KR20180124414A - Therapeutic use of biodegradable and biocompatible composite materials for dysuric diseases - Google Patents

Abstract

The present invention relates to a therapeutic use of a biodegradable and biocompatible composite material for treating dysuric diseases. More specifically, the present invention relates to a composition for the prevention or treatment of stress urinary incontinence and vesicoureteral reflux using a composite material obtained by mixing biodegradable and biocompatible materials. According to the present invention, an injectable filler, which contains a composite material obtained by mixing polylactic acid microparticles, which are biodegradable polymers, and hyaluronic acid hydrogel, which is a biocompatible polymer, is injected into the walls of the urethra, and compresses the urethra, thereby effectively preventing or treating stress urinary incontinence. The injectable filler is injected into the mucous membrane of the ureter and compressing the ureter, thereby effectively preventing or treating vesicoureteral reflux.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to biodegradable and biocompatible composite materials for dysuronic diseases,

The present invention relates to the use of biodegradable and biocompatible composite materials for the treatment of dysuria. More particularly, the present invention relates to a composition for preventing or treating stress urinary incontinence and vesicoureteral reflux of a composite material produced by combining a biodegradable polymer and a biocompatible polymer, and a method of manufacturing the same.

Urinary incontinence refers to a urinary incontinence disorder in which urination occurs through the urethra irrespective of its own will, and is a symptom frequently occurring in women with relatively short urethra, compared with men. The cause of urinary incontinence is various, but the ultimate cause is in the bladder or sphincter. Especially, the most common stress urinary incontinence is due to deficiency of pelvic support after aging or postpartum, or functional failure of cystoscope. . Urinary incontinence accounts for 10 to 40 percent of all women, and more than 200 million women worldwide suffer from incontinence. Stress urinary incontinence is the most common form of female urinary incontinence, accounting for about 50% of all urinary incontinence. Stress urinary incontinence is a disease in which urine leaks out of the urethra without shrinkage of the bladder due to sudden increase in urinary pressure, reaching a peak between the ages of 45 and 49 (about 65%), causing social problems or hygiene problems for many women do. For the treatment of the stress urinary incontinence described above, bladder neck suspension through the abdomen or vagina, midurethral sling operation using tape, sling operation, urethral submucosal injection, or artificial urethral sphincter implant, (Patent Literature 1 and Patent Literature 2), there is not yet a perfect treatment method that can be used for all urinary incontinence.

On the other hand, escoureteral reflux is a dysfunctional urinary disorder in which urinary tract stored in the bladder is reversed to the kidney through the ureter because the submucosal submucosal layer of the ureter of the ureter of the bladder is inherently deficient. Vesicoureteral reflux is most commonly detected by voiding urethrography during urination after urinary infection, and 20 to 30% of patients with total urinary tract infection have ureteric reflux. The treatment of vesicoureteral reflux is generally performed by antibiotic therapy and surgical treatment (Patent Document 3, Patent Document 4). Although the success rate is high (about 90%), there are disadvantages such as postoperative pain, complications (ureters perforation, ureterectomy, ureter obstruction, etc.) and long term admission.

One of the treatments for voiding dysfunction, such as urinary incontinence and vesicoureteral reflux, is an injection therapy using a bulking agent. For example, urinary incontinence is treated by increasing the resistance of the proximal urethra by injecting a filler into the submucosal tissue of the bladder neck and proximal urethra using an endoscope. In the case of reflux of the bladder, injection of a filler onto the ureter-bladder junction is performed to prevent the reflux by increasing the resistance of the ureteral inlet. These injections are noninvasive, easier to perform, less time-consuming, less painful, less costly, less hospitalized, and better procedure outcomes than conventional surgical procedures. These injections are widely practiced worldwide with safe procedures with few complications.

Currently, fillers used in these injections include Teflon paste, liquid silicone and collagen. However, these fillers have many problems such as adverse effects such as adverse effects due to migration of filler particles to other organs, inflammation reaction, foreign body reaction, necessity of re-operation due to reduction of filler volume, and the like. Therefore, it is urgent to develop injectable fillers suitable for injection therapy for the treatment of voiding dysfunction which does not cause side effects.

Korean Patent No. 10-563587 U.S. Published Patent Application No. 2005-0070930A1 U.S. Pat. No. 4,531,933 Korean Patent No. 10-0252584

Senuma Y et al., 2000, BioMaterials, 21, 1135.

It is an object of the present invention to provide a composition capable of preventing or treating a voiding disorder disease by using a composite material.

Another object of the present invention is to provide a method for producing a composite material capable of preventing or treating a dysuria disorder.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to " one embodiment " or " embodiment " means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment " or " an embodiment " in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

Unless defined otherwise herein, all scientific and technical terms used herein have the same meaning as commonly understood by one of skill in the art to which this invention belongs.

The inventors of the present invention have made intensive researches to examine the therapeutic effect of a composite material prepared by combining biodegradable polylactic acid microparticles and biocompatible hyaluronic acid polymers, and as a result, have found that hyaluronic acid hydrolysates Gel and polylactic acid spherical microparticles having a specific diameter, the present inventors have completed the present invention by confirming the excellent therapeutic effect on voiding disorder diseases, especially stress urinary incontinence and vesicoureteral reflux.

According to one embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating a voiding disorder disease comprising, as an active ingredient, a composite material comprising biodegradable polymer microparticles and a biocompatible polymer hydrogel.

In the present invention, the " dysuria disorder " means a disease in which urine is not easily excreted due to abnormalities in the body during urine discharge.

In the present invention, the urinary disorder disease is preferably one or more selected from the group consisting of urinary tract infection, enlarged prostate, urinary bladder, urinary incontinence, urinary incontinence, bladder neck sclerosis, urinary tract stricture, interstitial cystitis, , Urinary incontinence or vesicoureteral reflux, and most preferably, stress urinary incontinence or vesicoureteral reflux.

In the present invention, the term " incontinence " refers to a phenomenon in which urine is observed irrespective of the will of the patient. The prevalence rate is increasing with the increase in the elderly population due to the recent average life expectancy. Urinary incontinence is a type of urinary incontinence, which is caused by urinary incontinence without any triggering factors, such as stress incontinence that occurs when the urine is elevated due to coughing, urge incontinence, urge incontinence that can not be tolerated when urinating and urinary incontinence .

In the present invention, the term " vesicoureteral reflux " means a phenomenon in which the urine moves backwards from the bladder to the ureter. In general, vesicoureteral reflux is often caused by urinary abnormalities in children rather than adults. If the vesicoureteral reflux is not treated properly, kidney function and pyelonephritis may occur. In adults, vesicoureteral reflux may occur due to lower urinary tract obstruction, abnormal bladder function, rare urination, or constipation. Vesicoureteral reflux itself is not symptomatic, but urinary tract infection is accompanied by fever, urination, or frequent urination. Urinary tract infections are found in about 50% of infants, about 25% to 40% of infants, and about 5% of adults.

In the present invention, the "biodegradable polymer" is any substance that can be absorbed in vivo and can be used interchangeably with biocompatibility or bioabsorbability. The biodegradable material does not need to be chemically degraded or chemically degraded in vivo, but may be dispersed or dissolved in vivo in such a manner as to release a therapeutically effective amount of the drug for a significantly longer period of time than the normal in vivo lifetime of the drug Or < / RTI > In the present invention, the specific kind of the biodegradable polymer is not particularly limited, and examples thereof include polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA) At least one member selected from the group consisting of polyimides, polyimides, polyimides, polyimides, polyiminocarbonates, polyphosphoesters, polyanhydrides, polyorthoesters, polycaprolactones, polyhydroxybutyrates, polyamino acids, copolymers thereof and combinations thereof , And more preferably polylactic acid. In the present invention, the biodegradable material may be a homopolymer or copolymer, or any combination or blend of polymers.

In the present invention, the term " polylactic acid (PLA) " is a thermoplastic, chained polyester compound having biodegradability and bioactivity. The polylactic acid resin is a biodegradable resin made from starch as a raw material and has a small amount of heat generated by combustion and is easily hydrolyzed in soil or water and is completely decomposed into water and carbon dioxide by microorganisms in the soil. It does not have to go through the separate disposal process, which is environmentally friendly. Further, the melting point is sufficiently high to 120 to 180 占 폚, and exhibits excellent rigidity and transparency.

In the present invention, the composite material prepared by using the fine particles of irregularly shaped biodegradable polymer, which is not spherical, may not have a constant pressure output when used as a filler for injection. Therefore, in the present invention, the fine particles of the biodegradable polymer are preferably spherical.

If the diameter of the fine particles of the biodegradable polymer is less than 25 탆, the preservability in the tissue may be lowered after the tissue implantation of the composite material. If the diameter exceeds 250 탆, the inflammation reaction may occur in the tissue after the tissue implantation. Therefore, in the present invention, the fine particle diameter of the biodegradable polymer may be 25 to 250 탆.

More preferably, in the present invention, the biodegradable polymer includes fine particles having a diameter of 25 to 75 μm or less; Fine particles of more than 75 [mu] m but not more than 150 [mu] m; And fine particles having a diameter of more than 150 탆 and not more than 250 탆, and more preferably fine particles having a diameter of more than 75 탆 and not more than 150 탆, and more preferably fine particles having a diameter of 80 to 120 탆. And most preferably a fine particle having a diameter of 90 to 110 mu m.

In the present invention, the biocompatible polymer is preferably at least one selected from the group consisting of collagen, chitosan, gelatin, fibrin, hyaluronic acid and alginate, more preferably hyaluronic acid.

In the present invention, the term " hyaluronic acid (HA) " as used herein refers to a natural substance having biocompatibility that exists in many skin, articular fluid, cartilage and the like of animals. It is a component constituting a substrate in articular cartilage and is involved in the production of proteoglycan Is a glycoprotein complex of N-acetyl-D-glucosamine and D-glucuronic acid linked by a 1-4 glycosidic bond as a type of mucopolysaccharide. Since hyaluronic acid has a large amount of hydroxyl groups (-OH), it is hydrophilic and attracts a lot of water, so it has excellent moisturizing ability and is widely used in cosmetics such as lotion. It binds with water and becomes gel state. It is involved in lubrication of joints and flexibility of skin. It is also highly viscous and plays an important role in preventing invasion of bacteria or penetration of toxins into skin. If the skin lacks hyaluronic acid, the skin becomes dry, elastic and wrinkled. Because of this, hyaluronic acid can be used as a filler to prevent wrinkles or to fill in areas that have been turned off. The hyaluronic acid expressed in the microorganism has a diameter of 1.1 million daltons (1,100 kDa) or 3 million daltons (3,000 kDa) and can be obtained by a physical method such as ultraviolet ray, radiation or electron beam or by physical methods such as 1,4-butanediol diglycidyl ether , 4-Butandioldiglycidyl ether (BDDE), etc., physical and physiological properties are shown by cross-linking. Hyaluronic acid reacts with CD44 protein expressed in various epithelial cells to regulate various physiological functions and is found in many earthworms skin. Especially, since it is a component existing in human skin, there is no rejection reaction, and biocompatibility and caustic ratio are excellent. .

The hydrogel form of the biocompatible polymer necessary for the production of the composite material according to the present invention can be prepared by adding a crosslinking agent to hyaluronic acid, which is a biocompatible polymer. In the present invention, the "cross-linking agent" includes two or more reactive functional groups that form a covalent bond between two or more molecules. The crosslinking agent may be homobifunctional (i. E. Have the same two reactive ends) or heterobifunctional (i. E. Have two different reactive ends). The crosslinking agent to be added to hyaluronic acid includes a functional group of hyaluronic acid and a complementary functional group so that the crosslinking reaction proceeds. In the present invention, the crosslinking agent added to hyaluronic acid is 1,4-butanediol diglycidyl ether (BDDE), divinylsulfone (DVS), or 1-ethyl-3- Propyl) carbodiimide hydrochloride (EDC), or a combination thereof.

In the present invention, the term "hydrogel" may mean a three-dimensional network structure formed by crosslinking a hydrophilic polymer by covalent or non-covalent bonding. Due to the hydrophilic nature of the constituent material, absorbs and swells a large amount of water in an aqueous solution and an aqueous environment, but is not dissolved by the crosslinking structure. Accordingly, hydrogels having various shapes and properties can be produced depending on the constituent components and the manufacturing method, and generally have a property of intermediate property between liquid and solid since they contain a large amount of water.

In the present invention, when the viscoelasticity of the hydrogel of the biocompatible polymer is less than 100 Pa, there is a problem that the prepared composite material may have a reduced preservability in the body. When the viscosity exceeds 500 Pa, A problem may arise. Accordingly, in the present invention, the hydrogel of the biocompatible polymer may be a hydrogel having a viscoelasticity of 100 to 500 Pa (Pascal), the hydrogel having a viscoelasticity of 100 to 250 Pa or less; A hydrogel having a water content of not less than 250 Pa and not more than 350 Pa; The hydrogel may have at least one of hydrogels having a viscosity of more than 350 Pa and not more than 500 Pa, more preferably a hydrogel having a viscoelasticity of more than 250 Pa and not more than 350 Pa, more preferably a hydrogel having a viscoelasticity of 280 to 320 Pa, And a hydrogel having a viscoelasticity of 290 to 310 Pa.

In the present invention, the term " viscoelasticity " refers to a phenomenon in which both a property as a liquid (viscous) and a property (elasticity) as a solid when a force is applied to an object or a flow having an elastic deformation and a viscosity And all materials have viscoelastic properties. The hydrogel according to the present invention was injected into tissues and used for enhancing tissue volume or tissue correction, so that viscoelasticity was measured to confirm the physical properties of the hydrogel. Generally, the liquid has no restoring force (i.e., does not undergo elastic deformation) to the force causing the deformation. However, in the case of a colloidal solution or a thick polymer solution, the viscous property appears at the same time as it has a restoring force, thereby causing an elastic deformation. Pascal (Pa) used as a viscoelastic unit in the present invention is an SI derived unit against pressure. A Pascal is a unit that expresses the pressure when a force of one Newton per square meter is applied.

When the weight ratio of the fine particles of the biodegradable polymer and the hydrogel of the biocompatible polymer is included in a weight ratio exceeding the range of 1: 5 to 5: 1 in the production of the composite material according to the present invention, It is possible that the prevention or treatment effect of the urinary disturbance due to the increase of the pressure and the maintenance of the long-term effect may not occur.

Therefore, the fine particles of the biodegradable polymer and the hydrogel of the biocompatible polymer used in the composite material of the present invention are preferably contained in a weight ratio of 1: 5 to 5: 1, more preferably 1: 4 To 4: 1, more preferably from 1: 3 to 3: 1, most preferably from 1: 2 to 2: 1.

In the present invention, the composite material including the biodegradable polymer microparticles and the biocompatible polymer hydrogel may increase the oil leakage pressure and the urethral closure pressure by pressing the urethra or ureter.

The pharmaceutical composition according to the present invention may be in the form of capsules, tablets, granules, injections, ointments, powders or beverages according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. The pharmaceutical composition may be characterized in that it is intended for humans. The pharmaceutical composition may be formulated in the form of oral preparations such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterilized injection solutions according to a conventional method.

However, in the present invention, by using the pharmaceutical composition in the form of injectable filler, it can be injected into the urinary wall to urge the urethra, thereby effectively preventing or treating stress urinary incontinence. In addition, in the present invention, by injecting the injectable filling agent into the ureteral mucosa and compressing the ureter, the ureteral reflux of the bladder can be effectively prevented and treated.

The pharmaceutical compositions of the present invention may comprise a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be a binder, a lubricant, a disintegrant, an excipient, a solubilizing agent, a dispersing agent, a stabilizer, a suspending agent, a coloring matter, a perfume, etc. in the case of oral administration, A solubilizing agent, an isotonic agent, a stabilizer and the like may be mixed and used. In the case of topical administration, a base, an excipient, a lubricant, a preservative and the like may be used. Formulations of the pharmaceutical compositions of the present invention may be prepared in a variety of ways by mixing with a pharmaceutically acceptable carrier as described above. For example, oral administration may be in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. In the case of injections, they may be formulated in unit dosage ampoules or in multiple dosage forms have. Other, solutions, suspensions, tablets, capsules, sustained-release preparations and the like.

The term " therapeutically acceptable carrier " as used herein means a carrier that is acceptable for administration to an individual, such as a mammal, such as a carrier having acceptable mammalian safety for a given dosing schedule. The pharmaceutically acceptable carrier includes an aqueous diluent or solvent such as phosphate buffered saline, purified water, sterilized water, etc., and may be a non-aqueous diluent such as propylene glycol, polyethylene glycol, olive oil, . Further, if necessary, it may contain a wetting agent, a fragrance, a preservative, and the like.

Solid formulations for oral administration of the pharmaceutical compositions of the present invention include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate (Calcium carbonate), sucrose or lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witelsol, macrogol, tween 61, cacao paper, laurin, glycerol gelatin and the like can be used.

The route of administration of the pharmaceutical compositions according to the present invention may be, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, Sublingual or rectal. Oral or parenteral administration is preferred. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention may also be administered in the form of suppositories for rectal administration.

The pharmaceutical composition according to the present invention can be formulated into a parenteral dosage form. The parenteral dosage form may be a injectable filler or an external preparation for skin. The external skin preparation may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a drug-containing bandage, a lotion, or a combination thereof. The external preparation for skin may be a cosmetic product or a medicament for use in a composition for external use for skin such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickening agent, an ultraviolet absorber, a whitening agent, an antiseptic, Flavorings, coloring agents, various skin nutrients, or combinations thereof, and may be appropriately blended as required. The external preparation for skin may be a metal blocker such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridine, Extracts of fruits, various herbal medicines, drugs such as tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts thereof, vitamin C, magnesium ascorbic acid phosphate, glucoside ascorbic acid, arbutin, kojic acid, glucose, fructose , Trehalose and the like can also be appropriately blended. When formulating the composition in the form of a filler for injectable use, it is prepared using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant or the like which is usually used.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and any parenteral administration method may be used. The pharmaceutical composition of the present invention varies depending on various factors including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease to be prevented or treated. And the preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but may be suitably selected by those skilled in the art. Kg or 0.001 to 50 mg / kg, and the dose is not limited in any way to the scope of the present invention. The administration may be carried out once a day or divided into several times. The pharmaceutical compositions of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

According to another embodiment of the present invention, there is provided a method for producing a microparticle, comprising the steps of: dissolving a biodegradable polymer in a solvent to prepare fine particles; Crosslinking the biocompatible polymer to prepare a hydrogel; And mixing the fine particles and the hydrogel to produce a composite material. The present invention also relates to a method for producing a composite material for preventing or treating a voiding disorder disease.

In the present invention, the urinary disorder disease is preferably one or more selected from the group consisting of urinary tract infection, enlarged prostate, urinary bladder, urinary incontinence, urinary incontinence, bladder neck sclerosis, urinary tract stricture, interstitial cystitis, , Urinary incontinence or vesicoureteral reflux, and most preferably, stress urinary incontinence or vesicoureteral reflux.

In the present invention, the specific kind of the biodegradable polymer is not particularly limited, and examples thereof include polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA) At least one member selected from the group consisting of polyimides, polyimides, polyimides, polyimides, polyiminocarbonates, polyphosphoesters, polyanhydrides, polyorthoesters, polycaprolactones, polyhydroxybutyrates, polyamino acids, copolymers thereof and combinations thereof , And more preferably polylactic acid.

In the present invention, a specific method for producing the fine particles of the biodegradable polymer is not particularly limited, and for example, a method of preparing a fully dissolved continuous phase by adding polyvinyl alcohol into distilled water and stirring the mixture; Dissolving polylactic acid in a solvent to prepare a dispersed phase; Passing the prepared dispersed phase through the pores of the membrane to produce an emulsion; And centrifuging the emulsion to remove polyvinyl alcohol.

In the present invention, the diameter of the membrane pores is preferably 5 to 100 탆, more preferably 5 to 20 탆.

In the present invention, when the dispersed phase is passed through the pores of the membrane, the fine particle shape can be formed by performing dispersion at a pressure of 2 to 50 kPa, more preferably 5 to 20 kPa.

Further, in the present invention, the centrifugation of the emulsion can be carried out at a rotation speed of 200 to 5000 rpm, more preferably 500 to 2000 rpm.

In the present invention, the fine particles of the biodegradable polymer are preferably spherical, the spherical fine particles may have a diameter of 25 to 250 μm, fine particles having a diameter of 25 to 75 μm; Fine particles of more than 75 [mu] m but not more than 150 [mu] m; More preferably at least one of fine particles having a diameter of more than 150 μm but not more than 250 μm, more preferably fine particles having a diameter of more than 75 μm and not more than 150 μm, more preferably fine particles having a diameter of 80 μm to 120 μm And most preferably from 90 to 110 mu m.

In the present invention, the biocompatible polymer is preferably at least one selected from the group consisting of collagen, chitosan, gelatin, fibrin, hyaluronic acid and alginate, more preferably hyaluronic acid.

The method for preparing the hydrogel of the biocompatible polymer according to the present invention is not particularly limited. For example, the method may include the steps of dissolving sodium hydroxide in distilled water; Filtering the dissolved sodium hydroxide with a filter to prepare a sodium hydroxide solution; Completely dissolving hyaluronic acid in the prepared sodium hydroxide solution; Mixing the fully dissolved hyaluronic acid with a cross-linking agent and sufficiently mixing; Dividing the hyaluronic acid solution mixed with the cross-linking agent; And cross-linking the solution with the dispensed hyaluronic acid.

The diameter of the pores in the filter used for filtration of sodium hydroxide dissolved in the present invention may be 0.05-1 mu m, more preferably 0.1-0.5 mu m.

In the present invention, the filtered sodium hydroxide solution may be 0.1 to 5%, more preferably 0.5 to 2%.

In the present invention, the proportion of hyaluronic acid to be added to the sodium hydroxide solution may be 2 to 50 (wt / v)%, more preferably 5 to 15 (wt / v)%.

In the present invention, the dispensed amount of the hyaluronic acid solution mixed with the cross-linking agent may be 2 to 20 g, more preferably 8 to 12 g.

In the present invention, the time for which the hyaluronic acid solution dispensed in the incubator is crosslinked at room temperature may be 5 to 25 hours, more preferably 10 to 22 hours, and most preferably 16 to 22 hours.

In the present invention, the kind of the cross-linking agent is not particularly limited, and examples thereof include 1,4-butanediol diglycidyl ether (BDDE), divinylsulfone (DVS) 3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), or a combination thereof.

Also, in the present invention, the hydrogel of the biocompatible polymer may be a hydrogel having a viscoelasticity of 100 to 500 Pa (Pascal), the hydrogel having a viscoelasticity of more than 100 Pa and 250 Pa or less; A hydrogel having a water content of not less than 250 Pa and not more than 350 Pa; The hydrogel may have a viscoelasticity of more than 250 Pa but not more than 350 Pa, more preferably a hydrogel having a viscoelasticity of 280 to 320 Pa, and most preferably a viscoelastic May be a hydrogel of 290 to 310 Pa.

In the present invention, the fine particles of the biodegradable polymer and the hydrogel of the biocompatible polymer are preferably contained in a weight ratio of 1: 5 to 5: 1, more preferably 1: 4 to 4: 1, May be included in a weight ratio of 1: 3 to 3: 1, and most preferably in a weight ratio of 1: 2 to 2: 1.

According to the present invention, by injecting a filler for injection containing an active ingredient of a composite material prepared by combining polylactic acid microparticles as a biodegradable polymer and hyaluronic acid hydrogel as a biocompatible polymer into the urinary wall, It is possible to prevent or treat effectively, and by injecting the injectable filling agent into the ureteral submucosa, the ureter can be pushed to effectively prevent and treat the ureteric reflux.

1 shows the results of measurement of the pressure output of a composite material prepared by combining a hyaluronic acid hydrogel and various types of polylactic acid microparticles.
Fig. 2 shows the results of measurement of the pressure output of a composite material produced by combining hyaluronic acid hydrogel (viscoelasticity, unit Pa) and polylactic acid (particle diameter, unit μM) microparticles under various conditions.
Fig. 3 shows the results of measurement of the preservability of a composite material prepared using hyaluronic acid and polylactic acid (limited to spherical particles) under various conditions.
Fig. 4 shows the results of inflammation in tissues caused by composite materials prepared using hyaluronic acid and polylactic acid (limited to spherical particles) under various conditions.
FIG. 5 shows the result of measurement of urine leakage pressure using the composite material according to the present invention in an animal model in which a voiding disorder disease was induced.
FIG. 6 shows the result of measuring the urethral closure pressure using the composite material according to the present invention in an animal model in which a voiding dysfunction disorder has been induced.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Preparation Example  1. Hyaluronic acid (HA, Hyaluronic  Acid) Hydrogel  Produce

10 g of sodium hydroxide was dissolved in 1 L of distilled water, followed by filtration through a 0.2 탆 filter to prepare a 1% sodium hydroxide solution. 10 (wt / v)% hyaluronic acid was added to the prepared sodium hydroxide and completely dissolved. 1,4-butanediol diglycidyl ether (BDDE) was added to the completely dissolved hyaluronic acid and mixed thoroughly. The hyaluronic acid solution containing the cross-linking agent was dispensed into a flat container in an amount of 10 g, and the mixture was placed in an incubator and crosslinked at room temperature for 18 hours to prepare a highly elastic hydrogel.

Preparation Example  2. Spherical Polylactic acid ( PLA , Poly  Preparation of Lactic Acid Fine Particles

1 g of poly vinyl alcohol was added to 100 g of distilled water and stirred at room temperature to dissolve completely to prepare a continuous phase. 1 g of polylactic acid (PLA) was dissolved in 100 g of a dimethyl-chloride solvent to prepare a dispersed phase. The prepared dispersed phase was put into a pressure-tight tube assembled together with a membrane having a pore size of 10 mu m and dispersed at a pressure of 10 kPa to prepare an emulsion. The prepared emulsion was stirred at 1000 rpm for 1 hour and then polyvinyl alcohol was removed from the emulsion by centrifugation. In order to remove the dimethyl chloride solvent from the fine particles obtained by removing the polyvinyl alcohol, a vacuum state was formed by using a vacuum tube to completely remove the solvent to obtain polylactic acid fine particles.

compare Preparation Example  1. Irregular shape Polylactic acid ( PLA , Poly  Preparation of Lactic Acid Fine Particles

Polylactic acid microparticles were prepared by spraying. 0.5 g of poly vinyl alcohol was added to 100 g of distilled water and stirred at room temperature to dissolve completely to prepare a water phase. The oil phase was prepared by dissolving 1 g of polylactic acid (PLA) in 100 g of a dimethyl-chloride solvent. The prepared oil phase was placed in a 10 mL syringe and the syringe was fixed to the syringe pump. While the aqueous polyvinyl alcohol solution was stirred at 300 rpm, the aqueous solution was gradually dropped into the aqueous phase solution using a syringe pump to prepare fine particles. The polyvinyl alcohol was removed by centrifugation in order to obtain the prepared fine particles. In order to remove the dimethyl chloride solvent from the fine particles obtained by removing polyvinyl alcohol, a vacuum state was formed by using a vacuum tube to completely remove the solvent to obtain polylactic acid microparticles having irregular shapes.

Example  1. Manufacture of composite materials (1)

Hyaluronic acid hydrogel (hereinafter referred to as 'HA') was prepared in the same manner as in Preparation Example 1 except that the viscoelasticity of the HA was adjusted as shown in Table 1 in order to confirm the effect of the difference in viscoelasticity .

In addition, spherical and irregularly shaped polylactic acid microparticles (hereinafter referred to as PLA) were prepared in the same manner as in Preparation Example 2 and Comparative Preparation Example 1, respectively.

As described above, HA having various viscoelastic properties and spherical or irregularly shaped PLA were mixed at a weight ratio of 1: 1 to prepare a composite material.

Mixing ratio

200 Pa 300Pa 400 Pa Irregular shaped particle 1: 1 1: 1 1: 1 Spherical particle 1: 1 1: 1 1: 1

Experimental Example  1. Characteristics of composite materials (1)

The composite material prepared in Example 1 was charged into a syringe in an amount of 1 g, and centrifuged at 6000 rpm for 3 minutes to remove air bubbles in the composite material. After the air bubbles were removed, the syringe was placed on the pressure output device and the pressure output was measured at a speed of 12 mm. The results are shown in FIG.

As shown in FIG. 1, a composite material in which PLA particles are mixed with regulated particles is injected under a certain force, whereas a composite material in which irregular particles are mixed is not uniformly injected . In addition, it was confirmed that the hyaluronic acid hydrogels mixed together increased the pressure output as the viscoelasticity increased (200 Pa to 400 Pa).

Example  2. Manufacture of composite materials (2)

(Hereinafter referred to as 'HA') and spherical polylactic acid microparticles (hereinafter referred to as 'PLA') were prepared in the same manner as in Preparation Examples 1 and 2, except that the viscoelastic and spherical PLA Were prepared as shown in Table 2, and then mixed at a weight ratio of 1: 1 to prepare a composite material.

Mixing ratio

200 Pa 300Pa 400 Pa 20-50 탆 1: 1 1: 1 1: 1 70-100 탆 1: 1 1: 1 1: 1 170-200 탆 1: 1 1: 1 1: 1

The PLA particle diameter can be measured by dynamic light scattering (DLS). The dynamic light scattering method is a non-invasive method for measuring the size of nanoparticles in a dispersion. The intensity of the scattered light is measured with time in the particle suspension using Brownian motion, and the intensity of the scattered light is analyzed. And the average diameter of the particles from the above coefficients can be confirmed through the Stoch-Einstein equation.

The HA viscoelasticity can be measured using a texture analyzer (TAXT plus, Texture Technologies Corp.). The cross - sectional area and height of the disk - shaped hydrogel samples were measured using digital calipers. The 1/2 inch (12.7 mm) radius dacron tip was pressurized at a rate of 0.5 mm / sec. The hydrogel was held for 60 seconds to allow a 20% strain to occur, and then the tip was removed. Since the elastic modulus is defined as the ratio of the stress and strain of the elastic body within the elastic limit, when the small strain of about 0 to 2% occurs, the elasticity can be confirmed by measuring the change in the ratio of stress and strain. Pascal (Pa).

Experimental Example  2. Characteristics of composite materials (2)

2.1 Main input

The composite material prepared in Example 2 was charged into a syringe in an amount of 1 g, and centrifuged at 6000 rpm for 3 minutes to remove air bubbles in the composite material. After the air bubbles were removed, the syringe was placed on the pressure output device and the pressure output was measured at a speed of 12 mm. The results are shown in FIG.

As shown in FIG. 2, the main input of the composite material increased in proportion to the diameter of the PLA fine particles and the viscoelasticity of the HA hydrogel.

However, when the viscoelasticity of the HA is 400 Pa and the diameter of the PLA fine particles is 170 to 200 탆 (200 탆), the main input of the composite material is not maintained constant, but when the viscoelasticity of the HA is 200 to 300 Pa, It can be confirmed that the main input of the system is stably maintained. Particularly, the main input was stably maintained when the viscoelasticity of HA was 200 to 300 Pa, the diameter of PLA particles was 20 to 50 占 퐉 (50 占 퐉), and 70 to 100 占 퐉 (100 占 퐉).

Therefore, in the following experiment, experiments were performed using only HA having viscoelasticity of 200 Pa or 300 Pa.

2.2 Preservation of body

6-week-old rats (RAT) were prepared, intraperitoneally injected with zoletile (50 mg / kg) and rumun (10 mg / kg) and intraperitoneal anesthesia was performed. The anesthetized rats were rinsed with a razor to remove hair, and 0.4 mL of the composite material prepared in Example 2 was injected into the dorsal part. In order to confirm the preservability of the injected composite material, the volumes of the complex-injected tissues were measured at 1, 2, 4, and 6 weeks after injection of the composite material, and the results are shown in FIG. However, the tissue volume immediately after injection of the composite material was set at 100.

As shown in FIG. 3, it can be confirmed that the initial decomposition of the composite material is controlled by the viscoelasticity of the HA hydrogel. Compared to a composite material having viscoelasticity (200 Pa), which has a lower viscosities (300 Pa) I was able to confirm that it was excellent.

However, in the latter half of the injection, the preservation of the body was maintained in proportion to the particle diameter of PLA. Specifically, when the diameter of fine particles of PLA is in the range of 20 to 50 탆 (50 탆), the diameter of the fine particles is 70 to 100 탆 (100 탆) or 170 to 200 탆 (200 탆) It was confirmed that the preservability was kept high.

From these results, a composite material obtained by mixing a HA hydrogel having a viscoelasticity of 300 Pa and spherical PLA fine particles having a diameter of 100 占 퐉 or more can be stably injected into the body at a high main input, I can see that it can be.

2.3 Biocompatibility

In order to measure the biocompatibility of the composite material, composite material was injected into the rats, euthanized using carbon dioxide at 6 weeks, and tissues of the site where the composite material was injected were extracted. The extracted tissue was fixed with 4% paraformaldehyde for one day. The fixed tissues were prepared by using paraffin, cut into 4 ㎛ diameters, and attached to slides to make slides necessary for microscopic observation. After removing the paraffin, the slides were stained with hematoxylin and eosin, and the results were confirmed by an optical microscope. The results are shown in FIG.

As shown in Fig. 4, when the PLA particles had a diameter of 170 to 200 mu m (200 mu m) as a result of tissue staining, severe inflammatory reaction was induced in the tissues. In contrast, when the diameter of the fine particles was 20 to 50 탆 (50 탆) or 70 to 100 탆 (100 탆), no inflammation occurred in the tissue, and fibroblasts aggregated around the composite material, .

Therefore, from these results, a composite material obtained by mixing a HA hydrogel having a viscoelasticity of 300 Pa and spherical PLA fine particles having a diameter of 70 to 100 탆 (100 탆) can be stably injected into the body at a high main input , And it was confirmed that the best therapeutic effect on stress urinary incontinence and vesicoureteral reflux can be obtained by being maintained in the body for a long time after the injection.

Example  3. Manufacturing of composite materials (3)

HA hydrogel and PLA fine particles were prepared in the same manner as in Preparations 1 and 2 except that the viscoelasticity of the HA was set to 300 Pa and the PLA was spherically shaped and its particle diameter was controlled to 20 to 100 占 퐉. Then, the HA and PLA were mixed in the weight percentages shown in Table 3 below to prepare a composite material.

division HA content (%) PLA content (%) Comparative Example 1 100 0 Production Example 1 75 25 Production Example 2 50 50 Production Example 3 25 75 Comparative Example 2 0 100

Experimental Example  3. Therapeutic effect of dysuria

3.1 Leakage pressure  Measure

6-week-old rats were anesthetized by intraperitoneal anesthesia with a mixture of zoletile (50 mg / kg) and rumpun (10 mg / kg). Anesthetized rats were sacrificed after removal of hair to induce stress urinary incontinence. Rats in which stress urinary incontinence was induced were injected with 1 ml of the composite material prepared in the mixing ratio described in Example 3, divided into two portions on the wall of the urine, for 1 week, and at 1 week, 6 weeks and 12 weeks after injection, And the results are shown in Fig.

As shown in FIG. 5, HA and PLA were mixed at a ratio of 3: 1 or 1: 3 as in the case of the present invention (Comparative Examples 1 and 2) in which only HA or PLA was injected into rats in which stress urinary incontinence was induced In case of injecting the composite material (Preparation Example 1 or 3), it was confirmed that the leakage pressure was remarkably increased. In particular, when the composite material mixed with HA and PLA was mixed according to Production Example 2, And it was confirmed that it increased more remarkably.

3.2 Urethral closure pressure  Measure

The urinary bladder pressure was measured at 1, 6, and 12 weeks after the composite material according to Example 3 was injected into the urinary wall of the rats in which the stress incontinence induced according to Experimental Example 3.1 was induced.

As shown in FIG. 6, HA and PLA were mixed in a ratio of 3: 1 to 1: 3 as in the present invention, as compared with cases where only HA or PLA was injected into rats in which stress urinary incontinence was induced (Comparative Examples 1 and 2) It was confirmed that the urethral closure pressure was significantly increased when the composite material (Preparation Examples 1 to 3) was injected. In particular, when a composite material in which HA and PLA were mixed at a ratio of 1: 1 according to Production Example 2 was injected, Was significantly increased.

Through this, it was confirmed that the composite material in which HA and PLA were mixed at a ratio of 3: 1 to 1: 3 has an effect of improving or maintaining the urinary leakage pressure and the urethral closure pressure, and thus has a therapeutic effect on the stress urinary incontinence.

It will also be apparent to those skilled in the art that the composite material exhibits an improvement or maintenance effect of urinary leakage pressure and urethral closure pressure, thereby exhibiting therapeutic efficacy against vesicoureteral reflux.

All references, articles, publications and patents and patent applications cited herein are hereby incorporated by reference in their entirety. Accordingly, the spirit and scope of the following claims should not be limited to the description of the preferred embodiments described above.

Claims (19)

A pharmaceutical composition for preventing or treating a urinary disorder disease comprising a biodegradable polymer microparticle and a biocompatible polymer hydrogel-containing composite material as an active ingredient.
The method according to claim 1,
Wherein said urinary disorder disease is any one or more selected from the group consisting of urinary tract infection, prostate hyperplasia, overactive bladder, urinary incontinence, urinary stone, bladder neck sclerosis, urinary tract stricture, interstitial cystitis and vesicoureteral reflux A pharmaceutical composition.
The method according to claim 1,
Wherein said biodegradable polymer microparticles are spherical, wherein said biodegradable polymer microparticles are spherical.
The method according to claim 1,
Wherein the biodegradable polymer microparticles have a diameter of 20 to 250 占 퐉.
The method according to claim 1,
Wherein the viscoelasticity of the biocompatible polymer hydrogel is 100 Pa to 500 Pa.
The method according to claim 1,
Wherein the weight ratio of the microparticles and the hydrogel in the composition is 1: 3 to 3: 1.
The method according to claim 1,
Wherein the biodegradable polymer is selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, Wherein the pharmaceutical composition is any one selected from the group consisting of polyarse ester, polycaprolactone, polyhydroxybutyrate, polyamino acid, copolymers thereof, and combinations thereof.
8. The method of claim 7,
Wherein the biodegradable polymer is polylactic acid, wherein the biodegradable polymer is polylactic acid.
The method according to claim 1,
Wherein the biocompatible polymer is any one selected from the group consisting of collagen, chitosan, gelatin, fibrin, hyaluronic acid and alginate.
10. The method of claim 9,
Wherein said biocompatible polymer is hyaluronic acid, for the prevention or treatment of dysuria.
The method according to claim 1,
Wherein the composition is a filler for injection, for the prevention or treatment of dysuria.
Dissolving the biodegradable polymer in a solvent to prepare fine particles;
Crosslinking the biocompatible polymer to prepare a hydrogel; And
And mixing the fine particles and the hydrogel to produce a composite material.
13. The method of claim 12,
Wherein said urinary incontinence disease is any one or more selected from the group consisting of urinary tract infection, enlarged prostate, urinary bladder, incontinence, urinary incontinence, bladder neck scarring, urinary tract obstruction, interstitial cystitis and vesicoureteral reflux.
13. The method of claim 12,
Wherein the biodegradable polymer microparticles are spherical.
13. The method of claim 12,
Wherein the fine particles have a diameter of 20 to 250 mu m.
13. The method of claim 12,
Wherein the viscoelasticity of the hydrogel is 100 Pa (pascal) to 500 Pa.
13. The method of claim 12,
Wherein the weight ratio of the fine particles and the hydrogel in the composite material is 1: 3 to 3: 1.
13. The method of claim 12,
Wherein the biodegradable polymer is selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, Wherein the polymer is any one selected from the group consisting of polyorthoesters, polycaprolactones, polyhydroxybutyrates, polyamino acids, copolymers thereof, and combinations thereof.
13. The method of claim 12,
Wherein the biocompatible polymer is any one selected from the group consisting of collagen, chitosan, gelatin, fibrin, hyaluronic acid and alginate.

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