KR101937681B1 - Preparation Method of Sodium Hyaluronate Film by Solvent Casting and Hyaluronate Film Prepared Thereof - Google Patents

Abstract

The present invention relates to a process for producing a hyaluronate film by solvent casting and a hyaluronate film which can be used as a cosmetic mask pack, a patch for medicines, a medical device, a film-type adhesion inhibitor, and the like.
The method for producing a hyaluronate film by solvent casting comprises the steps of: (a) dissolving a hyaluronate in a solvent to prepare a hyaluronate solution of 0.6 to 2.9% by volume; And (b) drying the hyaluronate solution by injecting the hyaluronate solution into a mold at a relative humidity of 30 to 70% and a drying temperature of 30 to 50 ° C.
Since the hyaluronate film of the present invention is a dried film, unlike conventional liquid products, there is no fear of microbial contamination, and it is easy to produce / manage / distribute / treat, and also has excellent mechanical properties, , Packs for medical devices, patches, artificial skin, and the like.

Description

[0001] The present invention relates to a method for producing a hyaluronate film by solvent casting and a hyaluronate film prepared therefrom,

The present invention relates to a process for producing a hyaluronate film and a hyaluronate film produced therefrom, and more particularly to a process for producing a hyaluronate film by solvent casting, a mask pack for cosmetics, A patch for medical equipment, a film-type adhesion preventive, and the like.

Sodium hyaluronate (Hyaluronic Acid) is a mucopolysaccharide present in the dermis layer. It is known that N-acetyl-D-glucosamine and D-Glucuronic acid are β- 1, and 4 glucosidic bonds are linked by β-1,3 glucosidic bonds to form a hyaluronate chain.

Hyaluronate is a white powder but when dissolved in water it becomes a transparent liquid and has a high water retention due to a large number of hydroxyl groups (-OH) in the molecule and exhibits high viscoelasticity due to its large molecular weight of about 0.5 to 3.0 MDa .

In addition, hyaluronate is widely distributed in the connective tissues, epithelium and nervous tissues of human body, and has various physiological activities. As a biocompatible material, it has proven its effect on skin regeneration, moisturizing, elasticity maintenance and wrinkle improvement, The demand for antioxidant-related cosmetics, food, pharmaceuticals, and medical device fillers is rapidly increasing.

In the case of hyaluronate powder products in which moisture is minimized, intramolecular chains are connected very tightly and are less sensitive to changes in microorganisms or the external environment and thus are excellent in stability. However, when the moisture content in the hyaluronate powder is increased, The microbial contamination and the change of physical properties due to the decrease of the molecular weight are promoted, and the stability is drastically lowered.

Since most of the hyaluronate products currently on the market contain excessive amounts of water in the form of liquids, they must be manufactured in aseptic facilities during product commercialization due to microbial contamination or must use preservatives that are concerned with safety. There is a problem that it is difficult to expand it for industrial use.

Particularly, in the case of injections of joints or fillers mainly using hyaluronate, the content of hyaluronate in the product is as low as 1 ~ 3%, and most of 97 ~ 99% is water. Therefore, . In addition, since these products are very viscous, there is inconvenience that not only the suffering of the patient but also the bruising of the operation site due to the high pressure when the skin is injected in the form of an injection.

In terms of permeation or absorption of the skin, the low molecular weight and medium molecular weight hyaluronate are easier to penetrate or absorb the skin than the polymer, but the film is difficult to produce due to low mechanical properties. On the other hand, in the case of polymer hyaluronate, The coating effect and the mechanical properties are excellent and the film is easy to manufacture.

U.S. Patent No. 6,630,167 discloses a gel-type adhesion inhibitor using hyaluronate and a toxic crosslinking agent. Korean Patent Laid-Open Publication No. 2009-0012439 discloses a hyaluronic acid and a carboxymethylcellulose composite film on which a crosslinking activator and a crosslinking active agent And a method for producing the hyaluronic acid and carboxymethylcellulose complex derivative film in which an N-acylurea pendant form and an auto-crosslinked form are mixed.

However, since the above-mentioned patents did not completely remove the toxic crosslinking agent and the like after the production of the gel, there was a concern about the side effects thereof.

As a result of intensive efforts to solve the above problems, the present inventors have found that when a hyaluronic acid salt is dissolved in water or an aqueous ethanol solution and then dried under a specific temperature and humidity condition, the hyaluronate film having excellent mechanical properties and resistance to microbial contamination And the present invention has been completed.

It is an object of the present invention to prepare a film containing hyaluronate as an essential ingredient having excellent biocompatibility, moisture retention and anti-aging function and a process for producing the same.

It is another object of the present invention to provide a hyaluronate film having mechanical properties and high resistance to microorganisms, which is easy to produce, manage, distribute and process, and a method for producing the hyaluronate film.

It is still another object of the present invention to provide a mask pack for cosmetics, a patch, and an anti-adhesion agent for a medical device, which comprises a hyaluronate film having mechanical properties and high resistance to microorganisms.

In order to achieve the above object, the present invention provides a method for producing a hyaluronate, comprising: (a) dissolving a hyaluronate in a solvent to prepare a hyaluronate solution of 0.6 to 2.9% by volume; And (b) drying the hyaluronate solution prepared by solvent casting in a mold at a relative humidity of 30 to 70% and a drying temperature of 30 to 50 ° C. Of the present invention.

In the present invention, the hyaluronic acid salt has a molecular weight of 0.8 to 2.5 MDa, and the solvent is water or an aqueous solution of 0.01 to 29% by volume ethanol.

In the present invention, the drying of the hyaluronate solution is performed in a constant temperature and humidity chamber.

The present invention also provides a hyaluronate film produced by the above-mentioned method by solvent casting.

In the present invention, when the molecular weight of the hyaluronic acid salt is 1.2 to 2.5 MDa, the tensile strength is 151.5 to 300.0 MPa and the elongation is 45.5 to 65.0%.

The present invention also provides a mask pack for cosmetics comprising the hyaluronate film, a patch, and an anti-adhesion agent for a medical device.

Since the hyaluronate film of the present invention is a dried film, unlike conventional liquid products, there is no fear of microbial contamination, and it is easy to produce / manage / distribute / use, and also has excellent mechanical properties, It can be applied to various applications such as medicines, packs for medical devices, patches, and artificial skin.

1 is a photograph of a hyaluronate film prepared according to the molecular weight of a hyaluronic acid salt.
2 is a photograph of a hyaluronate film prepared according to the hyaluronate concentration.
Figures 3 and 4 are photographs of a hyaluronate film prepared according to a solvent.
Figure 5 is a photograph of a hyaluronate film prepared according to the drying method.

In the present invention, when a solvent casting method in which a hyaluronic acid salt is dissolved in water or an aqueous solution of ethanol and then dried under a specific temperature and humidity condition is used, the conventional liquid hyaluronic acid which is easily contaminated by microorganisms during production, storage, It has been confirmed that a hyaluronate film capable of solving the problems of acid salt products can be produced, and the present invention has been completed.

In the examples of the present invention, films were prepared under various conditions of the concentration of the hyaluronate and the drying method, and the mechanical strength for commercialization thereof and the stability against microorganisms were evaluated.

As a result, when a 1 to 2 volume% hyaluronate solution was dried at a relative humidity of 30 to 70% and a temperature of 30 to 50 ° C, a film having smooth surface and excellent mechanical strength and stability to microorganisms was produced I can confirm that I can do it. In addition, it was confirmed that the produced hyaluronate film has excellent mechanical properties and stability against moisture in the working room to such an extent that it can be easily formed into a desired shape with ordinary scissors or a knife.

Accordingly, in one aspect, the present invention provides a method for producing a hyaluronate, comprising: (a) dissolving a hyaluronate in a solvent to prepare a hyaluronate solution of 0.6 to 2.9% by volume; And (b) drying the hyaluronate solution prepared by solvent casting in a mold at a relative humidity of 30 to 70% and a drying temperature of 30 to 50 ° C. And a method for producing the same.

In the present invention, the hyaluronic acid salt is a salt in which hyaluronic acid is bound, and examples thereof include sodium hyaluronate, calcium hyaluronate, and potassium hyaluronate, but are not limited thereto.

Since the permeability, absorption and mechanical properties of the hyaluronic acid salt may vary depending on the molecular weight, it is preferable to use 0.8 to 2.5 MDa. When the molecular weight of the hyaluronate is less than 0.8 MDa, film formation is difficult. When the molecular weight exceeds 2.5 MDa, the hyaluronate has a problem of unevenness of the film surface due to high viscosity.

The solvent is used to dissolve the hyaluronate, and water or an aqueous solution of ethanol can be used. The content of ethanol in the aqueous ethanol solution is preferably 0.01 to 29% by volume. When water alone is used to dissolve hyaluronate, a great deal of attention is needed to prevent microbial contamination. However, when the hyaluronate is dissolved in an aqueous solution of ethanol, the effect of preventing microbial contamination But the film forming time can be shortened. However, when the ethanol content exceeds 29% by volume, there is a problem that the film becomes opaque or film is not formed as the molecular weight of the hyaluronic acid salt increases.

In the present invention, the concentration of the hyaluronate solution is preferably 0.6 to 2.9% by volume. When the concentration of the hyaluronate solution is less than 0.6 vol%, film formation is difficult. When the concentration of the hyaluronate solution exceeds 2.9 vol%, the film surface may become irregular due to high viscosity.

In the present invention, a step of injecting a hyaluronate solution for preparing a hyaluronate film into a mold and drying for 6 to 12 hours at a relative humidity of 30 to 70% and a drying temperature of 30 to 50 ° C is performed .

As long as the mold can accommodate the hyaluronate solution, it can be used without any particular limitation, and examples thereof include molds made of acrylic, glass or SUS.

In the present invention, drying can be performed without restriction as long as it can satisfy the above-mentioned relative humidity and drying temperature conditions, and it is preferably carried out in a constant temperature and humidity chamber. If the relative humidity or the drying temperature condition is out of the above range, the film may not be formed or a film of uneven surface may be formed.

The drying time can be appropriately adjusted within the above range depending on the concentration of the hyaluronate solution and the size of the mold.

In the present invention, the term "solvent casting" means a method widely used in the production of a film to completely dissolve an organic substance in a solvent, and then to blow the solvent to solidify the organic substance.

The present invention relates to a hyaluronate film produced by this method from a different viewpoint.

The hyaluronate film is characterized by excellent tensile strength and elongation. For example, when the molecular weight of the hyaluronate is 1.2 to 2.5 MDa, it has a tensile strength of 151.5 to 300.0 Mpa and an elongation of 45.5 to 65.0%. Thus, unlike conventional liquid hyaluronate products, there is no microbial contamination , Is easy to manage and use, and can be used for various purposes.

Accordingly, the present invention relates to a cosmetic mask pack, a patch, and an adhesion preventive agent for a medical device comprising the hyaluronate film from another viewpoint.

For reference, adhesion is usually seen during the healing process of inflammation, which means that the fibrous material that is entangled or massively precipitated when forming granulation tissue or scar is entangled. Generally, adhesion occurs at a frequency of 67 ~ 93% after laparotomy, and some of them are spontaneously decomposed. However, in most cases, after wound healing, adhesion occurs and causes various sequelae. An anti-adhesion agent is used to physically and chemically block the occurrence of adhesion with surrounding tissues such as covering or pharmacological action.

[Example]

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example 1: Determination of Film Formation Ability According to Molecular Weight Change

To confirm the film-forming ability according to the molecular weight of the hyaluronate, four kinds of sodium hyaluronate (Hi-Aqua ^TM , Jinwoo Bio) having a molecular weight of 0.3 MDa, 0.8 MDa, 1.2 MDa and 2.5 MDa And then dried in a constant temperature and humidity dryer (relative humidity: 50%, temperature: 40 ° C) for 6 to 24 hours to obtain a hyaluronate film. .

As a result, it was confirmed that in the case of the molecular weight of 0.3 MDa, a hole was formed in the middle portion and the film was not formed uniformly, but the film was well formed in the case of 0.8, 1.2 and 2.5 MDa. (Fig. 1)

In addition, when the concentration of hyaluronate was 1.0 vol%, the film-forming ability was the most excellent. In case of 0.5 vol%, the film was formed but the thickness was too small to be less than 0.005 mm, , And 3.0 volume%, the film surface was irregularly formed due to the high viscosity (FIG. 2).

Example 2: Determination of film formability according to solvent conditions

In order to confirm the film-forming ability according to the solvent conditions, 1.2 M sodium hyaluronate molecular weight was dissolved in an aqueous solution of 0-30 vol% ethanol at a concentration of 1.0 vol%, and each was injected into an acrylic mold, %, Temperature: 40 ° C) for 12 hours to prepare a hyaluronate film.

As a result, when the concentration of ethanol in the aqueous ethanol solution is 0 to 20% by volume, the transparent film is very well formed, whereas when the concentration of ethanol is 30% by volume, the hyaluronate is not dissolved well, (Fig. 3).

In addition, in the case of producing a film using only ordinary water, the film is intermittently contaminated with microorganisms and the film becomes opaque (FIG. 4). On the other hand, when the ethanol content is 5 vol% or more, .

It was also found that the film was formed in a shorter time as the concentration of ethanol in the aqueous ethanol solution increased. It was confirmed that when the purified water alone is used as the solvent, the drying time is required for about 24 hours, while the aqueous ethanol solution can be formed within 12 hours.

Example 3: Confirmation of film formability according to drying conditions

In order to confirm the film-forming ability according to the drying conditions, 1.2 M sodium hyaluronate having a molecular weight of 1.2 was dissolved in an aqueous 20 vol% ethanol solution at a concentration of 1.0 vol%, and each was injected into an acrylic mold, % And a temperature of 30 to 60 ° C) for 12 hours or more to prepare a hyaluronate film.

As a result, it was confirmed that the film was not formed even if it was dried for 24 hours or more at a relative humidity of 80% or more, and the film was formed within a shorter time (12 to 24 hours) as the temperature increased from 30 to 40 ° C .

For reference, in the case of hyaluronic acid salt, the molecular weight is decreased during long-term storage at a temperature of 50 占 폚 or more to lower the mechanical properties, so it is preferably carried out at 50 占 폚 or lower.

Comparative Example  1: Film using circulating drier Formability  Confirm

A hyaluronate film was prepared in the same manner as in Example 3, except that it was dried at 40 캜 for 12 hours or more using a circulation drier (Convection Oven) instead of the constant temperature and humidity dryer.

As a result, it was confirmed that the surface was wrinkled and a film containing some bubbles was formed.

Comparative Example  2: Film using a vacuum dryer Formability  Confirm

A hyaluronate film was prepared in the same manner as in Example 3, except that it was dried at 40 DEG C for 12 hours or more, using a vacuum dryer (~0.05 mbar) instead of the constant temperature and humidity dryer.

As a result, it was confirmed that a film having a wrinkled surface and dried at both ends was formed.

Experimental Example 1: Measurement of mechanical properties of a hyaluronate film

(Hi-Aqua ( ^TM ), Jinwai Bio) having a molecular weight of 1.2 MDa and 2.5 MDa was dissolved in a 20 vol% ethanol aqueous solution to a concentration of 1.0 vol%, and each was injected into an acrylic mold Followed by drying for 12 hours in a constant-temperature and constant-humidity dryer (relative humidity 50%, temperature 40 ° C) to produce a hyaluronate film.

In order to measure the mechanical properties of the hyaluronate film thus prepared, the film prepared using the TA-XT2i Texture Analyzer (Stable Micro System, UK) was cut into 3 cm x 5 cm, mounted on the tensile grips The tensile strength and elongation were measured, and the results are shown in Table 1.

Hyaluronate molecular weight 1.2MDa 2.5MDa Tensile Strength 151.5 300.0 Elongation (%) 45.5 65.0

From Table 1, it can be seen that the hyaluronic acid film has a tensile strength of 151.5 MPa, a stretch ratio of 45.5%, a tensile strength of a hyaluronate film of 2.5 MDa molecular weight, and an elongation percentage Was 65.0%, and the mechanical properties were excellent. As the molecular weight increased, both tensile strength and elongation increased.

Experimental Example 2: Measurement of microbial stability of hyaluronate film

The total number of bacteria means the number of bacteria that can develop in the standard agar medium among the bacteria present in the specimen. In order to evaluate the contamination stability of the hyaluronate film against the microorganism, the specimen and the standard agar medium were mixed and coagulated in petri dish and the number of living cells in the specimen was calculated from the number of colonies of the resulting bacteria after culturing.

(1) Preparation of reagents and reagents

- Standard agar medium (Plate count agar)

5.0g of Tryptone, 2.5g of Yeast extract, 1.0g of Dextrose and 15.0g of Agar were mixed with distilled water to make 1,000mL, adjusted to pH 7.0 ± 0.2, and sterilized at 121 ° C for 15 minutes.

- sterile physiological saline

And sterilized with a 0.9% sodium chloride aqueous solution at 121 캜 for 15 minutes.

(2) Test method

The hyaluronate solid film prepared using the molecular weight of 1.2 MDa prepared in Experimental Example 1 and the hyaluronic acid salt having the molecular weight of 1.2 MDa dissolved in the normal purified water at 1 vol% were each exposed to a general laboratory for 15 days.

Next, the hyaluronate film and 0.5 g of the hyaluronate solution were respectively dissolved in 50 mL of sterile physiological saline and then added to 250 mL of a sterile standard agar medium at 43 to 45 DEG C, followed by shaking.

Before the medium was hardened, the mixture was put in a Petri dish, cooled and solidified, and then cultured at 35 ± 1 ° C for 24 to 48 hours. The number of colonies produced after culturing was counted and the total number of bacteria was divided by 0.5 g. The results are shown in Table 2.

Before air exposure After air exposure Hyaluronate
solution Hyaluronate
film Hyaluronate
solution Hyaluronate
film Number of microorganisms (cfu / g) 0 0 1,500 ~ 5,000 0 to 8

From Table 2, it can be seen that the microorganism of the 1% hyaluronate solution after 15 days of exposure to air was about 1,500 to 5,000 cfu, whereas the hyaluronate film showed 0 to 8 cfu almost unchanged before exposure.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

(a) dissolving a hyaluronate having a molecular weight of 0.8 to 2.5 MDa in an aqueous solution of 0.01 to 29% by volume ethanol to prepare a hyaluronate solution of 0.6 to 2.9% by volume; And
(b) injecting the hyaluronate solution prepared above into a mold and drying in a thermo-hygrostat under the conditions of a relative humidity of 30 to 70% and a drying temperature of 30 to 50 ° C. Lt; / RTI >
delete delete delete A hyaluronate film produced by the method of claim 1.
The hyaluronate film according to claim 5, wherein the hyaluronic acid salt has a tensile strength of 151.5 to 300.0 Mpa and an elongation of 45.5 to 65.0% when the molecular weight of the hyaluronic acid salt is 1.2 to 2.5 MDa.
A mask pack comprising the hyaluronate film of claim 5.
An adhesion preventive agent comprising the hyaluronate film of claim 5.

Images