medical composition according to claim 3, wherein the content of repeating units derived from 4-hydroxybutyrate (4HB) is 0.1 to 60% by weight based on the total weight of the polyhydroxyalkanoate (PHA).

5. 2. The medical composition according to claim 1, wherein the polyhydroxyalkanoate (PHA) is poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer.

6. 2. The medical composition according to claim 1, wherein the polyhydroxyalkanoate (PHA) has a molecular weight of 10,000 to 1,200,000 g / mol.

7. 2. The medical composition according to claim 1, wherein the concentration of the polyhydroxyalkanoate (PHA) is 0.1 to 75% (w / v).

8. The medical composition according to claim 1, which is used for hemostasis.

9. The medical composition according to claim 1, which is used for wound healing.

10. The medical composition according to claim 1, which is used for tissue adhesion.

11. The medical composition according to claim 1, which is used to inhibit bacterial infection.

12. preparing a dispersed phase solution by dissolving polyhydroxyalkanoate (PHA) in a solvent; providing a continuous phase solution containing a surfactant; passing the dispersed phase solution through a membrane having a pore size of 10,000 nm or less to form an emulsion of dispersed phase particles dispersed in the continuous phase solution; and A method for preparing a medical composition, comprising the step of solidifying the emulsion to obtain polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less.

13. The method for producing a medical composition according to claim 12, wherein the content of the polyhydroxyalkanoate (PHA) contained in the dispersed phase solution is 0.01 to 5 wt % based on the total weight of the dispersed phase solution.

14. The method for producing a medical composition according to claim 12, wherein the dispersed phase solution is passed through the membrane at a pressure of 2.5 to 320 kPa.

15. preparing a dispersed phase solution by dissolving polyhydroxyalkanoate (PHA) in a solvent; providing a continuous phase solution containing a surfactant; mixing the dispersed phase solution and the continuous phase solution to form a premix emulsion; The premix emulsion is introduced into a high-pressure dispersing device to form an emulsion in which polyhydroxyalkanoate (PHA) particles having a particle size of 10,000 nm or less are dispersed; and A method for preparing a medical composition, comprising the step of solidifying the emulsion to obtain the polyhydroxyalkanoate (PHA) particles.

16. The method for producing a medical composition according to claim 15, wherein the pressure applied to the premix emulsion by the high-pressure dispersion device is 1 to 30 kpsi.

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