KR20060110915A - Method for preparing solid dispersion of sparingly water-soluble drug by using the supercritical antisolvent process - Google Patents

Abstract

A method for preparing solid dispersion of sparingly water-soluble drug by using the supercritical fluid process is provided to enhance homogenicity of the solid dispersion and improve solubility of sparingly water-soluble drug sufficient to be injected intravenously without non-aqueous solvents. The method for preparing solid dispersion of sparingly water-soluble drug comprises the steps of: (1) dissolving sparingly water-soluble drug, cyclodextrin derivatives and pharmaceutically acceptable additives in organic solvent to prepare the mixed solution; (2) spraying the mixed solution to the supercritical fluid to produce particles containing sparingly water-soluble drug, cyclodextrin derivatives and pharmaceutically acceptable additives; and (3) introducing the additional supercritical fluid into the particles to remove the organic solvent, wherein the sparingly water-soluble drug is paclitaxel, tacrolimus, cyclosporine, rapamycin or itraconazole; and the amount of cyclodextrin derivatives is 0.1 to 100 parts by weight based on 1 part by weight of sparingly water-soluble drug.

Description

METHOD FOR PREPARING SOLID DISPERSION OF SPARINGLY WATER-SOLUBLE DRUG BY USING THE SUPERCRITICAL ANTISOLVENT PROCESS}

1 is a schematic diagram showing the configuration of a supercritical fluid apparatus used in the present invention,

Figure 2 shows the differential scanning calorie curve of the paclitaxel powder used as a raw material,

Figure 3 shows the differential scanning calorie curve of the paclitaxel formulation (Example 1) according to the present invention,

Figure 4 shows the differential scanning calorie curve of the tacrolimus raw material used as a raw material,

Figure 5 shows the differential scanning calorie curve of the tacrolimus formulation (Example 16) according to the present invention,

Figure 6 shows the X-ray powder diffraction analysis of the paclitaxel raw material used as a raw material,

Figure 7 shows the X-ray powder diffraction analysis of the paclitaxel formulation (Example 1) according to the present invention,

8 shows the pharmacokinetic test results of the tacrolimus formulation (Example 13) and the control formulation Progrof (Progrof ^R , Fujisawa Pharmaceutical Co., LTD, Japan) according to the present invention,

9a shows the saturation solubility of paclitaxel preparations according to the present invention (Examples 1, 3, 6, 8, 13 and 15), paclitaxel preparations according to Comparative Examples 1 and 2, and paclitaxel original,

Figure 9b shows the saturation solubility of tacrolimus formulations according to the present invention (Examples 16, 18, 22 and 25), taclolimus formulations according to Comparative Example 3 and tacrolimus original.

The present invention relates to a method for preparing a solid dispersion of poorly soluble drug having improved solubility using a supercritical fluid process.

Paclitaxel (paclitaxel) is known to have an excellent effect on terminal cancer patients, such as terminal non-small cell lung cancer, ovarian cancer, breast cancer, but because it is very poorly soluble in water and low bioabsorption, it is a surfactant as ethanol and castor oil derivatives It has been prepared as an injection in liquid form with the Cremophor ^R EL. However, because Cremophor ^R EL can cause serious side effects, such as difficulty hoping, hypotension, angioedema, and uticaria, injections using it can only be used for long periods of drip injections and also prevent hypersensitivity. There are many inconveniences and risks, such as requiring hospitalization of patients for prior drug therapy.

Accordingly, US Pat. No. 5,681,846 discloses formulations that reduce the concentration of paclitaxel and castor oil derivatives and increase the content of alcohol. However, as in the patent, intravenous administration of high concentrations of alcohol causes side effects of the central nervous system. In addition, WO 00/01366 and WO 01/70220 mention liposome preparations of paclitaxel, but these preparations also have limitations in terms of manufacturing process difficulties, stability problems, and low drug content. . International patent application WO 98/30205 discloses an oil-in-water emulsion system using tocopherol as carrier oil, and US Pat. No. 5,407,683 contains paclitaxel dissolved in squalene as a solution in the absence of a surfactant. , A composition prepared by adding water soluble sucrose solution and then evaporating water. However, emulsions typically have a limited shelf life, are difficult to sterilize by heat or filters, and are problematic in terms of long-term stability due to their limitations as solutions.

Accordingly, the need for paclitaxel injection formulations, which have been secured by solving these problems, facilitates the manufacturing process and improves the solubility of paclitaxel so as not to use a large amount of castor oil derivatives, thereby increasing the safety.

In addition, tacrolimus is a macrolide immunosuppressive agent produced by fermentation of Streptomyces tsukubaensis . It is widely used for transplant rejection and graft-host disease. Such tacrolimus is in the form of white crystalline or crystalline powder, and is soluble in anhydrous ethanol, acetonitrile, dimethylformamide, ethanol, acetone and ether, but hardly soluble in water.

Due to this characteristic of tacrolimus, the currently available formulation of Progrof ^R (Fujisawa Pharmaceutical Co., LTD, Japan) injection contains 1 mg of tacrolimus per 1 ml of ampoule, 200 mg of derivative surfactant of castor oil (HCO-60), and U.S. Patent No. 5,260,301 discloses a tacrolimus containing solution formulation containing tacrolimus, a surfactant and a non-aqueous solvent, and European Patent No. 0483842 B1 discloses tacrolimus, an emulsifier, A solution formulation which improves the stability of tacrolimus by mixing an oil phase component and a solubilizer is disclosed.

However, these conventional tacrolimus formulations use non-aqueous solvents as solubilizers, and there is a risk of causing additional toxicity due to hemolysis and non-aqueous solvents when administered into human blood vessels.

Supercritical fluids, on the other hand, are incompressible fluids at temperatures and pressures above the critical point, and are characterized by unique characteristics not found in conventional organic solvents: high density close to liquid, low viscosity near gas and high diffusion coefficient, and very low surface tension. At the same time has excellent properties. In addition, supercritical fluids can continuously change their density from lean to near ideal gas to high density close to liquid density, so that the fluid's equilibrium (solubility), transport properties (viscosity, diffusion coefficient, thermal conductivity), molecules The clustering state can be adjusted.

Therefore, by using the ease of controlling the physical properties of the supercritical fluid, it is possible to obtain solvent properties comparable to various liquid solvents with one solvent. In particular, supercritical carbon dioxide has a low critical temperature of 31.1 ° C, which makes it suitable for thermally denatured substances such as drugs.It is non-toxic, non-flammable, very inexpensive, and can be recovered and reused to design several environmentally friendly processes. It has many advantages, such as the ability to do so, making it ideal for use in pharmaceuticals.

Using these unique properties of supercritical fluids, much research has recently been conducted in the fields of selective extraction of specific substances and analysis of substances through extraction. Also, supercritical fluids are used as solvents or anti-solvents. There is an active research on how to obtain recrystallized or fine particles by using).

Accordingly, the present inventors have developed a method for preparing a new formulation having improved solubility of poorly soluble drugs in a supercritical fluid process in order to solve poorly soluble drugs having poor water solubility of up to 10 μg / ml at room temperature. The composition of the poorly water-soluble drug prepared from the homogeneous to the ultra-fine level through a change in the temperature and pressure of the operation parameters of the supercritical fluid process or the composition of the additive used together to confirm that the solubility of the drug is improved Was completed.

It is an object of the present invention to provide a method for preparing a solid dispersion of poorly soluble drug having high water solubility, low toxicity and highly homogenized to an ultrafine level.

In order to achieve the above object, the present invention

1) dissolving a poorly soluble drug, a cyclodextrin derivative and a pharmaceutically acceptable additive in an organic solvent to prepare a mixed solution thereof;

2) spraying the mixed solution into a supercritical fluid to make contact with each other to produce particles of a mixture of poorly soluble drugs, cyclodextrin derivatives, and additives; And

3) It provides a method for producing a solid dispersion of poorly soluble drugs, comprising the step of removing the organic solvent used in the mixed solution by introducing a separate supercritical fluid.

The present invention also provides a formulation for intravenous administration comprising a solid dispersion of a poorly soluble drug obtained by the above method.

The basic principle of the production method is to dissolve poorly soluble drugs, cyclodextrin derivatives and pharmaceutically acceptable additives in an appropriate ratio and to dissolve in a suitable amount of an organic solvent, and sprayed through a nozzle to the reaction port equilibrated with a supercritical fluid After obtaining the particles by flowing a supercritical fluid several times to extract the organic solvent, and then remove the supercritical fluid to prepare a solid dispersion of poorly soluble drug homogeneous to the ultra-fine level.

Hereinafter, the solid dispersion of the poorly soluble drug according to the present invention will be described in detail for each step.

(Step 1) Preparation of a mixed solution of poorly soluble drugs, cyclodextrin derivatives and pharmaceutically acceptable additives

The poorly soluble drug is dissolved in the organic solvent together with the cyclodextrin derivative and the pharmaceutically acceptable additive.

In the present invention, the poorly soluble drug means a drug having a water solubility of up to 10 μg / ml or less at room temperature, and examples thereof include, but are not limited to, paclitaxel, tacrolimus, cyclosporin, rapamycin, and itraconazole.

Cyclodextrin derivatives used in the present invention are to improve the water solubility and stability of poorly soluble drugs, and include substituted α-, β- or γ-cyclodextrins having the structural formula of Formula 1.

Representative examples of the cyclodextrin derivatives include 2-hydroxypropyl-β-cyclodextrin, 2-hydroxyethyl-β-cyclodextrin, 2,6-dimethyl-β-cyclodextrin, sulfobutyl ether-7-β- Cyclodextrin, (2-carboxymethoxy) propyl-β-cyclodextrin, 2-hydroxyethyl-γ-cyclodextrin and 2-hydroxypropyl-γ-cyclodextrin, among which 2-hydroxypropyl- β-cyclodextrin is preferred. In the present invention, the cyclodextrin derivative is used in an amount of 0.1 to 100 parts by weight, preferably 1 to 50 parts by weight, based on 1 part by weight of the poorly soluble drug.

Pharmaceutically acceptable additives are added to increase the crystallinity and solubility of poorly soluble drugs, and hydrophilic polymers, surfactants and oil-soluble components and the like are used.

Examples of hydrophilic polymers usable in the present invention include polyvinylpyrrolidone (PVP), hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), hydroxyethylcellulose (HEC) and polyethylene glycol (PEG). ) And polyvinylpyrrolidone (PVP) is preferably used. In addition, the type and content of the hydrophilic polymer may be appropriately adjusted in consideration of the viscosity of the spray solution and the water solubility of the polymer itself, and preferably 0.1 to 100 parts by weight, more preferably 1 to 1 part by weight of the poorly soluble drug. To 50 parts by weight.

Representative examples of surfactants usable in the present invention include reaction products of natural or hydrogenated vegetable oils with ethylene glycol, such as polyoxyethylene glycolized natural or hydrogenated castor oil (trade name: Cremophor ^R (BASF) or HCO). ^R (Nikkol); Polyoxyethylene-sorbitan-fatty acid esters such as mono, trilauryl, palmityl, stearyl, oleyl esters (trade name: Tween ^R (ICI)); Polyoxyethylene stearic acid esters such as polyoxyethylene stearic acid ester (trade name: Myrj ^R (Uniqema)); polyoxyethylene-polyoxypropylene block copolymers (trade name: Poloxamer, Pluronic ^R (BASF) or Lutrol ^R ) (BASF)); Sodium dioctylsulfosuccinate or sodium lauryl sulfate; Phospholipids; Propylene glycol mono-fatty acid esters or propylene glycol di-fatty acid esters, for example propylene glycol dicaprylate, propylene glycol monocaprylate, propylene glycol dilaurate, propylene glycol Lycol isostearate, propylene glycol monolaurate and propylene glycol ricinorate, preferably propylene glycol monolaurate; Trans-esterification reaction products of natural vegetable oil triglycerides and polyalkylene polyols (trade name: Labrafil ^R M (Gattefosse)); Mono-, di- or mono / di-glycerides such as caprylic / capric acid mono- and di-glycerides (trade name: Imwitor ^R (Huls)); Sorbitan fatty acid esters such as sorbitan monolauryl, sorbitan monopalmityl and sorbitan monostearyl (trade name: Span ^R (Sigma)); Fatty acid esters of ascorbic acid, such as ascorbyl palmitate; And polyethylene glycol 15 hydroxystearate (trade name: Solutol ^R HS-15 (BASF)).

In the present invention, the surfactant is used in 1 to 100 parts by weight, preferably 5 to 80 parts by weight based on 1 part by weight of poorly soluble drug.

In the present invention, in addition to the hydrophilic polymer and the surfactant, a fat or oil component may be additionally used. The fat or oil component may be any pharmaceutically acceptable oil that is well mixed with the surfactant and can be emulsified in water stably. Representative examples of oil-soluble components usable in the present invention include fatty acid triglycerides, preferably coconut oil sold under the name Miglyol ^R as a middle fatty acid triglyceride; As ester compounds of fatty acids and monovalent alkanols, ester compounds of fatty acids having 8 to 20 carbon atoms and monovalent alkanols having 2 to 3 carbon atoms, for example isopropyl myristate, isopropyl palmitate, ethyl linoleate, ethyl oleate and the like ; Natural vegetable or animal oils such as corn oil, olive oil, soybean oil, fish oil and the like; Hydrocarbons such as squalene, squalane and the like; Tocopherols such as tocopherol, tocopherol acetate, tocopherol succinate, polyethylene glycol-1000-tocopherol succinate (TPGS), and the like.

In the present invention, the oleaginous component is used in an amount of 0 to 10 parts by weight, preferably 0 to 5 parts by weight, based on 1 part by weight of poorly soluble drug.

The organic solvent for preparing the mixed solution is an organic solvent having 1 to 6 carbon atoms that can dissolve homogeneously dissolving poorly soluble drugs, cyclodextrin derivatives and additives, the first organic that can dissolve poorly soluble drugs and cyclodextrin derivatives A mixed solvent of a second organic solvent capable of dissolving the solvent and the additive is used. For example, poorly soluble drugs and cyclodextrin derivatives are well soluble in ethanol, hydrophilic polymers are well soluble in acetone, and ethanol and acetone are well mixed with each other. It is possible to do Such a cosolvent system is advantageous in that the rate of generation of the solid dispersion particles can be controlled by the difference in the solubility of the supercritical carbon dioxide in each solvent in the supercritical fluid process of the supercritical carbon dioxide. First, a poorly soluble drug and a cyclodextrin derivative are dissolved in ethanol, and then acetone is mixed and other additives are completely dissolved therein to prepare a homogeneous solution. At this time, ethanol and acetone may be used by mixing in a weight ratio of 9: 1 to 5: 5, it is preferable to mix in a weight ratio of 7: 3. Methanol and isopropanol may be used instead of ethanol as the first organic solvent, and dichloromethane, chloroform, carbon tetrachloride, ethyl acetate, N, N-dimethylformamide, dimethyl sulfoxide instead of acetone as the second organic solvent. Said, tetrahydrofuran and the like can be used.

(Step 2) Generation of Particles Using a Supercritical Fluid Process

Supercritical fluids usable in the present invention include supercritical carbon dioxide (CO ₂ ), supercritical dinitrogen monoxide (N ₂ O), supercritical methane trifluoride, supercritical propane, supercritical ethylene, supercritical xenon, and the like. have.

According to one preferred embodiment of the present invention, the supercritical fluid process is performed using supercritical carbon dioxide having a critical point of 31.1 ° C. and 73.8 bar or more as the apparatus and supercritical fluid as shown in FIG. 1. First, carbon dioxide is injected into a reaction vessel made of stainless steel, pressurized and warmed, and waits until the carbon dioxide in the reaction vessel is in an equilibrium state by maintaining a supercritical state. Preferably, the temperature in the reaction vessel is 31 to 70 ° C and Maintain the pressure at 80-200 bar. At this time, it is preferable to use a injection pump (syringe pump) to maintain a constant pressure when pressurizing the carbon dioxide and to know the correct injection amount, and to use a circulating thermostat or a thermostat to maintain a constant temperature. When the reaction port is equilibrated to the supercritical state, a nozzle of the mixed solution of the poorly soluble drug, the cyclodextrin derivative, and the additive prepared in step 1) is injected into the reaction port using a small liquid pump capable of precise speed control. Via a constant rate, for example at a rate of about 0.1-1.0 ml / min if the volume of the reaction sphere is 92.4 cm 3. At this time, in order to prevent clogging of the nozzle, it is preferable to inject a small amount of co-solvent used for preparing the mixed solution before injection of the mixed solution, for example, about 3 to 4 ml. The higher the quality, the longer the cleaning time by the subsequent supercritical fluid.

In the above process, the organic solvent in the mixed solution sprayed through the nozzle is rapidly mixed into the supercritical carbon dioxide, the poorly soluble drug is precipitated together with the cyclodextrin and additives to produce particles mixed with each other at the molecular level.

In the above, a separate supercritical fluid may be injected to prevent the supercritical fluid in the reaction port from being saturated with the injection of the mixed solution.

(Step 3) Removal of the organic solvent using the supercritical fluid

Once the particles are produced, a particle washing process is required to introduce a supercritical fluid to remove the organic solvent in the particles. In the above process, the supercritical fluid is injected into the reaction port at a constant rate, for example, at a rate of about 5 to 15 ml / min when the volume of the reaction port is 92.4 cm 3, and the same rate as the injection rate is maintained to maintain the reaction port state at a constant pressure. Discharge through the outlet at speed. At this time, the back pressure regulator is connected to the outlet to maintain the constant pressure in the reaction port by adjusting the discharge rate. The outlet uses a membrane filter with a pore size of 0.22 μm to prevent particles from escaping.

If the organic solvent remains, when the temperature and pressure are lowered to collect the particles, the solvent is reprecipitated to re-dissolve the formed particles to form agglomeration, so the washing process should be performed sufficiently until all the solvent is removed. do. The amount of supercritical fluid for washing depends on the amount of solvent used and the size of the reaction port, preferably about 150 to 300 ml when the volume of the reaction port is 92.4 cm 3.

After the washing of the particles, the supercritical fluid supply to the reaction port is stopped and the supercritical fluid is discharged. At this time, if the discharge is made too fast, the produced particles may be damaged, so it is preferable to discharge slowly. After removing all of the supercritical fluid in the reactor, the particles are recovered from the base wall or bottom of the reactor.

As described above, according to the method for preparing a solid dispersion of the poorly soluble drug using the supercritical fluid process of the present invention, the supernatant solution is dissolved after dissolving the polymer and the drug in a general organic solvent that can be completely mixed with the supercritical fluid. When expanded through the nozzle in the critical fluid, the organic solvent is reduced in the solubility of the polymer and drug to be mixed with the supercritical fluid and the polymer and drug supersaturated and precipitated to form recrystallized particles. Here, the extraction speed of the organic solvent can be freely controlled by changing the type of the sprayed solution, the temperature and the pressure condition of the supercritical fluid, and thus the physical properties can be changed, resulting in the ultra-fine solid dispersion of the ultra fine level. The manufacturing and water solubility can be increased.

The solid dispersion of the poorly soluble drug prepared as described above can be usefully used for injectable preparations of poorly soluble drugs because the water solubility is improved so that it does not contain castor oil derivatives or is easily dissolved without recrystallization. It does not contain a non-aqueous solvent and has high solubility, so that the side effect of hemolysis such as hemolysis of the non-aqueous solvent at a higher concentration than the dilution concentration required for the administration of a poorly soluble drug during actual clinical administration, The same recrystallization is not produced, which reduces the administration time and improves patient compliance.

Hereinafter, the present invention will be described in more detail by the following examples. However, the following Examples are only for illustrating the present invention, and the scope of the present invention is not limited to these.

Example 1 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 1

To prepare a solid dispersion of paclitaxel for injection using a supercritical fluid process, a mixed solution of paclitaxel, a cyclodextrin derivative, and an additive for spraying was prepared in the composition and content as shown in Table 1 below.

First, Paclitaxel (Hanmi Pharm. Co., Ltd.) and 2-hydroxypropyl-β-cyclodextrin (Aldrich) were dissolved in ethanol, and then acetone was mixed and stirred, and then pladon (Plasdone) as a polyvinylpyrrolidone as a hydrophilic polymer was stirred. ^A homogeneous mixed solution was prepared by adding ^R C-30 (ISP) and Cremophor ^R ELP (BASF), Cremophor ^R RH40 (BASF), and Ascorbyl Palmitate (ROCHE). The internal temperature of the reactor with an internal diameter of 2.9 cm, height of 14 cm, and volume of 92.4 cm 3 was 35 ° C., and carbon dioxide was added with an injection pump (Isco ^R , model 260D) to bring the pressure inside the reaction port to 83 bar. After warming and pressing, it was maintained until equilibrium.

When the inside of the reaction port is in equilibrium, the mixed solution prepared above is injected into the reaction port using a small liquid pump (NP-AX-15, NIHON SEIMITSU KAGAKU CO., LTD.). In order to prevent, the mixed solvent used in the preparation of the mixed solution was sprayed into the reaction port by about 4 ml, and then the mixed solution prepared in advance was injected into the reaction port at a rate of 0.3 ml / minute. The mixed solution was injected into the reaction port and sprayed through a capillary nozzle in the reaction port, thereby producing a highly homogeneous solid dispersion in a supercritical state. In order to prevent supercritical carbon dioxide in the reaction port from being saturated with the mixed solution injection, the carbon dioxide in the reaction port was discharged through the exhaust port at the same rate while injecting new carbon dioxide into the reaction port at a rate of 10 ml / min.

After injecting the solution, use a back pressure regulator (TESCOM ^R , model 26-1723-24-194) while injecting new carbon dioxide into the reactor at a rate of 7 ml / min to remove the solvent dissolved in the supercritical carbon dioxide. At a rate the carbon dioxide in the reaction port was vented through the vent. The washing was carried out using 200 ml of carbon dioxide.

After the washing process, all of the carbon dioxide in the reaction port was discharged, and the solid dispersion of paclitaxel generated from the base wall and the bottom of the reaction port was recovered.

Example 2 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 2

Using the drug solution composition and supercritical conditions shown in Table 2 below, the solid dispersion of paclitaxel was prepared in the same manner as in Example 1.

Example 3 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 3

Using the drug solution composition and supercritical conditions shown in Table 3 below, the solid dispersion of paclitaxel was prepared in the same manner as in Example 1.

Example 4 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 4

Using the drug solution composition and supercritical conditions shown in Table 4 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 5 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 5

Using the drug solution composition and supercritical conditions as shown in Table 5 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 6 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 6

Using the drug solution composition and supercritical conditions shown in Table 6 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 7 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 7

To prepare a solid dispersion of paclitaxel in the same manner as in Example 1 using the drug solution composition and supercritical conditions as shown in Table 7 .

Example 8 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 8

Using the drug solution composition and supercritical conditions as shown in Table 8 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 9 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 9

Using the drug solution composition and supercritical conditions as shown in Table 9 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 10 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 10

Using the drug solution composition and supercritical conditions shown in Table 10 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 11 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 11

Using the drug solution composition and supercritical conditions as shown in Table 11 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 12 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 12

Using the drug solution composition and supercritical conditions shown in Table 12 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 13: Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 13

Using the drug solution composition and supercritical conditions shown in Table 13 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 14 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 14

Using the drug solution composition and supercritical conditions as shown in Table 14 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Example 15 Preparation of Paclitaxel Solid Dispersion by Supercritical Fluid Process 15

Using the drug solution composition and supercritical conditions as shown in Table 15 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Comparative Example 1 Preparation of Cyclodextrin-Free Paclitaxel Solid Dispersion by Supercritical Fluid Process

Using the drug solution composition and supercritical conditions shown in Table 16 , to prepare a solid dispersion of paclitaxel in the same manner as in Example 1.

Comparative Example 2: Preparation of Paclitaxel Solid Dispersion by Spray Drying Process

To prepare a solid dispersion of paclitaxel using a spray drying process, a solid dispersion of paclitaxel was prepared by a spray drying process with a drug solution composition as shown in Table 1 above.

First, Paclitaxel (Hanmi Pharm. Co., Ltd.) and 2-hydroxypropyl-β-cyclodextrin (Aldrich) were dissolved in ethanol, and then acetone was mixed and stirred, and then pladon (Plasdone) as a polyvinylpyrrolidone as a hydrophilic polymer was stirred. ^A homogeneous mixed solution was prepared by adding ^R C-30 (ISP) and Cremophor ^R ELP (BASF), Cremophor ^R RH40 (BASF), and Ascorbyl Palmitate (ROCHE). The mixed solution prepared above was spray dried using a spray dryer (Mini spray dryer B-191, BUCHI) to prepare a paclitaxel solid dispersion. At this time, spray drying conditions made the inlet temperature of a dryer 60 degreeC, and the outlet temperature 45-50 degreeC.

Example 16 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 1

In order to prepare a solid dispersion of tacrolimus for injection using a supercritical fluid process, a mixed solution of tacrolimus, a cyclodextrin derivative and an additive for spraying was prepared in the composition and content as shown in Table 17 below.

First, Tacrolimus (Concord Biotech Ltd.) and 2-hydroxypropyl-β-cyclodextrin (Aldrich) were dissolved in ethanol, and then acetone was mixed and stirred, and then Plasdone ^R as a hydrophilic polymer polyvinylpyrrolidone was stirred. Add C-30 (ISP) and surfactant Cremophor ^R ELP (BASF), Cremophor ^R RH40 (BASF) and Ascorbyl Palmitate (ROCHE), and d-α-tocopherol as oil-soluble component To prepare a homogeneous mixed solution. The internal temperature of the reactor with an internal diameter of 2.9 cm, height of 14 cm, and volume of 92.4 cm 3 was 35 ° C., and carbon dioxide was added with an injection pump (Isco ^R , model 260D) to bring the pressure inside the reaction port to 83 bar. After warming and pressing, it was maintained until equilibrium.

When the inside of the reaction port is in equilibrium, the mixed solution prepared above is injected into the reaction port using a small liquid pump (NP-AX-15, NIHON SEIMITSU KAGAKU CO., LTD.). In order to prevent, the mixed solvent used in the preparation of the mixed solution was sprayed into the reaction port by about 4 ml, and then the mixed solution prepared in advance was injected into the reaction port at a rate of 0.3 ml / minute. The mixed solution was injected into the reaction port and sprayed through a capillary nozzle in the reaction port, thereby producing a highly homogeneous solid dispersion in a supercritical state. In order to prevent supercritical carbon dioxide in the reaction port from being saturated with the mixed solution injection, the carbon dioxide in the reaction port was discharged through the exhaust port at the same rate while injecting new carbon dioxide into the reaction port at a rate of 10 ml / min.

After injecting the solution, use a back pressure regulator (TESCOM ^R , model 26-1723-24-194) while injecting new carbon dioxide into the reactor at a rate of 7 ml / min to remove the solvent dissolved in the supercritical carbon dioxide. At a rate the carbon dioxide in the reaction port was vented through the vent. The washing was carried out using 200 ml of carbon dioxide.

After the washing process, all of the carbon dioxide in the reaction port was discharged, and the solid dispersion of paclitaxel generated from the base wall and the bottom of the reaction port was recovered.

Example 17 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 2

Using the drug solution composition and supercritical conditions as shown in Table 18 , to prepare a solid dispersion of tacrolimus in the same manner as in Example 16.

Example 18 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 3

To prepare a solid dispersion of tacrolimus in the same manner as in Example 16, using the drug solution composition and supercritical conditions as shown in Table 19 .

Example 19 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 4

Using the drug solution composition and supercritical conditions as shown in Table 20 , to prepare a solid dispersion of tacrolimus in the same manner as in Example 16.

Example 20 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 5

To prepare a solid dispersion of tacrolimus in the same manner as in Example 16, using the drug solution composition and supercritical conditions as shown in Table 21 .

Example 21 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 6

Using the drug solution composition and supercritical conditions as shown in Table 22 , a solid dispersion of tacrolimus was prepared in the same manner as in Example 16.

Example 22 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 7

Using the drug solution composition and supercritical conditions as shown in Table 23 , to prepare a solid dispersion of tacrolimus in the same manner as in Example 16.

Example 23 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 8

To prepare a solid dispersion of tacrolimus in the same manner as in Example 16, using the drug solution composition and supercritical conditions shown in Table 24 below.

Example 24 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 9

To prepare a solid dispersion of tacrolimus in the same manner as in Example 16, using the drug solution composition and supercritical conditions shown in Table 25 below.

Example 25 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 10

To prepare a solid dispersion of tacrolimus in the same manner as in Example 16, using the drug solution composition and supercritical conditions as shown in Table 26 .

Example 26 Preparation of Tacrolimus Solid Dispersion by Supercritical Fluid Process 11

Using the drug solution composition and supercritical conditions as shown in Table 27 , a solid dispersion of tacrolimus was prepared in the same manner as in Example 16.

Comparative Example 3: Preparation of cyclodextrin-free tacrolimus solid dispersion by supercritical fluid process

To prepare a solid dispersion of tacrolimus in the same manner as in Example 16, using the drug solution composition and supercritical conditions as shown in Table 28 .

Test Example 1 Measurement of Saturation Solubility of Paclitaxel Solid Dispersion

In order to evaluate the saturation solubility of paclitaxel solid dispersion prepared using a supercritical fluid process according to the present invention, the solid dispersion prepared by the above examples, the solid dispersion prepared by Comparative Examples 1 and 2 and The following experiments were carried out using the raw materials of general paclitaxel.

Excess sample (approximately 8 mg as paclitaxel) and 2 ml of purified water were placed in a scintillation vial and mixed for 30 minutes at 10 × 100 rpm using a cute mixer (cute mixer CM-1000, EYELA) at room temperature. After leaving at room temperature for about 24 hours until the supersaturation state, it was filtered through a 0.45 ㎛ filter and the amount of paclitaxel dissolved by HPLC was measured. At this time, HPLC is Hitachi L-7100 by pump, Hitachi UV detector L-7400 by detector, Rheodyne 7725i by injector, Inertsil ^R ODS2 (C18) by column (5 μm, 4.6 mm × 15 cm, GL Science) was used and the amount dissolved at UV 228 nm wavelength was measured. A 50% acetonitrile aqueous solution was used as the mobile phase, the flow rate was 1.0 ml / min, and the injection volume was 20 µl.

The experimental results are shown in Figure 9a. From FIG. 9A, the case where paclitaxel solid dispersion obtained using cyclodextrin using the supercritical fluid process according to the present invention does not use cyclodextrin (Comparative Example 1), when a solid dispersion is used (Comparative Example 2), and It can be seen that the saturated solubility is significantly higher than that of the paclitaxel raw material itself.

Test Example 2 Measurement of Saturation Solubility of Tacrolimus Solid Dispersion

In order to evaluate the saturation solubility of the tacrolimus solid dispersion prepared using a supercritical fluid process according to the present invention, the solid dispersion prepared by the above examples, the solid dispersion prepared by Comparative Example 3, general tacrolimus Using the raw material of the following experiment was carried out.

Excess sample (about 8 mg as tacrolimus) and 2 ml of purified water were placed in a scintillation vial and mixed for 30 minutes at 10 × 100 rpm using a cute mixer (cute mixer CM-1000, EYELA) at room temperature. After leaving at room temperature for about 24 hours to escape the supersaturation state, it was filtered through a 0.45 ㎛ filter and the amount of tacrolimus dissolved by HPLC was measured. At this time, HPLC was performed by Hitachi L-7100 by pump, Hitachi UV detector L-7400 by detector, Rheodyne 7725i by injector, TSK gel ^R ODS-80TM by column (5 μm, 4.6 mm × 15 cm, TOSOH Co.) was used, and the amount dissolved at UV 220 nm wavelength was measured. In addition, an aqueous solution of distilled water: isopropyl alcohol: tetrahydrofuran (5: 2: 2) was used as the mobile phase, the flow rate was 0.4 ml / min, the column temperature was 50 ° C., and the injection volume was 20 μl. It was.

The experimental results are shown in Figure 9b. 9b shows that the tacrolimus solid dispersion obtained using cyclodextrins using the supercritical fluid process according to the present invention exhibits significantly higher saturation solubility than the tacrolimus raw material itself without using cyclodextrin (Comparative Example 3). Can be.

Test Example 3: Differential Scanning Calorimetry of Paclitaxel Solid Dispersion (DSC Analysis)

In order to compare the thermodynamic changes of the paclitaxel solid dispersion and the paclitaxel raw material prepared in Example 1 was tested as follows using DSC.

About 2 to 4 mg of the sample was weighed using a micro balance, the cap was sealed on an aluminum pan, which was used as a test sample, and the change of calorie with temperature was observed using the empty aluminum pan and the cap as a control. The rate of temperature increase was measured at 10 ° C. per minute and the instrument used for analysis was DLOS DSC from Rheometric Scientific.

The analyzed DSC pattern is shown in FIGS. 2 and 3.

In Figure 2, the paclitaxel raw material can be seen that a strong endothermic peak appears around 216 ℃, which is due to the melting point of paclitaxel is about 215 ℃.

On the other hand, in Fig. 3, in the case of the solid dispersion according to Example 1 of the present invention, the endothermic peak observed at the end of paclitaxel did not appear. From this, it can be seen that the paclitaxel solid dispersion prepared according to the present invention has a stable ultrafine level of high homogeneous state due to the change in molecular arrangement so as to show the thermodynamic change in DSC.

Test Example 4: Differential Scanning Calorimetry of Tacrolimus Solid Dispersion (DSC Analysis)

Except for using the tacrolimus solid dispersion and tacrolimus powder prepared in Example 13, DSC analysis was carried out in the same manner as in Test Example 3, the analyzed DSC pattern is shown in Figures 4 and 5.

In Figure 4, the tacrolimus powder can be seen that a strong endothermic peak appears in the vicinity of 122 ℃, this is due to the melting point of tacrolimus is about 125 ℃.

On the other hand, in Fig. 5, in the case of the solid dispersion according to Example 13 of the present invention, the endothermic peak observed in the tacrolimus powder did not appear. That is, it can be seen that the tacrolimus solid dispersion prepared according to the present invention has a stable ultrafine level of homogeneous state due to a change in molecular arrangement so as to show a thermodynamic change in DSC.

Test Example 5: X-ray powder diffraction analysis of paclitaxel solid dispersion

The crystal structures of the paclitaxel solid dispersion and the paclitaxel raw material prepared in Example 1 were analyzed using an X-ray powder diffractometer. At this time, M18XHF-SRA (Mac Science Co., LTD, Japan) was used as the X-ray powder diffractometer, and diffraction was recorded under conditions of Cu X-ray, 40 kv, 300 mA, and scan speed 15.

X-ray diffraction analysis results are shown in FIGS. 6 and 7.

6 shows that the paclitaxel raw material is in crystalline form, whereas the solid dispersion according to Example 1 is changed from crystalline to amorphous in the course of supercritical fluid treatment, and is thermodynamically stable.

Test Example 6: Pharmacokinetic Test of Tacrolimus Solid Dispersion

Comparison of bioavailability in intravenous administration with rats using the tacrolimus solid dispersion prepared in Example 13 as a test formulation and a prograb (Progrof ^R , Fujisawa Pharmaceutical co., LTD, Japan) injection as a control formulation was as follows. Was carried out.

As the experimental animals, five Sprague-Dawley male rats (250 g body weight and 14-15 weeks old) were used per sample, and the rats were solid feed for normal rats which were constant for 4 days or more under the same conditions. The animals were fed with water. Rats were fasted for more than 48 hours and used for the test. During fasting, water was allowed to drink freely.

Rats were administered intravenously with Example 13 and the control formulation in a 2 mg equivalent amount as tacrolimus per kg of rat body weight. Blood was collected before administration and at 0.02, 0.08, 0.25, 0.5, 1, 1.5, 2, 3, 5, 7 and 24 hours after administration, respectively.

To 200 µl of blood, 400 µl of a solution (0.2M ZnSO ₄ : MeOH = 2: 8 (v / v)) was added and mixed, followed by shaking. The solution was centrifuged at 12,000 rpm for 30 seconds, the supernatant was collected, filtered to 0.22 μm, and analyzed using LC-MS as follows. The results are shown in Table 29 and FIG. 8.

Column: Waters MS C18 (2.1 mm × 150 mm, with guard column)

Mobile phase: concentration gradient (65% MeOH to 95% MeOH)

Injection volume: 30 μl

Flow rate: 0.3 ml / min

Detection: SIR mode m / z: 826.7 (Na adduct)

From Table 29, it can be seen that the formulation prepared according to the present invention has a higher bioavailability than the control formulation.

Test Example 7 LD to Paclitaxel Solid Dispersion ₅₀ Measure

Comparative test of LD ₅₀ by intravenous administration with mouse using Taxi (Taxol ^R , Bristol-Myers Squibb Pharmaceutical Co., LTD, USA) injection as paclitaxel solid dispersion prepared in Example 1 as a test formulation and a control formulation It carried out as follows.

ICR mice (20 g) were used for the experiment, and a total of 50 mice were used, with 5 mice used at each dose in both groups. The doses of 83, 93, 100, 113 and 120 mg / kg were tested as paclitaxel for the paclitaxel solid dispersion of Example 1 as a test formulation and 28, 30, 35, 40 and 42 as paclitaxel for the control taxol. Doses of mg / kg were tested and the results are shown in Table 30 below.

As can be seen in Table 30, the paclitaxel solid dispersion according to the present invention can be seen that the safety is higher than the control preparation.

Using the supercritical fluid process as in the present invention, it is possible to prepare a solid dispersion of poorly soluble drugs, cyclodextrin derivatives and pharmaceutically acceptable additives at a very fine level of high homogeneity, and to add additives, drug combination ratios, and the like. It is possible to obtain a change in the physical properties of the drug, from which can be obtained injections with greatly improved solubility without using a non-aqueous solvent for poorly soluble drugs having a water solubility up to 10 μg / ㎖ at room temperature.

In addition, since the manufacturing process of the present invention uses a supercritical fluid such as carbon dioxide, which is nontoxic, nonflammable, very inexpensive, and can be recovered and reused, it can be usefully used for mass production of poorly soluble drug formulations.

Claims (13)

1) dissolving a poorly soluble drug, a cyclodextrin derivative and a pharmaceutically acceptable additive in an organic solvent to prepare a mixed solution thereof; 2) spraying the mixed solution into a supercritical fluid to make contact with each other to produce particles of a mixture of poorly soluble drugs, cyclodextrin derivatives, and additives; And 3) removing the organic solvent used in the mixed solution by introducing a separate supercritical fluid, a method for producing a solid dispersion of poorly soluble drugs. The method of claim 1 wherein the poorly soluble drug is selected from the group consisting of paclitaxel, tacrolimus, cyclosporin, rapamycin and itraconazole. The cyclodextrin derivative according to claim 1, wherein the cyclodextrin derivative is 2-hydroxypropyl-β-cyclodextrin, 2-hydroxyethyl-β-cyclodextrin, 2,6-dimethyl-β-cyclodextrin, sulfobutyl ether-7- at least one member selected from the group consisting of β-cyclodextrin, (2-carboxymethoxy) propyl-β-cyclodextrin, 2-hydroxyethyl-γ-cyclodextrin, and 2-hydroxypropyl-γ-cyclodextrin. How to. The method of claim 3, wherein the cyclodextrin derivative is used in an amount of 0.1 to 100 parts by weight based on 1 part by weight of the poorly soluble drug. The supercritical fluid of claim 1 wherein the supercritical fluid is selected from the group consisting of supercritical carbon dioxide (CO ₂ ), supercritical dinitrogen monoxide (N ₂ O), supercritical trimethane trifluoride, supercritical propane, supercritical ethylene and supercritical xenon Characterized in that the method. The method of claim 1 wherein the supercritical fluid is contacted with the mixed solution at a temperature of 31-70 ° C. and a pressure of 80-200 bar. The method of claim 1 wherein the pharmaceutically acceptable additive comprises a hydrophilic polymer, a surfactant or an oily component. The method of claim 7, wherein the hydrophilic polymer is polyvinylpyrrolidone (PVP), hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), hydroxyethylcellulose (HEC) and polyethylene glycol (PEG) At least one selected from the group consisting of a method. 10. The method of claim 8, wherein the hydrophilic polymer is used in an amount of 0.1 to 100 parts by weight based on 1 part by weight of the poorly soluble drug. 8. The composition of claim 7, wherein the surfactant is selected from the reaction products of natural or hydrogenated vegetable oils with ethylene glycol; Polyoxyethylene-sorbitan-fatty acid esters; Polyoxyethylene stearic acid esters; Polyoxyethylene-polyoxypropylene block copolymers; Sodium dioctylsulfosuccinate or sodium lauryl sulfate; Phospholipids; Propylene glycol mono-fatty acid esters or propylene glycol di-fatty acid esters; Trans-esterification reaction products of natural vegetable oil triglycerides with polyalkylene polyols; Mono-glycerides, di-glycerides, mono / di-glycerides; Sorbitan fatty acid esters; Fatty acid esters of ascorbic acid; And polyethylene glycol 15 hydroxystearate. The method of claim 10, wherein the surfactant is used in an amount of 1 to 100 parts by weight based on 1 part by weight of the poorly soluble drug. 8. The oil-based component of claim 7, wherein the oil-soluble component is selected from fatty acid triglycerides; Ester compounds of fatty acids and monovalent alkanols; Natural vegetable or animal oils; Hydrocarbons; And at least one member selected from the group consisting of tocopherols. 13. The method of claim 12, wherein the oleaginous component is used in an amount of 0 to 10 parts by weight relative to 1 part by weight of the poorly soluble drug.

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