Method for manufacturing composite filament comprising atelocollagen and composition comprising same

Abstract

The present invention relates to a method for manufacturing a composite filament for a filler comprising collagen and a composition comprising same, wherein the composition exhibits improved dispersibility in a solvent. Specifically, the filler composition of the present invention has enhanced filler retention and excellent storability. When formulated as an injectable preparation, the filler composition exhibits improved dispersibility and allows the amount and site of injection to be easily controlled so that the filler composition does not form lumps or aggregates after filler injection and is easy to control according to the intention of the practitioner.

Description

Method for producing composite filaments containing atelocollagen and composition containing the same

[0001] The present invention relates to a method for producing a composite filament containing atelocollagen and a composition containing the same, wherein the composition has improved dispersibility in a solvent.

[0002] As life expectancy increases and interest in beauty grows across all age groups, interest in various plastic surgeries, procedures, and cosmetics is also increasing, leading to rapid technological growth in related fields.

[0003] Recently, there has been a significant increase in interest in cosmetic procedures that provide immediate effects immediately after the procedure, compared to cosmetics, and do not require surgery or anesthesia, such as plastic surgery. Accordingly, new technologies for various procedures, such as fillers, botox, laser treatment, and radiofrequency treatment, are being actively developed.

[0004] Fillers have been developed from the first generation fillers that used collagen extracted from animals, the second generation fillers that use hyaluronic acid, the third generation fillers that use substances that do not easily break down in the body, such as calcium fillers or PMMA, and the fourth generation fillers that use biodegradable polymers such as polycaprolactone.

[0005] Collagen fillers have been known for many years, but due to problems such as allergic reactions, the next generation of fillers was developed repeatedly, but the problem of allergic reactions was solved by using atelocollagen, which is collagen extracted from pigs and treated with enzymes to remove the telopeptides at both ends of the collagen that cause immune reactions.

[0006] However, even if these allergic reactions are overcome, collagen fillers generally have the problem of showing a short maintenance period compared to PCL fillers or polymer fillers.

[0007] Korean Patent No. 1796914 relates to a polymer composite for a filler comprising collagen and dialdehyde starch, and discloses a polymer composite for a biocompatible filler comprising collagen (COL), dialdehyde starch, and heparin, characterized in that the collagen is crosslinked by an imine bond with dialdehyde starch, and the heparin is interconnected with the collagen by an amide bond.

[0008] Korean Patent No. 2527685 relates to a silicone oil-in-water composition useful as an injectable filler and a scaffold for collagen growth, the filler composition comprising: (a) 1 to 79 wt% of silicone having a viscosity of at least 12500 centistokes (cSt) at 25°C; (b) 20 to 98 wt% of water; and (c) 0.005 to 10 wt% of a thickener, wherein the filler composition is a pharmaceutically acceptable emulsion, the silicone is dispersed in water as droplets having an average diameter of 2000 microns or less, and the thickener is biodegradable when implanted subcutaneously in a human body to provide a temporary scaffold for collagen growth between the silicone droplets.

[0009] Japanese Patent No. 7054954 relates to a polycaprolactone microsphere filler containing collagen peptides and a method for producing the same. The technology discloses a method for providing a cosmetic filler that is excellent in the properties of repairing or increasing volume of soft tissues such as cheeks, breasts, nose, lips, and buttocks and improving wrinkles by encapsulating collagen peptides in polycaprolactone microspheres and, when injected into a living body, rapidly exhibiting a collagen formation effect and exhibiting high tissue repair properties, as well as maintaining the effect for a long period of time.

[0010] In this way, development of various fillers that utilize collagen or have the effect of promoting collagen formation is still actively in progress, and the inventors of the present invention have completed an invention regarding a collagen filler composition that has an excellent filler maintenance effect due to delayed decomposition when injected into the body, while also having excellent dispersibility within the injection and being easy to control during the procedure, after repeated experiments and research.

[0011] The purpose of the present invention is to provide a filler composition having an effect of delaying the rate of decomposition in the body and thus improving the filler retention effect.

[0012] The purpose of the present invention is to provide a filler composition having excellent storage properties and improved dispersibility when manufactured as an injection.

[0013] In order to solve the above problems, the present invention provides a method for manufacturing a composite filament for a filler, comprising the steps of: (a) adding atelocollagen, hyaluronic acid, and polylactic acid to a solvent in a weight ratio of 1: 0.1 to 0.5: 0.1 to 0.5 to prepare a mixed solution, (b) distilling the mixed solution prepared in step (a) under reduced pressure to form a viscosity of the mixed solution of 500 to 800 cPs, (c) electrospinning the mixed solution obtained through step (b) to form continuous fibers and filaments of 10 to 30 μm (micrometers), (d) vacuum-spraying carbodiimide onto the surface of the filament prepared in step (c) to surface-coat it, and (e) freeze-drying the surface-coated filament prepared in step (d).

[0014] In the present invention, the average diameter of the continuous fiber manufactured in step (c) may be 1 to 5 ㎛ (micrometers).

[0015] In the present invention, a step of sterilizing the surface-coated filament that has undergone the step (e) and storing it in a sealed state at a temperature of 3 to 5°C for 24 hours may be further included.

[0016] The present invention also provides a filler composition comprising 5 to 20 parts by weight of a composite filament for filler, 0.05 to 20 parts by weight of a carrier, 0.05 to 20 parts by weight of a pain reliever, and 100 parts by weight of a vehicle for injection.

[0017] In the filler composition according to the present invention, the carrier may include at least one selected from the group consisting of sodium carboxymethyl cellulose, sodium alginate, gelatin, albumin, collagen, sodium hyaluronic acid, dextran, hydroxyethyl cellulose, hypromellose, glycerin, sorbitol, and propylene glycol.

[0018] In the filler composition according to the present invention, the pain reliever may include at least one selected from the group consisting of lidocaine, bupivacaine, lignocaine, ropivacaine, cocaine, tetracaine, amethocaine, amylocaine, benzydamine, cinchocaine, levobupivacaine, mepivacaine, oxybuprocaine, prilocaine, procaine, proparacaine, and salts thereof.

[0019] In the filler composition according to the present invention, the injection vehicle may include at least one selected from the group consisting of water for injection, saline solution, polyalcohols, and fatty acids.

[0020] The filler composition according to the present invention has an improved filler retention effect.

[0021] The filler composition according to the present invention has excellent storage properties and has improved dispersibility when manufactured into an injection.

[0022] The filler composition according to the present invention has the characteristics of being easy to control according to the intention of the practitioner without lumping or clumping after filler injection, as it is easy to control the injection amount and injection location by injection.

[0023] Figure 1 is a graph showing changes in skin volume of SD rats over time after filler injection in examples and comparative examples.

[0024] Hereinafter, each component of the present invention will be described in more detail so that a person having ordinary skill in the art to which the present invention pertains can easily practice it; however, this is only an example, and the scope of the rights of the present invention is not limited by the following contents.

[0025] As used herein, the terms "preferred" or "preferably" refer to embodiments of the invention that have particular advantages under specific conditions. However, other embodiments may also be preferred under the same or different conditions. Furthermore, the fact that one or more preferred embodiments are preferred does not imply that other embodiments are not useful, nor does it exclude other embodiments within the scope of the invention.

[0026] The term "comprises" as used herein is used to list materials, compositions, devices, and methods useful in the present invention, but is not limited to the listed examples.

[0027] Collagen is a major protein component of the extracellular matrix and is found in large quantities in soft tissues such as skin, tendons, and blood vessels, as well as hard tissues such as bones and teeth. It accounts for about one-third of the total protein in mammals and plays a role in forming the basic structure of tissues and organs through the assembly of cells.

[0028] As we age, the amount of collagen fibers that make up most of the dermis decreases, the activity of fibroblasts that synthesize collagen decreases, and the synthesis of new collagen fibers also decreases, resulting in wrinkles and loss of elasticity.

[0029] Aging skin loses the ability to support fibers like collagen and retain moisture. This leads to a loss of elasticity, which makes wrinkles difficult to repair. Furthermore, as skin cell activity declines, the production of collagen, a protein that makes up the dermis, accelerates. Dermal collagen is further destroyed by collagenase, a collagen-degrading enzyme, and elastinase, an enzyme that breaks down elastin.

[0030] In order to solve the above-described problem, the composite filament for filler according to the present invention, which includes collagen, has the effect of forming volume in the skin when injected into the filler, filling in sunken skin areas, and improving elasticity.

[0031] In particular, the composite filament for filler according to the present invention has a collagen cross-linking agent applied to the surface, and after insertion into the body, it is formed into a denser tissue through cross-linking over time, thereby providing skin volume immediately after injection, and at the same time, it forms a solid cross-linking structure in a settled state, thereby having an improved maintenance effect.

[0032] Specifically, the method for manufacturing a composite filament for filler comprising collagen according to the present invention comprises the following steps:

[0033] (a) a step of preparing a mixed solution by adding atelocollagen, hyaluronic acid, and polylactic acid to a solvent in a weight ratio of 1: 0.1 to 0.5: 0.1 to 0.5;

[0034] (b) a step of distilling the mixed solution prepared in step (a) under reduced pressure to form the viscosity of the mixed solution to 500 to 800 cPs;

[0035] (c) a step of forming continuous fibers by electrospinning the mixed solution that has gone through step (b) and forming them into filaments of 10 to 30 μm (micrometers);

[0036] (d) a step of vacuum-spraying carbodiimide onto the surface of the filament manufactured in step (c) to form a surface coating; and

[0037] (e) A step of freeze-drying the surface-coated filament manufactured in step (d).

[0038] Below, the composition of each step is described in more detail.

[0039] The above step (a) is a step of preparing a mixed solution by adding atelocollagen, hyaluronic acid, and polylactic acid to a solvent at a predetermined ratio.

[0040] The above solvent is a solution adjusted to pH 4.5 to 6 by mixing 95% concentration ethanol and acetic acid, and is used so that the weight of the solute is 1 to 10 parts by weight based on 100 parts by weight of the solvent.

[0041] Atelocollagen has a very low level of immune response, few side effects, and its molecular weight is smaller than that of regular collagen, making it easy for the skin to absorb.

[0042] The above atelocollagen can be a biocompatible atelocollagen, and can have a molecular weight of 300,000 Da or less.

[0043] The above hyaluronic acid may be used having a weight average molecular weight of 2.5 MDa to 3.5 MDa, and the polylactic acid may be specifically poly-L-lactic acid (PLAA, Poly-L-Lactic Acid), and may be used having a weight average molecular weight of 2.0 MDa to 4.0 MDa.

[0044] In the present invention, in particular, a composite filament is manufactured by mixing atelocollagen, hyaluronic acid, and polylactic acid in a weight ratio of 1: 0.1 to 0.5: 0.1 to 0.5. However, if hyaluronic acid and polylactic acid are not included or the ratio is lower than the above ratio, there is a problem that the decomposition speed of the filament is accelerated and the degree of improvement in the filler retention effect is not great, and if it is included in a ratio higher than the above ratio, the collagen crosslinking effect by the crosslinking agent on the surface to be post-treated is minimal, so there is a problem that the volume forming effect after filler injection is low.

[0045] The above step (b) is a step for removing the solvent from the mixed solution to produce a high-viscosity solution. In this step, the solvent is removed by reduced pressure distillation, thereby producing a solution with a viscosity of 500 to 800 cPs without damage or deformation of the components such as collagen and hyaluronic acid.

[0046] If the viscosity is lower or higher than the above, there is a problem in manufacturing the mixed solution into a continuous fiber by electrospinning.

[0047] At this time, the diameter of the continuous fibers manufactured is formed to be 1 to 5 ㎛ (micrometers), and if the thickness is thicker than the above, there is a problem of reduced dispersibility when manufacturing a filler composition including composite filaments. In this case, additional excipients or dispersants may be added and used.

[0048] In the above step (c), the continuous fiber formed by electrospinning is formed into filaments of 10 to 30 μm (micrometers), preferably, 12 to 28 μm (micrometers), more preferably, 15 to 25 μm (micrometers). At this time, in order to cut into filaments of the corresponding length, the fibers may be dispersed in purified water and ultrasonic waves may be applied, or a laser may be used to cut into filaments while moving at intervals of the filament length.

[0049] In the above step (d), the surface of the obtained filament is coated by vacuum spraying carbodiimide. This can be done by spraying carbodiimide on the surface of the filament in a vacuum environment such as a vacuum chamber without a carrier such as a solvent or dispersant, and the coating can be done using a vacuum spraying device.

[0050] The above carbodiimide is a cross-linking agent for collagen, and is injected into the body in a state in which it is coated on the surface of the composite filament, cross-linking collagen in the body to form a firmer and more superior volume.

[0051] The amount of carbodiimide used is sprayed onto the surface of the composite filament in an extremely small amount using the vacuum spraying method described above to prevent unnecessary reactions in the body, and 0.001 to 0.01 parts by weight is used based on 100 parts by weight of the filament.

[0052] The above step (e) is a step of freeze-drying the surface-coated filament, which is a step to remove all solvent remaining inside and on the surface of the composite filament and to firmly bond the carbodiimide. Through this step, the composite filament for filler is completed.

[0053] If necessary, a step of sterilizing the surface-coated filament and storing it in a sealed state at a temperature of 3 to 5°C for 24 hours may be further included. Through this process, the adhesion between the composite filament surface and the carbodiimide can be further improved, and even when manufactured and stored as a filler composition, the effect can be maintained for a long period of time without deterioration, thereby improving the storage period.

[0054] The composite filament described above can be manufactured into a filler composition in an injectable formulation. Specifically, the filler composition can be manufactured into a filler composition comprising 5 to 20 parts by weight of the composite filament for filler described above, 0.05 to 20 parts by weight of a carrier, 0.05 to 20 parts by weight of a pain reliever, and 100 parts by weight of an injectable vehicle.

[0055] Any carrier that can be added to an injectable filler composition may be used, and for example, may include at least one selected from the group consisting of sodium carboxymethyl cellulose, sodium alginate, gelatin, albumin, collagen, sodium hyaluronic acid, dextran, hydroxyethyl cellulose, hypromellose, glycerin, sorbitol, and propylene glycol.

[0056] The pain reliever may also be any agent that can be added to an injectable filler composition, and may include, for example, one or more selected from the group consisting of lidocaine, bupivacaine, lignocaine, ropivacaine, cocaine, tetracaine, amethocaine, amylocaine, benzydamine, cinchocaine, levobupivacaine, mepivacaine, oxybuprocaine, prilocaine, procaine, proparacaine, and salts thereof.

[0057] Hereinafter, the present invention will be described in more detail based on examples, but this is only an exemplary description for understanding the present invention, and the scope of the present invention is not limited or restricted to the following examples.

[0058] [Example]

[0059] Atelocollagen, hyaluronic acid, and polylactic acid were mixed in a weight ratio of 1: 0.2: 0.2 based on the actual weight of each component, and the mixture was added to 5 L of a solution adjusted to pH 5 by mixing 95 wt% ethanol and acetic acid to prepare a mixed solution. At this time, the content was adjusted to 1 to 10 parts by weight based on 100 parts by weight of the solvent, and the mixture was mixed.

[0060] The above-mentioned mixed solution was distilled under reduced pressure to form a viscosity of 600 cPs, and then electrospinning (10 kV, drum rotation speed 15 rpm, accumulation distance 8 cm) was performed to form continuous fibers with an average diameter of 1 ㎛ (micrometer), and a laser cutter was used to manufacture filaments with an average length of 20 ㎛ (micrometer).

[0061] The surface of the manufactured filament was coated by vacuum spraying carbodiimide, and a very small amount of 0.005 parts by weight based on 100 parts by weight of the filament was vacuum sprayed to coat the surface, and this was freeze-dried to remove all remaining solvent and dispersion medium.

[0062] The obtained composite filament was sterilized and stored in a sealed state at a temperature of 3 to 5°C for 24 hours, and then mixed with water for injection, a carrier (sodium carboxymethyl cellulose), and a pain reliever (lidocaine) to prepare a filler composition.

[0063] At this time, the mixing ratio was prepared by mixing 12 parts by weight of composite filament, 5 parts by weight of carrier, and 1 part by weight of pain reliever based on 100 parts by weight of injection water.

[0064] [Comparative Example 1]

[0065] Use commercially available collagen filler (Company A)

[0066] [Comparative Example 2]

[0067] Use commercially available collagen filler (Company B)

[0068] [Comparative Example 3]

[0069] Use commercially available collagen filler (Company C)

[0070] [Comparative Example 4]

[0071] In the above example, when manufacturing a composite filament, the same method was used except that instead of adding atelocollagen, hyaluronic acid, and polylactic acid in a weight ratio of 1:0.2:0.2, only atelocollagen of the same weight was used.

[0072] [Comparative Example 5]

[0073] In the above example, when manufacturing a composite filament, the same method was used except that the mixing ratio of atelocollagen, hyaluronic acid, and polylactic acid was added at a weight ratio of 1:1:1 instead of 1:0.2:0.2.

[0074] [Experimental Method]

[0075] 1. Toxicity test

[0076] To investigate the effects of the composition of the present invention on SD rats, a toxicological evaluation was conducted. An equal volume of saline solution was injected as a control group.

[0077] After 8 weeks, blood tests were performed on SD rats in each group to examine hematopoiesis, hepatotoxicity, and nephrotoxicity through CBC, OT / PT (Oxaloacetic transaminase / Pyruvic transaminase), and BUN / Cr (Blood urea nitrogen / Creatinine) analysis.

[0078] White blood cells (6.0-14.0)(10 3 / mm 3 )Red blood cells (7.5~8.4)(10 6 / mm 3 )Hemoglobin (13~16) (g / dL)Hematocrit (40~50) (%)MCV (Mean Corpuscular Volume) (㎛ 3)MCH(Mean Corpuscular Hemoglobin)(pg)MCHC(Mean Corpuscular Hemoglobin Concentration)(g / dL)Platelet(900~1200)(10 3 / mm 3 )Control group 6.098.8115.147.653.216.930.91018Example 6.418.9916.246.950.117.032.0922

[0079] OT(20~40)PT(80~200)BUN(11~16)Cr(0.2~0.4)Control35.5103.611.30.26Example36.2106.412.50.31

[0080] Referring to Tables 1 and 2 above, it can be confirmed that the filler composition of the example is not toxic compared to the control group using saline solution. 2. Evaluation of filler retention effect

[0081] 100 μL (microliter) of the control group, examples, and comparative examples 1 to 5 were injected into the dorsal area of ​​SD rats, and the volume change at the injection site was measured for 50 weeks using PRIMOS CR (Canfield). The measurement results are shown in Fig. 1 and Table 3.

[0082] (Unit: mm 3 ) Week 4 Week 8 Week 25 Week 50 Control group 000-5 Example 2 4 4 2 5 2 2 6 0 1 9 1 Comparative example 1 2 5 0 1 8 3 1 0 4 4 9 Comparative example 2 1 6 7 1 1 6 8 6 2 4 Comparative example 3 1 3 2 1 2 9 2 3 1 Comparative example 4 2 5 6 2 3 1 1 6 2 5 6 Comparative example 5 1 5 7 1 6 6 1 5 2 1 0 3

[0083] 3. Expert Panel Test: Ten filler treatment experts (plastic surgeons, dermatologists, or filler treatment clinic doctors) were recruited, and a test was conducted in which 1 cc of filler composition was injected into the model's nasolabial fold area using an injection practice model capable of filler injection.

[0084] When injecting, ease of use of the injection (related to syringe injection and viscosity of the injection), ease of injection into the target treatment area (control of injection volume and injection location), whether the filler is maintained in the injection location after injection (observation after 5 minutes), and satisfaction after the procedure (evaluation by the practitioner) were comprehensively evaluated on a scale of 1 to 10 for each item, and the average score for each item is shown in the table below.

[0085] (Unit: mg / g) Injection properties Ease of administration Position maintenance Effectiveness Satisfaction with administration Example 8899 Comparative example 18988 Comparative example 28755 Comparative example 39532 Comparative example 48888 Comparative example 55383

[0086] Fig. 1 is a graph showing changes in skin volume of SD rats over time after filler injection of examples and comparative examples. Referring to Tables 1 to 4 and Fig. 1, it can be confirmed that the filler composition of the examples does not have toxicity to animals, and the filler maintenance effect is maintained for a long period of time compared to commercially available collagen fillers.

[0087]

[0088] *In addition, it can be confirmed that the filler composition of the embodiment is convenient for the practitioner to use, and that the procedure can be easily performed according to the practitioner's intention during the procedure.

[0089] Meanwhile, it can be confirmed that the commercially available collagen fillers, Comparative Examples 1 to 3, have a filler maintenance effect in that the change in volume is not large even at the initial stage compared to the same injection amount, and the volume decreases rapidly over time.

[0090] In the case of Comparative Example 4, since a filament using only atelocollagen is used without utilizing hyaluronic acid or PLA polymer, the initial volume is excellent, but the volume gradually decreases, and it can be confirmed that the volume decreases to about 1 / 5 times at 50 weeks, so it can be confirmed that the filler maintenance effect is somewhat reduced.

[0091] In the case of Comparative Example 5, although it does not form a high volume, it maintains a similar volume until the 25th week, and the decrease compared to the initial volume is not large even until the 50th week, but there was an expert opinion that the ease of procedure of the filler composition, in particular, the dispersibility of the filaments within the filler composition is poor and the viscosity of the composition is high, making it difficult to use and control the injection, and thus the ease of procedure is greatly reduced, as shown in Table 4.

[0092] Therefore, as in the examples, it can be confirmed that the filler composition according to the present invention, compared to existing collagen fillers, has a filler maintenance effect, that is, a volume maintenance effect, that is, a long-term maintenance effect, and has properties suitable for use as an injection, and has characteristics suitable for use in filler treatment.

Claims

1. (a) A step of preparing a mixed solution by adding atelocollagen, hyaluronic acid, and polylactic acid to a solvent in a weight ratio of 1: 0.1 to 0.5: 0.1 to 0.5; (b) a step of distilling the mixed solution prepared in step (a) under reduced pressure to form the viscosity of the mixed solution to 500 to 800 cPs; (c) a step of forming continuous fibers by electrospinning the mixed solution that has gone through step (b) and forming them into filaments of 10 to 30 μm (micrometers); (d) a step of vacuum-spraying carbodiimide onto the surface of the filament manufactured in step (c) to form a surface coating; and (e) a step of freeze-drying the surface-coated filament manufactured in step (d); A method for manufacturing a composite filament comprising:

2. In paragraph 1, A method for manufacturing a composite filament, wherein the average diameter of the continuous fiber manufactured in the above step (c) is 1 to 5 ㎛ (micrometers).

3. In paragraph 1, A method for manufacturing a composite filament, further comprising a step of sterilizing the surface-coated filament that has undergone the above step (e) and storing it in a sealed state at a temperature of 3 to 5°C for 24 hours.

4. A composition comprising 5 to 20 parts by weight of a composite filament for filler manufactured according to paragraph 1, 0.05 to 20 parts by weight of a carrier, 0.05 to 20 parts by weight of a pain reliever, and 100 parts by weight of a vehicle for injection.

5. In paragraph 4, A composition comprising at least one carrier selected from the group consisting of sodium carboxymethyl cellulose, sodium alginate, gelatin, albumin, collagen, sodium hyaluronic acid, dextran, hydroxyethyl cellulose, hypromellose, glycerin, sorbitol, and propylene glycol.

6. In paragraph 4, A composition comprising at least one selected from the group consisting of lidocaine, bupivacaine, lignocaine, ropivacaine, cocaine, tetracaine, amethocaine, amylocaine, benzydamine, cinchocaine, levobupivacaine, mepivacaine, oxybuprocaine, prilocaine, procaine, proparacaine, and salts thereof.

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