KR101118199B1 - Polymeric micelle composition for solubilizing tacrolimus - Google Patents

Abstract

The present invention relates to a polymer micelle composition for solubilizing tacrolimus, and more particularly, to an amphiphilic tacrolimus containing an amphiphilic block copolymer composed of a hydrophilic block and a hydrophobic block and a carboxylic acid end group. The present invention relates to a polymer micelle composition capable of solubilizing tacrolimus by forming a stable polymer micelle in an aqueous solution using a polylactic acid derivative or a mixture thereof.

Tacrolimus, amphiphilic block copolymers, polylactic acid derivatives, micelles, solubilization

Description

Polymer micelle composition for solubilizing tacrolimus

The present invention relates to a polymer micelle composition for solubilizing tacrolimus, a poorly soluble drug, and more particularly, an amphiphilic block copolymer composed of a hydrophilic block and a hydrophobic block in tacrolimus, a poorly soluble drug, and carboxylic acid. A polymer micelle composition capable of solubilizing tacrolimus by forming a stable polymer micelle in an aqueous solution using a polylactic acid derivative containing a terminal group or a mixture thereof.

Tacrolimus is a drug known as FK506. It is a macrolide lactone compound with a molecular weight of 882.05. It has immunosuppressive activity and is a prophylactic inhibitor for transplantation and atopic eczema in organ transplant patients. It was commercialized. However, tacrolimus has a very low solubility in water of 1 to 2 ㎍ / ml, so that the absorption rate is very low during oral administration, and the change between individuals is very severe.

Oral Prograf®, a commercially available formulation, is a capsule containing tacrolimus and contains excipients such as lactose, hydroxypropyl methylcellulose, croscarmellose sodium salt and magnesium stearate. Absolute bioavailability is about 17 ± 10% when Prograf® capsules are administered to kidney transplant patients. This relatively low bioavailability is due to the low solubility of tacrolimus in water.

Due to these problems, several attempts have been made to orally formulate tacrolimus. In particular, there have been several studies of high-solid composites. In particular, Yamashita et al. Have known a method for the preparation of a novel tacrolimus formulation and the results thereof on the method of not using dichloromethane as an organic solvent in a tacrolimus high-solid composite using HPMC. [International Journal of Pharmacetics 267 (2003) 79-91. The solid-solid complex may be used orally, but not for injection.

Korean Patent No. 177158 discloses tacrolimus-containing solution formulations, which compositions include pharmaceutically acceptable surfactants including tacrolimus, polyoxyethylene hydrogenated castor oil and ethanol, propylene glycol, glycerin and There is a known immunosuppressive solution formulation containing a pharmaceutically acceptable non-aqueous solvent selected from the group consisting of polyethylene glycol.

On the other hand, Prograf® for intravenous injection is a surfactant in which 5 mg of tacrolimus is dissolved in 1 ml of dehydrated ethanol 80.0% (v / v) containing 200 mg of HCO-60 (polyoxyl 60 hydrogenated castor oil). It is diluted in 0.9% aqueous sodium chloride solution or 5% dextrose aqueous solution and injected at a concentration of about 0.2 mg / ml. Surfactants of polyoxyl castor oils have the disadvantage of side effects such as hypersensitivity reactions when administered to humans, and must be diluted and used for 4 to 24 hours to be instilled and used in a PVC pipe used as an infusion set. There is a disadvantage that a plasticizer such as eluted. In addition, non-aqueous solvents such as ethanol, propylene glycol, glycerin and polyethylene glycol should be handled with caution as they cause hemolysis and local irritation upon injection.

Tacrolimus ointment (Protopic®) has been developed with a formulation containing 0.03% or 0.1% tacrolimus in a base consisting of mineral oil, paraffin, propylene carbonate, white petrolatum and white wax.

Therefore, while increasing the solubility of tacrolimus, there is little side effect on the human body, the situation is required to develop a formulation containing tacrolimic anhydride that is easy to apply to the human body.

On the other hand, the present applicant is a stable micelle formation in an aqueous solution containing a poorly soluble drug carrier using a polylactic acid derivative having a carboxylic acid terminal group, and an amphiphilic block copolymer and a polylactic acid derivative having a carboxylic acid terminal group to solubilize poorly soluble materials The polymer composition has been applied for a patent (Domestic Patent Application No. 2002-63955, 2002-75787). However, no mention is made of poorly soluble substances having a solubility in water of 1 to 2 µg / ml, such as tacrolimus, as the poorly soluble substance.

 Accordingly, the present inventors have studied to solubilize tacrolimus, and as a result, an amphiphilic block copolymer composed of a hydrophilic block and a hydrophobic block in tacrolimus, a poorly soluble drug, a polylactic acid derivative containing a carboxylic acid end group or The present invention has been accomplished by using the mixture thereof to establish an optimum solubilization composition with improved solubility of 0.2 mg / ml or more.

Accordingly, the present invention provides a polymer micelle composition comprising 0.1 to 10.0 wt% of an immunosuppressive agent, tacrolimus, and 90 to 99.9 wt% of an amphiphilic block copolymer, a polylactic acid derivative having a carboxy end group, or a mixture thereof. The purpose is to provide.

The present invention provides a polymer micelle composition comprising 0.1 to 10.0% by weight of tacrolimus, an immunosuppressive agent, and 90 to 99.9% by weight of an amphiphilic block copolymer, a polylactic acid derivative having a carboxy terminal group, or a mixture thereof. It features.

Hereinafter, the present invention will be described in detail.

The present invention forms a stable polymer micelle in an aqueous solution by using an amphiphilic block copolymer composed of a hydrophilic block and a hydrophobic block, a polylactic acid derivative containing a carboxylic acid end group, or a mixture thereof in tacrolimus, a poorly soluble drug. The present invention relates to a polymer micelle composition capable of solubilizing tacrolimus.

Referring to the composition of each of the polymer micelle composition according to the invention in detail as follows.

Tacrolimus, an effective drug of the present invention, is a poorly soluble substance having a solubility in water of 1 to 2 ㎍ / ml, and is mainly used for the cytokine gene which plays an important role in the immune response by acting at the initial stage of T cell activation. It is a drug that effectively suppresses immunity by inhibiting expression. The preferred content of tacrolimus for use as the polymer micelle composition of the present invention is 0.1 to 10% by weight, wherein when the tacrolimus is less than 0.1% by weight, the desired effect cannot be obtained. Gradually decreased so that the reconstitution liquid could not remain stable for a long time.

Amphiphilic block copolymer of the present invention is an AB-type double block copolymer composed of a hydrophilic block (A) and a hydrophobic block (B), in which the hydrophobic block forms a core and the hydrophilic block forms a shell. To form a polymer micelle of the form.

Hydrophilic block (A) is a water-soluble polymer, specifically, polyalkyleneglycol or derivatives thereof, polyvinylalcohol, polyvinylpyrolidone or polyacrylamide, etc. Preferably monomethoxypolyalkyleneglycol, monoacetoxypolyethyleneglycol, polyethyleneglycol, polyethylene-co-propyleneglycol and polyvinylpyrrolidone ( polyvinylpyrolidone). The number average molecular weight of the hydrophilic block is preferably 1000 to 10,000 Daltons, and more preferably 1,000 to 5,000 Daltons.

The hydrophobic block (B) is a polymer that is insoluble in water, has excellent biocompatibility, and is biodegradable, and specifically, polyester, polyanhydride, polyamino acid, and polyorthoester ( polyorthoester or polyphosphazine, and the like, preferably polylactide, polyglycolide, polycaprolactone, polydioxane-2-one, polydioxane-2-one, Polylactic-co-glycolide, polylactic-co-dioxane-2-one, polylactic-co-caprolactone ) And polyglycolic-co-caprolactone. In addition, the hydroxy terminal of the hydrophobic block (B) may be substituted with a fatty acid group, benzoate and the like, the fatty acid group may be a butyric acid group, propionic acid group, acetic acid group, stearic acid group, palmitic acid group and the like. It is preferable that the said hydrophobic block (B) is 500-5,000 Daltons in number average molecular weight.

The composition ratio of the hydrophilic block and the hydrophobic block of the amphiphilic block copolymer is preferably from 2 to 8: 8 to 2 (W / W), more preferably from 4 to 7: 6 to 3 (W / W). The ratio not only satisfies the solubility of the amphiphilic block copolymer in water but also sufficiently secures the hydrophobicity of the core so that the drug can be encapsulated well.

In addition, a polylactic acid derivative including a carboxy end group may be used instead of the amphiphilic block copolymer, or a mixture of an amphiphilic block copolymer and a polylactic acid derivative including a carboxy end group may be used.

Polylactic acid derivatives comprising carboxylic acid end groups include one or more carboxylic acid groups or carboxylic acid alkali metal salts at one end thereof, and the other end groups are hydroxy, acetoxy, benzoyloxy and decanoyloxy. (decanoyloxy), palmitoyloxy (palmitoyloxy) and alkoxy (alkoxy) consists of one selected from the group consisting of. The polylactic acid derivatives include D, L-polylactic acid, copolymers of D, L-lactic acid and glycolic acid, copolymers of D, L-lactic acid and caprolactone, copolymers of D, L-lactic acid and mandelic acid, and D Preference is given to those selected from the group consisting of copolymers of, L-lactic acid and 1,4-dioxan-2-one. The carboxylic acid group or carboxylic acid alkali metal salt acts as a hydrophilic group in an aqueous solution of pH 4 or more to form a polymer micelle in the aqueous solution. The carboxylic acid alkali metal salt is in the form of a metal ion salt which is a monovalent metal ion of sodium, potassium or lithium. The number average molecular weight of the polylactic acid derivative is preferably from 500 to 2,500 daltons. The molecular weight of the polylactic acid derivative may be achieved by appropriately adjusting the reaction temperature, time, and the like during preparation.

In the present invention, the preferred content of the amphiphilic block copolymer, a polylactic acid derivative including a carboxy end group or a mixture thereof is 90 to 99.9% by weight. In the case of the mixture of the amphiphilic block copolymer and the polylactic acid derivative, since both tacrolimus is soluble in both polymers, both polymers may be present in any composition ratio.

In this composition, tacrolimus is physically associated with the hydrophobic block portion of the polymer and is located in the hydrophobic core of the micelle formed by the polymer in an aqueous solution, and the particle size of the tacrolimus-containing polymer micelle is 10 to 200 nm. Range.

Tacrolimus in the composition of the present invention is soluble in water to a concentration of 0.2 to 10 mg / ml, it is preferable to dissolve up to 5 mg / ml in consideration of stability in an aqueous solution.

The method for obtaining the polymer micelle composition for solubilizing tacrolimus is as follows.

Tacrolimus and the polymer of the present invention are dissolved in a low boiling organic solvent, that is, dichloromethane, ethanol, methanol, acetone, acetonitrile, etc. Polymer micelles containing mousse are produced. Excipients such as polysaccharide, mannitol, sorbitol, lactose, trehalose, sucrose and the like can be added to the solution and dried by a method such as lyophilization or spray drying to obtain a tacrolimus-containing solid polymer micelle composition.

The solid polymer micelle composition may be formulated in oral dosage forms such as tablets, capsules, granules, powders, and solutions, or may be reconstituted and injected into water for injection, physiological saline, or 5% dextrose solution. Intravenous injection has no hemolysis and can be injected within a short time, so there are no side effects such as hypersensitivity reactions. It can also be applied to skin or eyes in the form of aqueous solutions, gels, and ointments.

The effective dose of tacrolimus varies depending on the age, disease, etc. of the patient, but can generally be administered at a dose of 0.001-100 mg, preferably 0.01-50 mg, more preferably 0.1-20 mg per day.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Preparation Example 1 Synthesis of D, L-Polylactic Acid (PLA-COOH)

 100 g of D, L-lactic acid was placed in a 250 ml three-necked round bottom flask, equipped with a stirrer, heated in an oil bath heated at 80 ° C. and reacted with an aspirator at 25 mmHg for 1 hour while being present in excess. Moisture was removed.

The reaction temperature was raised to 160 ° C., the reaction was carried out under reduced pressure to 10 mmHg for 12 hours, and the reaction was terminated. Then, 1 L of distilled water was added to the resulting reaction product to precipitate a polymer.

The precipitated polymer was added again to distilled water to remove the low molecular weight polymer dissolved in an aqueous solution of pH 4 or lower, and then the precipitated polymer was added to 1 L of distilled water again, and sodium hydrogen carbonate was added to confirm that the polymer was almost completely dissolved. A small amount was added to confirm that the pH of the aqueous solution was 6-8. At this time, the polymer insoluble in water was removed by centrifugation or filter separation.

A small amount of 1N hydrochloric acid solution was added thereto, and the aqueous solution was further adjusted to pH 2 to precipitate the polymer in the aqueous solution. The precipitated polymer was washed twice with distilled water, separated, and dried under reduced pressure to obtain a very viscous liquid. The number average molecular weight of the polymer was determined by ¹ H-NMR spectrum and was 1140 Daltons.

Preparation Example 2 Synthesis of Carboxylic Acid Salt of Polylactic Acid Derivative

The D, L-polylactic acid (number average molecular weight 1140 Daltons) of Preparation Example 1 was dissolved in acetone, placed in a round bottom flask, agitated and slowly stirred at room temperature, followed by slowly adding an aqueous sodium hydrogen carbonate solution (1N). Neutralized.

A small amount of acetone solution was taken and diluted in a large amount of distilled water to confirm that the pH was 7, and then anhydrous magnesium sulfate was added to remove excess water, followed by filtration. Acetone was evaporated with a solvent evaporator to obtain a white solid. This was again dissolved in anhydrous acetone and filtered to remove the material insoluble in anhydrous acetone and acetone was evaporated to obtain a white solid D, L-polylactic acid sodium salt (yield 96%). When the produced polymer was dissolved in water, the pH of the aqueous solution was 6.5 to 7.5.

Preparation Example 3 Monomethoxy Polyethylene Glycol-Polylactide (mPEG-PLA) Block Copolymerization (Type A-B)

5.0 g of monomethoxy polyethylene glycol (weight average molecular weight 2,000 Daltons) was placed in a two-necked 100 ml round bottom flask and heated to 130 ° C. for 3 to 4 hours under reduced pressure (1 mmHg) to remove moisture. The reaction flask was filled with dried nitrogen, and 0.1 wt% (10.13 mg, 25 mmol) of lactide was added to the reaction catalyst, Stationus Octoate (Sn (Oct) ₂ ), using a syringe, followed by stirring for 30 minutes. After depressurizing (1 mmHg) at 130 ° C. for 1 hour, the solvent (toluene) in which the catalyst was dissolved was removed. 10.13 g of purified lactide was added, followed by heating at 130 ° C. for 18 hours. The resulting polymer was dissolved in methylene chloride and added to diethyl ether to precipitate the polymer. The obtained polymer was dried in a vacuum oven for 48 hours.

The number average molecular weight of the said monomethoxy polyethyleneglycol polylactide (mPEG-PLA) was 2,000-1,700 daltons. In addition, it was found that the AB type using ¹ H-NMR.

Examples 1 to 3 and Comparative Example 1: Polymer micelle composition comprising tacrolimus and D, L-polylactic acid sodium salt

Next, tacrolimus and D, L-polylactic acid sodium salt were completely dissolved by adding 2 mL of methylene chloride, and the methylene chloride was removed using a round bottom flask, and purified water was added thereto. Thereafter, the mixture was filtered using a filter having a pore size of 200 nm, and then lyophilized to prepare a polymer micelle composition containing tacrolimus in powder form.

division Ingredient (%) Tacrolimus Polylactic acid sodium salt
(Number average molecular weight 1,140 daltons) Example 1 One 99 Example 2 5 95 Example 3 10 90 Comparative Example 1 15 85

Examples 4 to 6 and Comparative Example 2: Polymer micelle composition comprising tacrolimus and monomethoxy polyethylene glycol-polylactide (mPEG-PLA)

Next, tacrolimus and monomethoxypolyethyleneglycol-polylactide were completely dissolved by adding 2 mL of methylene chloride, and the methylene chloride was removed using a round bottom flask, and purified water was added thereto. After melting, the mixture was filtered using a filter having a pore size of 200 nm, and then lyophilized to prepare a polymer micelle composition containing tacrolimus in powder form.

division Ingredient (%) Tacrolimus mPEG-PLA
(Number average molecular weight 2000-1,700 Daltons) Example 4 One 99 Example 5 5 95 Example 6 10 90 Comparative Example 2 15 85

Examples 7 to 8 and Comparative Example 3: Polymer micelle composition comprising tacrolimus and D, L-PLA-COONa / mPEG-PLA

Next, tacrolimus, D, L-polylactic acid sodium salt and monomethoxypolyethyleneglycol-polylactide were completely dissolved by adding 2 mL of methylene chloride, and the methylene chloride was dissolved in a round bottom flask. After removing the solution, the resultant was completely dissolved by adding purified water, filtered using a filter having a pore size of 200 nm, and then lyophilized to prepare a polymer micelle composition containing tacrolimus in powder form.

division Ingredient (%) Tacrolimus Sodium polylactic acid salt (number average molecular weight 1,140 daltons) mPEG-PLA
(Number average molecular weight 2,000-1,700 Daltons) Comparative Example 3 20 40 40 Example 7 10 45 45 Example 8 5 48 47

Test Example 1: Solubilization Confirmation

100 mg of each of the lyophilized compositions in Examples 1 to 8 and Comparative Examples 1 to 3 were taken into a vial, dissolved in purified water, and dissolved in a total amount of 1 mL. The solution was filtered through a filter having a pore size of 200 nm, and the concentration of tacrolimus was measured by HPLC using the analysis conditions of Table 4 below. After storage for 24 hours at 25 ℃ to determine the concentration of the drug by measuring the concentration of the drug and the results are shown in Table 5 below.

Inclusion amount (mg) = concentration of tacrolimus (mg / ml) × 1 ml (total amount of solution)

division Condition Eluent 52% acetonitrile / 48% water column CN, inner diameter 4.6 mm, length 25 cm (SUPELCOSIL, USA) Detection wavelength 207 nm Flow rate 1.5 mL / min Temperature 50 ℃ Injection volume 10 μl Retention time 3.4 minutes

division Tacrolimus Concentration (mg / mL) Inclusion Rate (%) 0 hours 24 hours Example 1 One 100 100 Example 2 5 100 100 Example 3 10 100 100 Comparative Example 1 15 100 70 Example 4 One 100 100 Example 5 5 100 100 Example 6 10 100 100 Comparative Example 2 15 100 80 Comparative Example 3 20 100 75 Example 7 10 100 100 Example 8 5 100 100

As shown in Table 5, it is possible to effectively solubilize tacrolimus up to 10% by weight of D, L-polylactic acid sodium salt or monomethoxypolyethylene glycol-polylactide (mPEG-PLA) polymer At this time, the drug was confirmed that the solubility is maintained stable for 24 hours at 25 ℃ storage conditions.

Formulation Example 1 Tablet Preparation

Tablets containing the polymer micelle composition according to the present invention are prepared by the following method.

250 g of polymer micelle composition

Lactose 175.9 g

180 g potato starch

32 g of colloidal silicic acid

10% gelatin solution

Potato Starch 160 g

50 g of talc

5 g of magnesium stearate

The polymer micelle composition was mixed with lactose, potato starch and colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding potato starch, talc and magnesium stearate to it was tableted.

Formulation Example 2: Preparation of Injection

Injection solution containing the polymer micelle composition according to the present invention was prepared by the following method.

1 g of polymer micelle composition

0.6 g sodium chloride

0.1 g of ascorbic acid

Distilled water

The polymer micelle composition, sodium chloride and ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

Formulation Example 3: Preparation of Ointment

Ointment containing the polymer micelle composition according to the present invention was prepared by the following method.

4 g of polymer micelle composition

5 g of diglyceryl monostearate

5 g of behenyl alcohol

1 g of aluminum monostearate

8 g liquid paraffin

73 g of white petrolatum

3 g of diclofenac sodium

1 g ammonium chloride

The components were stirred uniformly at 90-95 ° C, then stirred and cooled to 35 ° C to prepare an ointment.

Formulation Example 4 Preparation of Eye Drops

The preparation method of eye drops containing the polymer micelle composition according to the present invention is as follows.

10 mg of polymer micelle composition

Carbopol 934 20 mg

Triethanolamine appropriate amount

2 mg of paraoxine benzoate

1 g of sterile purified water

Methyl paraoxine benzoate was added to sterile purified water, heated, dissolved, and cooled to dissolve the polymer composition. Carbopol 934 was added thereto, mixed with a high speed stirrer, dispersed, and left to stand to remove air. Triethanolamine was added dropwise thereto while being careful not to let air enter, thereby stirring the eyedrops.

Formulation Example 5 Preparation of Eye Drops

The preparation method of an eye drop containing the polymer micelle composition according to the present invention is as follows.

0.2 g of polymer micelle composition

0.1 g of benzalkonium chloride

5 g sodium chloride

6.2 g of boric acid

1 g Tyroxapol

Diluted hydrochloric acid

1000 ml of sterile purified water

Sodium chloride and boric acid were added to the polymer micelle composition in order to dissolve it, and benzalkonium chloride and thyroxapol dissolved in sterile purified water were added thereto and stirred. Dilute hydrochloric acid was added to adjust pH. Sterilization was performed using a 0.45 microfilter.

As described above, the polymer micelle composition according to the present invention can improve the solubility of tacrolimus in water by solubilizing tacrolimus in an optimal state, there is no hemolysis phenomenon in human administration, and within a short time May be administered.

Claims (9)

Tacrolimus from 0.1 to 10.0% by weight, Amphiphilic block copolymer comprising monomethoxypolyalkylene glycol as hydrophilic block and polylactide as hydrophobic block, polylactic acid derivative which is D, L-polylactic acid or an alkali metal salt thereof, or a mixture thereof 90 to 99.9% by weight Polymeric micelle composition, characterized in that it contains. The polymer micelle composition according to claim 1, wherein the hydrophilic block has a number average molecular weight of 1,000 to 10,000 Daltons, and the hydrophobic block has a number average molecular weight of 500 to 5,000 Daltons.  The polymer micelle composition according to claim 1, wherein the amphiphilic block copolymer has a composition ratio of the hydrophilic block and the hydrophobic block of 2 to 8: 8 to 2 (weight ratio). The polymer micelle composition according to claim 1, wherein the polylactic acid derivative has a number average molecular weight of 500 to 2,500 daltons. delete An injectable agent comprising the composition of any one of claims 1 to 4. An oral dosage form comprising the composition of any one of claims 1 to 4. A topical skin preparation comprising the composition of any one of claims 1 to 4. An eye drop comprising the composition of any one of claims 1 to 4.