KR102581434B1 - Solution used for production of composition of hyaluronic acid filler and composition of hyaluronic acid filler using the same - Google Patents

Abstract

The present invention relates to a solution used in the production of a hyaluronic acid filler composition and a hyaluronic acid filler composition using the same. The hyaluronic acid filler composition prepared using the solution according to the present invention is capable of combining calcium ions present in body fluids and It is effective in preventing calcification and the resulting nodules, which are typical side effects that occur when combined, so it can be safely used not only as a facial filler injected in small quantities, but also as a filler injected in large quantities to increase tissue volume in the chest and buttocks. there is.

Description

Solution used in the production of hyaluronic acid filler composition and hyaluronic acid filler composition using the same {SOLUTION USED FOR PRODUCTION OF COMPOSITION OF HYALURONIC ACID FILLER AND COMPOSITION OF HYALURONIC ACID FILLER USING THE SAME}

The present invention relates to a solution used in the production of a hyaluronic acid filler composition and a hyaluronic acid filler composition using the same, and more specifically, to hyaluronic acid in which the total content of phosphate is adjusted to suppress calcification in the body caused by the hyaluronic acid filler composition. It relates to a gel cleaning solution and a hyaluronic acid filler composition using the same.

As people age, wrinkles appear due to the effects of ultraviolet rays, facial expression changes, gravity, etc., and the volume of related tissues decreases along with loss of fat tissue, which is called aging. Additionally, various factors can cause scars where the skin becomes depressed.

Fillers were developed to improve wrinkles, skin depression, and tissue volume reduction. Fillers are complementary materials or contents that are injected or inserted into body parts where volume is desired to increase, such as wrinkles or dimpled scars. The injection of filler temporarily restores a smooth, youthful appearance to the skin and tissues.

When injected into the skin or under the skin, fillers must maintain continuity for a certain period of time and have the functionality of inducing a soft, smooth, and natural appearance. When injected into the body, side effects such as allergic reactions, swelling, nodules, and granulomas must be minimized. do.

Hyaluronic acid (HA) (also known as hyaluronan), which is used as the main ingredient in filler, is a type of glycosaminoglycans (GAGs) and is a major component of the extracellular matrix. Because it is widely distributed in animal tissues, it provides the advantage of significantly reducing the risk of allergic reactions. Hyaluronic acid is present in abundance in several different layers of the skin, and its main functions include, for example, correcting hydration of the skin, assisting in the organization of the extracellular matrix, acting as a filler material, and tissue regeneration mechanism. It has complex functions such as being involved in

However, despite these excellent safety and efficacy, hyaluronic acid filler compositions can bind to calcium ions in the body, causing precipitation and causing calcification reactions, causing side effects such as nodules in the area where the filler was injected. This problem can be especially noticeable when large amounts of filler are injected for the purpose of breast or buttock plastic surgery. The larger the amount injected, the higher the probability of it combining with calcium ions present in body fluids, which also increases the occurrence of nodules due to calcification.

The present inventors were looking for a way to reduce side effects such as calcification and the resulting nodules due to filler injection, and found a filler in which the total content of phosphate in Phosphate Buffered Saline (PBS) used in filler production was controlled. In the case of the composition, the present invention was completed by confirming that calcification was significantly reduced in reaction with an analogous biological solution.

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Domestic Patent Publication No. 10-2006-0127897 (published on December 13, 2006) Domestic Patent Publication No. 10-2020-0077449 (published on June 30, 2020)

The purpose of the present invention is to provide a hyaluronic acid gel washing solution with adjusted total phosphate content and a hyaluronic acid filler composition using the same to suppress calcification in the body, which is a major side effect that occurs when hyaluronic acid filler injection.

The present invention provides a solution used in the production of a hyaluronic acid filler composition, which contains phosphate, has a content of 0.1mM to less than 10mM, and is used to suppress calcification in the body caused by the hyaluronic acid filler composition. do.

The solution may additionally contain sodium chloride.

The solution may be for washing the hyaluronic acid gel.

The phosphate content of the solution may be 1mM to 7mM.

The phosphate in the solution may include one or more selected from the group consisting of monohydrogen phosphate and dihydrogen phosphate.

In one embodiment of the present invention, a hyaluronic acid filler composition comprising a hyaluronic acid gel washed with the above solution is provided.

The hyaluronic acid filler composition may include cross-linked hyaluronic acid or a salt thereof.

The hyaluronic acid filler composition may be a monophasic hyaluronic acid gel with a MoD of 1% to 13%.

The pH of the hyaluronic acid filler composition may be 5.5 to 8.5.

The osmotic pressure of the hyaluronic acid filler composition may be 200 mOsmol/kg to 400 mOsmol/kg.

The hyaluronic acid filler composition may additionally include an anesthetic agent.

The hyaluronic acid filler composition may be administered in small doses of less than 0.1 mL to 10 mL.

The hyaluronic acid filler composition may be administered in a large volume of 10mL to 200mL.

The hyaluronic acid filler composition may be used to improve wrinkles and tissue volume reduction in soft tissues selected from the group consisting of the face, neck, chest, buttocks, arms, armpits, hands, legs and feet, or to treat wounds, scars or stretch marks. .

The hyaluronic acid filler composition prepared using the solution according to the present invention has the effect of preventing calcification and the resulting nodules, which are typical side effects that can occur when injected in vivo by binding to calcium ions present in body fluids, and can be injected in small quantities. It can be safely used not only as a facial filler, but also as a filler injected in large quantities to increase tissue volume in the chest and buttocks.

Figure 1 shows the results of confirming whether an analogous biological solution with a composition similar to body fluid is suitable for detecting a calcification reaction.
Figure 2 shows the reaction of Phosphate Buffer adjusted to have different pH (weight ratio of monohydrogen phosphate and dihydrogen phosphate) and total phosphate content according to preliminary embodiments of the present invention (Preliminary Experiment Examples 1 to 5) with a similar biological solution. This is a preliminary result of observing the calcification reaction after treatment.
Figure 3 shows hyaluronic acid gel being washed using Phosphate Buffered Saline (PBS) having a total phosphate content of 1, 5, and 7mM, respectively, according to one embodiment of the present invention (Experimental Examples 1 to 3), but no anesthetic was added. This is the result of observing the calcification reaction for 14 days after reacting a hyaluronic acid filler composition with an analogous biological solution.
Figure 4 shows a hyaluronic acid filler composition in which the hyaluronic acid gel was washed using Phosphate Buffered Saline (PBS) with a total phosphate content of 10mM according to an embodiment of the present invention (Comparative Experiment Example 1), but no anesthetic was added. This is the result of observing the calcification reaction for 14 days after reacting with a similar biological solution.
Figure 5 shows a hyaluronic acid filler composition in which the hyaluronic acid gel was washed using Phosphate Buffered Saline (PBS) with a total phosphate content of 5mM according to another embodiment of the present invention (Experimental Example 4), but no anesthetic was added. This is the result of observing the calcification reaction for 14 days after reacting with a similar biological solution.
Figure 6 shows that according to another embodiment of the present invention (Experimental Example 4), the hyaluronic acid gel was washed using Phosphate Buffered Saline (PBS) with a total phosphate content of 5mM, but the hyaluronic acid filler composition to which an anesthetic agent was added was similar. This is the result of observing the calcification reaction for 14 days after reacting with the biological solution.
Figure 7 shows the results of observing the calcification reaction through (a) the naked eye and (b) a microscope for 14 days after reacting a commercially available filler composition (Bv) with a biological-like solution.
Figure 8 shows the results of observing the calcification reaction through (a) the naked eye and (b) a microscope for 14 days after reacting a commercially available filler composition (Cs) with a biological-like solution.
Figure 9 shows the results of observing the calcification reaction (a) with the naked eye and (b) with the microscope for 14 days after reacting a commercially available filler composition (Nv) with a biological-like solution.
Figure 10 shows the results of visual observation of the calcification reaction for 14 days after reacting a commercially available filler composition (CN°4) with a biological-like solution.

Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. In general, the nomenclature used herein is well known and commonly used in the art.

Conventional hyaluronic acid-based filler compositions had the problem of causing side effects such as nodules in the area where the filler was injected. Accordingly, as a result of collecting literature information to solve the above side effects, it was confirmed that calcification can occur when calcium is bound to the carboxyl group of an amino acid and then phosphate is added. Based on the results of literature analysis, the present inventors observed the calcification reaction after reacting a currently commercially available hyaluronic acid filler composition with an analogous biological solution. As a result, it was confirmed that a white crystalline substance was generated, although the aspect was different for each product, and the crystals formed. As a result of analyzing the components using SEM-EDS (scanning electron microscope, LEO SUPRA55, Carl zeiss, Germany), it was confirmed that the crystal component was calcium phosphate. Accordingly, the present inventors confirmed that calcification due to the reaction between the hyaluronic acid filler composition and the biological-like solution is suppressed by optimizing the total content of phosphate in Phosphate Buffered Saline (PBS) used in manufacturing the hyaluronic acid filler composition, and thus, the present invention was completed.

Hereinafter, the present invention will be described in detail.

Solutions used in the preparation of hyaluronic acid filler compositions

The present invention provides a solution used in the production of a hyaluronic acid filler composition, which contains phosphate, has a content of 0.1mM to less than 10mM, and is used to suppress calcification in the body caused by the hyaluronic acid filler composition. to provide.

The solution according to the invention may additionally comprise sodium chloride.

The solution according to the invention can be used for the preparation of hyaluronic acid filler compositions, in particular for washing hyaluronic acid gels.

First, the solution according to the present invention contains phosphate, the content of which needs to be adjusted to prevent calcification in the body. The content of phosphate in the composition is characterized as being 0.1mM to less than 10mM, and is more preferably 1mM to 7mM, but is not limited thereto. At this time, when the phosphate content in the composition is less than 0.1mM, there is a problem that the NaOH used to prepare the cross-linked hyaluronic acid gel is not completely washed away, and when the phosphate content in the composition is more than 10mM, calcification is not sufficiently prevented. There is a problem that does not exist.

Meanwhile, the phosphate may be one or more types of hydrogen phosphate, and preferably may include at least one selected from the group consisting of monohydrogen phosphate and dihydrogen phosphate. The one or more types of hydrogen phosphate may exist independently in the form of In the case of a hydrogen salt, it can be expressed as (Na,K) ₂ HPO ₄ ·xH ₂ O or (Na,K)H ₂ PO ₄ ·xH ₂ O, respectively. In order to adjust the pH of the final hyaluronic acid filler composition in one aspect of the present invention, when the monohydrogen phosphate is in an anhydrous form and the dihydrogen phosphate is in a dihydrate form, the weight ratio of the monohydrogen phosphate and dihydrogen phosphate is It is preferably 1:5 to 28:1, but is not limited thereto. At this time, when the ratio of monohydrogen phosphate increases, there is a problem that the pH increases, and when the ratio of dihydrogen phosphate increases, there is a problem that the pH decreases. This is because the final hyaluronic acid filler composition is different from the pH of the human body. Accordingly, there is a problem that pain may occur when administered into the body.

Next, the solution according to the present invention contains sodium chloride, which plays a role in regulating the osmotic pressure of the final hyaluronic acid filler composition. The concentration of sodium chloride in the composition is preferably 0.33 to 1.46 (w/v)%, more preferably 0.47 to 1.16 (w/v)%, but is not limited thereto.

Hyaluronic acid filler composition

The present invention provides a hyaluronic acid filler composition comprising a hyaluronic acid gel washed with the above solution.

The hyaluronic acid filler composition comprises hyaluronic acid or a salt thereof, preferably cross-linked hyaluronic acid or a salt thereof. The concentration of hyaluronic acid or its salt in the hyaluronic acid filler composition is preferably 1.0 (w/v)% to 3.0 (w/v)%, and 1.5 (w/v)% to 2.5 (w/v)%. It is more preferable, but is not limited thereto.

The hyaluronic acid filler composition is preferably a monophasic hyaluronic acid gel with a MoD (MoD (%) = number of moles of crosslinker/number of moles of repeating units of hyaluronic acid disaccharide or its salt) Х100) of 1% to 13%, but is not limited thereto. No.

On the other hand, when the average particle size of the cross-linked hyaluronic acid or its salt is small, the reaction with the biological solution becomes active due to the high surface area, which may accelerate the calcification reaction. Accordingly, large particle sizes such as biphasic fillers with an average particle size (D50) of 250 to 900 ㎛ are desirable in that they can effectively suppress calcification in the body, but there is a risk of the filler composition moving after injection due to low cohesion. There is. Therefore, in the present invention, it is preferable that the average particle size (D50) is monophasic with an average particle size (D50) of 50 ㎛ to 500 ㎛, but is not limited thereto.

The pH of the hyaluronic acid filler composition according to the present invention is preferably about pH 5.5 to 8.5, which is a range that is physiologically acceptable and can suppress calcification in the body, but is not limited thereto.

The osmotic pressure of the hyaluronic acid filler composition according to the present invention is a physiologically acceptable range and may be non-limitingly in the range of about 150 mOsmol/kg to 500 mOsmol/kg, and is more preferably about 200 mOsmol/kg to 400 mOsmol/kg, but is limited thereto. It doesn't work.

The hyaluronic acid filler composition according to the present invention may further include an anesthetic, and the concentration of the anesthetic in the filler composition may be an amount effective in relieving pain that can be felt when injected into the body. Moreover, since the inclusion of an anesthetic in the filler composition lowers the pH, it is necessary to optimize the pH by adjusting the weight ratio of the monohydrogen phosphate and dihydrogen phosphate. The concentration of the anesthetic (preferably a local anesthetic) in the filler composition may be 0.1 to 5.0 (w/v)%, preferably 0.2 to 1.0 (w/v)%, and 0.3 (w/v)%. This is more preferable, but is not limited to this. Specifically, the local anesthetics include ambucaine, amolanone, amylocaine, benoxinate, benzocaine, betoxycaine, and biphenamine. (biphenamine), bupivacaine, butacaine, butamben, butanilicaine, butethamine, butoxycaine, carticaine, chlorine Chloroprocaine, cocaethylene, cocaine, cyclomethycaine, dibucaine, dimethysoquine, dimethocaine, diperodone ( diperodon, dicyclonine, ecgonidine, ecgonine, ethyl chloride, etidocaine, beta-eucaine, euprosine (euprocin), fenalcomine, formocaine, hexylcaine, hydroxytetracaine, isobutyl p-aminobenzoate, leucinocaine mesylate, levoxadrol, lidocaine, mepivacaine, meprylcaine, metabutoxycaine, methyl chloride, myrtecaine , naepaine, octacaine, orthocaine, oxethazaine, parethoxycaine, phenacaine, phenol, piperocaine ( piperocaine, piridocaine, polidocanol, pramoxine, prilocaine, procaine, propanocaine, proparacaine, propy Propipocaine, propoxycaine, psuedococaine, pyrrocaine, ropivacaine, salicyl alcohol, tetracaine, tolycaine ), trimecaine, zolamine, and salts thereof, but lidocaine is preferred, and lidocaine hydrochloride is more preferred, but is not limited thereto.

The hyaluronic acid filler composition according to the present invention may optionally include a buffer, preservative, isotonic agent, antioxidant, emulsifier, wetting agent, etc., but is not limited thereto, and may include other pharmaceutically acceptable ingredients.

The hyaluronic acid filler composition according to the present invention may further include other substances or combinations of substances that provide beneficial effects when administered to a subject. Such beneficial substances include, but are not limited to, antioxidants, anti-itch agents, anti-cellulite agents, anti-scar agents, anti-inflammatory agents, anesthetics, irritant-relieving agents, vasoconstrictors, vasodilators, anti-hemorrhagic agents such as hemostatic agents or anti-inflammatory agents. Includes anti-fibrotic agents, exfoliants, firming agents, anti-acne agents, colorants, anti-staining agents, or moisturizers.

The hyaluronic acid filler composition according to the present invention may be sterilized, for example, by autoclave or by heat, pressure, gamma irradiation, etc. In certain embodiments of any of the above aspects, a composition placed subcutaneously or intradermally may provide immediate 'lifting' due in part to its cohesive or viscoelastic properties. This smoothes lines and folds on the skin, giving the appearance of skin rejuvenation.

The hyaluronic acid filler composition according to the present invention can be used as a facial filler injected in a small volume of about 0.1 mL to less than 10 mL, and can also be used as a filler injected in a large volume of 10 mL to 200 mL, but is not limited thereto. In particular, the hyaluronic acid filler composition according to the present invention can effectively suppress calcification in the body by optimizing the total content of phosphate, so it can be safely used as a filler injected in large quantities to increase tissue volume such as the breasts and buttocks. There is.

The hyaluronic acid filler composition according to the present invention is used for improving wrinkles and tissue volume reduction targeting soft tissues selected from the group consisting of the face, neck, chest, buttocks, arms, armpits, hands, legs and feet, or for treating wounds, scars or stretch marks. It is characterized by being acceptable, but is not limited to this. At this time, the scar may be a post-surgical scar, an old scar, or a newly developed scar, and the wound may be an acute or chronic wound, including a burn.

The hyaluronic acid filler composition according to the present invention provides a method of treating soft tissue disease by administering it to an individual. As used herein, “treating” means reducing or eliminating the cosmetic or clinical symptoms of a soft tissue pathology characterized by soft tissue defects, defects, diseases, and/or disorders in an individual; or delaying or preventing the onset of cosmetic or clinical symptoms of a condition characterized by soft tissue defects, defects, diseases, and/or disorders in an individual. The effectiveness of the filler composition may be measured by observing one or more cosmetic, clinical symptoms, and/or physiological indicators associated with the condition. Improvement of soft tissue defects, diseases, and/or disorders may also be indicated by reduced need for concurrent therapy. Those skilled in the art will know the relevant symptoms or indicators associated with a particular soft tissue defect, disease, and/or disorder and will know how to determine whether an individual is a candidate for treatment with a hyaluronic acid filler composition according to the present invention.

The term “hyaluronic acid” used in the present invention refers to microbial-derived glycosaminoglycans (GAGs) used to formulate fillers mainly used to reduce facial wrinkles and enhance volume. This is because the polymer chains are easily broken down by enzymes and free radicals found in humans, so the retention time of non-denatured hyaluronic acid is several days. Therefore, to improve residence time in the human body, the linear chains of hyaluronic acid are usually cross-linked with a cross-linker such as BDDE.

The term “calcification” used in the present invention refers to the abnormal deposition of calcium salts, such as calcium phosphate (Ca ₃ (PO ₄ ) _2, CaHPO ₄ ), amorphous amino acids/calcium phosphate, etc., in biological tissues. This is a phenomenon in which biological tissue hardens, and can be seen as a type of side effect that can occur when filler containing excessive amounts of phosphate is administered into the body.

As used herein, the term “administering” refers to any mode of delivery that provides a subject with the composition that potentially produces a clinically, therapeutically, or experimentally beneficial result. The actual delivery mode used to administer the composition to a subject will be determined by those skilled in the art, including, but not limited to, the type of dermatopathy, the location of the dermatopathy, the cause of the dermatopathy, the severity of the dermatopathy, the degree of relief desired, and the degree of relief desired. The period of relief achieved, the specific composition used, the secretion rate of the specific composition used, the pharmacodynamics of the specific composition used, the nature of other compounds included in the specific composition used, the specific route of administration, the specific characteristics, the medical history and risks of the subject. The determination may be made by taking into account factors such as age, weight, general health, etc., or any combination thereof.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Verification of similar biological solutions

The composition of the Simulated Body Fluid (Biosesang, Korea) used in the present invention is as follows: Na ⁺ 142.0mM, K ⁺ 5.0mM, Ca ²⁺ 2.5mM, Mg ²⁺ 1.5mM, Cl ^- 125.0mM, HCO ^3- 27.0mM, HPO ₄ ^2- 1.0mM, SO ₄ ^2- 0.5mM, Ca/P 2.5mM in Tris Buffer(pH 7.4).

In order to check whether the analogous biological solution prepared to be equivalent to the electrolyte concentration of body fluid was appropriately prepared, H ₂ O (negative control group), CaCl ₂ (positive control group), NaH ₂ PO _{4.2H 2} O (test group 1) ), 1.0g each of Na ₂ HPO ₄ (test group 2) was taken and mixed with 9.0g of biosimilar solution, and then treated with ultrasound (Elamsonic P, ELMA) for 10 minutes to accelerate the reaction speed (Sonication conditions: Pulse mode, Temp.: 35℃ power: 100, frequency: 80, kHz: 35).

As a result, as shown in Figure 1, a calcification reaction was confirmed in CaCl ₂ , a positive control group, and it was confirmed that the pseudo-biological solution was appropriately prepared. In particular, a calcification reaction was confirmed in the test group Na ₂ HPO ₄ , that is, monohydrogen phosphate, and it was confirmed that phosphate is an influential factor that can cause a calcification reaction in the body.

Preliminary verification of pH (weight ratio of monohydrogen phosphate and dihydrogen phosphate) and total phosphate content that can inhibit calcification in the body in Phosphate Buffer

As shown in Table 1, a Phosphate Buffer Stock solution was prepared by varying the weight ratio of Na ₂ HPO ₄ and NaH ₂ PO ₄ .2H ₂ O so that the total content of phosphate at each pH was 100mM, and then diluted three times to obtain phosphate. The total content of was adjusted to 10mM to 100mM, and then 1.0g of each Phosphate Buffer was mixed with 9.0g of similar biological solution. Afterwards, it was left at room temperature for 20 minutes or treated with ultrasound (Elamsonic P, ELMA) for 10 minutes to accelerate the reaction rate (Sonication conditions: Pulse mode, Temp.: 35°C power: 100, frequency: 80, kHz: 35).

manufacturing company
(CAS-No) Preliminary comparative experiment example 1 Spare
Experimental Example 1 Spare
Experimental Example 2 Spare
Experimental Example 3 Spare
Experimental Example 4 Spare
Experimental Example 5 Preliminary comparative experiment example 2 Phosphate Buffer Sodium monohydrogen phosphate
(anhydride) Merck
(7558-79-4) 0
g/g 2.32
g/g 5.42
g/g 9.39
g/g 12.22
g/g 13.51
g/g 14.20
g/g Sodium dihydrogen phosphate
(dihydrate) Sigma Aldrich
(13472-35-0) 15.60
g/g 13.06
g/g 9.65
g/g 5.29
g/g 2.18
g/g 0.76
g/g 0
g/g 3rd order 984.40 g 984.63 g 984.93 g 985.33 g 985.61 g 985.73 g 985.80 g pH 4.33 6.13 6.49 7.00 7.52 8.05 9.21

As a result, as shown in Figure 2, it was confirmed that the calcification reaction intensified as the pH and total phosphate content of the Phosphate Buffer increased. Therefore, there is a need to minimize the total content of phosphate, and the pH also needs to be maintained similar to that of the human body.

Verification of hyaluronic acid filler composition by total content of phosphate in Phosphate Buffered Saline (PBS) capable of inhibiting calcification

As shown in Table 2, a hyaluronic acid filler composition was prepared by washing the hyaluronic acid gel using Phosphate Buffered Saline (PBS) with different total contents of phosphate. 1.0 g of the finally prepared hyaluronic acid filler composition was mixed with 9.0 g of the biosimilar solution, and the calcification reaction was observed at 37°C for 14 days.

manufacturing company
(CAS-No) Experimental Example 1
(Phosphate 1mM) Experimental Example 2
(Phosphate 5mM) Experimental Example 3
(Phosphate 7mM) Comparative Experiment Example 1
(Phosphate 10mM) Phosphate Buffered
Saline
(PBS) Sodium monohydrogen phosphate
(anhydride) Merck
(7558-79-4) 0.1073 mg/mL 0.5364 mg/mL 0.7510 mg/mL 1.0728 mg/mL Sodium dihydrogen phosphate
(dihydrate) Sigma Aldrich
(13472-35-0) 0.0381 mg/mL 0.1907 mg/mL 0.2670 mg/mL 0.3814 mg/mL sodium chloride Sigma Aldrich (7647-14-5) 8.6482 mg/mL 8.1806 mg/mL 7.9469 mg/mL 7.5962 mg/mL 3rd order 991.2064 mg/mL 991.0923 mg/mL 991.0351 mg/mL 990.9496 mg/mL Hyaluronic acid filler composition cross-linked
Sodium Hyaluronate 24mg/mL 24mg/mL 24mg/mL 24mg/mL pH 6.88 7.28 7.22 7.19 osmotic pressure 215 230 231 274

As a result, as shown in Figure 3, no calcification reaction was observed at all in the filler compositions finally prepared with the total phosphate content in Phosphate Buffered Saline (PBS) of 1mM, 5mM, and 7mM, respectively, for 14 days. Meanwhile, as shown in Figure 4, in the case of the filler composition finally manufactured with a total phosphate content of 10mM in Phosphate Buffered Saline (PBS), a significant calcification reaction was observed at 14 days. Therefore, it was confirmed that the solution of the present invention preferably contains a total phosphate content of less than 10mM, and more preferably less than 7mM.

Meanwhile, as shown in Table 3, the hyaluronic acid gel was washed using Phosphate Buffered Saline (PBS) with the total content of phosphate fixed at 5mM, and then a hyaluronic acid filler composition containing or not containing lidocaine was prepared. 1.0 g of the finally prepared hyaluronic acid filler composition with or without lidocaine was mixed with 9.0 g of the biosimilar solution, and then the calcification reaction was observed at 37°C for 14 days.

manufacturing company
(CAS-No) Experimental Example 4
(Phosphate 5mM) Experimental Example 5
(Phosphate 5mM) Phosphate
Buffered
Saline
(PBS) Sodium monohydrogen phosphate
(anhydride) Merck
(7558-79-4) 0.5364 mg/mL 0.5364 mg/mL Sodium dihydrogen phosphate
(dihydrate) Sigma Aldrich
(13472-35-0) 0.1907 mg/mL 0.1907 mg/mL sodium chloride Sigma Aldrich
(7647-14-5) 8.1796 mg/mL 8.1796 mg/mL 3rd order 991.0933 mg/mL 991.0933 mg/mL Hyaluronic acid
filler
composition Cross-linked sodium hyaluronate 24mg/mL 24mg/mL Lidocaine hydrochloride
(monohydrate) Delta Synthetic
Co., Ltd.
(73-78-9) N.A. 3.2 mg/mL pH 7.01 6.83 osmotic pressure 285 323

[Comparative example]

1.0 g each of Bv (Company D, Korea), Cs (Company B, Korea), Nv (Company M, Korea), and CN°4 (Company H, Korea) currently available on the domestic market were used to produce 9.0 g of biosimilar solution. After mixing, the calcification reaction was observed visually or through a microscope for 14 days at 37°C.

As a result, as shown in Figures 7 to 10, calcification reaction was confirmed in all four commercially available products. Based on this, it is expected that all four commercially available products were washed using Phosphate Buffered Saline (PBS) with a total phosphate content exceeding 7mM.

As described above, the present invention has been described through the above embodiments, but the present invention is not necessarily limited thereto, and various modifications and implementations are of course possible without departing from the scope and spirit of the present invention. Accordingly, the scope of protection of the present invention should be interpreted as including all embodiments falling within the scope of the patent claims attached to the present invention.

Claims (18)

delete delete delete delete delete A hyaluronic acid filler composition that inhibits calcification in the body, comprising a hyaluronic acid gel washed with a solution containing 1mM to 5mM phosphate,
The hyaluronic acid filler composition is characterized in that it further comprises an anesthetic agent at a concentration of 0.2 to 1.0 (w/v)%.
According to clause 6,
A hyaluronic acid filler composition, characterized in that the hyaluronic acid filler composition includes cross-linked hyaluronic acid or a salt thereof.
According to clause 6,
The hyaluronic acid filler composition is characterized in that it is a monophasic hyaluronic acid gel with a MoD of 1% to 13%,
The MoD value is a hyaluronic acid filler composition, characterized in that it is derived as the number of moles of crosslinker/the number of moles of repeating units of hyaluronic acid disaccharide or its salt Х 100.
According to clause 6,
A hyaluronic acid filler composition, characterized in that the pH of the hyaluronic acid filler composition is 5.5 to 8.5.
According to clause 6,
A hyaluronic acid filler composition, characterized in that the osmotic pressure of the hyaluronic acid filler composition is 200 mOsmol/kg to 400 mOsmol/kg.
delete According to clause 6,
The hyaluronic acid filler composition is characterized in that it is for small dose administration of less than 0.1 mL to 10 mL.
According to clause 6,
A hyaluronic acid filler composition, characterized in that the hyaluronic acid filler composition is for large-scale administration of 10 mL to 200 mL.
According to clause 6,
The hyaluronic acid filler composition is used to improve wrinkles and tissue volume reduction in soft tissues selected from the group consisting of the face, neck, chest, hips, arms, armpits, hands, legs, or feet, or to treat wounds, scars, or stretch marks. A hyaluronic acid filler composition.
The hyaluronic acid filler composition according to claim 6, wherein the solution further comprises sodium chloride.
The hyaluronic acid filler composition according to claim 6, wherein the phosphate contains at least one selected from the group consisting of monohydrogen phosphate and dihydrogen phosphate.
i) preparing a solution containing 1mM to 5mM of phosphate;
ii) washing the hyaluronic acid gel with the solution prepared above; and
iii) mixing the washed hyaluronic acid gel with an anesthetic agent at a concentration of 0.2 to 1.0 (w/v)%;
A method for producing a hyaluronic acid filler composition containing suppressed calcification in the body.
A hyaluronic acid filler composition that inhibits calcification in the body prepared by the manufacturing method of claim 17.