KR100426636B1 - Injectable Gel Type Lipid Composition And Preparation Method Thereof - Google Patents

Abstract

The present invention relates to compositions on injectable gels and methods for their preparation, and more particularly, to phospholipid compounds, drugs, polymers and water, wherein the composition on gels is a topical drug delivery system by slowly releasing drugs. It is useful for.

Description

Injectable Gel Type Lipid Composition And Preparation Method Thereof

The present invention is a gel type composition (gel type); Gel phase compositions comprising drugs; Gel phase formulations; And it relates to a method for producing the composition and formulation, more specifically, the gel phase composition is 10 to 99% by weight of at least one phospholipid compound, 0.001 to 50% by weight of the drug, at least one biocompatible polymer 3 Up to weight% and from 0.5 to 90 weight% of water.

When the drug is administered for the purpose of treatment, systemic administration using various routes such as oral administration, intravenous injection, and subcutaneous injection is mainly used. However, since such systemic administration has many side effects, researches on a local drug delivery system that can locally administer drugs only to the affected area have recently been actively conducted to eliminate such side effects. .

The topical drug delivery system has been developed to show the maximum effect of a small amount of drug at a desired location, and can maintain high local concentration and eliminate side effects of systemic administration.

As a topical drug delivery system, a biocompatible polymer is usefully used, and various formulations using the same are manufactured. Currently, sustained-release formulations using polymeric microspheres have been developed and marketed in the treatment of periodontal disease (Korean Patent No. 1996-16882; Korean Patent No. 2001-86908), and hyaluronic acid formulations on gels have been developed in local anesthetic delivery systems. have. (US Pat. No. 5,990,096: Formulations containing hyaluroninc acid). Non-steroidal anti-inflammatory drugs (NSAIDs) and phospholipid complexes have been actively developed to overcome gastrointestinal disorders upon oral administration. (US Pat. No. 4,687,766: Pharmaceutical preparation for the therapeutic treatment of rheumatic disease; US Pat. No. 5,955,451: Methods of enhancing the therapeutic activity of NSAIDS and compositions of zwitterionic phospholipids useful therein).

Hyaluronic acid formulations on gels have been developed for the treatment of rheumatoid arthritis. Hylan or Hyaluronan is known to reduce joint pain because of its viscoelastic nature. In addition, hyaluronic acid has been used in a variety of materials, such as sutures, adhesion preventing gel (adhesion preventing gel) because of its excellent biocompatibility and biodegradability.

Phospholipids are mostly included in the components of cell membranes, and are highly biocompatible and present in a hydrated form mixed with water. However, hydrated phospholipids easily disperse into the structure of multilamellar vesicles when placed in excess water, and easily erosion when hydrated phospholipid pastes are present in body fluids when using drug delivery systems. ), There is a problem that the encapsulated drug is rapidly released.

Hydrated phospholipid can form various kinds of phases by various factors such as the type of phospholipid, amount of water, temperature, pressure, and salt concentration. Among them, the saturated biophosphorylated saturated phosphatidyl choline has a lamellar crystalline phase, a lamella gel phase, a lamellar liquid phase, and a ripple. phases, hexagonal phases, and cubic phases. Phospholipids (hydrated phospholipids), which are hydrated by water, are generally viscous white pastes, depending on the type of phase. A viscous white paste containing the drug in the hydrated phospholipid and the drug formed therein can be effectively used in the drug delivery system.

Accordingly, the present inventors have tried to solve the above problems, to prepare a composition containing a specific content of the biocompatible polymer in order to use the hydrated phospholipids as a drug delivery system, the erosion effect is suppressed even if the composition is immersed in excess water The present invention has been completed by finding out that it effectively releases the drug to the topical area.

It is an object of the present invention to provide a composition on a gel which can be used as a topical drug delivery system and a method for producing the same.

Figure 1a shows after immersing in a water the gel composition of the present invention using a biocompatible polymer hyaluronic acid.

(Composition: Egg phosphatidylcholine: hyaluronic acid: water = 1000: 1: 500 (weight ratio))

Figure 1b shows after immersing the gel-like composition containing no biocompatible polymer in water.

(Composition: Egg phosphatidylcholine: water = 1000: 500 (weight)

Figure 2a shows after vortexing for two minutes after immersing the gel composition of the present invention, characterized in that the biocompatible polymer hyaluronic acid and tetracycline as a drug comprising tetracycline.

(Egg phosphatidylcholine: tetracycline: hyaluronic acid: water = 1000: 75: 1: 500 (weight ratio))

FIG. 2B shows the gel phase composition containing no tetracycline as a drug without biocompatible polymers and then vigorously stirred for 2 minutes after immersion in water.

(Composition: Egg phosphatidylcholine: tetracycline: water = 1000: 75: 500 (weight ratio))

3 is a graph showing the drug release rate of the gel phase composition containing tetracycline as the drug.

; Gel phase formulation consisting of egg phosphatidylcholine / water

; Gel phase formulation consisting of egg phosphatidylcholine / hyaluronic acid / water

; Tetracycline formulation in aqueous solution (control)

In order to achieve the above object, the present invention provides a composition on a gel containing phospholipids, drugs, biocompatible polymers and water.

In the present invention, the step of adding water to the residue obtained by dissolving the phospholipid compound and the drug in an organic solvent and then removing the organic solvent (step 1), adding a biocompatible polymer aqueous solution to the mixture prepared in step 1 It provides a method for producing a composition on a gel consisting of (step 2). In the above method, in order to prepare on the injectable gel, two syringes are connected with three-way stoppers, and the piston of the syringe is pushed more than 100 times in both directions to prepare a uniform gel.

Hereinafter, the present invention will be described in detail.

Specifically, the composition on the injectable gel of the present invention comprises 10 to 99% by weight of one or more phospholipid compounds, 0.001 to 50% by weight of drug, 3% by weight or less of biocompatible polymer on one species and 0.5 to 90% by weight of water. It consists of%.

Specifically describing each composition of the present invention, the phospholipid compound is hydrated when mixed with water to exhibit a lamellar structure and encapsulates a drug in the lamellar structure. Phospholipid compounds usable in the present invention may be used alone or in combination, as long as the phospholipid compounds commonly used in the art.

Preferably, a phosphatidylcholine compound, a phosphatidylethanolamine compound, a phosphatidylserine compound, or a phosphatidinic acid compound selected from saturated or unsaturated C ₆ to C ₂₂ Or a mixture of two species is preferred, and more preferably phosphatidylcholine is used.

The phosphatidylcholine-based compound of the present invention is egg phosphatidylcholine (egg phosphatidylcholine, egg lecitin), soya phosphatidylcholine (soya lecitin), dipalmitoyl phosphatidylcholine, dimyristoyl phosphatidylcholine (dimichoyl phosphatidyl thiophosphate) ), Saturated phosphatidylcholine, such as distearyl phosphatidylcholine, diylaidoyl phosphatidylcholine, dioleyl phosphatidylcholine, dipalmitolylphosphatidylcholine (dipalmitoleylyldiphosphylyl phosphatidyldiphosphidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl phosphatidyl unsaturated phosphadidylcholine, such as phosphatidylcholine), may be used, preferably saturated phosphadidylcholine is used.

The phosphatidylethanolamine compound of the present invention is egg phosphatidylethanolamine (egg phosphatidylethanolamine), soy phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine (dipalmitoylphosphatidylethanolamine), dimyristoyl phosphatidyl ethanolamine didi ethanol amine didi ethanolamine ), Saturated phosphatidyl ethanolamines such as dilaurylphosphatidylethanolamine, and distearyl phosphatidylethanolamine, and dielaidoylphosphatidylethanolamine, and dioleyl phosphatidyl ethanol amine Unsaturated phosphatides such as dipalmitoleylphosphatidylethanolamine, and dimyristoleylphosphatidylethanolamine Diethanolamine is used.

The phosphatidylserine-based compounds of the present invention are egg phosphatidylserine (egg phosphatidylserine), soybean phosphatidylserine, dipalmitoylphosphatidylserine, dimyristoyl phosphatidylserine (dimiristoylphosphatidylserine dimethyl lauryl phosphatidylserine) ), And saturated phosphatidylserine such as destearyl phosphatidylserine, diylaidoyl phosphatidylserine, dioleyl phosphatidylserine, dipalmitolyl phosphatidylserine, and dipalmitoleylphosphatidylserine. Unsaturated phosphatidylserine such as stoleyl phosphatidylserine can be used.

The phosphatidic acid-based compounds of the present invention are egg phosphatidic acid (egg phosphatidic acid), soy phosphatidic acid (soya phosphatidic acid), dipalmitoyl phosphatidic acid (dipalmitoylphosphatidic acid), dimyristoyl phosphatidic acid (dimiristoylphosphatidic acid) ), Saturated phosphatidic acids such as dilaurylphosphatidic acid, and distearylphosphatidic acid, and dielaidoylphosphatidic acid, and dioleyl phosphatidic acid. Unsaturated phosphatidic acid, such as (dioleylphosphatidic acid), dipalmitoleyl phosphatidic acid, and dimyristoleyl phosphatidic acid, can be used.

The content of the phospholipid compound used in the present invention is included in 10 to 99% by weight based on the total composition, preferably 40 to 70% by weight when showing the maximum effect. At this time, if the content of the phospholipid is too small beyond the above content, it is undesirable because it forms a colloidal form such as a liposome or a multilayer thin film vesicle (mitilamellar vesicle (MLV)).

In particular, in the present invention, a biocompatible polymer is added to suppress the rapid release of the drug when the hydrated phospholipid compound is immersed in water. The biocompatible polymer may control the release rate of the drug by adjusting the viscosity of the formulation and maintaining the morphology of the formulation for a long time when immersed in water. In other words, when the biocompatible polymer is not contained, the phospholipid formulation spontaneously forms the colloidal system in a large amount of aqueous solution, but the gel state is maintained when the polymer is contained. As the biocompatible polymer of the present invention, hyaluronic acid and salts thereof, chitosan and salts thereof, alginic acid and salts thereof may be used, and hyaluronic acid is preferably used.

As the drug of the present invention, both water-soluble, fat-soluble or amphoteric drugs may be used. Specifically, drugs which may be used in the present invention may be antiviral agents, steroid-based anti-inflammatory drugs, and non- Non-steroidal anti-inflammatory drugs, treatment for periodontal disease, antibiotics, antifungals, vitamins, hormones, prostaglandin, prostacyclin, anticancer agents, anti-metabolic agents, mitotics, choline Sex drugs, adrenaline antagonists, anticonvulsants, antianxiety agents, thermostatics, antidepressants, anesthetics, analgesics, anabolic steroids, estrogens, progesterons, glycosaminoglycans, polynucleotides (polynucleotide) can be used as an immunosuppressant or immune promoter.

In addition to the above composition, additives may be additionally used only by those skilled in the art, and the content thereof is contained in the total composition content of 5 wt% or less. After the drug is selected, the additive is selected if necessary due to its characteristics, and the additives used may be variously added as antioxidants, triglycerides, monoglycerides, fatty acids, ester derivatives of fatty acids, alcohol derivatives of fatty acids, and polymers. have.

The present invention also provides a method for preparing a composition on a gel containing a drug.

Dissolving the phospholipid and the drug in an organic solvent and then adding water to the residue obtained by removing the organic solvent (step 1), and adding a biocompatible polymer aqueous solution to the mixture prepared in step 1 (step 2). Provided are methods for preparing a composition on a gel.

In particular, the organic solvent is preferably alcohol, chloroform, benzene, toluene, acetonitrile or a mixed solvent thereof.

In addition, the alcohol is preferably ethanol, propanol, isopropyl alcohol, butanol or a mixed solvent thereof, more preferably ethanol.

The formulation may be prepared by mixing a biocompatible polymer aqueous solution with hydrated phospholipids, including drugs.

The formulations according to the invention also exhibit a sustained release pattern for the drug they contain, which is particularly advantageous for drug delivery systems (see FIG. 2A ).

In particular, when the formulation according to the present invention is used in the drug delivery system, subcutaneous injection, intraoral administration, injection in a periodontal pocket, intranasal administration, intraarticular injection, etc., may be used as an external ointment.

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

However, the following examples are illustrative of the present invention, and the content of the present invention is not limited by the examples.

Example 1 Preparation of a Gel Phase Formulation Containing a Drug

After dissolving phospholipids together with drugs in organic solvents such as ethanol, the ethanol is heated to about 40 ℃ in a nitrogen environment or evaporated under low pressure and blown out to make viscous gel by adding water to the mixture of phospholipids and drugs. Put into syringe. A hyaluronic acid aqueous solution was put in one syringe, and two syringes were connected with a three-way stop cork to prepare a uniform mixed liquid. Based on the composition of the present invention and carried out by a conventional lipid mixer method.

Example 2 Preparation of a Composition on a Gel Containing Hyaluronic Acid According to Water Content

Inject 10 g of egg phosphatidylcholine into the syringe with a phospholipid compound, add 1 mL of an aqueous solution of hyaluronic acid into the other syringe, connect the two syringes with a three-way stopper, and then rotate the piston of the syringe 100 times in both directions. Pushed over to prepare a uniform gel. The used egg phosphatidylcholine was purchased from Sigma, and used as an ingredient obtained from egg yolk containing about 60% of egg phosphatidylcholine. Inject the prepared gel (gel) into a syringe and put water of 100, 250, 500, 750, 1000 and 2000 weight ratios for the composition in another syringe and connect it with a three-way control stopper, and then mix the gel 100 times or more. Was prepared. For each formulation, the hardness and injectability of the gel according to the water content per 1 g of egg phosphatidylcholine and 1 mg of hyaluronic acid are shown in the table below.

Water (w / v) Hardness of gel Usable needle (gauge) 100 Very strong <24 250 Strong <26 500 Strong ≤26 750 Rather strong <28 1000 weakness All possible 2000 Very weak All possible

Compositions on injectable gels were readily prepared using egg phosphatidylcholine, hyaluronic acid salt, water, and the physical properties of the gel were best when the weight ratio of egg phosphatidylcholine, hyaluronic acid salt and water was 1000: 1: 250-750.

Example 3 Preparation of Gel Containing Tetracycline and Hyaluronic Acid

As in Comparative Example 1 below, 1 g of egg phosphatidylcholine was dissolved in 1 mL of ethanol with 75 mg of tetracycline as a phospholipid compound, and then ethanol was heated to about 40 ° C. in a nitrogen environment or evaporated under low pressure. This mixture was injected into a syringe. The egg phosphatidylcholine is the same as used in Example 2. 0.5 mL of an aqueous solution of hyaluronic acid was added to another syringe, and two syringes were connected with three-way adjusting stoppers, and the piston of the syringe was pushed 100 times in both directions to prepare a uniform gel.

Comparative Example 1 Preparation of Tetracycline-Containing Gel

1 g of egg phosphatidylcholine, a phospholipid, and 75 mg of tetracycline were completely dissolved in 1 mL of ethanol, and then depressurized to remove ethanol. The mixture obtained after depressurization was placed in a syringe, and 0.5 mL of water was injected into another syringe, and then connected with a three-way adjusting stopper. The piston of each syringe was pushed 100 or more times in both directions to prepare a composition on gel containing uniform tetracycline.

Example 4 Preparation of Gels Containing Diclofenac and Hyaluronic Acid

In the preparation method presented in Example 3, except that instead of tetracycline, diclofenac is added Gels were prepared in the same manner as in Example 3.

Example 5 Preparation of Gels Containing Bupivacaine and Hyaluronic Acid

In the preparation method presented in Example 3, instead of tetracycline, a gel was prepared in the same manner as in Example 3 except that the bulk bakane was added.

Experimental Example 1 Physical Stability Comparison of Tetracycline-Containing Gel in Aqueous Solution

In order to compare the physical stability on the aqueous solution of the gel containing no hyaluronic acid salt prepared in Comparative Example 1 and the gel of the present invention containing the hyaluronic acid salt prepared in Example 3 was carried out as follows.

Each prepared gel was injected into a 6-well plate by injecting 200 mg of each syringe into a 1 mL syringe without needles, and then 3 mL of water was added thereto, followed by vigorous stirring to observe the physical stability of the gel. It was.

Figure 1aAndFigure 2aof The formulation on the gel comprising hyaluronic acid prepared in Example 3 is the formulation on the gel without hyaluronic acid prepared in Comparative Example 1, while the release occurs only on the surface.1bAndFig 2bThe gel disappeared and most of it was dispersed in water.

Experimental Example 2 Biocompatibility Experiment

In order to determine the biocompatibility of the composition on the gel prepared in the above example, an animal model was prepared as described below and the biocompatibility was observed after administering the gel. i ) Animal Model of Air Pouch

20 g body weight (BALB / c mouse) was used, and the male was not distinguished. During the experimental period, all the animals were kept in a constant feed in the same environment. A syringe equipped with a 10 μm strainer (Milipore) is used to repeatedly inject 3 to 5 mL of air into the center of the mouse for 1 week at intervals of 2 to 3 days to artificially remove the air pouch. Generated.

ii) biocompatibility testing

Tissues were observed pathologically for 3 weeks after injection into the subcutaneous, air-pouch or pericardium of the animal model (BALB / c mouse). Very mild inflammation was observed for the first three days on all gels, but normal tissue was observed on the first week. Therefore, the formulation of the present invention is judged to be excellent in biocompatibility.

Experimental Example 3 Drug Release Experiment

In order to determine the stability after administering the drug in the gel form of the present invention, the drug release experiment was carried out as follows.

i) in biological in vitro ( in vitro) Drug release experiment

100 mg of the composition on the gel prepared in Comparative Example 1 and Example 3 were respectively placed in a dialysis membrane bag (MWCO = 3500, Spectra / Por ^R , Spectrum Medical Industries, Inc. Laguna Hills, CA. USA) and closed on both sides. ) And then immersed in 10 mL of phosphate buffered saline (pH = 7.4) to perform a release experiment in a vortex bath (37 ° C). Phosphate buffer solution outside the dialysis membrane bag was taken at regular intervals and the released drug was quantified by fluorescence. (λ _ex = 330 nm, λ _em = 450 nm). The release experiment was conducted using tetracycline aqueous solution as a control.

As a result, as shown in Figure 3 , the control group was released more than 80% of the drug within 10 hours, while the liquid formulation according to the present invention showed a sustained release form, the drug was released slowly over 120 hours.

ii) within living organisms (in vivo) Drug release experiment

500 mg of the formulation prepared in Example 3 in the synovial membrane cavity of the animal model prepared in Experimental Example 2 was carried out in vivo ( in vivo) release experiment for tetracycline. In order to confirm the presence of tetracycline in the tissue, the tissue was observed with a UV lamp in the dark, and then the yellow fluorescence of tetracycline was observed. As a result, yellow fluorescence was observed until day 6, but was not observed from day 7. Thus, the drug appeared to be sustained for about 6 days under conditions in vivo ( in vivo) .

The inventors have devised a method for preventing such erosion of the formulation and can actually reduce its speed. That is, biocompatible hyaluronic acid was added to the aqueous solution when preparing the hydrated phospholipid. Hyaluronic acid-added formulations were observed to erode only on the surface when added to water, whereas the formulations were easily broken when not added. It was also observed that injection can be easily performed even when hyaluronic acid was added. As such, the physical properties of the formulations were improved, but the drug encapsulation was not impeded at all. The formulations thus prepared were excellent in biocompatibility as a result of animal testing and continued to release the drug at topical sites for about a week.

As described above, since the formulation according to the present invention is a gel phase capable of releasing the drug at the topical site for 6 days, it can be used very effectively as an injectable topical drug delivery system. Such a property may be applied to the treatment of arthritis, and may be applied to pain treatment for delivering local anesthetics, periodontal disease treatment for delivering local antibiotics, fungal disease treatment for delivering antifungal agents, and trauma treatment.

Claims (10)

Phosphatidylcholine compounds that are saturated C ₆ to C ₂₂ ; One drug selected from the group consisting of tetracycline, diclofenac and bupivacaine; And a composition comprising water, wherein the composition on a gel comprises a biocompatible polymer selected from the group consisting of hyaluronic acid, chitosan and alginic acid The gel phase composition of claim 1, wherein the biocompatible polymer is included in an amount of 0.0001 to 3% by weight. delete The gel phase composition of claim 1, wherein the biocompatible polymer is hyaluronic acid. delete delete delete delete Dissolving one drug selected from the group consisting of saturated C ₆ to C ₂₂ phosphatidylcholine-based compounds and tetracycline, diclofenac and bupivacaine in alcohol, and then adding water to the residue obtained by removing the alcohol (step) One), Method for producing a biocompatible polymer aqueous solution selected from the group consisting of hyaluronic acid, chitosan and alginic acid to the mixture prepared in step 1 (step 2). delete