Filler composition and method for performing filler procedure using same

Abstract

The present invention relates to a filler composition injected subcutaneously and a method for performing a filler procedure using same. The filler composition includes an aqueous solution containing a photoinitiator and an uncrosslinked photocrosslinkable hyaluronic acid polymer having a photoreactive functional group. Prior to being injected into the skin, the filler composition is in a solution state including the uncrosslinked photocrosslinkable hyaluronic acid polymer and thus can be injected into the skin at a low pressure that can be applied by hand. In addition, the filler composition can be quickly formed into a desired shape or form concurrent with or following injection into the skin, and can stably maintain the formed shape or form through photocrosslinking.

Description

Filler composition and filler treatment method using the same

[0001] The present invention relates to a filler composition that can be injected subcutaneously and a filler treatment method using the same.

[0002] Dermal fillers are medical devices injected subcutaneously into facial wrinkles to temporarily improve wrinkles and restore skin volume through physical repair without any pharmacological action. Fillers can be classified based on the properties of their raw materials and their duration of action. Specifically, fillers can be divided into non-absorbable and absorbable fillers based on whether they are absorbed by the body.

[0003] The most commonly used fillers are hyaluronic acid (HA)-based absorbable fillers. These fillers are highly safe, can be used without skin reaction testing, and are easily degraded by enzymes. HA fillers are composed of cross-linked particles, which have fixed physical properties. Depending on the injection site, the properties of the filler used need to be tailored. Generally, fillers with good shape retention have high viscosity, requiring high injection pressure, which can be painful during the procedure. Furthermore, existing fillers are already cross-linked hydrogels, making it difficult to incorporate active ingredients such as peptides, proteins, and cells into the filler or deliver them over long periods of time.

[0004] In order to be applied as an ideal filler, the following characteristics are required: 1) easy injection with little force, 2) rapid molding into a desired shape, 3) easy control of physical properties, 4) decomposition by enzymes, and 5) usable as a carrier for delivering effective substances. However, to date, no filler that satisfies all of these characteristics has been developed.

[0005] The main purpose of the present invention is to provide a filler composition that is easy to inject with little force, can be quickly molded into a desired shape, has easy control of mechanical properties, is decomposable by a decomposing enzyme, can be used as a carrier capable of delivering an effective substance, and can be injected into the skin or living tissue.

[0006] Another object of the present invention is to provide a filler treatment method using the filler composition.

[0007] To achieve the above object, the present invention provides a filler composition to be injected subcutaneously, comprising: an uncrosslinked photocrosslinkable hyaluronic acid polymer having a photoreactive functional group; and a photoinitiator.

[0008] In addition, the present invention provides a filler composition in which, when the photocrosslinkable hyaluronic acid polymer is photocrosslinked to form a hydrogel, an active substance therein is released from the hydrogel.

[0009] In addition, the present invention provides a filler treatment method comprising: a first step of preparing a filler composition comprising a photocrosslinkable hyaluronic acid polymer, a photoinitiator, and a solvent; a second step of injecting the filler composition subcutaneously into a predetermined shape, molding the filler composition into a predetermined shape after injection, or mixing an effective substance therein if necessary; and a third step of crosslinking the photocrosslinkable hyaluronic acid polymer by irradiating the molded filler composition with light.

[0010] According to the filler composition of the present invention and the filler treatment method using the same, since it is in a solution state containing a photocrosslinkable hyaluronic acid polymer in an uncrosslinked state before being injected into the skin, it can be injected into the skin with a low pressure that can be pressed by hand, and it is possible to quickly form it into a desired shape or form simultaneously with or after injection into the skin, and the form or form formed through photocrosslinking can be stably maintained.

[0011] In addition, by controlling the light irradiation time for photocrosslinking of the photocrosslinkable hyaluronic acid polymer, the strength of the filler injected into the skin can be varied, and if removal is required after photocrosslinking, it can be easily decomposed by hyaluronic acid decomposing enzyme (Hyaluronidase, HAase).

[0012] In addition, since it contains a hyaluronic acid polymer in an uncrosslinked state before being injected into the skin, it can be uniformly mixed with the effective substance and injected into the skin, and after being injected into the skin, it can be photocrosslinked to induce sustained release of the effective substance, thereby serving as a carrier capable of delivering the effective substance.

[0013] Figures 1a and 1b are images showing the results after enzyme treatment of the filler composition of Example 1 cured in an in vitro environment.

[0014] Figures 2a and 2b are images and graphs showing the results after enzyme treatment of the filler composition of Example 1 cured in an in vivo environment.

[0015] FIGS. 3a and 3b are images and graphs measuring the volume change over time of the filler composition of Example 1 cured in an in vivo environment without treatment with HAase.

[0016] Figure 4 shows an image of a mold used in a moldability test (a) and images of a commercial hyaluronic acid hydrogel filler composition and a filler composition of Example 1 injected subcutaneously into the back of a hairless mouse (b).

[0017] Figure 5 shows images of the filler compositions of Examples 1, 2, and 3 after photocuring.

[0018] Figure 6 is a graph and images showing the results of measuring the sustained release ability of the active substance (Rho-Dex) released after in vitro photocuring of the filler compositions of Examples 2 and 3.

[0019] Figures 7a and 7b are graphs showing the results of measuring the pH and viscosity of the filler compositions of Examples 6 to 9 before and after moist heat sterilization.

[0020] Figure 8 is a drawing for explaining an experimental process for testing the curing performance of a photocurable filler.

[0021] Figure 9 is a photograph showing the results of a performance test of fillers using photocurable hyaluronic acids of Examples 10 to 12 and 14.

[0022] Figure 10 is a photograph of a hydrogel formed after photocuring of fillers using photocurable hyaluronic acids of Examples 10 to 14, respectively.

[0023] Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings. The present invention can be modified in various ways and can take various forms, and thus specific embodiments are illustrated in the drawings and described in detail in the text. However, this is not intended to limit the present invention to a specific disclosed form, but should be understood to include all modifications, equivalents, and substitutes included in the spirit and technical scope of the present invention. In describing each drawing, similar reference numerals are used to indicate similar components. In the attached drawings, the dimensions of structures are shown larger than actual size to ensure clarity of the present invention.

[0024] While terms like "first" and "second" may be used to describe various components, these components should not be limited by these terms. These terms are used solely to distinguish one component from another. For example, without departing from the scope of the present invention, a first component could be referred to as a "second component," and similarly, a second component could also be referred to as a "first component."

[0025] The terminology used in this application is only used to describe specific embodiments and is not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this application, it should be understood that the terms "comprise" or "have" indicate the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but do not preclude the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0026] Meanwhile, unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning within the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0027]

[0028] A filler composition according to an embodiment of the present invention may include a photocrosslinkable hyaluronic acid polymer, a photoinitiator, and a solvent.

[0029] The photocrosslinkable hyaluronic acid polymer may have a photoreactive functional group. For example, the photocrosslinkable hyaluronic acid polymer may include a photoreactive functional group such as a methacyrlate group (MA), a 4-pentenoate group (PA), or the like.

[0030] In one embodiment, the photocrosslinkable hyaluronic acid polymer may include a repeating unit of the following chemical formula 1. For example, the photocrosslinkable hyaluronic acid polymer may include a single repeating unit of the following chemical formula 1, or may include one or more other repeating units together with the repeating unit of the following chemical formula 1.

[0031] [Chemical Formula 1]

[0032]

[0033] In the above chemical formula 1, R1 to R 10 Each of which may be independently one selected from the group consisting of a hydroxyl group, a carboxyl group, a photoreactive functional group, etc., and the photoreactive functional group may include at least one selected from the group consisting of glycidyl methacrylate, methacrylate, acrylate, pentyl acrylate, 4-pentenoate, diazirines, aryl azide, benzophenones, norbornene, maleimide, vinyl ester, thiol, vinyl sulfone, etc. For example, the R1 to R 10 One or more of the selected groups may independently be a methacrylate functional group or a 4-pentenoate functional group, and the remainder may be a hydroxy or carboxyl group.

[0034] In one embodiment, the photocrosslinkable hyaluronic acid polymer may include the methacrylate functional group or the 4-pentenoate functional group at a degree of substitution of about 140% or less, for example, about 10 to 140%, so that it can be decomposed using hyaluronidase (HAase) after photocrosslinking. Here, a degree of substitution of 100% means that one methacrylate functional group and one 4-pentenoate functional group are substituted per repeating unit of Chemical Formula 1, and since there are four substitutable -OH groups per repeating unit before substitution, the maximum degree of substitution is 400%.

[0035] In one embodiment, the photocrosslinkable hyaluronic acid polymer may include both the methacrylate functional group and the 4-pentenoate functional group. In this case, in one embodiment, the photocrosslinkable hyaluronic acid polymer may include the methacrylate functional group and the 4-pentenoate functional group in a molar ratio of about 9:1 to 7:3. When the molar ratio of the 4-pentenoate functional group is less than 9:1, the photocrosslinkable hyaluronic acid polymer may have a problem of being easily broken by external impact after crosslinking, and when the molar ratio of the 4-pentenoate functional group is greater than 7:3, the photocrosslinkable hyaluronic acid polymer may not have sufficient strength after crosslinking, and thus may have a problem of not being able to maintain its shape.

[0036] In one embodiment, when the molecular weight of the photocrosslinkable hyaluronic acid polymer is about 1000 kDA or more, the photocrosslinkable hyaluronic acid polymer may include the photoreactive functional group at a degree of substitution of about 20% or more and about 100% or less. On the other hand, when the molecular weight of the photocrosslinkable hyaluronic acid polymer is less than 1000 kDA, even if the photoreactive functional group is included at a degree of substitution of about 50% or more and about 140% or less, it may be decomposed by a hyaluronic acid decomposing enzyme.

[0037] In one embodiment, the photocrosslinkable hyaluronic acid polymer may include a methacrylate group and 4-pentenoate as the photoreactive functional group, and may also include only the methacrylate group.

[0038] In one embodiment, the weight average molecular weight of the photocrosslinkable hyaluronic acid polymer may be, but is not limited to, about 100 to 3000 kDa.

[0039] In one embodiment, the photocrosslinkable hyaluronic acid polymer may exist in an uncrosslinked state before being injected subcutaneously, and may be photocrosslinked to form a hydrogel after being injected subcutaneously. According to an embodiment of the present invention, when the filler composition includes the photocrosslinkable hyaluronic acid polymer in an uncrosslinked state, compared to a conventional filler composition including a crosslinked hyaluronic acid polymer, the filler composition may have a lower viscosity even when the concentration of the hyaluronic acid polymer is increased, and as a result, the filler composition may be injected subcutaneously with less force, thereby reducing pain to the recipient. In one embodiment, the filler composition may include the photocrosslinkable hyaluronic acid polymer in a concentration of about 0.5 to 20 w / v%, for example, about 0.5 to 5 w / v%. For example, when the photocrosslinkable hyaluronic acid polymer has a molecular weight of about 500 to 3000 kDa, the filler composition may contain the photocrosslinkable hyaluronic acid polymer at a concentration of about 0.5 to 2 w / v%, and when the photocrosslinkable hyaluronic acid polymer has a molecular weight of about 100 to 500 kDa, the filler composition may contain the photocrosslinkable hyaluronic acid polymer at a concentration of about 5 to 10 w / v%. In one embodiment, the filler composition may have a viscosity of about 50 to 1500 cP, for example, about 100 to 500 cP, at 25°C, but is not limited thereto.

[0040] Meanwhile, the filler composition according to an embodiment of the present invention may further include one or more active substances selected from the group consisting of peptides, proteins, drugs, polymers, cells, etc. Since the conventional filler composition includes a hyaluronic acid polymer that is crosslinked and in the form of a hydrogel, there is a disadvantage in that it is difficult to deliver the active substance into the human body by mixing it together with the hydrogel. However, the filler composition according to an embodiment of the present invention includes a hyaluronic acid polymer in an uncrosslinked state before being injected subcutaneously, so that the active substance can be uniformly mixed with the hyaluronic acid polymer in an uncrosslinked state, and by photocrosslinking after subcutaneously injecting the same, the active substance can be included in the human body for a long period of time or be slowly released.

[0041] In one embodiment, for photocrosslinking of the photocrosslinkable hyaluronic acid polymer, the active ingredient may be included in a proportion of about 50% or less, for example, about 0.1 to 30%, of the weight of the photocrosslinkable hyaluronic acid polymer.

[0042] The photoinitiator can absorb light energy to initiate a crosslinking reaction between the photoreactive functional groups. In one embodiment, any water-soluble material that can be activated by visible light to form free radicals can be used as the photoinitiator without limitation. For example, the photoinitiator is an α-hydroxyketone series derivative (irgacure 2959, irgacure 184, irgacure 651, irgacure 369, irgacure 907, etc.), a phosphine series derivative (TPO, TPO-Na, LAP, BAPO, BAPO-ONa, BAPO-OLi, etc.), an azo-initiator (2,2'-azobis[2-methyl-N-(2-hydroxyethyl) promionamide] (VA-086)), an eosin-Y, riboflavin (B2), camphorquinone, an erythrosine (Erythrosine), Rose bengal, WSPI (1,4-bis(4-N,N-bis(6-N,N,N-trimethylammonium)hexyl)amino)-styryl)-2,5-dimethoxybenzene tetraiodide), BDEA (2,5-bis-[4-(diethylamino)-benzylidene]-cyclopentanone), P2CK (3,3'-((((1E,1'E)-(2-oxocyclopentane-1,3-diylidene)bis(methanylidene))bis(4,1-phenylene))bis(methylase indyl))dipropanoate)(P2CK (3,3'-((((1E,1'E)-(2-oxocyclopentane-1,3 diylidene)bis(methanylylidene))bis(4,1-phenylene))bis(methylazanediyl))dipropanoate)) and G2CK (sodium 2,2'-((((1E,1'E)-(5-methyl-2-oxocyclohexane-1,3-diylidene)bis(methanylylidene))bis(4,1-phenylene))bis(methylazanediyl))diacetate)), Lithium benzoyl(phenyl)phosphinate, Magnesium benzoyl(phenyl)phosphinate, etc. May include:

[0043] In one embodiment, the photoinitiator may be a material having a chemical structure represented by Chemical Formula 2 below. The photoinitiator having the chemical structure represented by Chemical Formula 2 is water-soluble and can be activated by visible light having a wavelength of about 400 to 500 nm. Meanwhile, the photoinitiator of Chemical Formula 2 below may have a higher molar extinction coefficient and lower cytotoxicity compared to other photoinitiators.

[0044] [Chemical Formula 2]

[0045]

[0046] In the above chemical formula 2, R1 can be H or F, and R2 can be Li + or Mg 2+ It could be.

[0047] In one embodiment, the photoinitiator may be included at a concentration of about 0.01 to 1 w / v%, for example, about 0.05 to 0.15 w / v%.

[0048] In another embodiment, LAP (Lithium phenyl-2,4,6-trimethylbenzoylphosphinate) may be used as the photoinitiator.

[0049] The solvent may further include at least one selected from the group consisting of water, saline, phosphate buffered saline, trisodium citrate buffer, tris-HCl buffer, HEPES buffer, etc.

[0050] In one embodiment, since the methacrylate group (MA) and 4-pentenoate group (PA) bound within the photocrosslinkable hyaluronic acid polymer may change the pH of the composition when separated due to physical factors during storage, a solvent capable of buffering the pH may be used. In one embodiment, the solvent may be a phosphate buffered saline solution having a concentration of about 2 to 20 mM, a trisodium citrate buffer solution, a tris-HCl buffer solution, a HEPES buffer solution, or the like, which may be used alone or in combination of two or more thereof.

[0051] In another embodiment, water can be used as the solvent.

[0052] In one embodiment, the filler composition may further include a sugar alcohol such as mannitol or sorbitol. When the filler composition further includes a sugar alcohol, after performing a moist heat sterilization process on the filler composition, the pH, viscosity, gelation point, etc. of the composition may be stably maintained compared to before performing the moist heat sterilization process.

[0053] According to the filler composition of the present invention, since it has a low viscosity solution state including an uncrosslinked hyaluronic acid polymer before being injected into the skin, it can be injected into the skin without pain to the recipient, and it is possible to quickly shape it into a desired shape or form simultaneously with or after injection into the skin, and the shape or form formed through photocrosslinking can be stably maintained.

[0054] In addition, by controlling the light irradiation time for photocrosslinking of the photocrosslinkable hyaluronic acid polymer, the strength of the filler injected into the skin can be varied, and if removal is required after photocrosslinking, it can be easily decomposed by hyaluronic acid decomposing enzyme.

[0055] In addition, since it contains a hyaluronic acid polymer in an uncrosslinked state before being injected into the skin, it can be uniformly mixed with the effective substance and injected into the skin, and after being injected into the skin, it can be photocrosslinked to induce sustained release of the effective substance, thereby serving as a carrier capable of delivering the effective substance.

[0056]

[0057] Hereinafter, a filler treatment method using a filler composition according to an embodiment of the present invention will be described.

[0058] A filler treatment method according to an embodiment of the present invention may include a first step of preparing a filler composition including a photocrosslinkable hyaluronic acid polymer, a photoinitiator, and a solvent; a second step of injecting the filler composition subcutaneously into a predetermined shape or molding the filler composition into a predetermined shape after injection; and a third step of crosslinking the photocrosslinkable hyaluronic acid polymer by irradiating the molded filler composition with light.

[0059] In the first step, the filler composition may be prepared by dissolving the photocrosslinkable hyaluronic acid polymer and the photoinitiator in a solvent. Alternatively, the filler composition may be prepared by additionally dissolving an effective substance in addition to the photocrosslinkable hyaluronic acid polymer and the photoinitiator. Since the filler composition is substantially the same as described above, a detailed description thereof will be omitted.

[0060] In the second step, the filler composition can be injected subcutaneously using an injection device such as a syringe. In this case, a mold having an opening of a desired shape is placed on the skin, and the filler composition is injected into the opening at a location so that the filler composition is injected subcutaneously into a shape corresponding to the opening. Alternatively, the filler composition can be injected subcutaneously and then molded into a desired shape using a mold or the like.

[0061] According to the filler treatment method of the present invention, since the filler composition includes an uncrosslinked photocrosslinkable hyaluronic acid polymer, it can have low viscosity even if it contains a relatively high concentration of photocrosslinkable hyaluronic acid polymer, and as a result, the pressure for injecting the filler composition can be lowered, thereby alleviating pain of the recipient, and the shape of the injected filler composition can be easily molded, and it can be injected subcutaneously in a state where the photocrosslinkable hyaluronic acid polymer and the effective substance are uniformly mixed.

[0062] In the third step, the filler composition injected subcutaneously may be cured by irradiating light to crosslink the photocrosslinkable hyaluronic acid polymers. At this time, the wavelength and intensity of light for the photocrosslinking may vary depending on the type of photoinitiator included in the composition, and light with a wavelength of about 320 to 600 nm may be used.

[0063] According to the filler treatment method of the present invention, by controlling the intensity and time of light irradiated for the photocrosslinking, the mechanical properties of the cured filler composition, i.e., strength, degree of curing, etc., can be controlled, thereby enabling the use of a filler composition having the same composition in various parts and applications. In addition, the effective substance can be made to remain in the cured filler composition for a long time or be released relatively uniformly.

[0064] Meanwhile, the filler treatment method according to an embodiment of the present invention may further include a step of injecting a hyaluronic acid decomposing enzyme into the cured filler composition to decompose the cross-linked hyaluronic acid polymer.

[0065] The above photocured HA filler can be easily decomposed by hyaluronic acid decomposing enzymes, like conventional HA fillers.

[0066]

[0067] Hereinafter, specific examples of the present invention will be described in detail. However, the following examples are merely some embodiments of the present invention, and the scope of the present invention is not limited to the following examples.

[0068]

[0069] [Example 1]

[0070] A filler composition of Example 1 was prepared by dissolving 1 w / v% of photocurable hyaluronic acid (HAMA-PA) having a weight average molecular weight (Mw) of 1000 kDa, in which 40% of the hydroxyl groups of hyaluronic acid are substituted with photocurable groups such as methacrylate (MA) and pentaacrylate (PA) groups, and 0.1 w / v% of lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), a photoinitiator, in water.

[0071]

[0072] [Examples 2 and 3]

[0073] Filler compositions of Examples 2 and 3 were prepared by mixing the filler composition of Example 1 and an aqueous solution containing Rho-Dex (rhodamine-labeled dextran) having a molecular weight of 70 kDa at a concentration of about 4 w / v% in a volume ratio (v:v) of 9:1 and 7:3, respectively. Rho-Dex is an organic substance having a molecular weight similar to that of albumin protein (66 kDa) and was used as a model effective substance.

[0074]

[0075] [Experimental Example 1] Viscosity Experiment

[0076] The viscosity of the filler composition of Example 1 and the commercial hyaluronic acid hydrogel filler composition was measured using a rheometer (MCR 302, Anton Paar) at 25°C and a shear rate of 300 / s. As a result, the viscosity of the filler composition of Example 1 was confirmed to be about 200 to 1500 cP, which is significantly lower than that of the commercial hyaluronic acid hydrogel filler composition having a viscosity of about 2000 cP.

[0077]

[0078] [Experimental Example 2] Resolution Experiment

[0079] To confirm the decomposition ability of the filler composition of photo-cured Example 1, decomposition experiments were conducted in vitro and in vivo.

[0080] Figure 1 is an image showing the results after the enzymatic treatment of the filler composition of Example 1 cured in an in vitro environment. In the in vitro experiment, 0.1 mL of the filler composition of Example 1 contained in an experimental vessel was irradiated with light of a wavelength of 400-500 nm at ~1 J / cm. 2 (30 mW / cm 2 , 35 s) and hardened, and then treated with hyaluronic acid decomposing enzyme (HAase) to confirm the decomposition ability.

[0081] Referring to Fig. 1a, the photocurable filler composition not treated with HAase was not decomposed and maintained its shape even after 6 hours, but it was confirmed that both the photocurable filler compositions treated with 150 U of HAase and 750 U of HAase were completely decomposed within 6 hours.

[0082] Referring to Fig. 1b, the photocurable filler composition having a weight average molecular weight (Mw) of 100 kDa and 5 w / v% that was not treated with HAase was not decomposed and maintained its shape even after 48 hours, but it was confirmed that all photocurable filler compositions treated with 750 U of HAase were completely decomposed within 48 hours.

[0083] Figures 2a and 2b are images and graphs showing the results after enzyme treatment of the filler composition of Example 1 cured in an in vivo environment. For the in vivo experiment, 0.1 mL of the filler composition of Example 1 was injected subcutaneously into the dorsal area of ​​SKH-7 hairless mice and photocured. After about 1 day, 150 U of HAase and 500 U of HAase were injected into the cured filler composition, and the size of the filler over time was measured using a vernier caliper.

[0084] Referring to FIGS. 2a and 2b, in the case of fillers without HAase, the shape was maintained without decomposition for 8 days, but in the case of fillers with 150 U of HAase and 500 U of HAase, the volume decreased by about 50 to 80% compared to the initial volume by the second day, and it was confirmed that the higher the HAase concentration, the greater the volume decrease.

[0085]

[0086] FIGS. 3a and 3b are images and graphs measuring the volume change over time of the filler composition of Example 1 cured in an in vivo environment without treatment with HAase.

[0087] Referring to FIGS. 3a and 3b, when 0.1 mL of the filler composition of Example 1 was injected subcutaneously into the dorsal surface of a hairless mouse and maintained without treatment with HAase after photocuring, it was confirmed that a volume of at least 83% of the initial volume was maintained after about 30 days.

[0088]

[0089] [Experimental Example 3] Moldability Experiment

[0090] The moldability (formability) of existing commercial hyaluronic acid-based fillers and photocurable HA fillers (HA photo-filler) was confirmed through animal experiments using mice.

[0091] Figure 4 shows (a) an image of a mold used in a moldability test and (b) images of fillers injected subcutaneously into the dorsal area of ​​a hairless mouse using a commercial hyaluronic acid hydrogel filler composition and the filler composition of Example 1. Figure 4b shows images of fillers formed by placing the star-shaped mold illustrated in Figure 4a on the mouse skin and then injecting the filler subcutaneously into the skin where the mold is located.

[0092] Referring to FIG. 4, in the case of a commercial hyaluronic acid hydrogel filler composition, when the mold was removed after injection, the shape was not maintained and the composition spread under the skin, but in the case of the filler composition of Example 1, it was found that the star-shaped shape was maintained even when the mold was removed after photocuring.

[0093] And, as a result of checking the shape 1 day after injection, it was observed that the filler using the commercial hyaluronic acid hydrogel filler composition swelled subcutaneously and a change in the volume of the filler was observed, but the filler using the filler composition of Example 1 was found to maintain its shape stably without a change in volume.

[0094]

[0095] [Experimental Example 4] Drug loading and release ability experiment

[0096] Figure 5 shows images of the filler compositions of Examples 1, 2, and 3 after photocuring.

[0097] Referring to Fig. 5, it can be confirmed that photocuring effectively occurs even when the filler composition and the Rho-Dex aqueous solution are mixed in a volume ratio ranging from 10:0 to 7:3.

[0098]

[0099] Figure 6 is a graph and images showing the results of measuring the sustained release ability of the active substance (Rho-Dex) released after in vitro photocuring of the filler compositions of Examples 2 and 3.

[0100] Referring to FIG. 6, when the filler compositions of Examples 2 and 3 were photocured, it was found that the active substance was slowly released from the filler for more than 24 hours, and even after 24 hours, the filler was not decomposed and its shape was maintained.

[0101]

[0102] [Examples 4 and 5]

[0103] The photocurable fillers of Examples 4 and 5 were prepared by adding 10 mM PBS and 15 mM PBS, respectively, to the filler composition of Example 1, and then sterilized by moist heat at 121°C for 15 minutes and stored in an incubator at 80°C for 2 weeks.

[0104]

[0105] [Experimental Example 5] Solution Stability Test

[0106] As described in Table 1 below, the pH of the filler compositions after moist heat sterilization was measured to be 7.07 for the filler composition of Example 4 (10 mM PBS), and 6.98 for the filler composition of Example 5 (15 mM PBS). In addition, after storage at 80°C for one week, the pH of the filler compositions of Examples 4 and 5 was measured to be 6.56 and 6.65, respectively, and after storage at 80°C for two weeks, the pH of the filler compositions of Examples 4 and 5 was measured to be 6.48 and 6.63, respectively, confirming that this is within the range of pH 5.5 to 8.5, which is the filler specification allowed for incorporation into the body. From these results, it can be confirmed that the pH of the filler composition can be relatively stably maintained when a buffer solution is used even under harsh conditions of 80°C.

[0107] After sterilization of the buffer solution, store at pH 80℃ for 1 week, then store at pH 80℃ for 2 weeks, then store at pH 10mM PBS7.076.566.4815mM PBS6.986.656.63

[0108] [Examples 6 to 9]

[0109] Mannitol and sorbitol were added to the filler composition of Example 1 according to Table 2 below and sterilized by moist heat at 121°C for 20 minutes.

[0110] Stabilizer added amount (weight %) Example 6 Mannitol 1 Example 7 Mannitol 4.3 Example 8 Sorbitol 1 Example 9 Sorbitol 4.3

[0111] [Experimental Example 6] Filler composition containing sugar

[0112] In Example 1, stabilizers such as mannitol and sorbitol were added to the composition to confirm its stability during the sterilization process. After sterilization by moist heat at 121°C for 20 minutes, pH, viscosity, and crosslinking performance (gelation point) were measured, and the results are shown in Table 3 and Figures 7a and 7b.

[0113]

[0114]

[0115] Referring to Table 3, Figures 7a and 7b, in terms of pH, it was found that the pH decreased after sterilization compared to before sterilization in all filler compositions. However, the filler compositions of Examples 7 to 9, particularly the filler composition of Example 9 containing 4.3% sorbitol, showed less change in pH.

[0116] In the case of viscosity, the filler composition not containing sugar alcohol decreased by up to 60.6% after sterilization compared to before sterilization, but the filler composition of Example 7 containing 4.3% mannitol and the filler composition of Example 9 containing 4.3% sorbitol decreased by only 19.0% and 27.2%, respectively, showing relative stability.

[0117] In the case of the gelation point, the filler composition not containing sugar alcohol changed by +6.5 after sterilization compared to before sterilization, but the filler compositions of Examples 6 to 9 changed only by +0.5, -0.4, +1.8, and +2.6, showing relative stability.

[0118]

[0119] <Examples 10 to 13>

[0120] In order to compare the performance as a filler according to the molar ratio (MA:PA) of the photocurable groups, methacrylate functional group (MA) and 4-pentenoate functional group (PA), in photocurable hyaluronic acid (HAMA-PA), an experiment was conducted to test the photocrosslinking performance as shown in Fig. 8. The filler solution used the same polymer concentration and photoinitiator material as in Example 1.

[0121] PDMS molds were filled with photocurable hyaluronic acid filler solutions having MA:PA ratios of 10:0 (Example 10), 9:1 (Example 11), 7:3 (Example 12), 5:5 (Example 13), and 0:10 (Example 14) (see a in FIG. 8), covered with rat skin on top (see b in FIG. 8), and then photocured by external irradiation with visible light (405 nm) (see b in FIG. 8), and the curing characteristics of the fillers were analyzed.

[0122]

[0123] Experimental Example 7

[0124] Figure 9 is a photograph showing the results of a performance test of fillers using photocurable hyaluronic acids of Examples 10 to 12 and 14.

[0125] Referring to FIG. 9, photocurable hyaluronic acid (HAMA) (Example 10), in which only MA with a relatively short cross-linking length was introduced as a photocurable group, formed a hard hydrogel after curing, which was easily broken by external impact. On the other hand, photocurable hyaluronic acid (HAPA) (Example 14), in which only PA with a relatively long cross-linking length was introduced as a photocurable group, was found to not have sufficient strength after curing and thus could not maintain its shape. However, photocurable hyaluronic acid (HAMA-PA) (Examples 11 and 12) in which both MA and PA were introduced formed a hydrogel with excellent mechanical strength, which was restored to its original shape even when an external impact (after compression) was applied.

[0126] Figure 10 is a photograph of a hydrogel formed after photocuring of fillers using photocurable hyaluronic acids of Examples 10 to 14, respectively.

[0127] Referring to Figure 10, it was confirmed that photocurable hyaluronic acid possesses excellent mechanical performance when it contains MA and PA in a molar ratio of 9:1 to 7:3. Since the injected filler is exposed to various external stimuli, such excellent mechanical performance is an important factor in maintaining the filler volume for a long period of time.

[0128]

[0129] Below, a method for synthesizing a novel photoinitiator of chemical formula 2 is described in detail.

[0130]

[0131] <Example 15-1> Preparation of photoinitiator

[0132] A first mixture was prepared by adding dimethyl phenylphosphonate (about 187 mg, about 1.1 mmol) to benzoyl chloride (about 140 mg, about 1 mmol) and 2-butanone (about 10 mL), and stirring the mixture at room temperature for about 24 hours under nitrogen. A second mixture was prepared by adding lithium bromide (about 95 mg, about 1.1 mmol) to a 2-butanone (about 10 mL) solution. The first mixture and the second mixture were mixed, heated at about 60 °C for about 20 minutes, and cooled to room temperature to produce a reactant. The reactant was washed sequentially with 2-butanone and diethyl ether, and then dried under vacuum to obtain a photoinitiator for bioprinting, which was named LBP (about 237 mg, about 94% yield).

[0133] 1 H NMR (600 MHz, DO): 8.09 (d, 2H), 7.61-7.65 (m, 2H), 7.58 (t, 1H), 7.49 (t, 1H), 7.46-7.42 (m, 4H). 13 C NMR (151 MHz, DO): 212.39, 211.61,135.72, 135.43, 134.53, 133.54, 132.66, 131.91, 131.89, 131.70, 131.64, 129.18, 129.16, 128.83, 128.46, 128.36.31 P NMR (243 MHz, DO):18.37. HRMS (ESI MS) m / z: calculated: 252.13 found:253.06 ([M+H]+ detected)

[0134]

[0135] <Example 15-2> Preparation of photoinitiator

[0136] A photoinitiator for bioprinting was prepared in the same manner as in Example 10-1, except that 4-fluorobenzoyl chloride (about 158 ​​mg, about 1 mmol) was used instead of the benzoyl chloride in Example 10-1, and was named LFBP.

[0137] 1 H NMR (600 MHz, DO): 8.18-8.17 (m, 2H), 7.64-7.61 (m, 2H), 7.50 (t, 1H), 7.44-7.42 (m, 2H), 7.17 (t, 2H). 13 C NMR (151 MHz, DO): 210.53, 209.74, 167.03, 165.34, 133.41, 132.53, 132.27, 132.21, 131.99, 131.97, 131.95, 131.93, 131.71, 131.65, 128.46, 128.39. 31 P NMR (243 MHz, DO):18.59. HRMS (ESI MS) m / z: calculated,270.12 found:271.05 ([M+H]+ detected)

[0138]

[0139] <Example 15-3> Preparation of photoinitiator

[0140] A photoinitiator for bioprinting was prepared in the same manner as in Example 10-1, except that magnesium bromide (about 203 mg, about 1.1 mmol) was used instead of the lithium bromide in Example 10-1, and was named MBP.

[0141] 1 H NMR (600 MHz, DO): 8.14-8.13 (d, 2H), 7.70-7.65 (m, 2H), 7.65-7.60 (t, 1H), 7.44-7.42 (t, 2H), 7.51-744 (m, 4H).

[0142]

[0143] Although the present invention has been described above with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various modifications and changes may be made to the present invention without departing from the spirit and scope of the present invention as set forth in the claims below.

Claims

1. In a filler composition injected subcutaneously, A filler composition comprising: an uncrosslinked photocrosslinkable hyaluronic acid polymer having a photoreactive functional group; and a photoinitiator.

2. In paragraph 1, The photocrosslinkable hyaluronic acid polymer is a filler composition comprising a repeating unit of the following chemical formula 1: [Chemical Formula 1] In the above chemical formula 1, R1 to R 10 Some of the photoreactive functional groups independently of each other include at least one selected from the group consisting of glycidyl methacrylate, methacrylate, acrylate, pentyl acrylate, 4-pentenoate, diazirines, aryl azides, benzophenones, norbornene, maleimide, vinyl ester, thiol and vinyl sulfone, and the rest are independently hydroxy or carboxyl groups.

3. In paragraph 2, A filler composition wherein the photoreactive functional group comprises a methacrylate functional group or a 4-pentenoate functional group.

4. In paragraph 3, A filler composition, characterized in that the photocrosslinkable hyaluronic acid polymer comprises both the methacrylate functional group and the 4-pentenoate functional group.

5. In paragraph 4, A filler composition, characterized in that the photocrosslinkable hyaluronic acid polymer comprises the methacrylate functional group and the 4-pentenoate functional group in a molar ratio of 9:1 to 7:

3.

6. In paragraph 3, A filler composition wherein the photocrosslinkable hyaluronic acid polymer comprises the methacrylate functional group or the 4-pentenoate functional group with a substitution degree of 10 to 140%, and can be decomposed by a hyaluronic acid decomposing enzyme after photocrosslinking.

7. In paragraph 6, A filler composition wherein the photocrosslinkable hyaluronic acid polymer has a weight average molecular weight of 100 to 3000 kDa.

8. In paragraph 7, A filler composition comprising 0.5 to 20 w / v% of the photocrosslinkable hyaluronic acid polymer, and having a viscosity of 50 to 1500 cP at 25°C.

9. In paragraph 1, The photoinitiators are irgacure 2959, irgacure 184, irgacure 651, irgacure 369, irgacure 907, TPO, TPO-Na, LAP, BAPO, BAPO-ONa, BAPO-OLi, 2,2'-azobis[2-methyl-N-(2-hydroxyethyl) promionamide] (VA 086)), Eosin-Y, Riboflavin (B2), Camphorquinone, Erythrosine, Rosebengal, 1,4-bis(4-(N,N-bis(6-N,N,N-trimethylammonium)hexyl)amino)-styryl)-2,5-dimethoxybenzene tetraiodide) (1,4-bis(4-(N,N-bis(6-(N,N,N-trimethylammonium)hexyl)amino)-styryl)-2,5-dimethoxybenzene tetraiodide)), 2,5-bis-[4-(diethylamino)-benzylidene]-cyclopentanone) (2,5-bis-[4-(diethylamino)-benzylidene]-cyclopentanone)), 3,3'-((((1E,1'E)-(2-oxocyclopentane-1,3-diylidene)bis(methanylylidene))bis(4,1-phenylene))bis(methylazanediyl))dipropanoate) (3,3'-((((1E,1'E)-(2-oxocyclopentane-1,3-diylidene)bis(methanylylidene))bis(4,1-phenylene))bis(methylazanediyl))dipropanoate) and sodium 2,2'-((((1E,1'E)-(5-methyl-2-oxocyclohexane-1,3-diylidene)bis(methanylylidene))bis(4,1-phenylene))bis(methylazanediyl))diacetate) (sodium 2,2'-((((1E,1'E)-(5-methyl-2-oxocyclohexane-1,3-diylidene)bis(methanylylidene))bis(4,A filler composition comprising at least one selected from the group consisting of lithium benzoyl(phenyl)phosphinate, lithium benzoyl(phenyl)phosphinate, and magnesium benzoyl(phenyl)phosphinate.

10. In paragraph 1, The photoinitiator is a filler composition comprising a compound having a chemical structure represented by the following chemical formula 2: [Chemical Formula 2] In the above chemical formula 2, R1 is H or F, and R2 is Li + or Mg 2+ am.

11. In paragraph 9 or 10, A filler composition wherein the concentration of the photoinitiator is about 0.01 to 1 w / v%.

12. In paragraph 1, A filler composition further comprising one or more active substances selected from the group consisting of peptides, proteins, drugs, and cells.

13. In paragraph 10, A filler composition, wherein the effective substance is included in a proportion of 0.1 to 50% of the weight of the photocrosslinkable hyaluronic acid polymer.

14. In paragraph 13, A filler composition wherein, when the photocrosslinkable hyaluronic acid polymer is photocrosslinked to form a hydrogel, the active substance is released from the hydrogel.

15. In paragraph 1, The filler composition further comprises a solvent comprising at least one selected from the group consisting of water, saline, phosphate buffered saline, trisodium citrate buffer, tris-HCl buffer, and HEPES buffer.

16. In paragraph 15, A filler composition, wherein the solvent comprises at least one selected from the group consisting of phosphate buffered saline, trisodium citrate buffer, tris-HCl buffer, and HEPES buffer having a concentration of 2 to 20 mM.

17. In paragraph 15, Further comprising sugar alcohol dissolved in the above solvent, A filler composition wherein the sugar alcohol comprises mannitol or sorbitol.

18. In paragraph 15, A filler composition comprising the sugar alcohol in an amount of 1 to 5 wt%.

19. A first step of preparing a filler composition comprising a photocrosslinkable hyaluronic acid polymer, a photoinitiator, and a solvent; A second step of injecting the filler composition into the subcutaneous layer in a predetermined shape, or molding it into a predetermined shape after injection, or mixing an effective substance therein if necessary; and A filler treatment method, comprising a third step of irradiating the molded filler composition with light to crosslink the photocrosslinkable hyaluronic acid polymer.

20. In paragraph 19, A filler treatment method, wherein the filler composition further comprises one or more active substances selected from the group consisting of peptides, proteins, drugs, and cells.

21. In paragraph 20, A filler treatment method for releasing the effective substance from the cross-linked filler composition.

22. In paragraph 19, The photocrosslinkable hyaluronic acid polymer has a photoreactive functional group with a degree of substitution of 10 to 140% and an average molecular weight of 100 to 3000 kDa, The above filler treatment method further comprises a step of injecting a hyaluronic acid decomposing enzyme into the cross-linked filler composition to decompose the cross-linked hyaluronic acid polymer.

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