KR20220103304A - Method for producing amifostine inhalant formulation using wet ball mill process - Google Patents

Abstract

The present invention relates to a method for manufacturing an amifostine inhalation formulation using a wet ball mill process, and when the amifostine inhalation formulation is manufactured by the method of the present invention, the inhalation efficiency of amifostine can be significantly increased. Therefore, the method of the present invention is expected to be usefully used for manufacturing drugs for protection against chemotherapy or radiation treatment or for treatment of lung diseases in an inhalation formulation.

Description

Method for producing amifostine inhalant formulation using wet ball mill process

The present invention relates to a method for preparing an amifostine inhalation formulation using a wet ball mill process.

Amifostine trihydrate is produced by DNA binding chemotherapeutic agents, including alkylating agents (eg cyclophosphamide) and platinum-containing agents (eg cisplatin). It has been used as a therapeutic adjuvant to reduce normal cell destruction. In addition, amifostine trihydrate can prevent nephrotoxicity induced by cisplatin, and patients with advanced ovarian cancer or non small cell lung cancer or head and neck cancer who have received radiation therapy. It is known to be effective in reducing the incidence of xerostomia in patients with head and neck cancer.

In the case of inhaled formulations administered to the lungs, the drug is capable of local action in the lungs and can increase bioavailability due to the liver first pass effect and large absorption surface area as well as reduce the single dose. There are advantages that can be When amifostine is prepared in an inhaled formulation for the protection of normal cells during radiation therapy for lung cancer patients and administered to the lungs, the amount reaching the target tissue, the lung, increases, so bioavailability can be increased, and efficacy can be exhibited even with a small amount. have. In addition, when amifostine is administered to the lungs in an inhaled formulation, side effects of gastrointestinal disturbances can be reduced because it does not pass through the gastrointestinal tract.

On the other hand, Korea Patent No. 1516057 discloses 'an inhalation formulation for the treatment of lung disease and a method for manufacturing the same' comprising a polylactide glycolide (PLGA)-based biocompatible polymer and a pulmonary disease treatment agent, and Korean Patent Publication No. 2020 No. -0063078 discloses 'a method for producing an inhalation formulation for the treatment of lung disease with improved inhalation efficiency of pirfenidone using an excipient', but the method for producing amifostine inhalation formulation using the wet ball mill process of the present invention There is nothing described about

The present invention was derived from the above requirements, and the present inventors prepared fine particles of the amifostine component by varying solvent conditions (polar and non-polar) using a wet ball mill process, and then As a result of analysis, drug release amount and drug delivery rate, it was confirmed that the particle size decreased and the drug release amount and drug delivery rate increased in all solvent conditions compared to the raw amifostine raw material. In addition, the present invention was completed by confirming that when a non-polar solvent was used in the wet ball mill process, it was possible to prepare amifostine fine powder having a homogeneous and stable crystal form compared to using a polar solvent.

In order to solve the above problems, the present invention provides a method for producing an amifostine inhalation formulation comprising the step of pulverizing a mixture in which amifostine is dispersed in a non-polar solvent using a wet ball mill process. provides

In addition, the present invention provides an amifostine inhalation formulation prepared by the above method.

In addition, the present invention provides an anti-cancer or radioprotective agent composition comprising the amifostine inhalation formulation as an active ingredient.

The amifostine inhalation formulation prepared by the method of the present invention can be efficiently delivered to the deep lung, thereby increasing the amount of drug reaching the target tissue, the lung. In addition, the amifostine inhalation formulation of the present invention can have a local action in the lungs and increase bioavailability, and since it does not pass through the gastrointestinal tract, it is effective in reducing side effects of gastrointestinal disturbances.

1 shows amifostine inhalation formulations (Examples 1 to 3) prepared using a jet mill process using a scanning electron microscope (SEM), and amifostine inhalation prepared using a wet ball mill process. These are the results of observation of the formulations (Examples 4 to 7) and the raw amifostine raw material (Comparative Example 1) without processing. Examples 4 and 5: Using polar solvents (methanol, ethanol, respectively), Examples 6 and 7: Using non-polar solvents (chloroform, toluene, respectively).
2 shows amifostine inhalation formulations prepared using a jet mill process (Examples 1 to 3), amifostine inhalation formulations prepared using a wet ball mill process (Examples 4 to 7), and raw Ami It is the result of performing thermogravimetric analysis (TGA) on the postine raw material (Comparative Example 1). Examples 4 and 5: Using polar solvents (methanol, ethanol, respectively), Examples 6 and 7: Using non-polar solvents (chloroform, toluene, respectively).
3 shows amifostine inhalation formulations prepared using a jet mill process (Examples 1 to 3), amifostine inhalation formulations prepared using a wet ball mill process (Examples 4 to 7), and raw Ami It is the result of performing differential scanning calorimetry (DSC) on the postine raw material (Comparative Example 1). Examples 4 and 5: Using polar solvents (methanol, ethanol, respectively), Examples 6 and 7: Using non-polar solvents (chloroform, toluene, respectively).
4 shows amifostine inhalation formulations prepared using a jet mill process (Examples 1 to 3), amifostine inhalation formulations prepared using a wet ball mill process (Examples 4 to 7), and raw Ami This is the result of performing powder X-ray diffraction (PXRD) on the postine raw material (Comparative Example 1). Examples 4 and 5: Using polar solvents (methanol, ethanol, respectively), Examples 6 and 7: Using non-polar solvents (chloroform, toluene, respectively).

In order to achieve the object of the present invention, the present invention provides an amifostine inhalation formulation comprising the step of pulverizing a mixture in which amifostine is dispersed in a non-polar solvent using a wet ball mill process. A manufacturing method is provided.

In the method according to an embodiment of the present invention, the non-polar solvent may be preferably chloroform or toluene, but is not limited thereto.

In the method according to one embodiment of the present invention, the method for preparing the amifostine inhalation formulation includes the steps of dispersing amifostine by adding and dispersing amifostine to chloroform or toluene, which are non-polar solvents; and performing a wet ball mill process on the mixture in which the amifostine is dispersed using a ball having a diameter of 2 to 4 mm at a rotation speed of 350 to 450 rpm and a rotation condition of 4 to 6 minutes.

More specifically, the step of dispersing by adding amifostine to chloroform or toluene as a non-polar solvent; and performing a wet ball mill process on the mixture in which the amifostine is dispersed using a ball having a diameter of 3 mm at a rotation speed of 400 rpm and rotation conditions for 5 minutes; but is not limited thereto.

The present invention also provides an amifostine inhalation formulation prepared by the above method.

The amifostine inhalation formulation of the present invention may have a particle size of 5 to 7 μm, but is not limited thereto.

The present invention also provides a chemoprotective agent or radioprotective agent composition containing the amifostine inhalation formulation as an active ingredient.

The anticancer agent or radioprotective agent composition of the present invention includes an inhaled formulation of amifostine, and it has been reported that amifostine is effective in protecting tissue damage caused by administration of an anticancer agent or radiation through several studies. For example, amifostine is carboplatin, carmustine, chlormethine, cisplatin, cyclophosphamide, fluorouracil, lomustine ), melphalan (melphalan) or oxidopamine (oxidopamine) has been reported to have a protective effect against tissue damage induced by cytotoxic agents such as. In addition, it is known that a number of normal tissues have a radiation-protective effect when irradiated with radiation after pretreatment of amifostine in the abdominal cavity of a mouse (Drugs, 1995, 50(6), 1001-1031).

Hereinafter, an embodiment of the present invention will be described in detail. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

Preparation Example 1. Preparation of amifostine inhalation formulation using a jet mill process

In order to prepare a dry powder inhalation formulation containing amifostine trihydrate (TIANJIN ZHONGRUI PHARMACEUTICAL CP., LTD), the jet mill process was performed using an A-O JET MILL machine (JS-TECH) under the conditions shown in Table 1 below. In the jet mill process, Examples 1 to 3 were prepared by varying the number of processes, and 1 g of amifostine trihydrate was used.

Jet Mill Process Conditions Example 1 Example 2 Example 3 G nozzle (MPa) 0.4 0.4 0.4 P nozzle (MPa) 0.6 0.6 0.6 Number of repetitions (times) One 2 3

Preparation Example 2. Preparation of amifostine inhalation formulation using a wet ball mill process

In order to prepare a dry powder inhalation formulation containing amifostine trihydrate, a wet ball mill process was performed using a ball mill machine (FRITSCH) under the conditions shown in Table 2 below. In the wet ball mill ball, Examples 4 to 7 were prepared with the same ball diameter, amifostine trihydrate:ball volume ratio, rotation speed and rotation time conditions, but with different dispersion solvents, and Examples 4 and 5 used a polar solvent In Examples 6 and 7, a relatively non-polar solvent was used. The volume ratio of amifostine trihydrate:solvent is 1:4, and the volume ratio of amifostine trihydrate:ball:solvent is 5 ml (1 g):25 ml:20 ml.

Wet Ball Mill Process Conditions Example 4 Example 5 Example 6 Example 7 menstruum methanol ethanol chloroform toluene Ball diameter (mm) 3 3 3 3 Armifostine: Ball
volume ratio 1:5 1:5 1:5 1:5 Rotational speed (rpm) 400 400 400 400 Rotation time (min) 5 5 5 5

Example 1. Particle Size Distribution Analysis

Amifostine inhalation formulations prepared using the jet mill process (Examples 1 to 3), amifostine inhalation formulations prepared using the wet ball mill process (Examples 4 to 7), and raw amifostine The particle size distribution of the trihydrate raw material (Comparative Example 1) was evaluated through laser diffraction particle size measurement using a mastersizer 3000 particle size analyzer (Malvern). In the particle size analyzer, when a laser is irradiated to a particle, the light source collides with the particle and diffracts, reflects, and refracts. At this time, the angle is determined according to the size or type of the particle, and the particle size can be inferred by measuring the angle. The angle was measured through the angle of diffraction by irradiating a laser to the tube after dispersing the particles in an insoluble solvent and allowing them to flow through a transparent tube.

As a result, as shown in Table 3 below, the particle sizes (Dv 50) of Examples 1 to 3 prepared using the jet mill process and Examples 4 to 7 prepared using the wet ball mill process were processed. It was confirmed that it was about 12 to 16 times smaller than the raw material of amifostine trihydrate (Comparative Example 1). In addition, in order for the inhalation formulation to be delivered to the lungs, it is suitable that the particle size is about 5 μm. Since the particle size of Examples 1 to 7 is about 5 μm, it was found that the inhalation formulation can be delivered to the lungs.

In addition, the span value in Table 3 below means the range of the particle size distribution, and was derived by an equation such as Span=(D90-D10)/D50. The particles of Examples 4 and 5 exhibited relatively large span values compared to the particles of Examples 6 and 7. Therefore, it was found that a homogeneous amifostine inhalation formulation could be obtained by performing the wet ball mill process using a non-polar solvent.

Particle Analysis Comparative Example 1 Example 1 Example 2 Example 3 Span 1.7 1.7 1.6 2.1 Dv 10 28.0 2.9 2.4 2.3 Dv 50 89.2 7.2 5.8 5.5 Dv 90 181.0 15.4 11.7 13.7 Example 4 Example 5 Example 6 Example 7 Span 3.8 2.5 1.9 1.7 Dv 10 1.1 1.6 2.0 2.3 Dv 50 6.0 5.8 6.0 5.4 Dv 90 23.6 16.3 13.5 11.3

(Unit: μm)

Example 2. Morphological Analysis of Amifostine Inhaled Formulation Using Scanning Electron Microscopy

Amifostine inhalation formulations prepared using a jet mill process (Examples 1-3), Amifostine inhalation formulations prepared using a wet ball mill process (Examples 4-7), and crude amifostine 3 For morphological analysis of the hydrate raw material (Comparative Example 1) particles, it was observed using an Ultra plus scanning electron microscope (SEM, Scanning Electron Microscope, Zeiss).

As a result, it was confirmed that the particles of Examples 1 to 3 and Examples 6 and 7 were similar to the crystalline form of the particles of Comparative Example 1 ( FIG. 1 ). The particles of Examples 4 and 5 were expected to have a change in crystal form due to hydrate drop-off (dehydration reaction) by using a polar solvent in the wet ball mill process.

Example 3. Analysis of Inhalation Efficiency of Amifostine Inhalation Formulation

Amifostine inhalation formulations prepared using a jet mill process (Examples 1-3), Amifostine inhalation formulations prepared using a wet ball mill process (Examples 4-7), and crude amifostine 3 In order to evaluate the inhalation efficiency of the hydrate raw material (Comparative Example 1), it was analyzed using ACI (Andersen cascade impactor, Copley).

Specifically, after filling the capsules with the particles of Examples 1 to 7 and the particles of Comparative Example 1, put them in an inhalation device, install them in the ACI equipment, mount the inhaler on the stage, and then inhale the suction with a decompression pump below the stage did. At this time, the stage simulates the lungs of a person, and the suction pressure of the decompression pump can simulate the suction power of a person.

MMAD (Mass Median Aerodynamic Diameter) described in Table 4 below means the particle size corresponding to the median value when the drug particle size deposited from stages 1 to 6 is plotted as a cumulative graph, and GSD ( geometric standard deviation) means the geometric standard deviation of aerodynamic particle size.

In addition, FPF (Fine Particle Fraction, %) means the drug delivery rate to the lungs, ED (Emitted Dose, %) means the ratio of the drug released from the capsule, FPF and ED were derived by the following formula.

ED(%): {(Amount of drug filled in capsule)-(Amount of drug remaining in capsule after inhalation)}/(Amount of drug filled in capsule)×100

FPF(%): (Amount of particles deposited from stage 1 to 6)/{(Amount of drug filled in capsule)-(Amount of drug remaining in capsule after inhalation)}×100

Since the effective particle delivery amount of drug particles can be known through the FPF (%) value, a large FPF value means a high drug delivery rate, and a large ED value means a large amount of drug release from the capsule. At this time, when the size of the inhalation particle is 5 μm or less in terms of aerodynamic size, it is considered as a size that can be delivered to the deep lung.

Analysis of Inhalation Efficiency of Amifostine Inhalation Formulation MMAD (μm) GSD ED (%) FPF (%) Comparative Example 1 Measurable Measurable 97.1 Measurable Example 1 4.0 1.9 93.1 28.8 Example 2 3.8 1.9 87.8 26.4 Example 3 3.5 1.7 76.6 24.3 Example 4 3.3 5.6 97.4 51.1 Example 5 2.8 4.4 96.5 58.7 Example 6 3.6 5.4 97.8 49.5 Example 7 3.7 4.6 98.0 46.8

As can be seen in Table 4, in Comparative Example 1, it was impossible to measure the FPF value and the inhalation efficiency could not be measured, so it was found that the unprocessed amifostine trihydrate raw material itself was almost impossible to deliver through the lungs. could

In addition, among the inhalation formulations of Examples 1 to 3, the ED and FPF values are the highest in Example 1 and the lowest in Example 3, so the ED and FPF values decrease as the number of repetitions of the jet mill process increases. confirmed that In addition, since the inhalation formulations of Examples 4 to 7 had high ED and FPF values, it was expected that the amifostine inhalation formulation prepared using the wet ball mill process could be sufficiently delivered to the deep lungs.

Example 4. Analysis of Chemical Properties of Amifostine Inhalation Formulation

Amifostine inhalation formulations prepared using a jet mill process (Examples 1-3), Amifostine inhalation formulations prepared using a wet ball mill process (Examples 4-7), and crude amifostine 3 To analyze the chemical properties of the hydrate raw material (Comparative Example 1), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), powder X-ray diffraction (Powder X-ray diffraction) , PXRD) and Karl fisher analysis were performed.

4-1. thermogravimetric analysis

As a result of thermogravimetric analysis, when Examples 1 to 3 and Examples 6 and 7 were compared with Comparative Example 1, the weight reduction section according to the temperature increase showed a similar tendency to each other, and the pyrolysis temperature was also similar. Specifically, Comparative Example 1, Examples 1, 2, 3, 6 and 7 showed a similar weight (%) reduction at around 100 ° C., whereas Examples 5 and 7 showed a similar decrease in weight (%) at around 100 ° C due to hydrate dropout (dehydration reaction). It was confirmed that different weight (%) reduction trends appeared (FIG. 2). Through this, it was found that the particles of Examples 1 to 3 and Examples 6 and 7 according to the present invention had chemical properties similar to those of the amifostine trihydrate raw material (Comparative Example 1).

4-2. Differential scanning calorimetry analysis

As a result of differential scanning calorimetry analysis, it was confirmed that the particles of Comparative Example 1 and Examples 1 to 3 all showed similar heat flow. Specifically, as shown in FIG. 3 , it was confirmed that the peak appearing near 100° C. of Comparative Example 1 was an endothermic peak of the hydrate, and the Tm (melting point) of the drug was approximately 145° C., Examples 1 to 3 are Comparative Examples It was confirmed that the hydrate endothermic peak was shown at a position similar to that of 1, and the Tm was also similar.

In addition, the particles of Examples 6 and 7 also showed an endothermic peak of hydrate at a similar position compared to Comparative Example 1, and the Tm of the drug was about 140 to 145 ° C. It was confirmed that a heat flow similar to that of Comparative Example 1 appeared. On the other hand, in the particles of Examples 4 and 5, the hydrate endothermic peak disappeared near 100° C., which was found to be due to hydrate drop-off (dehydration reaction). Through this, it was found that the particles of Examples 1 to 3 and Examples 6 and 7 had chemical properties similar to those of the amifostine trihydrate raw material (Comparative Example 1).

4-3. Powder X-Ray Diffraction

As a result of powder X-ray diffraction analysis, it was confirmed that the PXRD analysis results showed that the patterns were similar to each other when Examples 1 to 3 and Examples 6 and 7 were compared with Comparative Example 1. On the other hand, in Examples 4 and 5, it was confirmed that the main peak observed in Comparative Example 1 disappeared or had a shifted peak in the PXRD analysis pattern ( FIG. 4 ).

4-4. Karl fisher analysis

As a result of Karl fisher analysis, it was confirmed that Examples 1 to 3 and Examples 6 and 7 were in the form of crystals having a water content of amifostine trihydrate compared to Comparative Example 1. On the other hand, it was confirmed that Examples 4 and 5 prepared using a polar solvent when performing the wet ball mill process were crystalline forms having a lower water content than amifostine trihydrate (Table 5). Through this, when performing the wet ball mill process, it is possible to prepare amifostine inhalation formulations with excellent stability if a non-polar solvent is used, and if a polar solvent is used, amifostine inhalation formulations, an unstable crystal form vulnerable to moisture due to dehydration It was found that this can be manufactured.

Karl fisher analysis Comparative Example 1 Example 1 Example 2 Example 3 Karl fisher (%) 21.8 22.6 23.7 23.7 Example 4 Example 5 Example 6 Example 7 Karl fisher (%) 12.1 12.4 22.6 22.7

In summary, when the amifostine inhalation formulation is prepared using the jet mill process and the wet ball mill process, inhalable fine particles having the amifostine trihydrate crystalline form can be prepared, and in particular, the amifostine inhalation formulation can be prepared using the wet ball mill process. The effect of increasing the amount of amifostine delivered to the lungs was excellent when the postin inhalation formulation was prepared. In addition, it was found that when a non-polar solvent was used rather than a polar solvent in the wet ball mill process, it was possible to prepare the amifostine inhalation formulation in a stable state without modification of the crystalline form.

Claims (6)

A method for producing an amifostine inhalation formulation, comprising the step of pulverizing a mixture in which amifostine is dispersed in a non-polar solvent using a wet ball mill process. The method of claim 1, wherein the non-polar solvent is chloroform or toluene. The method of claim 1,
Dispersing amifostine by adding to chloroform or toluene as a non-polar solvent; and
Comprising the step of performing a wet ball mill (wet ball mill) process using the mixture in which the amifostine is dispersed using a ball having a diameter of 2 to 4 mm at a rotation speed of 350 to 450 rpm and a rotation condition of 4 to 6 minutes A method for preparing an amifostine inhalation formulation, characterized in that The amifostine inhalation formulation prepared by the method of any one of claims 1 to 3. 5. The amifostine inhalation formulation according to claim 4, wherein the inhalation formulation has a particle size of 5-7 μm. An anticancer or radioprotective agent composition comprising the amifostine inhalation formulation of claim 4 as an active ingredient.

Images