KR20200011133A - Pharmaceutical formulation containing of bosentan - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing bosentan. According to the present invention, the pharmaceutical composition containing bosentan has excellent lung-targeting drug delivery properties while enabling the drug toxicity of bosentan to be minimized by the organic combination of bosentan and formulation ingredients, and has a longer drug action duration while reducing the number of times of drug administration, thereby being able to be usefully used as an inhalant formulation.

Description

PHARMACEUTICAL FORMULATION CONTAINING OF BOSENTAN}

FIELD OF THE INVENTION The present invention relates to pharmaceutical formulations containing bosentane, and more particularly to pharmaceutical formulations containing bosentane that can be administered to the lungs of an individual via inhalation.

The present invention relates to a composition for preventing or treating pulmonary arterial hypertension, including a pharmaceutical preparation containing bosentan.

Bosentane is represented by Formula 1, 4-tert-butyl-N- [6- (2-hydroxy-ethoxy) -5- (2-methoxy-phenoxy) -2- (pyrimidine-2- Il) -pyrimidin-4-yl] -benzenesulfonamide is a compound with the chemical name of Endothelin Receptor antagonist (ERA) which prevents the decrease in motor performance due to pulmonary hypertension. It is known from 0526708 A1 and is a representative pulmonary hypertension therapeutic currently marketed under the product name Tracleer tablet (Accelion). It is primarily a therapeutic agent for patients with pulmonary hypertension functional class II, III, or IV, which is administered twice daily (morning and evening).

[Formula 1]

The major side effect of bosentane is liver cell damage caused by the induction of cytochrome P2C9 P450 enzymes. Hepatic dysfunction that should stop bosentan occurs in more than 1% of patients with an aminotransferase of more than five-fold increase and bilirubin more than two-fold increase. Other side effects include headaches, nasopharyngitis, flushing and lower extremity edema. Also, in combination with sildenafil, bosentan requires a 50% reduction in the concentration of sildenafil to increase the dose of sildenafil, and also requires dose control by increasing the metabolism of warfarin. These side effects are mainly due to interactions with other drugs, bypass and systemic circulation.

In addition, bosentane is taken twice a day because of its short half-life. This twice-daily administration is inconvenient to take the medication, not only reduce the patient's compliance with the medication, but also miss the appropriate time to increase the possibility of side effects is inevitably needs a solution. By the way, bosentan is a drug having a large limit as an oral dosage form because it is difficult to achieve the convenience of taking it by improving the absorption or dispersibility by improving the solubility because it is very difficult to dissolve in water.

On the other hand, the anatomical physiological characteristics of the lungs should be good in the lung area to which the inhaled drug is delivered, that is, transpulmonary delivery efficiency. Moreover, such transpulmonary delivery efficiency is considered more important because pulmonary hypertension can be effective only when it is delivered and absorbed up to the alveolar level of respiratory bronchioles. The transpulmonary delivery efficiency of the inhaled particles is determined by the physicochemical properties of the particle size distribution, surface energy, shape, surface shape, crystallinity, moisture content, and the like, but hardly any such studies have been conducted with respect to bosentane.

Accordingly, the present inventors have made intensive efforts to solve the drawbacks of the above bosentane drug, and thus completed the present invention by changing the formulation of bosentane into an inhalation formulation and formulating a suitable formulation.

KR 10-2014-0089919 KR 10-2014-0065036

An object of the present invention is a microparticle of bosentan hydrate; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It is to provide a pharmaceutical formulation.

An object of the present invention is a microparticle of bosentan hydrate; And pulmonary arteries comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It is to provide a pharmaceutical aerosol composition for preventing or treating hypertension.

An object of the present invention is a microparticle of bosentan hydrate; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It is to provide a dry powder delivery device containing a pharmaceutical formulation.

Object of the present invention is bosentan hydrate; And micronizing any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It provides a method for producing a pharmaceutical formulation for inhalation comprising a.

The present invention is based on the development of a safe and effective pharmaceutical formulation of bosentan that can deliver an amount effective to exert potent biological activity in these target tissues while minimizing bosentan associated toxicity. In addition, the present invention shows the surprising pharmacokinetic properties of the formulated bosentan. As discussed in more detail below, bosentane delivered directly to the lung produces significantly higher drug concentrations in lung tissue and the amount of drug in lung tissue is higher than predictable from previous oral and intravenous studies. It can show good therapeutic effect without causing toxicity to tissue. In addition, the organic binding of the carrier components can have a longer duration of action while reducing the number of administration of the drug can be useful in inhalation formulations.

As one embodiment for solving the above problems, fine particles of bosentan hydrate; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol Provide a pharmaceutical formulation.

Inhaled pharmaceutical formulations of the present invention deliver bosanthane hydrates directly to the lungs, but are non-toxic and provide longer duration of action at lower doses compared to other dosage forms of bosentan, such as oral or intravenous dosage forms. Indicates. Sedimentation occurs mainly when very small particles moving with the inhaled airflow face the physiological surface as a result of random diffusion inside the airflow. The inhaled pharmaceutical formulations of the present invention are suitable for maximizing their deposition by sedimentation in the alveoli, in order to achieve the desired therapeutic efficacy.

Pharmaceutical inhalation refers to pharmaceutical inhalable through the respiratory tract, nasal cavity, or the like, including respirable particles or droplets containing bosentan.

Bosentan hydrate (hereinafter, bosentan) is represented by Formula 1, 4-tert-butyl-N- [6- (2-hydroxy-ethoxy) -5- (2-methoxy-phenoxy)- 2- (pyrimidin-2-yl) -pyrimidin-4-yl] -benzenesulfonamide.

[Formula 1]

It is currently marketed under the name Tracleer tablet (Acelion) and is used only as an oral dosage.

Bosentane microparticles in the present invention may be particles having an average median diameter in the range of 0.1 to 10 μm, preferably in the range of 1 to 5 μm. The bosentane microparticles can be produced, for example, by spray drying or jet milling.

The carrier includes any one or more selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol.

As the carrier, it is preferable to use microparticles prepared by spray drying, jet milling, sieving, or the like.

Preferably the carrier may be mannitol. Specifically, the mannitol has an advantage in that it is easy to adjust the stabilization of the crystal structure and the shape of the particle surface even in the manufacture according to the spray drying method, and also to remove the bronchial mucus. The weight ratio of such bosentane and mannitol is 5: 1 to 1: 5, preferably 3: 1 to 1: 3, more preferably 2: 1 to 1: 2, most preferably 1: 1. Within the above preferred range, Zeta-potential has the largest absolute value and shows relatively stable surface energy, and most preferably shows the best and stable surface energy at a ratio of 1: 1, which is suitable for pharmaceutical preparations for inhalation. Moreover, the drug delivery to the pulmonary bronchiole is very good with the smallest mass median aerodynamic diameter (MMAD) and highest fine particle fraction (FPF%) values in the gastric ratio.

The use of the mannitol carrier and the setting of the weight ratio show a remarkable effect in comparison with the use of a simple bosentane undifferentiated compound or other carriers. Effect.

In addition, where the carrier consists of a blend of two different carriers, each carrier consists of particles of a different size range, measured by average particle diameter. In one embodiment, the carrier consists of two different carriers, namely a first carrier and a second carrier (carrier). The first carrier consists of particles having a diameter of 0.1 to 10 μm, preferably 0.5 to 5 μm, and the second carrier consists of particles having a diameter in the range of about 5-300 μm, preferably 10-200 μm. The weight ratio of the combined weight of the bosentane and the first carrier to the weight of the second carrier is 1:15 to 1:30, preferably 1:17 to 1:23, and more preferably 1:19.

Most preferably, the first carrier is mannitol and the second carrier is lactose. Like the salpin bar, mannitol has an advantage of easily stabilizing the crystal structure and controlling the surface shape of the particles and removing the bronchial mucus. Lactose maintains homogeneity of the dispersion of bosentan microparticles to ensure homogeneity of capacity and improves flow characteristics, making it easy to handle and eject when used as an inhalation formulation. Furthermore, it can help the bosentane fine particles reach the lungs easily during inhalation. Preferably the lactose is milled lactose, such as lactose sold under the granulac® 200 brand, or lactose sold under the trademarked lactose, such as the inhalac® 250 brand.

The amount of bosentane in the inhaled pharmaceutical formulation is about 0.01% to 20% (w / w) based on the total weight of the composition. In one embodiment, the amount is about 0.1% to 4% (w / w), and in other embodiments about 2.5%.

Inhalable pharmaceutical formulations of the present invention may further comprise additional pharmaceutically acceptable excipients. For example, it may further include a material such as a diluent, stabilizer, adjuvant, antioxidant, adjuvant, propellant, or vehicle. Dry powders such as, but not limited to, hydrocarbons and fluorocarbon propellants, compressed gases, sterile liquids, water, buffered saline, ethanol, suspending agents, antioxidants (eg, ascorbic acid or glutathione), chelating agents, low molecular weight proteins , Or suitable mixtures thereof.

Inhaled pharmaceutical formulations of the present invention are suitable for use in either dry powder inhaler devices (DPIs) or compressed metered dose inhalers (pMDIs), preferably in dry powder inhaler devices. Drug particles are lightly compressed into a frangible matrix contained within a delivery device (dry powder inhaler). In operation, the delivery device abolishes some of the drug particles from the matrix and disperses them into the inhaled breath that delivers the drug particles to the airways. Alternatively, the drug particles may be free flowing powders contained within a reservoir in a delivery device (dry powder inhaler). The reservoir can be an integral chamber inside the device, or a capsule, blister or similar capacity reservoir inserted into the device prior to operation. In operation, the device disperses some of the drug particles from the reservoir and disperses them into inhaled breath that delivers the drug particles to the airways.

An object of the present invention is a microparticle of bosentan hydrate; And pulmonary arteries comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It is to provide a pharmaceutical aerosol composition for preventing or treating hypertension.

Aerosol composition means an aerosolizable composition suitable for producing respirable particles or droplets containing bosentane. It is preferably a dry powder suitable for administration via a dry powder inhalation device.

The present invention provides methods and compositions for the treatment and prophylaxis in a subject in need thereof for pulmonary hypertension, comprising administering a pharmaceutical aerosol composition. The present invention may be administered in an effective amount of about 0.125 mg to 125 mg, preferably 1.25 mg to 62.5 mg, but is not limited to once per week, twice a week, three times a week, four times a week, Pulmonary hypertension can be treated by administering five times a week, six times a week, and seven times a week. This minimizes the side effects of drug administration by reducing the frequency of administration compared to the conventional oral administration method.

The aerosol composition of the present invention can be administered in combination with one or more additional therapeutic agents. That is, the aerosol formulations of the invention may be administered alone or in combination with one or more additional therapies, each of which may be administered by the same or different routes, such as orally, intravenously, and the like.

The target tissue of the aerosol composition of the invention is the lung and the therapeutic level lasts at least 12 hours, 24 hours, 2 days, 3 days, 4 days or more after delivery.

In one embodiment, the amount of bosentane composition in the aerosol composition is 20, 40, 50, 100, 125, or 250 μg.

An effective amount of the composition of the present invention reduces or ameliorates the progression, severity, and / or duration of pulmonary hypertension (PAH) or one or more symptoms of pulmonary hypertension, prevents the development of pulmonary hypertension, resulting in the regression of pulmonary hypertension, To prevent the onset or onset of one or more symptoms associated with pulmonary hypertension, to prevent the severity or onset of one or more symptoms of pulmonary hypertension, or to develop or progress pulmonary hypertension, or to prevent other therapies (eg, prophylactic or therapeutic agents), or May enhance or improve the therapeutic effect. Also in connection with the treatment of pulmonary arterial hypertension, a therapeutically effective amount can inhibit or reduce the proliferation of vascular cells, inhibit or reduce smooth muscle cell proliferation, reduce hypertrophy of the pulmonary canal or improve FVC or FEV1. .

"Treatment" refers to the severity, duration, or progression of pulmonary arterial hypertension, or the reduction of one or more symptoms associated with pulmonary arterial hypertension.

"Prevention" means the prevention of recurrence, development, progression or onset of one or more symptoms of pulmonary hypertension resulting from the administration of a composition identified in accordance with the methods of the present invention or the combination of a known therapy for a disease or disorder with such a compound. Refers to.

An object of the present invention is a microparticle of bosentan hydrate; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It is to provide a dry powder delivery device containing a pharmaceutical formulation.

The dry powder delivery device, for example Accuhaler®, Conix ^TM, Rotahaler®, TwinCaps®, XCaps®, FlowCaps®, Turbuhaler®, NextHaler®, CycloHaler®, Revolizer ^TM, Diskhaler®, Diskus®, Spinhaler, Handihaler®, Microdose inhaler, It is contained in a dry powder inhaler (DPI) device selected from GyroHaler®, OmniHaler®, Clickhaler®, Duohaler® (Vectura), and ARCUS® Inhaler (Civitas Therapeutics). The present invention provides a DPI device containing the dry powder composition described herein. The device is for example selected from the group consisting of XCaps, FlowCaps, Handihaler, TwinCaps, Aerolizer®, Plastiape® RS01 Model 7, and Plastiape® RS00 Model 8.

Pulmonary delivery is preferably achieved by inhalation of the aerosol through the mouth and throat into the lungs. In one embodiment, the aerosol is delivered to the airways via the oral cavity.

The present invention also provides fine particles of bosentan hydrate; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol Provided is a method of treating pulmonary hypertension by administering a pharmaceutical formulation inhaled to a patient in a pharmaceutically effective amount.

The present invention also provides microparticles of the bosentane hydrate in the manufacture of a medicament for the treatment of pulmonary hypertension; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol Provides the use of pharmaceutical preparations.

The present invention also relates to microparticles of bosentane hydrate in the manufacture of a medicament for the treatment of pulmonary hypertension for use in the treatment of pulmonary hypertension; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It provides a composition comprising a pharmaceutical formulation.

Object of the present invention is bosentan hydrate; And micronizing any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol It provides a method for producing a pharmaceutical formulation for inhalation comprising a.

Particle sizes can be obtained from conventional polishing methods, such as by grinding in air-jet mills, ball mills or vibratory mills, from wet polishing, microprecipitation, spray drying, lyophilization or subcritical or supercritical solutions. Recrystallization can be reduced to the desired microparticulates. Jet milling or grinding in this context refers to the micronization of dry drug particles by mechanical means. In certain embodiments, it is desirable to include a carrier material that will be co-micronized with bosentan.

In one embodiment bosentan fine particles in the size range of 0.1 to 10 or 1 to 5 μm are produced by a jet milling method. The carrier may also be prepared together or separately prepared through spray drying, jet milling, sieving, or the like, followed by mixing.

Spray drying generally involves preparing a solution, slurry, or suspension of a drug, atomizing the solution, slurry, or suspension to form particles, and then evaporating the solution, slurry, or suspension medium to form the particles. Include. Solutions, slurries or suspensions may be formed in subcritical or supercritical conditions. The evaporation step can be accomplished by raising the temperature of the atmosphere in which spraying occurs therein, by reducing the pressure, or by a combination of both.

In one embodiment, a powder formulation comprising bosentane spray-drys an aqueous dispersion of bosentane and mannitol to form a dry powder consisting of agglomerated particles of bosentane having a size suitable for pulmonary delivery, as described above. Are prepared. Aggregated particle size may be adjusted (increased or reduced) to target either the deep lung or the upper respiratory area, such as the upper bronchial region or the nasal mucosa. This can be achieved, for example, by increasing the concentration of bosentane in the spray-dried dispersion or by increasing the droplet size produced by the spray dryer.

Bosentane and mannitol microparticles can then be prepared by mixing with lactose microparticles.

That is, the method for preparing the inhaled pharmaceutical formulation may preferably include the following steps.

(a) micronizing bosentan hydrate and mannitol together; And

(b) mixing the bosentan hydrate and mannitol microparticles prepared by micronization in step (a) with lactose microparticles.

In the step (a), the micronization step may be a spray drying method, and the weight ratio of bosentane hydrate and mannitol is 5: 1 to 1: 5, preferably 3: 1 to 1: 3, more preferably 2: 1 to 1: 2, most preferably 1: 1.

In the mixing of step (b), the weight ratio of bosentan hydrate and mannitol microparticles to lactose microparticles is 1:15 to 1:30, preferably 1:17 to 1:23, and more preferably 1:19. .

The pharmaceutical composition containing bosentan according to the present invention has excellent drug delivery characteristics to target the lungs while minimizing drug toxicity of bosentan by organic bonding of bosentan and formulation components, and has a longer action while reducing the frequency of drug administration. It can have a duration and can be usefully used as an inhalation formulation.

Figure 1 shows a scanning electron micrograph of the bosentan and mannitol-containing microparticles.
Figure 2 shows a scanning electron micrograph of the microparticles mixed with bosentan and mannitol-containing microparticles and lactose microparticles (carrier).
Figure 3 shows the results of differential scanning calorimetry of bosentan-mannitol fine particles.
4 shows the results of confirming the aerosol performance of bosentan-mannitol fine particles.
Figure 5 shows the results of confirming the aerosol performance of the microparticles mixed with bosentan and mannitol-containing microparticles and lactose microparticles (carrier).
Figure 6 shows the dissolution rate test results of bosentan-mannitol fine particles.
Figure 7 shows an in vivo experimental schedule for confirming the effect of the treatment of pulmonary hypertension of bosentan-mannitol microparticles.
Figure 8 shows a parasternal short axis image of the ventricles confirmed in vivo experiments to confirm the effect of pulmonary hypertension treatment.
Figure 9 shows the results of confirming the change in morphology of the pulmonary artery and the wall thickness ratio in the in vivo experiment to confirm the effect of pulmonary hypertension treatment.

Hereinafter, the configuration and effects of the present invention through the embodiments will be described in more detail. However, the following examples are merely to illustrate the present invention is not limited to the contents of the present invention.

Example 1 Preparation of Micronization Bosentan

Example 1-1. Preparation of Undifferentiated Bosentane by Spray Drying

Bosentan hydrate (Hanmi Pharm Co. Ltd., Seoul, Korea) was ground by ultrasonication for 20 minutes, dissolved in ethanol and sprayed with 1%, 3%, and 5% drug concentration solutions (SD 1%, respectively). SD 3% and SD 5%) were obtained. Borsentan was spray dried using EYELA SD-1000 (Rikakikai Co., Ltd., Japan) under the following conditions: inlet temperature 110 ° C .; outlet temperature 65-75 ° C .; nozzle size 0.4 mm; feeding rate 10 mL / min; atomization air pressure 200 kPa; And blower rate 0.30 m ³ / min. The spray dried bosentane microparticles prepared were stored in glass vials containing silica gel at −20 ° C. until use.

The bosentane microparticles produced are round smooth amorphous particles.

Example 1-2. Preparation of Undifferentiated Bosentane by Jet-milling

Jet-mill bosentan hydrate microparticles were prepared using A-O JET MILL (J S Tech Co., Ltd., Korea). Prepared according to 0.2, 0.3 and 0.4 MPa air pressure (JM 0.2, JM 0.3 and JM 0.4) and other parameters were performed identically under the following conditions: feeding rate 250%; feeding vibration 40 Hz; And pushing air pressure of 0.5 MPa. The prepared jet-mill bosentan microparticles were stored in glass vials containing silica gel at −20 ° C. until use.

The prepared bosentane microparticles have a smaller particle size compared to ordinary bosentane.

Example 2 Preparation of Undifferentiated Bosentane Containing Bosentane Hydrate and Mannitol

Similar to Example 1-1, bosentane hydrate and D-mannitol were dissolved in 70% ethanol at a ratio of 3: 1, 1: 1, 1: 3 (w / w), respectively, and the total powder concentration was 1% ( w / v) was obtained (referred to as SDBM 3: 1, SDBM 1: 1, SDBM 1: 3 after preparation, respectively). Then, bosentane and mannitol-containing microparticles were prepared and stored by spray drying under the same conditions as in Example 1-1.

< Example  3> dry powder inhaler base Carrier  Produce

Milled lactose (granulac® 200) and filled lactose (inhalac® 250) (respectively referred to as M-SDBM, S-SDBM, respectively) were used to prepare carrier-based bosentan microparticles.

In order to break up larger aggregates with relatively identical particle size distributions, the starting materials of lactose were separated using 75 μm and 150 μm sieves and only particles were left between the two.

The microparticles prepared in Example 2 in the ratio of 1: 1 and each lactose in a ratio of 1:19 were mixed with a Turbula mixer (Impandex Inc., Maywood, NJ, USA) at 72 rpm for 15 minutes in a glass vial. Stored.

< Example  4> Undifferentiated Bossentan  Physical and chemical property evaluation

Scanning electron microscope (SEM), particle size and particle distribution, surface charge, water content, true density, surface area, and differential scanning calorimetry (DSC) were measured to evaluate the physicochemical properties.

Example  4-1. Scanning electron microscope ( SEM ) analysis

 Bosentane samples were placed on carbon tape and platinum coated using a Hummer VI sputtering device. Then, it was observed using a scanning electron microscope (ZEISS-GEMINI LEO 1530, Zeiss, Germany).

The results are shown in FIGS. 1 and 2.

1 shows the results of scanning electron microscopy analysis of bosentan and mannitol-containing microparticles prepared in Example 2. FIG. As a result, when observed macroscopically, they all showed a rough and irregular crumpled shape, while the microscopic surfaces showed slightly different shapes from each other.

FIG. 2 shows a scanning electron micrograph of lactose carrier-based bosentan microparticles prepared in Example 3. FIG. As a result, the group using milled lactose or filled lactose confirmed non-spherical and irregular shapes. In terms of size, the groups using milled lactose or filled lactose were polydiperse.

Example  4-2. Particle size and particle distribution, surface charge, water content, true density, surface area, differential scanning calories ( DSC )

Particle size, distribution and surface charge were determined using a dynamic light scattering technique (Zetasizer Nano ZS, Malvern Instruments, UK, measurement range of 0.3 nm-10.0 μm). After adding 3 mg of sample to 10 mg distilled water, the suspension was vortexed for 20 seconds and left for 1 hour. The average particle size and particle distribution were then expressed as Z-means, polydispersity index.

Water content was quantified using Karl Fischer titration (736 GP Titrino, Metrohm, Switzerland). After dissolving the sample in methanol, the sample solution was filled with Hydranal ^® Composite and injected into the titration cell. Residual water in the sample was checked through the above instrument.

True density was determined using a pycnometer under helium gas, and the temperature was repeated 10 times at 27 ° C.

Surface area was analyzed using an accelerated surface area and porosimetry analyzer (ASAP 2000, Micrometrics Instrument Corp., GA, USA). Samples were degassed for 90 minutes at 90 ° C. under nitrogen before analysis and the surface area was calculated using Brunauer-Emmet-Teller (BET) equation.

The above results are summarized in Table 1.

TABLE 1

As confirmed in Table 1, all of the generated microparticles were confirmed to have a stable surface energy having a large absolute value Z-potential (ZP). In particular, it was confirmed that the fine particles mixed in a 1: 1 ratio showed the best Z-potential and corresponded to the most suitable mixing ratio.

Differential scanning calorimetry was measured using a differential scanning calorimeter (DSC 2910, TA Instruments, DE, USA). Samples were placed in a DSC aluminum pan and scanned at 30 ° C. to 180 ° C. at 10 ° C./min after sealing.

The results are shown in FIG. As confirmed in FIG. 3, the thermogram of the bosentan-mannitol microparticles showed a melting peak of mannitol while no melting peak of bosentan was found. This shows that amorphous bosentane and crystalline mannitol form a particle together, indicating that the formulation is suitable for delivery to the lungs.

Example  4-3. carrier -base Bosentan  Particle Size of Fine Particles

Particle size and distribution of lactose carrier-based bosentan microparticles were dispersed using a sample in isopropyl alcohol and then laser diffraction particle sizing by wet dispersion method (Mastersizer 2000, Malvern Instruments, Worcestershire, UK).

The results are shown in Table 2.

TABLE 2

As confirmed in Table 2, when using milled lactose (granulac® 200) or filled lactose (inhalac® 250), it showed a relative polydispersity particle size distribution, it was confirmed that D _v10 is 75 μm or less. That is, it was confirmed that it corresponds to the fine particles to be delivered to the lungs and lungs deep lung level.

Example 5 Aerosol Performance Verification

According to the USP Chapter <601> description of aerosols, the aerosol performance of the fine particles prepared in Examples 2 or 3 was determined by 8-stage non-viable Andersen Cascade Impactor (ACI) (TE-20-800, TISCH Environmental, Inc.). , OH, USA) and Handihaler® (Boehringer Ingelheim, Germany) DPI devices. 60 L / min was operated and measured before using a flow meter (DFM 2000, COPLEY scientific, Nottingham, UK). ACI stage collection plates were precoated with silicone oil to prevent particle bounce and re-entrainment. At 20 mg the powder equivalent was loaded into a hydroxypropyl methylcellulose hard capsule. The capsule was placed in the Handihaler® and firmly inserted into the mouthpiece suitable for the induction port. The particles were aerosolized at 60 L / min for 4 seconds and the amount of drug deposited in each stage was quantified by HPLC analysis. The cut-off diameters represented by -1, 0, 1, 2, 3, 4, 5 and 6 stages at 60 L / min were 8.6, 6.5, 4.4, 3.3, 2.0, 1.1, 0.54, and 0.25 μm, respectively. .

From the above, ED%, FPF%, MMAD, and GSD were used as aerosol performance indicators.

Mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) were calculated using a web-based tool (www.mmadcalculator.com/andersen-impactor-mmad.html) and all the above experiments were repeated three times.

The results using the microparticles prepared in Example 2 are shown in Figure 4 and Table 3.

TABLE 3

As seen in FIG. 4 and Table 3, the mixing of bosentane and mannitol significantly improved the MMAD and FPF%. In particular, the smallest MMAD and the highest FPF% values were found in the microparticles of bosentan and mannitol in a 1: 1 ratio. In addition, it was confirmed that all of the prepared microparticles showed enhanced aerosol performance than bosentan single microparticles.

The results using the microparticles prepared in Example 3 are shown in Figure 5 and Table 4.

TABLE 4

As shown in FIG. 5 and Table 4, when using lactose as a carrier, it was confirmed that the aerosol performance which was enhanced in the mixing with mannitol was well maintained, and that lactose was a suitable factor as a carrier.

< Example  6> elution evaluation

Franz cell (FCDS-900C; Labfine Instruments) was used to perform elution evaluation by implementing lung air-liquid interface for inhalation drugs.

Specifically, the medium of the receptor phase was used at 37 ° C PBS pH 7.4 (tween # 80 5% w / w; 12 mL; 37 ° C), and a cellulose filter (pore size: 0.45 μm; Advantec) was used as the membrane. It was.

The amount of drug transferred to medium by loading and eluting the bosentane-mannitol microparticles prepared in Example 2 corresponding to about 5 mg was analyzed by HPLC.

HPLC analysis was performed using HPLC (Ultimate 300 series HPLC, Thermo Fisher Scientific), and the analysis conditions were column: Luna L11 250X4.60mm, 5 μm, mobile phase: acetonitrile and buffer (0.1% triethylamine solution, pH 2.5) at a ratio of 45:55 (v / v), flow rate: 1.5 mL / min, injection volume: 10 μL, detector: UV 220 nm.

The results are shown in FIG.

As confirmed in FIG. 6, it was confirmed that the combination of conditions in which the butene is in an amorphous state and conditions including mannitol having high solubility shows a significantly improved dissolution rate compared to that of the general bosentane.

Moreover, it was confirmed that the dissolution rate increases with the ratio of mannitol.

That is, it was confirmed that the microparticles produced as a result of the co-spray drying of bosentan and mannitol exhibited significantly enhanced aerosol performance and dissolution rate, and the microparticles were rapidly eluted after being delivered deep to the pulmonary bronchiole as a dry powder inhalant. It can be seen to show effectively enhanced bioavailability.

< Example  7> Confirmation of pulmonary hypertension treatment effect

Pulmonary hypertension treatment effect was confirmed using the pharmaceutical preparation according to the present invention.

Example  7-1. Pulmonary arterial hypertension ( PAH Manufacture of Induced Animal Models and Drug Administration

Pulmonary hypertension was induced by 8-week-old male SD-rats fed free diet at 50 ± 10% relative humidity and room temperature at 12-hour night / day cycles and intraperitoneally injected at 50 mg / kg.

The experimental group was set up as shown in Table 5 below to perform the experiment.

TABLE 5

 As confirmed in the table above, negative control group (NC), positive control group (PC, pulmonary arterial hypertension), normal bosentane oral administration group (OR), general bosentane inhalation administration group (IR), bosentan-mannitol inhalation of the present invention Administration group (ISD, 1: 1 SDBM), and pulmonary hypertension uninduced general bosentan inhalation administration group (HIR) were set up.

Oral administration was administered drug suspensions using oral maximal and for inhalation administration to rat airways of anesthetized animals using Dry Powder Insufflator ™ (Model DP-4; Penn-Century, Inc.) for rats.

In the case of the experiment was carried out according to the schedule shown in the schematic diagram of Figure 7, the experiment was carried out for three weeks.

Example  7-2. Pharmacokinetic Properties

After administration of bosentan, the pharmacokinetics 륵 measurements for 6 hours are shown in Table 6.

TABLE 6

 As confirmed in Table 6, the bosentan-mannitol inhalation group showed an AUC value about 10 times higher than oral administration. Compared to oral administration, inhalation administration can avoid the hepatic bypass of the drug, and has the advantage of increasing the bioavailability due to the advantages of the large surface area of the lung, large blood flow, thin epithelial cells, and the like. Among them, the bosentan-mannitol inhalation group, which had very high aerosol performance and dissolution rate, showed the highest effect.

Example  7-3. Check survival rate

After completing the experimental schedule, the survival rate was confirmed.

Both the negative control group and non-induced pulmonary hypertension normal bosentan inhalation group (HIR) survived. On the other hand, the positive control group showed a survival rate of about 60% and the normal bosentane oral group (OR) showed a slightly increased survival rate of about 70%. In the case of the bosentan-mannitol inhalation administration group of the present invention (ISD, 1: 1 SDBM), only one animal died after the completion of the experimental schedule, showing high survival rate with excellent treatment effect for high pulmonary hypertension. there was.

Example  7-4. Long-term weighing to determine the effectiveness of treatment

After the experiment, the weight ratio of liver, lung, heart, and right ventricle (RV) to body weight (BW) and the right ventricle to left ventricular and ventricular wall (LV + S) were measured and the results are shown in Table 7.

TABLE 7

Pulmonary hypertension is a disease that causes myocardium to exercise excessively to maintain cardiac output due to continuous blood flow resistance of the pulmonary artery. As a result, pulmonary hypertension is a typical symptom of pulmonary hypertension. appear.

As confirmed in the above table, when looking at the RV / (LV + S) indicating the right ventricular hypertrophy which is a representative symptom of pulmonary hypertension, the positive control group showed a significantly larger value than the negative control group, and the bosentan- according to the present invention. The experimental group administered with mannitol inhalation group (ISD, 1: 1 SDBM) showed the best improvement of right ventricular hypertrophy.

Example  7-5. Confirmation of treatment effect by confirming heart shape through echocardiography

Immediately before the end of the experiment, the heart morphology was observed through echocardiography and the ventricular morphology parameters were measured and are shown in FIGS. 8 and 8.

TABLE 8

Eccentricity is an indicator of the deformation of the circular left ventricular axial phase into elliptical shape due to the increase in the right ventricular pressure. It is expressed as the ratio of the long axis and the short axis of the left ventricular lumen during contraction.

As a result of the experiment, it was confirmed that the best Eccentricity value appeared in the experimental group administered the bosentan-mannitol inhalation administration group (ISD, 1: 1 SDBM) according to the present invention.

In addition, a decrease in pulmonary blood flow due to pulmonary hypertension can lead to congestion of the right heart, which leads to an increase in the right ventricular internal diameter (RVIDd) and excessive myocardial exercise to maintain cardiac output in the event of overload of the ventricles. This increases the right ventricular posterior wall end diastole (RVPWd).

As a result, the experimental group administered the bosentan-mannitol inhalation administration group (ISD, 1: 1 SDBM) according to the present invention reduced the right ventricular internal diameter (RVIDd) and the right ventricular posterior wall end diastole (RVPWd) close to the negative control group. Showed.

Example 7-6. Confirmation of treatment effect through histopathology

At the end of the experiment, lung tissue samples were stained with H & E to evaluate the morphology of the muscular pulmonary artery, and the results are shown in FIG. 9.

As shown in Figure 9, the positive control group induced pulmonary hypertension was confirmed that the muscle layer (pink area) present in the blood vessel muscles significantly compared to the negative control group.

In addition, when checking the thickness, the thinnest vessel wall thickness ratio was confirmed when the experimental group administered the bosentan-mannitol inhalation administration group (ISD, 1: 1 SDBM) according to the present invention.

From the above results, the excellent pulmonary arterial hypertension prevention and treatment effect of the pharmaceutical preparation according to the present invention was confirmed.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto.

Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (16)

Microparticles of bosentan hydrate; And inhalation comprising any one or more carriers selected from arabinose, glucose, fructose, ribose, mannose, sucrose, trehalose, maltose, starch, dextran, lactose and mannitol Pharmaceutical preparations. The pharmaceutical formulation for inhalation according to claim 1, wherein the carrier is mannitol microparticles. The pharmaceutical formulation for inhalation according to claim 1, wherein the pharmaceutical formulation for inhalation is microparticles prepared by micronizing bosentan hydrate and mannitol together. The pharmaceutical formulation for inhalation according to claim 3, wherein the weight ratio of bosentan hydrate and mannitol is 5: 1 to 1: 5. The pharmaceutical formulation for inhalation according to claim 4, wherein the weight ratio of bosentan hydrate and mannitol is 1: 1. The pharmaceutical preparation for inhalation according to claim 1, wherein the carrier comprises mannitol fine particles as the first carrier and lactose as the second carrier. The pharmaceutical preparation for inhalation according to claim 6, wherein the weight ratio of the combined weight of the bosentane and the first carrier and the weight of the second carrier is 1:15 to 1:30. The pharmaceutical preparation for inhalation according to claim 6, wherein the weight ratio of bosentan hydrate and the first carrier is 1: 1, and the weight ratio of the sum of the bosentan and the first carrier and the weight of the second carrier is 1:19. The inhalation pharmaceutical formulation of claim 1, wherein the inhalation pharmaceutical formulation is for use in a dry powder inhaler device or a compression metered dose inhaler. A pharmaceutical aerosol composition for preventing or treating pulmonary arterial hypertension, comprising the pharmaceutical preparation for inhalation according to any one of claims 1 to 9. The pharmaceutical aerosol composition of claim 10, wherein the aerosol composition is delivered orally to the respiratory tract. The method of claim 11, wherein the aerosol composition is administered orally in an amount of 0.125 mg to 125 mg, pharmaceutical aerosol composition for preventing or treating pulmonary arterial hypertension. 10. A dry powder delivery device comprising the pharmaceutical formulation for inhalation of claim 1. (a) micronizing bosentan hydrate and mannitol together; And
(B) a method for producing a pharmaceutical formulation for inhalation comprising the step of mixing the micronized bosentan hydrate and mannitol microparticles prepared in step (a) with lactose microparticles. The method of claim 14, wherein the micronization step in step (a) is a spray drying method, and the weight ratio of bosentan hydrate and mannitol is 5: 1 to 1: 5. The method of claim 14, wherein the weight ratio of bosentan hydrate and mannitol microparticles to lactose microparticles in the mixing of step (b) is 1: 15 to 1: 30.

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