KR20170005265A - multilayered coated microparticle for tissue reconstruction and method for production thereof - Google Patents

Abstract

By producing multilayer coated microparticles for tissue reconstruction with different crosslinking degree using various ingredients such as hyaluronic acid, the present invention reduces an injecting pressure when objects are injected by operating the outer layer of the tissue reconstruction with low crosslinking degree and delays disintegration when the objects enter the body by operating the inner layer of the tissue reconstruction with high crosslinking degree. So, the retention time for the tissue reconstruction is lengthened. Also, the present invention was completed as the invention improves main problems of existing tissue reconstruction like injection pressure, shape maintenance and the retention time by increasing the shape maintenance with high elasticity of the inner layer.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a multilayered coated microparticle for tissue reconstruction and method for producing the same,

The present invention is a clone of hyaluronic acid-based tissue, wherein the structure of the clone is composed of an inner particle and an outer coating layer, the inner particle is a particle-shaped material formed through a lot of crosslinking reaction, It is a gel layer consisting of a small number of cross-linked hyaluronic acids to lower the injection pressure when injecting the clone into the body. When the tissue clone of this composition is introduced into the body, it is discharged even under a weak pressure, so that partial correction can be performed. Internal particles have a high degree of crosslinking and are advantageous in morphogenesis and the decomposition enzyme action is difficult. . By doing so, it relates to cloning of hyaluronic acid microparticle tissues which can advantageously help repair tissue in the body, such as the skin.

Non-Animal Stabilized Hyaluronic Acid Gel, a biodegradable, non-anesthetic stabilized hyaluronic acid gel, is a wrinkle-removing injectable product that removes wrinkles and lips, and is approved by the US FDA on December 12, 2003 . Medicis Pharmaceuticals' proprietary gel-type filler Resilane is also a hyaluronic acid formulation that focuses on improving wrinkles around the mouth and nose while Botox focuses on improving wrinkles on the forehead area. Hyaluronic acid has also been released in more than 30 countries since its introduction in Europe in 1996, mainly based on hyaluronic acid extracted from the crests of birds such as chickens. Juvederm from Elleran also removes wrinkles by injecting hyaluronic acid into the face. In addition, Anica Therapeutics of the United States has been authorized to release Eleven, a skin filler for wrinkles and wound healing, from the FDA. Many of these materials have been used as skin fillers for wrinkle and wound healing, but many products have been approved by the US Food and Drug Administration (FDA) for the development of products that use proven hyaluronic acid. Hyaluronic acid is a biomolecular substance of linear form of millions and tens of millions of Dalton lengths as discrete units of N-acetyl-D-glucosamine and D-glucuronic acid as discrete units, and is used in many applications such as ocular vitreous fluid, joint synovial fluid, exist. Due to its excellent biocompatibility and viscoelasticity, hyaluronic acid is widely used for medical and medical purposes such as postoperative adhesion inhibitor, shaping aid, wrinkle improving agent, ophthalmic surgery aid, joint function improver, drug delivery material and eye drop, . However, by itself, it is easily decomposed in vivo or under acid and alkali conditions and its use is limited. Therefore, efforts to develop structurally stable hyaluronic acid derivatives have been made widely. As one of the hyaluronic acid derivatives, hyaluronic acid bridges in which hyaluronic acids are mutually covalently bonded using a crosslinking agent have excellent biocompatibility, physical stability and biodegradability, and a large number of manufacturing methods have been proposed in connection therewith.

However, hyaluronic acid bridges (hyaluronic acid derivatives) produced by the methods disclosed in these patents have a difficulty in finding a compromise between ease of implantation in the body and prolongation of morphogenesis and maintenance in the body. That is, when the degree of crosslinking of hyaluronic acid is lowered to apply a small pressure in the injection process using a syringe, the crosslinking product is easily decomposed in the body to shorten the maintenance period. In order to extend the retention period of the hyaluronic acid, Is increased to inject a hyaluronic acid crosslinked material into the body, which makes it difficult to perform delicate procedures because of high pressure.

Accordingly, it is an object of the present invention to separate a crosslinked part having functional properties in the process of producing a hyaluronic acid crosslinked product, that is, to separate the crosslinked part and the injection pressure into the body, And a method of producing the hyaluronic acid tissue. BACKGROUND OF THE INVENTION

In order to repair the tissue, it is necessary to apply a low injection pressure during the process of injecting hyaluronic acid bridges (hyaluronic acid derivatives) into the body, and the bridged products introduced into the body require proper shape formation and a long maintenance period as long as possible do.

However, lowering the degree of crosslinking of hyaluronic acid for low injection pressure makes it difficult to form the crosslinked material in the body, and it is easy to decompose and shorten the holding period. Therefore, there is a demand for a method for producing a hyaluronic acid crosslinked product which is easy to be injected and has excellent functionality in the morphogenesis and maintenance period.

Hyaluronic acid should have a high degree of crosslinking in order to exhibit proper morphogenesis and a long maintenance period in the body as a clone of tissue. In order to do this, a large amount of hyaluronic acid reactant and cross-linking agent must be added to the reaction solution, and by setting the reaction temperature and time in order to optimize the cross-linking reaction, it is possible to increase the number of cross-linkages between hyaluronic acid molecules or molecules.

When the internal microparticles are produced with a high degree of crosslinking, injecting only the internal particles using a syringe may be difficult to inject due to a large elasticity of the particles and a large friction between the particles, thereby greatly increasing the injection pressure. In order to lower the injection pressure of the hyaluronic acid crosslinking agent in such a situation, a method of reducing the inter-particle elasticity and the frictional force is needed. As a means to be considered for this purpose, the shape of the internal fine particles is rounded to reduce intergranular friction, and each internal fine particle is wrapped with a crosslinked cross-linked substance in a coating form to reduce the external elasticity of each internal fine particle, The lubrication effect can be exhibited by the existence of a soft viscous layer between the layers. If necessary, the crosslinked layer for coating is not limited to one layer, but may be two or more layers. The degree of crosslinking of the crosslinked layers for this coating can be lowered to 1% or less for better lubrication effects.

In the present invention, the principle of application of the invention is described using a hyaluronic acid component as a representative component. However, the present invention is not limited to these components, and various crosslinkable components can be applied to the present invention. It can be manufactured in various forms using various crosslinkable components and utilized in various areas, so that the application area is not limited to the reproduction of the tissue number.

In the present invention, since the hyaluronic acid fine particles produced through the process for producing the multi-layer coated hyaluronic acid crosslinked product distinguished by function are injected into the body, the lower layer is softened due to the less crosslinked water in the outer layer, It is possible to provide a material and a manufacturing method thereof that can be used variously in cosmetic molding and other tissue restoration areas by exposing the inner layer having a large number of crosslinked water in a short period of time after the introduction into the body, have.

1 is a scanning electron micrograph of the coated microparticles prepared in Preparation Example 2 according to the present invention.

The present invention provides a method for producing a multi-layer coated hyaluronic acid microparticle tissue replica.

PREPARATION EXAMPLE 1. Preparation of Internal Fine Particles Having High Bridges >

In order to achieve the above object, the present invention provides a method for producing polysaccharide polymer microparticles,

1) A natural polymer polysaccharide is dissolved in distilled water or an aqueous alkali solution to prepare an aqueous solution having a concentration of 1 to 12% by weight, 1,4-butanediol diglycidyl ether (BDDE) is added as a crosslinking agent, and sodium hydroxide solution or potassium hydroxide solution Adjusting the pH of the aqueous solution to 12 to 14;

2) dropping the aqueous polymeric polysaccharide solution of step 1) into a stirred oil layer using various syringe devices to form a water in oil emulsion (w / o);

3) continuing the stirring at a temperature of 1 to 95 ° C. for 10 minutes to 24 hours so as to maintain the crosslinking reaction in the emulsion state in the step 2), thereby proceeding a crosslinking reaction between the polysaccharide polymers to prepare fine particles;

4) removing the oil layer using a filter to obtain fine particles, and then washing the crosslinking agent, unreacted material and oil remaining in the fine particles with ethanol;

≪ Preparation Example 2: Preparation of coated microparticles >

1) Mixing the fine particles obtained in 4) of Production Example 1 with the crosslinked aqueous solution of 1) above. At this time, the amount of the polysaccharide as a crosslinking component is as small as 1/2 to 1/100 of the amount charged in step 1) and the amount of the crosslinking agent is in a range of 1/1 to 1/100 Inject a small amount;

2) preparing the coated hyaluronic acid fine particles obtained in step 4) of the second step by repeating the steps 2) to 4) of Preparation Example 1;

EXPERIMENTAL EXAMPLE 1. Morphology Analysis of Coated Fine Particles [

To observe the form of the material prepared in Preparation Example 2, about 50 mg of the product was immobilized on an aluminum stove and platinum coated on the products for 3 minutes under a vacuum of 0.1 torr and a high voltage (10 kV) -4800 FE-SEM) After the sample is placed in the body of the equipment, the shape of the product is analyzed and shown in FIG.

EXPERIMENTAL EXAMPLE 2 Measurement of Viscoelasticity of Coated Fine Particles [

The coated microparticles prepared in Preparation Example 2 were sterilized at a concentration of 50 mg / ml using physiological saline, and then rheometer was used to confirm the rheological properties. The complex viscosity at a frequency of 0.02 to 1 Hz viscosity was measured. The composite viscosity measured at a frequency of 0.02 Hz of the prepared sample showed an average of 7,500,000 cP.

EXPERIMENTAL EXAMPLE 3 Measurement of Injection Performance of Coated Microparticle-Filled Syringes [

In order to measure the scanning performance, the tension meter was set to a compression strength measuring mode. The injectate containing the multilayer coated hyaluronic acid microparticles was filled into the syringe. A needle having a standard of 27 1 / 2G was inserted into the injection section of the syringe. The syringe was placed on a measuring plate of a tensile tester and fixed. After operating the tensile tester, the plunger of the syringe was pushed at a speed of 1 mm / sec until the injection in the syringe was completely pushed out, and the protrusion pressure at that time was measured.

In this case, the range of the protrusion pressure was about 0.5 ~ 3N.

Claims (12)

Duplication of particulate form of tissue coated with multiple layers of different degrees of crosslinking using ingredients capable of crosslinking, including polysaccharides. The method of claim 1, wherein the polysaccharide is selected from the group consisting of hyaluronic acid, hyaluronic acid salt, poly-gamma-glutamic acid, polygamat glutamate, agar, alginic acid, alginate Carrageenan, furcellaran, pectin, arabic gum, karaya gum, tragacanth gum, ghatti gum, But are not limited to, guar gum, locust bean gum, psyllium seed gum, gleatin, chitin, dextran, xanthane gum, chitosan chitosan, chondroitin-4-sulfate, chondroitin-6-sulfate, beta-glucan, carboxymethyl cellulose, heparin and starch ). ≪ / RTI > The method of claim 1, wherein the number of layers is more than one but is not limited thereto. The method of claim 1, wherein the cross-linking degree in the coating-functioning layer can be configured with a constant or gradient change. 2. The method of claim 1, wherein the coat comprises both a full or partial application to the immediate immediate preceding layer. The method of claim 1, wherein the degree of crosslinking can range from 0.01% to 70%. The method for producing the internal fine particles according to claim 1, wherein the internal fine particles are formed by a droplet method, a spray drying method, a suspension polymerization method, a lump fracturing method, and the like. The method according to claim 1, wherein the shape of the cross-sectional area of the microparticles may be a polygonal shape such as a circle, an ellipse, a triangle, a rectangle, a pentagon, a hexagon, a deformed shape thereof, and an irregular shape. [7] The method according to claim 7, wherein, when the fine particles are prepared by the gravimetric method in the dropping method, that is, the aqueous crosslinking reaction solution is dropped into an organic solvent to produce fine particles. , Ethanol, propanol, isopropanol, butanol, isobutanol, pentanol, hexanol, benzene, benzyl alcohol, ethyl acetate, gluform, methylene chloride, 1,2-dichloroethane, diethyl ether, acetonitrile, corn Oil, soybean oil, rice bran oil, grape seed oil, canola oil, sunflower oil, and olive oil. 3. The method of claim 1, wherein the crosslinking reaction temperature is maintained at a temperature of 1 to 95 占 폚, but is not limited thereto. The manufacturing method according to claim 1, wherein the fine particles are produced but coated with a layer having a different degree of crosslinking are not limited to the particulate form, and may be a gel, a sponge, an irregular particle, a microparticle, It can also be applied to objects with a three-dimensional shape. The method of claim 1, wherein each layer can be prepared by combining and mixing one or more components.

Images