WO2017018717A1 - Dermal filler hydrogel composition - Google Patents

Abstract

The present invention relates to: a dermal filler hydrogel composition facilitating the control of mechanical properties and having improved mechanical properties and durability; and a preparation method therefor. The composition of the present invention is used so as to be applied, as a filler, to body parts requiring high mechanical properties, and if the method of the present invention is used, the mechanical properties of a dermal filler hydrogel, to be prepared, can be easily controlled.

Description

Hydrogel composition for dermal filler

The present invention relates to a hydrogel composition for dermal filler that is easy to control mechanical properties, mechanical properties and durability is improved.

As the individual's interest in beauty increases day by day, the cosmetic surgery field is also undergoing great development. In addition to surgical procedures that result in relatively large cosmetic changes, injections of botulinum toxin (eg, the brand name “Botox”), fillers, and the like, which can benefit from simple injection treatments such as wrinkle removal, are becoming commonplace. It is becoming. Dermal fillers are agents that are injected into a patient, for example to improve facial lines and reduce wrinkles, and unlike Botox, which uses the principle of paralyzing facial muscles that cause wrinkles, Use the principle.

As a filler material, hyaluronic acid-based compounds that are biocompatible materials have been frequently used.

Hyaluronic acid (HA), also known as hyaluronan, is a non-sulfated glycosaminoglycan that is widely distributed in binding, epithelial and nervous tissues throughout the human body. Hyaluronic acid is abundant in many different layers of skin and includes, for example, the ability to ensure good hydration, to assist the organization of the extracellular matrix, and to act as a filler; And complex functions such as those involved in tissue regeneration mechanisms. However, with aging, the amount of hyaluronic acid, collagen, elastin, and other matrix polymers present in the skin decreases. For example, repeated exposure to ultraviolet light, for example, from the sun allows dermal cells to reduce their production of hyaluronan as well as to increase their rate of degradation. Loss of such materials leads to, for example, wrinkles, holes, water loss, and / or other undesirable conditions that contribute to aging. Injectable dermal fillers are successfully used for skin aging. Dermal fillers can replace lost endogenous matrix polymers or treat / promote the function of existing matrix polymers to treat these skin conditions.

Conventional fillers made of hyaluronic acid are Restylane, Ivoire, Cugel, Juvederm Perfectha, Teosial, Rofilan, Elevess. , Ellavie (Elavie), Glytone (Glytone), Princess (Princess), etc., but long-term use is difficult, there is a problem of low mechanical properties (G ') to use for purposes such as raising the nose. Therefore, inorganic fillers such as polymethyl methacrylate, calcium hydroxyl apatite, polylactic acid, polyalkyllimide and the like are used as filler materials for long-lasting fillers.

While using hyaluronic acid, a material having excellent biocompatibility, mechanical properties (G ′) can be easily controlled, high mechanical properties can be exhibited, and there is a need to develop a filler material having improved sustainability.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

The present inventors earnestly researched to develop a hydrogel composition for dermal filler that is easy to control mechanical properties, and improved mechanical properties and durability. As a result, when the alginic acid-bound hyaluronic acid is crosslinked by a divalent cation, the present invention is completed by elucidating that a hydrogel for dermal filler having improved mechanical properties and durability can be prepared as compared to a conventional hyaluronic acid hydrogel. It became.

Accordingly, an object of the present invention is to provide a hydrogel composition for dermal filler.

Still another object of the present invention is to provide a method for preparing a dermal filler hydrogel.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

The present invention to solve the above problems,

And an ion crosslinkable hyaluronic acid hydrogel comprising hyaluronic acid, alginic acid bound to the hyaluronic acid, and an ion crosslinking agent.

The ion crosslinking agent provides a hydrogel composition for dermal filler that is a divalent cation or a salt thereof.

In addition, the present invention to solve the above problems,

(a) reacting a linker with alginic acid or hyaluronic acid as a first reactant to covalently link the linker to a carboxyl group of alginic acid or hyaluronic acid;

(b) reacting the reaction product of step (a) with a second reaction material, hyaluronic acid or alginic acid, to obtain an alginic acid-linked hyaluronic acid reformer linked through a linker; And

(c) adding a divalent cation or a salt thereof to the resulting alginic acid-linked hyaluronic acid reformer to form a crosslinking product to obtain an ionically crosslinkable hyaluronic acid hydrogel; To provide.

In addition, the present invention to solve the above problems,

Provided is a composition for reinforcing mechanical properties of a chemical crosslinked hyaluronic acid filler comprising an ion crosslinkable hyaluronic acid hydrogel comprising:

(a) hyaluronic acid;

(b) alginic acid bound to hyaluronic acid; And

(c) divalent cations or salts thereof as ionic crosslinkers.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a hydrogel composition for dermal filler with improved persistence and mechanical properties (G ′).

(b) The present invention provides a method for producing a hydrogel for dermal filler.

(c) The present invention provides a composition for reinforcing mechanical properties of chemical cross-linked hyaluronic acid filler.

(d) The composition of the present invention can be applied to parts of the body where high mechanical properties are required.

(e) Using the method of the present invention, it is possible to easily adjust the mechanical properties of the hydrogel for dermal filler produced.

1 shows a theoretical explanation for its hydrogel formation in the presence of alginic acid-grafted hyaluronic acid (HGA) and calcium ions. Hyaluronic acid (HA) was first modified with ethylenediamine (NH2-HA). NH2-HA was then reacted with alginic acid (AL) through the carbodiimide chemistry.

2A shows an image of a hyaluronic acid (HA) or alginic acid-grafted-hyaluronic acid (HGA) solution after mixing, in the presence (+) or absence (-) condition of calcium ions.

2B shows an image of HGA after injection through a syringe needle.

FIG. 2C shows the change in storage modulus (G ′, filled representation) and loss modulus (G ″, empty representation) of HGA1 (circle) and HA (square) solutions in the presence of calcium ions.

2D shows the change in storage modulus dependent on hyaluronic acid content.

FIG. 2E shows storage modulus change dependent on calcium concentration in alginic acid-grafted hyaluronic acid gel ([HGA1] = 2 wt.%).

2F shows the gelation time of HGA gel.

FIG. 2G shows the storage modulus of the HGA gel ([HGA1] = 2 wt%, [CaSO4] = 29.7 Mm) depending on the weight of the alginic acid molecule.

3A shows the change in dry weight of both hyaluronic acid / alginic acid mixed gels or alginic acid-grafted-hyaluronic acid gels after treatment with hyaluronidase at 37 ° C. for 2 weeks.

3B shows a cross-sectional SEM image of the alginic acid / hyaluronic acid mixed gel.

Figure 3c is two weeks as it hyaluronidase azepin culture ([polymer] = 2% by weight, hyaluronic acid / alginate (mg / mg) = 1, [Ca ^{2 +]} = 29.7 mM, in 37 ℃ [The hyaluronidase dehydratase; = 100 μg / ml) and cross-sectional SEM images of alginic acid-grafted hyaluronic acid gels are shown.

4A shows the ¹ H-NMR spectra of hyaluronic acid, alginic acid and alginic acid-grafted hyaluronic acid.

4B shows the results of DMMB analysis for quantitative graft efficacy of alginic acid-grafted hyaluronic acid.

Figures 5a-5b shows the results of comparison of hydrogel properties according to the molecular weight of hyaluronic acid used in the ion-crosslinked alginic acid-linked hyaluronic acid reformer. Figure 5a shows the properties of the hydrogel when using an alginic acid-linked hyaluronic acid reformer containing a hyaluronic acid of 1000 kD. Figure 5b shows the properties of the hydrogel when using an alginic acid-linked hyaluronic acid reformer containing a hyaluronic acid of 1000 kD. 5A and 5B show the concentration of the alginic acid-binding hyaluronic acid modifier.

6a-6c show the results of physical property changes when the ion-crosslinked alginic acid-bonded hyaluronic acid hydrogel is mixed with a conventional chemical crosslinked hyaluronic acid filler. Figure 6a shows the physical properties when the existing filler cugel Aqua S (BNC Korea) and alginic acid-linked hyaluronic acid hydrogel containing hyaluronic acid having an average molecular weight of 1000 kD at 50:50. FIG. 6B shows the physical properties of the existing filler cugel Aqua S and an alginic acid-linked hyaluronic acid hydrogel including hyaluronic acid having an average molecular weight of 1500 kD at 50:50. Figure 6c shows the physical properties when the mixture of the existing filler cugel S (BNC Korea) and alginic acid-linked hyaluronic acid hydrogel containing hyaluronic acid having an average molecular weight of 1500 kD at 50:50. 6A to 6C show the types and concentrations of hydrogels used.

Figure 7 shows the cytotoxicity test results according to the mixing weight ratio when the ion-crosslinked alginic acid-linked hyaluronic acid hydrogel is mixed with the conventional chemical cross-linked hyaluronic acid filler.

Figure 8 shows the results of the change in physical properties (elastic coefficient) according to the mixing weight ratio when the ion-crosslinked alginic acid-linked hyaluronic acid hydrogel is mixed with the conventional chemically cross-linked hyaluronic acid filler.

According to an aspect of the present invention, the present invention includes a hyaluronic acid, an alginic acid bonded to the hyaluronic acid, and an ion crosslinkable hyaluronic acid hydrogel comprising an ion crosslinking agent, wherein the ion crosslinking agent is a divalent cation or a salt thereof It provides a hydrogel composition for dermal filler, characterized in that.

The present inventors earnestly researched to develop a hydrogel composition for dermal filler that is easy to control mechanical properties, and improved mechanical properties and durability. As a result, when the alginic acid-bound hyaluronic acid is crosslinked by a divalent cation, it has been found that a hydrogel for dermal filler having improved mechanical properties and durability can be prepared as compared with a conventional hyaluronic acid hydrogel.

As used herein, the term "dermal filler" is used for improving or treating wrinkles, depressions, scars, etc. formed on the skin of a subject, and specifically, for improving wrinkles such as forehead, brow, and nasolabial folds, It may mean a biocompatible material for injection into the dermis for the purpose of raising the nose, increasing the skin area volume, improving the scar area, and the like.

As used herein, the term "chemical crosslinking hyaluronic acid filler" may refer to a conventional hyaluronic acid filler prepared using a chemical crosslinking agent, and may be any hyaluronic acid filler prepared by adding a chemical crosslinking agent. Commercially available filler products Restylane, Ivoire, Cutegel, Juviderm Perfectha, Teosial, Rofilan, Elevess For example, Elavie (Elavie), Glytone (Glytone), Princess (Princess) and the like.

The hydrogel composition for dermal fillers according to the present invention may further include a chemically cross-linked hyaluronic acid filler, through which the mechanical properties of the dermal filler can be significantly improved.

In this case, the weight ratio of the ion crosslinkable hyaluronic acid hydrogel and the chemical crosslinked hyaluronic acid filler may be 1: 1-9, more specifically 1: 1.

In the hydrogel composition for dermal filler of the present invention, hyaluronic acid into which alginic acid is introduced is crosslinked by a divalent cation to form a hydrogel. For example as specifically Ca ^{2 +,} Ba ^{2 +,} Cu ^{2 +,} Fe ² 2, such as ^+, and Mg ^{2 +,} causing cross-linking by the interaction with a cation is introduced into hyaluronic acid, alginate, a hydrogel To form. The hydrogel may be adjusted to have higher mechanical properties (G ′) than the hydrogel formed from a simple mixture in which hyaluronic acid and alginic acid are not bound.

In one embodiment of the present invention, the divalent cation of the present invention may be calcium, barium or magnesium ions. The calcium, barium or magnesium ions of the present invention may be provided in the form of sulfate or chloride salts with respect to the alginic acid bound hyaluronic acid. Specifically, for example, divalent cations can be introduced by adding calcium chloride, calcium sulfate, barium chloride, magnesium chloride, magnesium sulfate, and the like.

In one embodiment of the present invention, the composition of the present invention may have a mechanical property (G ′) of 10-3000 Pa. More specifically, it may have a mechanical property of 20-3000 pa, even more specifically 500-3000 pa, even more specifically 500-2500 pa, even more specifically 1000-2500 pa, even more specifically 1500-2500 pa. .

In one embodiment of the present invention, the composition of the present invention may have a mechanical property of 1500-2500 pa, the composition may be used as a dermal filler composition for skeletal reinforcement. In the case of the conventional hyaluronic acid filler, there was a problem of low mechanical properties to use for skeleton reinforcement, when using the hydrogel of the present invention, it can be used for skeleton reinforcement due to the improved mechanical properties. As used herein, the term "skeletal reinforcement" may refer to a use for applying a deformation to a facial contour including a nose, a cheekbone, and a jawline, and in the case of the skeletal reinforcement dermal filler, a filler having a purpose of removing wrinkles is higher. May require mechanical properties.

As used herein, the term mechanical property (G ′) may mean shear storage modulus or elastic modulus, and specifically, the sugar storage modulus is a shear test that transmits a force pushing the sample. It may mean the force required to apply a constant deformation to the sample, the elastic modulus may refer to the force required to apply a constant deformation to the sample by shear test by the force to pull the sample.

In one embodiment of the present invention, the alginic acid of the present invention may have a molecular weight of 10000-1000000 g / mol, the hyaluronic acid of the present invention may have a molecular weight of 10000-2500000 g / mol. More specifically, the alginic acid of the present invention may have a molecular weight of 20000 to 500000 g / mol, more specifically, may be used 50000 to 500000 g / mol, and more specifically 100000 to 300000 g / mol. have. In addition, the hyaluronic acid of the present invention may be used having a molecular weight of 100000 to 2500000 g / mole, more specifically 300000 to 2500000 g / mol can be used, and more specifically 500000 to 1500000 g / mol can be used. .

In one embodiment of the present invention, the weight ratio of hyaluronic acid and alginic acid of the present invention may be 4: 1 to 1: 2. The weight ratio of the hyaluronic acid and the alginic acid described above may mean a ratio of the total weight of each of the hyaluronic acid and the alginic acid included in the hydrogel composition. The combined composition ratio of hyaluronic acid and alginic acid included in the hydrogel composition of the present invention is one of the factors that can control the mechanical properties (G ') of the hyaluronic acid gel, to achieve the required physical properties according to the site and purpose of use of the dermal filler For the hyaluronic acid and alginic acid can be appropriately adjusted to the combined composition ratio, the weight ratio of the hyaluronic acid and alginic acid in the composition ratio can be suitably used in the range of 4: 1 to 1: 2. More specifically, hyaluronic acid and alginic acid may be used in a weight ratio of 2: 1 to 1: 1.5, and even more specifically, 2: 1 to 1: 1.

The divalent cation of the present invention can be used by adjusting the amount in a desired range in order to achieve the desired mechanical properties in the preparation of the hydrogel. Depending on the type of divalent cation and its salt form, an appropriate amount may be selected and mixed in consideration of solubility and the like.

In one embodiment of the invention, the divalent cation salt of the invention may be 5 mM to 100 mM calcium sulfate.

In another embodiment of the invention, the divalent cation salt of the invention may be 1 mM to 50 mM calcium chloride.

When using calcium chloride, it is possible to produce a hydrogel having high mechanical properties with only a small amount of use compared to the case of using calcium sulfate.

In one embodiment of the present invention, the coupling of hyaluronic acid and alginic acid of the present invention may be a covalent bond.

In one embodiment of the present invention, the covalent bond of the present invention may be made through a linker covalent-formable with the carboxyl group of alginic acid and the carboxyl group of hyaluronic acid. The covalent bond of the present invention may be a direct bond between hyaluronic acid and alginic acid, or may be a bond through a linker as described above. In the linker of the present invention, one molecule forms a bond with both hyaluronic acid and alginic acid, thereby binding the hyaluronic acid and alginic acid. More specifically, the linker of the present invention may use any linker known in the art including at least two functional group moieties capable of covalent bond formation with a carboxyl group included in hyaluronic acid and a carboxyl group included in alginic acid. .

In one embodiment of the invention, the linker of the invention is diamine, divinylsulfone, 1,4-butanediol, diglycidyl ether (BDDE) and glutaraldehyde, carbodiimide, hydroxysuccine It is selected from the group consisting of imide, imido ester, maleimide, haloacetyl, disulfide, hydroazide, alkoxyamine. More specifically, for example, diamine may be used as the linker of the present invention.

In one embodiment of the present invention, the composition of the present invention may further comprise an analgesic agent. The hydrogel composition for dermal filler of the present invention is injected into the dermis via topical injection, and may cause short-term pain at the injected site. In order to alleviate such pain, an analgesic may be additionally injected, but by using a hydrogel composition including the analgesic, the same effect may be achieved by only one injection.

In one embodiment of the invention, the analgesic agent of the present invention, lidocaine, mepivacaine, bupivacaine, procaine, chloroprocaine, ethidocaine, prilocaine diclonin, hexylcaine, procaine, cocaine, Ketamine, morphine, pramoxin, propofol, phenol, naloxone, meperidine, butorpanol, pentazosin, morphine-6-glucuronide, codeine, dihydrocodeine, diamorphine, dextrosepropoxyphene, fetidine , Fentanyl, alfentanil, alphaprodine, buprenorphine, dextromoramide, diphenoxylate, dipiphanone, heroin (diacetylmorphine), hydrocodone (dihydrocodeinone), hydromolphone (die Hydromorphinone), levopanol, meptazinol, methadone, methopone (methyldihydromolpinone), nalbuphine, oxycodone (dihydrohydroxycodeinone), oxymolphone (dihydrohydroxymorphinone), phenadoxone , Phenazosin, remy Any one selected from titanyl, tramadol, tetracaine, and the group consisting of pharmaceutically acceptable salts may be higher. More specifically, lidocaine can be used.

The composition of the present invention may be prepared as a pharmaceutical composition.

Compositions of the present invention may be prepared by mixing with one or more additional pharmaceutically acceptable aqueous carriers or excipients. Topical pharmaceutical or cosmetic pharmaceutical compositions, including, for example, pharmaceutically acceptable carriers or diluents, such as buffered saline, as well as dermatologically or pharmaceutically acceptable carriers, vehicles or mediums. It may include other components typically used in. As used herein, the term "dermatologically or pharmaceutically acceptable" may mean that the composition or ingredient described above does not cause excessive toxicity, incompatibility, allergic reactions, etc. upon skin contact.

The compositions of the present invention may optionally include other pharmaceutically acceptable ingredients, including but not limited to buffers, preservatives, isotonicity modifiers, salts, antioxidants, osmolality modifiers, emulsifiers, wetting agents and the like.

Pharmaceutically acceptable buffers are buffers that can be used to prepare the hydrogel compositions disclosed herein provided that the formulations obtained are pharmaceutically acceptable. Non-limiting examples of pharmaceutically acceptable buffers may include acetate buffers, borate buffers, citrate buffers, neutral buffered saline, phosphate buffers and phosphate buffered saline. Any concentration of pharmaceutically acceptable buffer can be useful for formulating the hydrogel compositions disclosed herein, provided that the effective amount of active ingredient can be calculated in consideration of this effective concentration of buffer. Physiologically acceptable buffer concentrations range, for example, from about 0.1 mM to about 900 mM, but are not limited thereto. The pH of the pharmaceutically acceptable buffer can be adjusted provided that the agent obtained is pharmaceutically acceptable. If desired, acids or bases may be used to adjust the pH of the hydrogel composition. Any buffered pH level may be useful for formulating pharmaceutical compositions, provided that a therapeutically effective amount of active matrix polymer ingredient can be calculated in consideration of this effective pH level. Physiologically acceptable pH is, for example, in the range of about pH 5.0 to about pH 8.5, but not limited thereto. For example, the pH of the hydrogel compositions disclosed herein can be about 5.0 to about 8.0, or about 6.5 to about 7.5, about 7.0 to about 7.4, or about 7.1 to about 7.3.

Pharmaceutically acceptable preservatives may include sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene. Pharmaceutically acceptable preservatives include benzalkonium chloride, chlorobutanol, thimerosal, phenylmercury acetate, phenylmercury nitrate, stabilized oxychloro composition, such as PURITE® (California) Allergan Inc., Irvine, Ltd.) and chelating agents, such as DTPA or DTPA-bisamide, calcium DTPA, and CaNaDTPA-bisamide.

Pharmaceutically acceptable isotonicity modifiers useful in the hydrogel compositions disclosed herein include, for example, sodium chloride and potassium chloride; And salts such as glycerine, but are not limited to these. The composition may be provided as a salt and may be formed by a number of acids, including but not limited to hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid, and the like. Salts tend to be more soluble in aqueous or other protic solvents than the corresponding free base forms. It is understood that these and other materials known in the pharmaceutical arts can be included in the hydrogel compositions disclosed herein.

Furthermore, if necessary, the composition may further comprise any ingredient conventionally used for cosmetic purposes, such as improvement of skin aging, such as wrinkles, or in the field of treatment of neuromuscular related diseases.

The composition of the present invention is an injectable composition, which may be used in the form of being administered to the skin region of a subject using an injection device having a fine needle. As used herein, the term "injection device having a fine needle" may refer to an injection device having a needle of, for example, about 21 gauge to about 32 gauge. The route of administration of the hydrogel composition to a subject may typically be determined based on the cosmetic and / or clinical effects desired by the subject and / or operator, and the body part or region in which the composition is administered. have. The compositions disclosed herein may be administered by any means known to those of skill in the art, including needled syringes, pistols (eg, hand-compressed pistols), catheters by topical or direct surgical implantation. It is not limited to this. The hydrogel compositions disclosed herein can be administered into a skin region, such as, for example, the dermal or subcutaneous region.

The suitable dosage of the hydrogel composition of the present invention can be determined in various ways by factors such as the mode of administration, the subject's age, weight, sex, time of administration and site of administration. On the other hand, the dosage of the hydrogel composition of the present invention may be preferably 0.001-100 mg / kg (body weight) per serving.

According to another aspect of the present invention, the present invention provides a method for preparing a hydrogel composition for dermal filler comprising the following steps:

(a) reacting a linker with alginic acid or hyaluronic acid as a first reactant to covalently link the linker to a carboxyl group of alginic acid or hyaluronic acid;

(b) reacting the reaction product of step (a) with a second reaction material, hyaluronic acid or alginic acid, to obtain an alginic acid-linked hyaluronic acid reformer linked through a linker; And

(c) adding a divalent cation or a salt thereof to the resulting alginic acid-linked hyaluronic acid reformer to form a crosslinking to obtain an ionically crosslinkable hyaluronic acid hydrogel.

Step (a) of the present invention is a step of covalently linking a linker to a carboxyl group of alginic acid or hyaluronic acid by reacting the linker with alginic acid or hyaluronic acid, and the linker used is the same as the linker which may be included in the hydrogel composition. As used herein, the term “first reactant” may refer to a substance that first reacts with a linker in hyaluronic acid or alginic acid to form a bond. The linker of the present invention may be used in an amount capable of binding to all of the carboxyl groups included in the first reactant.

Step (b) of the present invention is a step of coupling the hyaluronic acid and alginic acid through the linker by reacting a linker-bound hyaluronic acid or alginic acid with a second reactant. As used herein, the term “second reactant” may refer to a compound other than the compound selected as the first reactant among hyaluronic acid or alginic acid. As used herein, the term "alginic acid-linked hyaluronic acid modifier" may refer to hyaluronic acid in a state in which alginic acid is bound. By controlling the weight ratio of hyaluronic acid and alginic acid, it is possible to easily adjust the mechanical properties of the resulting hydrogel, and in particular, it is possible to obtain a hydrogel having higher mechanical properties (G ') than the conventional hyaluronic acid hydrogel. .

Step (c) of the present invention is a step of adding a divalent cation or a salt thereof to the resulting alginic acid-linked hyaluronic acid reformer to cause a crosslinking reaction to gel. The crosslinking reaction occurs by the interaction of the divalent cation added with alginic acid, thereby obtaining a hydrogel.

In one embodiment of the present invention, after the step (c), the present invention may further comprise the step of mixing the obtained crosslinked hyaluronic acid hydrogel with a chemical cross-linked hyaluronic acid filler; As can be seen from the results of the following examples, the mechanical properties of the dermal filler can be significantly improved.

In this case, the mixed weight ratio of the ion crosslinkable hyaluronic acid hydrogel and the chemical crosslinked hyaluronic acid filler may be 1: 1-9, and more specifically 1: 1.

In one embodiment of the present invention, the divalent cation of the present invention may be calcium, barium or magnesium ions. The calcium, barium or magnesium ions of the present invention can be used in the form of sulfate or chloride salts with respect to alginic acid bound hyaluronic acid. Specifically, for example, it may be used in the form of calcium chloride, calcium sulfate, barium chloride, magnesium chloride, or magnesium sulfate compound.

In one embodiment of the present invention, the present invention further comprises the step of adding an analgesic agent to the hydrogel of the present invention after the step (c).

"Method for producing a hydrogel composition for dermal filler" according to the present invention relates to a method for producing a "hydrogel composition for dermal filler" which is another embodiment of the present invention, the overlapping content is used, and excessive In order to avoid complexity, description of duplicate contents will be omitted.

According to another aspect of the present invention, the present invention provides a composition for reinforcing mechanical properties of a chemical cross-linked hyaluronic acid filler comprising an ion-crosslinkable hyaluronic acid hydrogel comprising:

(a) hyaluronic acid;

(b) alginic acid bound to hyaluronic acid; And

(c) divalent cations or salts thereof as ionic crosslinkers.

In the case of the conventional chemical crosslinking-hyaluronic acid filler, in addition to the problems caused by using the chemical crosslinking agent as described above, there was a difficulty in preparing a hydrogel having high mechanical properties. When using the composition of the present invention, it can be used for the purpose of reinforcing the mechanical properties of the chemical cross-linked hyaluronic acid filler. This has the advantage that it is possible to easily change the mechanical properties while using a known material used as a composition for the conventional dermal filler as it is, it can raise the limit of the mechanical properties that can be achieved by chemical cross-linked hyaluronic acid filler There is an advantage.

In this case, the weight ratio of the ion crosslinkable hyaluronic acid hydrogel and the chemical crosslinking-hyaluronic acid filler may be 1: 1-9, more specifically 1: 1.

In one embodiment of the present invention, the hyaluronic acid of the present invention may have a molecular weight of 500000 g / mol to 2500000 g / mol.

In one embodiment of the present invention, the divalent cation of the present invention may be calcium, barium or magnesium ions.

In one embodiment of the invention, the divalent cations of the invention may be added in the form of chloride salts.

In one embodiment of the invention, the chloride salt of the invention may be calcium chloride.

The composition for reinforcing the mechanical properties of the chemical cross-linked hyaluronic acid filler according to the present invention uses the hydrogel composition for dermal filler, which is one embodiment of the present invention described above, and the overlapping content is omitted so as to avoid excessive complexity of the present specification. do.

Hereinafter, the present invention will be described in more detail with reference to Examples.

These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example

Experimental Materials and Methods

Example 1 Preparation of Alginic Acid-Linked Hyaluronic Acid Modifiers

For the introduction of amine groups, hyaluronic acid (molecular weight 600000-850000; Lifecore) was reacted with ethylenediamine (Sigma-Aldrich). Synthesis of NH2-hyaluronic acid was synthesized by a conventionally known method using an EDC / NHS reaction. In order to suppress the crosslinking between hyaluronic acid chains during the reaction, an excess of 10 times molar ratio of ethylenediamine was added. And it reacted for 20 hours at room temperature. The solution was dialyzed and treated with activated charcoal, filtered through a 0.22 μm filter for sterilization and lyophilized. Alginic acid was combined with NH 2 -hyaluronic acid via a carbodiimide chemistry according to a conventionally known method (see FIG. 1).

Example 2: Nuclear Magnetic Resonance Spectroscopy

Hyaluronic acid, alginic acid and alginic acid-binding hyaluronic acid modifiers were analyzed by ¹ H NMR spectroscopy (Bruker avance 500 MHz) at 70 ° C. Samples were dissolved in D ₂ O at 3 mg / ml.

Example 3: Dimethyl Methylene Blue (DMMB) Assay

DMMB analysis was performed to quantify the alginic acid / hyaluronic acid content in the alginic acid-binding hyaluronic acid reformer. That is, 16 mg of DMMB was dissolved in 25 ml of ethanol, filtered with filter paper, and 100 ml of 1 M guanidine hydrochloride containing 0.17 M of sodium formate and 1 ml of formic acid was mixed with the filtered DMMB. . The solution was mixed with deionized water to a total volume of 500 ml. Each sample was diluted with deionized water and 0.1% by weight solution was obtained. 1 ml of DMMB solution was added to 100 μl of each sample and mixed vigorously for 30 minutes. The cultured samples were centrifuged at 12000 g for 10 minutes to precipitate the complex. The supernatant was removed and dried at room temperature for 30 minutes. The pellet was dissolved in 1 ml of decomplexation solution. A decomplexed solution was prepared with 50 mM sodium acetate buffer (pH 6.8) containing 10% 1-propanol and 4 M guanidine hydrochloride. After 30 minutes of mixing, 100 μl of each sample was transferred to a 96-well plate. Absorbance was measured at 656 nm using a spectrophotometer (SpectraMax M2; Molecular Devices).

Example 4: Enzyme Stability Test

Alginic acid-linked hyaluronic acid modifier (HGA1) was dissolved in PBS and mixed with calcium sulfate using two glass plates with 1 mm thick spacers to obtain gel ([polymer] = 2 wt%, [CaSO ₄ ] = 29.7 mM). Gel discs were obtained using a punch (10 mm diameter), fed into a PBS solution containing hyaluronidase in a concentration range of 0 to 100 μg / ml and incubated at 37 ° C. for 24 hours. Gel discs were frozen and lyophilized before measuring dry weight. In addition, a physical mixture of hyaluronic acid and alginic acid was used as a control ([polymer] = 2 wt%, [CaSO4] = 29.7 mM). After treatment with hyaluronidase, a cross-sectional image of the gel was observed using a scanning electron microscope (S-4800 UHR FE-SEM; Hitachi).

Example 5 Rheological Measurement

Viscoelastic properties of ion-crosslinked alginic acid-linked hyaluronic acid gels were measured using a rotary rheometer with a cone-and-plate (20 mm diameter plate, 4 ° cone angle) coneand-plate fixture (Bohlin Gemini 150). . A 150 μm gap opening was set at the top of the cone and plate, and the operating temperature was set constant at 37 ± 0.1 ° C.

Alginic acid-binding hyaluronic acid modifiers were dissolved in PBS. The 2 wt% polymer solution was mixed with a CaSO ₄ slurry in an amount dependent on the alginic acid content of the polymer solution (0.42 g of CaSO ₄ per 1 g of alginic acid could be mixed). Mechanical properties of ion-crosslinked hydrogels were measured using a rotary rheometer (Bohlin Gemini 150). Measurement was made at 0.5 Hz and the temperature was kept constant at 37 ± 0.1 ° C.

Example 6: Statistical Analysis

All data are presented as mean ± standard deviation (n = 6). Statistical analysis was performed using Student's t-test. ^* P-values <0.05 and ^** P <0.01 were considered statistically significant.

Example  7: Ion crosslinking  Alginic acid-linked hyaluronic acid On the reformer  Comparison of Hydrogel Properties by Hyaluronic Acid Molecular Weight

Alginic acid-bound hyaluronic acid modifiers synthesized using hyaluronic acid having a molecular weight of 1000000 g / mol or 1500000 g / mol were dissolved in PBS. 2 wt% of the polymer solution was mixed with a CaSO ₄ slurry in an amount depending on the alginic acid content of the polymer solution (0.42 g of CaSO ₄ per 1 g of alginic acid could be mixed). Mechanical properties of ion-crosslinked hydrogels were measured using a rotary rheometer (Bohlin Gemini 150). Measurement was made at 0.5 Hz and the temperature was kept constant at 37 ± 0.1 ° C.

Example  8: Chemical crosslinking  Hyaluronic acid With filler Ion crosslinking  Changes in Physical Properties of Alginate-Linked Hyaluronic Acid Modifiers

(1) Two syringes were used to combine the alginate-linked hyaluronic acid reformer synthesized with commercial gelling filler CULEL Aqua S or Cugel S of Korea BNC Co., Ltd. and hyaluronic acid having a molecular weight of 1000000 g / mol or 1500000 g / mol. And mixed. The 2 wt% polymer solution was mixed with a CaSO ₄ slurry in an amount dependent on the alginic acid content of the polymer solution (0.42 g of CaSO ₄ per 1 g of alginic acid could be mixed). Mechanical properties of ion-crosslinked hydrogels were measured using a rotary rheometer (Bohlin Gemini 150). Measurement was made at 0.5 Hz and the temperature was kept constant at 37 ± 0.1 ° C.

(2) In addition, as a commercially available molding filler, the mixed weight ratios of Humidics' elavies and ion-crosslinkable alginic acid-bonded hyaluronic acid hydrogels were respectively set to 9: 1. The elastic modulus was measured after mixing at 7: 3 and 5: 5.

Example  9: Chemical crosslinking  Hyaluronic acid With filler Ion crosslinking  Cytotoxicity Testing of Alginic Acid-Binding Hyaluronic Acid Modifier Mixtures

First, the alginic acid-bonded hyaluronic acid was ion-crosslinked with calcium sulfate (60 mM), and then a mixture was prepared at various mixing ratios with Elavier of Humedix, a chemical cross-linked hyaluronic acid filler (Elabie: Alginic acid-bonded hyaluronic acid). = 9: 1, 7: 3, 5: 5).

Next, DDC / F containing ATug5 mouse chondrogenic cell line (ECACC) 10ug / ml human transferring (Sigma-Aldrich), 3 × 10 ^-8 M sodium selenite (Sigma-Aldrich), 5% FBS, 1% antibiotics Cells were cultured using -12 culture. The cells were cultured at 37 ° C. and 5% CO ₂ , and 5000 cells were cultured in each well in a Transwell 0.4um polyester membrane 12-well plate (Costar). A mixture of the chemically crosslinked hyaluronic acid fillers and the ionic crosslinked alginic acid-binding hyaluronic acid fillers prepared in various mixing ratios of 300 ul was prepared on the membrane, immersed in the culture medium, and subjected to EZ-cytox assay to evaluate cytotoxicity. .

Experiment result

1. Characterization of Alginic Acid-Linked Hyaluronic Acid

Alginic acid-linked hyaluronic acid (HGA) was designed and synthesized by the preparation of ion crosslinkable hyaluronic acid. Ie, hyaluronic acid (HA) was first modified with ethylenediamine (NH ₂ -HA) and conjugated with alginic acid with carbodiimide (HA-g-AL) (see Figure 1). Various alginic acid-bound hyaluronic acid samples were synthesized. Samples were synthesized with the weight ratios of hyaluronic acid and alginic acid being 1: 1, 2: 1 and 4: 1 (hereinafter referred to as HGA1, HGA2, and HGA4, respectively).

Alginic acid-bound hyaluronic acid (HGA) was confirmed by ¹ H-NMR spectroscopy (see FIG. 4A). The δ peak area of 4.2-4.4 increased after modification with ethylenediamine, indicating successful introduction of ethylenediamine to hyaluronic acid (NH ₂ -HA). After NH ₂ -hyaluronic acid modification of alginic acid, specific peaks of alginic acid and hyaluronic acid can be seen on the spectra at δ = 4.2-4.7 and δ = 2.5-2.6 (NCOCH ₃ ), respectively. The graft efficiency of HA-g-AL was determined by calculating the polymer content from the spectrum (HGA1-93.7%, HGA2 = 82.6%, HGA4 = 94.7%). Binding efficiency was also measured by dimethylmethylene blue (DMMB) analysis (see Figure 4b). Hyaluronic acid was not attached to the DMMB dye; Therefore, only the alginic acid content of HA-g-AL was determined from the above analysis (HGA1 = 90.4 ± 4.9%, HGA2 = 86.0 ± 7.9%, HGA4 = 83.6 ± 9.0%). The results found above were in good agreement with those obtained from the ¹ H-NMR spectrum.

2. Hydrogel Formation of Alginic Acid-Linked Hyaluronic Acid

When the hyaluronic acid solution was mixed with calcium ions ([hyaluronic acid] = 2% by weight, [CaSO ₄ ] = 30 mM), no hydrogel was formed. In contrast, in the presence of calcium ions, alginic acid-binding hyaluronic acid could form a gel ([HGA1] = 2 wt.%, [CaSO4] = 30 mM) (see FIG. 2A). Most of the toxicity-related issues in hyaluronic acid-based hydrogels arise from the use of chemical crosslinkers rather than from polymer-related issues [37, 38]. Toxicity-related issues in typical hyaluronic acid gels generated by chemical crosslinking reagents after implantation in vivo can be overcome by the use of ion crosslinkable hyaluronic acid-based hydrogels. Ion-crosslinked alginic acid-linked hyaluronic acid hydrogels can be injected through a cylinder with a 23-G needle (see FIG. 2B). This is also attractive in terms of enabling minimally invasive delivery of drugs and / or cells to the body.

3. Characterization of Alginic Acid-Linked Hyaluronic Acid Hydrogels

The viscoelastic and gelling behavior of the alginic acid-bound hyaluronic acid gels were investigated using a rotary rheometer (see FIGS. 2C-2G). When the HGA1 solution was mixed with calcium ions, the storage modulus (G ') was higher than the loss modulus (G ") at various frequencies (Fig. 2c), indicating that the mixture builds a hydrogel structure through ion crosslinking. In contrast, the hyaluronic acid solution does not form a hydrogel against calcium addition, and then examines the effect of the alginic acid content on the mechanical properties of the ion-crosslinked alginic acid-linked hyaluronic acid gel (see FIG. 2D). Hydrogels were prepared on constant polymer and calcium concentrations for all samples ([polymer] = 2 wt%, [CaSO ₄ ] = 30 mM) The weight ratios between hyaluronic acid and alginic acid varied. As the weight ratio of hyaluronic acid to 1 increased from 1 to 4, the mechanical properties of the gel changed significantly from 2.5 ± 0.2 kPa to 0.075 ± 0.001 kPa. The change in the elasticity of the gel changes with calcium concentration (see Fig. 2e) As the calcium sulfate concentration increases from 7.4 mM to 30 mM, the G 'value of the alginic acid-bound hyaluronic acid gel is 0.3 ± 0.1. The kPa increased from 2.7 ± 0.6 kPa, but no further increase was observed when the calcium sulfate concentration increased to 60 mM.The gelation time of the alginic acid-linked hyaluronic acid gel was significantly affected by the alginic acid content in the gel. The shear modulus of the alginic acid-linked hyaluronic acid gel depends on the alginic acid molecular weight (see Fig. 2g), as the alginic acid molecular weight increases from 50000 g / mol to 250000 g / mol. The G 'of the ion crosslinked gel also increased from 0.04 ± 0.003 to 2.4 ± 0.2 kPa Alginate-linked hyaluronic acid gels made with 50000 g / mol or 100000 g / mol alginic acid showed poor mechanical properties, even calcium Even in the presence of ions, it is very important to control the mechanical properties of the polymer scaffold in relation to cellular behavior (eg adhesion, proliferation and migration) [39-41]. In addition, mechanical properties are also important for the regulation of stem cell differentiation, which has an immediate effect on tissue regeneration [41–44]. The mechanical properties of alginic acid-binding hyaluronic acid gels, which can be very attractive for use in tissue engineering, are readily controllable through control of polymer composition and calcium ion concentration.

4. Enzyme Stability of Alginic Acid-Linked Hyaluronic Acid Hydrogels

Alginic acid-binding hyaluronic acid hydrogels were treated with hyaluronidase to test stability for in vitro enzymatic reactions. A slight decrease in gel weight was observed on the two week incubation time. Gel weight was dependent on enzyme concentration (see FIG. 3A). The hydrogels maintained their initial weight of approximately 80% after treatment with 100 μg / ml hyaluronidase for 2 weeks compared to control conditions without enzymes. Alginic acid-binding hyaluronic acid gels maintained a well packed porous structure after enzymatic treatment (see FIG. 3C). In contrast, gels prepared from a simple mixture of hyaluronic acid and alginic acid showed a significant reduction in gel weight during culturing even in the presence of 1 μg / ml hyaluronidase. Regardless of the hyaluronidase content, a weight loss of approximately 40% was observed. The weight loss is similar to that expected based on the hyaluronic acid content on the gel. Hyaluronic acid / alginic acid mixed gel cross-sectional images after treatment with hyaluronidase showed significant gel loss by enzymatic digestion (FIG. 3B). Hyaluronic acid was cleaved into small oligosaccharide fragments by the action of hyaluronidase [45, 46]. However, hyaluronic acid was rapidly degraded in the absence of crosslinking or chemical modification by endogenous hyaluronidase in vitro [47]. During the development of new ECMs from delivered cells, fast gel degradation can be detrimental to the regeneration of many types of tissues, as the gels must provide a 3-D structural environment and maintain their integrity as scaffolds. Hyaluronic acid / alginic acid mixed hydrogels do not form fully interpenetrating polymer network structures, since only alginic acid in the mixture participates in gel formation when calcium ions are added. Hyaluronic acid in the mixed gels was mostly degraded entirely by the action of hyaluronidase (see Figure 3). On the other hand, the degradation of hyaluronic acid in the alginic acid-binding hyaluronic acid was limited by the covalent conjugation of the alginic acid chain leading to slow gel degradation by hyaluronidase. This may make these gels more suitable than mixed gels for in vivo applications.

5. Ion crosslinking  Alginic acid-linked hyaluronic acid On the reformer  Comparison results of hydrogel properties according to the molecular weight of hyaluronic acid used

The viscoelastic behavior of alginic acid-linked hyaluronic acid hydrogels having different hyaluronic acid molecular weights was investigated using a rotary rheometer (see FIGS. 5A to 5B). When the HGA solution was mixed with calcium ions, the alginic acid-bonded hyaluronic acid hydrogel synthesized from the high molecular weight hyaluronic acid showed the highest storage modulus (G '). For example, when the hyaluronic acid molecular weight increased from 1000000 g / mol to 1500000 g / mol compared to the same polymer concentration of 2%, the G 'value of the alginic acid-linked hyaluronic acid gel increased from 480 Pa to 600 Pa. All samples tested produced hydrogels at a constant alginic acid / calcium concentration ratio. This means that at the same alginic acid molecular weight and calcium ion concentration, the hyaluronic acid molecular weight has an important effect on the physical properties of the alginic acid-linked hyaluronic acid gel. In addition, as the concentration of the alginic acid-bound hyaluronic acid reformer decreased, the G 'value also decreased.

6. Chemical crosslinking  Hyaluronic acid On filler Ion crosslinking  Alginic acid-linked hyaluronic acid Modifying  Observation results of physical property change when mixed

(1) After mixing alginic acid-bonded hyaluronic acid hydrogels having different hyaluronic acid molecular weights with Cugel Aqua S of BNC Korea Co., Ltd., a commercially available filler product, changes in viscoelastic behavior were investigated using a rotary rheometer (see FIG. 6A). To 6b). When Cue Gel Aqua S and Alginate-linked Hyaluronic Acid gel with a molecular weight of 1000000 g / mol are mixed at 50:50 (weight ratio) (expressed as a 2% gel), it is more than twice the G 'value of conventional products. It was. When the alginic acid-bonded hyaluronic acid gel having a hyaluronic acid molecular weight of 1500000 g / mol was mixed with the cugel Aqua S at 50:50 (weight ratio) (marked as a 2% gel), the physical properties increased by three times or more. . In addition, the change of viscoelastic behavior after mixing the gel gel and alginic acid-bonded hyaluronic acid gel of BNC Korea Co., Ltd. was investigated using a rotary rheometer (see FIG. 6C). When the alginic acid-bonded hyaluronic acid gel having a hyaluronic acid molecular weight of 1500000 g / mol is mixed with Cugel S at 50:50 (weight ratio) (marked as a 2% gel), the physical properties of about 1.5 times increase compared to conventional products. Confirmed. This means that by simply mixing the alginic acid-bonded hyaluronic acid gel with the existing molding filler product, the physical properties of the existing product can be easily improved, and the physical properties change not only by adjusting the mixing ratio of the gel but also by controlling the molecular weight of the hyaluronic acid used in the synthesis. Means that it can be facilitated. Existing products need to increase the amount of chemical crosslinking agent to improve the properties of the filler gel, but there is concern about the potential toxicity. However, when the alginic acid-bonded hyaluronic acid modifier is used, since the crosslinking is formed only with calcium ions, the gel is formed, and thus the physical properties of the gel can be variously controlled without concern for safety.

(2) FIG. 8 shows the results of changes in the physical properties (elastic coefficient) according to the mixing weight ratio when the ion-crosslinkable alginic acid-bonded hyaluronic acid hydrogel is mixed with the conventional chemically crosslinked hyaluronic acid filler. It was confirmed that the modulus of elasticity of the ella crosslinked alginic acid-bonded hyaluronic acid hydrogel mixture tended to increase as the content of the ionic crosslinked alginic acid-bonded hyaluronic acid hydrogel increased. When the alginic acid-bonded hyaluronic acid hydrogel was mixed at a ratio of 5: 5, it was confirmed that the physical properties (elastic coefficient) increased by about 35% than when the elavie was used alone (FIG. 8).

7. Chemical crosslinking  Hyaluronic acid With filler Ion crosslinking  Alginic acid-linked hyaluronic acid Modifier  Cytotoxicity test results of the mixture

Figure 7 shows the cytotoxicity test results according to the mixing weight ratio when the ion-crosslinked alginic acid-linked hyaluronic acid hydrogel is mixed with the conventional chemical cross-linked hyaluronic acid filler.

This resulted in 96% or more cell survival when using elavie, a conventional chemical crosslinking hyaluronic acid filler product alone, and nearly 100% in the case of ionic crosslinking alginic acid-linked hyaluronic acid hydrogel without using a chemical crosslinking agent. It was confirmed. In addition, even in the case of a mixture of elavie and ionic crosslinked alginic acid-binding hyaluronic acid hydrogel showed a high cell viability of more than 91%, it was confirmed that the mixture does not significantly affect the cytotoxicity (Fig. 7) .

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

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According to the present invention, it is possible to provide a filler having excellent mechanical properties without using a biotoxic chemical crosslinking agent, which can replace a commercially available hyaluronic acid filler, and as a filler to a body part requiring high mechanical properties. It can be usefully applied.

Claims (20)

1. And an ion crosslinkable hyaluronic acid hydrogel comprising hyaluronic acid, alginic acid bound to the hyaluronic acid, and an ion crosslinking agent.

   The ion crosslinking agent is a divalent cation or a salt thereof dermal filler hydrogel composition.
2. The method of claim 1,

   The composition is a hydrogel composition for dermal filler further comprising a chemical cross-linked hyaluronic acid filler.
3. The method of claim 2,

   A weight ratio of the ion crosslinkable hyaluronic acid hydrogel and the chemical crosslinked hyaluronic acid filler is 1: 1-9.
4. The method of claim 1,

   The divalent cation is calcium, barium or magnesium ions hydrogel composition for dermal filler.
5. The method of claim 1,

   The composition is a hydrogel composition for dermal filler for skeletal reinforcement.
6. The method of claim 1,

   The alginic acid has a molecular weight of 10000-1000000 g / mole, the hyaluronic acid has a molecular weight of 10000-2500000 g / mole hydrogel composition for dermal fillers.
7. The method of claim 1,

   The weight ratio of the hyaluronic acid and alginic acid is 4: 1 to 1: 2 hydrogel composition for dermal filler.
8. The method of claim 1,

   The divalent cation salt is 5 mM to 100 mM calcium sulfate hydrogel composition for dermal filler.
9. The method of claim 1,

   The divalent cation salt is 1 mM to 50 mM calcium chloride hydrogel composition for dermal filler.
10. The method of claim 1,

    The bond is a hydrogel composition for dermal filler is a covalent bond made through a covalent bond-formable linker with a carboxyl group of alginic acid and a carboxyl group of hyaluronic acid.
11. The method of claim 10,

    The linker is diamine, divinylsulfone, 1,4-butanediol, diglycidyl ether (BDDE) and glutaraldehyde, carbodiimide, hydroxysuccinimide, imidoester, maleimide, haloacetyl Hydrogel composition for dermal filler that is selected from the group consisting of, disulfide, hydroazide, alkoxyamine.
12. The method of claim 1,

    The composition is lidocaine, mepivacaine, bupivacaine, procaine, chloroprocaine, ethidocaine, prilocaine diclonin, hexylcaine, procaine, cocaine, ketamine, morphine, pramoxin, propofol, phenol, Naloxone, meperidine, butorpanol, pentazosin, morphine-6-glucuronide, codeine, dihydrocodeine, diamorphine, dextrosepropoxyphene, pettidine, fentanyl, alfentanil, alphaprodine, butane Prenorphine, dextrosemoramid, diphenoxylate, dipipanone, heroin, hydrocodone, hydromolone, levophanol, meptazinol, methadone, methopone, nalbuphine, oxycodone, oxymolone, phenadoxone, phenazosin , Remifentanil, tramadol, tetracaine and any one or more analgesics selected from the group consisting of pharmaceutically acceptable salts thereof.
13. (a) reacting a linker with alginic acid or hyaluronic acid as a first reactant to covalently link the linker to a carboxyl group of alginic acid or hyaluronic acid;

    (b) reacting the reaction product of step (a) with a second reaction material, hyaluronic acid or alginic acid, to obtain an alginic acid-linked hyaluronic acid reformer linked through a linker; And

    (c) adding a divalent cation or a salt thereof to the resulting alginic acid-linked hyaluronic acid reformer to form a crosslinking product to obtain an ionically crosslinkable hyaluronic acid hydrogel; .
14. The method of claim 13,

    After the step (c), mixing the obtained ion-crosslinkable hyaluronic acid hydrogel with a chemical cross-linked hyaluronic acid filler; further comprising a method for producing a hydrogel composition for dermal filler.
15. The method of claim 14,

    The ion-crosslinkable hyaluronic acid hydrogel and the chemically cross-linked hyaluronic acid filler mixing weight ratio of 1: 1-9 is a method for producing a hydrogel composition for dermal filler.
16. The method of claim 13,

    The divalent cation is calcium, barium or magnesium ion method for producing a hydrogel composition for dermal filler.
17. The method of claim 13,

    The divalent cation is a method for producing a hydrogel composition for dermal filler is added in the form of calcium chloride salt.
18. The method of claim 13,

    After the step (c), the hydrogel in lidocaine, mepivacaine, bupivacaine, procaine, chloroprocaine, ethidocaine, prilocaine diclonin, hexylcaine, procaine, cocaine, ketamine, Morphine, pramoxin, propofol, phenol, naloxone, meperidine, butorpanol, pentazosin, morphine-6-glucuronide, codeine, dihydrocodeine, diamorphine, dextropropoxyphene, pettidine, fentanyl , Alfentanil, alphaprodine, buprenorphine, dextromeramide, diphenoxylate, dipipanone, heroin, hydrocodone, hydromolphone, levopanol, meptazinol, methadone, methopone, nalbuphine, oxycodone , Addition of any one or more analgesics selected from the group consisting of oxymolphone, phenadoxone, phenazoxin, remifentanil, tramadol, tetracaine and their pharmaceutically acceptable salts; The method of reoyong hydrogel composition.
19. A composition for reinforcing mechanical properties of a chemical crosslinked hyaluronic acid filler comprising an ion-crosslinkable hyaluronic acid hydrogel comprising:

    (a) hyaluronic acid;

    (b) alginic acid bound to hyaluronic acid; And

    (c) divalent cations or salts thereof as ionic crosslinkers.
20. The method of claim 19,

    A composition for reinforcing mechanical properties of a chemical crosslinking hyaluronic acid filler having a weight ratio of the ion crosslinkable hyaluronic acid hydrogel and the chemical crosslinking hyaluronic acid filler is 1: 1-9.

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