KR20220065960A - Method for granulating powder medium having increased solubility and ease of handling - Google Patents

Abstract

The present invention relates to a method for granulating a powder medium having increased solubility and ease of handling and a granulated medium prepared by the method, and more specifically, relates to a method for preparing a granulated medium having optimal particle shape and size and high solubility by optimizing the type, content and process conditions of additives as granulation conditions while performing fluid bed granulation using a fluid bed granulator or dry granulation using a roller compactor for a powder medium; and a granulated medium having increased solubility and ease of handling prepared by the method.

Description

Method for granulating powder medium having increased solubility and ease of handling

The present invention relates to a method for granulating a powder medium having improved solubility and ease of handling, and to a granulated medium prepared by the method.

With the rapid growth of the biopharmaceutical market, the cell culture medium manufacturing technology, which is a core technology used in the bio industry, which is a core technology in the stages of research, development, and production of biopharmaceuticals, is a genetically modified drug, gene that creates high added value through cell culture. It is expected to become a key driving force in the treatment, biopharmaceutical and cell therapy (stem cell) industries.

Almost all of the biopharmaceutical production medium used in Korea is manufactured abroad and imported into Korea. With the growth of domestic biopharmaceutical companies, the demand for media is continuously increasing, so it is necessary to reduce dependence on foreign companies' products and develop new markets through localization of media.

The domestic medium development level is at the level of adding some raw materials to a commercial medium, and while the domestic biopharmaceutical production technology dominates the global market, core material technology-based industries such as culture medium manufacturing are at an early stage. development is needed

Recently, according to the safety problem caused by using a medium containing animal-derived components (eg, fetal bovine serum) and the difference in cell growth by lot, the development of a serum-free chemical composition culture medium in which all compositions are clearly identified is a global trend. is becoming

Animal cell culture for the production of therapeutic agents requires a mixture-type medium containing carbohydrates, amino acids, lipids, vitamins, inorganic salts, and various growth factors necessary for cell growth. Suitable. However, in dry mixing of raw powders of different sizes, the uniformity of each position in the lot may decrease, and in the case of trace elements added in small amounts in ppm units, it is very difficult to uniformly disperse and mix them in the powder medium mixture.

In order to solve the problem of frequent aggregation between particles when dissolving powder medium and the problem of dust generated when handling powder in large quantities, it is essential to introduce powder granulation technology that can improve solubility and minimize the generation of dust from medium raw materials. However, in Korea, there is no technology for pulverizing, mixing, or formulating powder medium, or technology development is very poor.

Accordingly, as a result of the present inventors' efforts to establish the granulation technology of the powder medium, the type, content and The present invention was completed by preparing a granulation medium having an optimal particle shape and size and high solubility by optimizing process conditions.

Republic of Korea Patent No. 10-1617858 Republic of Korea Patent No. 10-1899175

An object of the present invention is to achieve optimum particle shape and size and high solubility through optimization of the type, content and process of additives in fluid bed granulation and dry granulation processes for granulation of powder medium with increased solubility and ease of handling. It is to provide a method for producing a granulation medium having

Another object of the present invention is to provide a granulated medium having increased solubility and ease of handling through the granulation method of the powder medium according to the present invention.

In order to achieve the above object, the present invention

a) preparing a powder medium aqueous solution by dissolving the powder medium in a solvent; and

b) supplying the powder medium aqueous solution and additives to a fluidized bed granulator for granulation; provides a granulation method of a powder medium comprising.

Also, the present invention

a) preparing a powder medium aqueous solution by dissolving the powder medium in a solvent; and

b) supplying the powder medium aqueous solution and additives to a fluidized bed granulator for granulation;

The additive provides any one or two or more selected from the group consisting of saponin, polyethylene glycol, and polyvinyl pyrrolidone, a granulation method of a powder medium.

Also, the present invention

a) preparing a powder medium aqueous solution by dissolving the powder medium in a solvent; and

b) supplying the powder medium aqueous solution and additives to a fluidized bed granulator for granulation;

The granulating step is,

i) fluidizing the powder particles by high-pressure hot air rising from the bottom of the vessel of the fluidized bed granulator;

ii) the granulation fluid is sprayed onto the powder particles by a bottom spray to shape the granules by a surface tension effect; and

iii) drying the granules; consists of,

The additive provides any one or two or more selected from the group consisting of saponin, polyethylene glycol, and polyvinyl pyrrolidone, a granulation method of a powder medium.

Also, the present invention

Provided is a granulated medium having an average particle size of 200 to 350 μm and a solubility of 95 to 100%, prepared according to the fluidized bed granulation method according to the present invention.

Also, the present invention

a') mixing the powder medium and additives; and

b') supplying a mixture of the powder medium and additives to a roller compactor to granulate; provides a granulation method of a powder medium comprising.

Also, the present invention

a') mixing the powder medium and additives; and

b') supplying a mixture of the powder medium and additives to a roller compactor to granulate;

The additive is

It provides a granulation method of any one or two or more selected from the group consisting of saponin, hydroxypropyl cellulose (HPC) and polyvinyl pyrrolidone (Polyvinyl pyrrolidone).

Also, the present invention

a') mixing the powder medium and additives; and

b') supplying a mixture of the powder medium and additives to a roller compactor to granulate;

The granulating step is,

i') quantitatively feeding from the hopper of the roller compactor to the screw feeder;

ii') rolling the powder particles by a screw feeder to form a compact;

iii') forming a flake by pressing the compacted material on the upper and lower rolls; and

iv') forming granules of a certain size by crushing the flakes with a stainless steel scrap; consists of,

The additive is

It provides a granulation method of any one or two or more selected from the group consisting of saponin, hydroxypropyl cellulose (HPC) and polyvinyl pyrrolidone (Polyvinyl pyrrolidone).

In addition, the present invention

It provides a granulated medium, which is prepared according to the dry granulation method according to the present invention, has an average particle size of 500 to 900 μm and a solubility of 98 to 100%.

Since the granulation method of the powder medium according to the present invention has an optimal particle shape and size and high solubility, it has improved solubility and ease of handling.

Through this method, it is possible to produce a fast and uniform cell culture solution by increasing dispersion and solubility during cell culture production, and to minimize dust generation through granulation of powder medium, and it is possible to manufacture optimal cell culture solution by minimizing powder loss .

1 is a graph showing the volume particle size distribution of a granulated medium for each sample according to an embodiment of the present invention.
2 is a graph showing the volume particle size distribution of the granulated medium for each process condition according to an embodiment of the present invention.
3 is a graph showing the solubility of a granulated medium for each type of granulation according to an embodiment of the present invention.
4 is a graph showing the solubility of granulated media for each type of additive in fluid bed granulation according to an embodiment of the present invention.
5 is a graph showing the solubility of the granulated medium by additive type and process conditions in dry granulation according to an embodiment of the present invention.
6 is a graph showing the degree of dust generation of the powder medium and the granulated medium of the medium according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

the present invention

a) preparing a powder medium aqueous solution by dissolving the powder medium in a solvent; and

b) supplying the powder medium aqueous solution and additives to a fluidized bed granulator for granulation; provides a granulation method of a powder medium comprising.

In the method, the medium is any mixture of components that maintain or support the in vivo growth of cells as a cell culture medium, and may include all components or additional components necessary to maintain or support the in vivo growth of cells. . The medium may be a mammalian cell, insect cell, plant cell or bacterial culture medium.

In the method, the granulating step,

i) fluidizing the powder particles by high-pressure hot air rising from the bottom of the vessel of the fluidized bed granulator;

ii) the granulation fluid is sprayed onto the powder particles by a bottom spray to shape the granules by a surface tension effect; and

iii) drying the granules; may include.

In the method, the fluidized bed granulator changes the particle layer in the product container to a fluid state by the upward flow of liquid or gas, and generally high-speed upwardly moving air ejects the particles in the bed to form an 'expansion bed' to operate the fluidization process.

The fluidized bed granulator is a wet granulation and bed fluidization step, in which air is heated to a required temperature and air is blown into the product container with powder to control the airflow, thereby making the bed particles behave like a fluid. , ② As a granulation fluid addition step, if there is a spray nozzle for liquid on the top and the granulation fluid (liquid provided for granulation) is pumped to the nozzle while the bed is completely fluidized, the liquid is evenly sprayed on the fluidized powder and wetted. As the particles collide, they form granules, and ③ dry granulation step. When the particles become granules to a desired size, turn off the spray nozzle and continue to flow hot fluidizing air into the bed to dry all the formed granules. granulate

In the method, the additive is preferably any one or two or more selected from the group consisting of saponin, polyethylene glycol, and polyvinyl pyrrolidone, and various additive materials other than the additive are used for granulation. can be applied.

In the above method, the concentration of the additive is preferably 1/6 to 1/4 of the concentration of the aqueous powder medium solution, and more preferably 1/5.

In the method, the final fluid pressure of the fluidized bed granulator is preferably 0.45 to 0.65 bar, more preferably 0.45 to 0.62.

Also, the present invention

a') mixing the powder medium and additives; and

b') supplying a mixture of the powder medium and additives to a roller compactor to granulate; provides a granulation method of a powder medium comprising.

In the method, the medium is any mixture of components that maintain or support the in vivo growth of cells as a cell culture medium, and may include all components or additional components necessary to maintain or support the in vivo growth of cells. . The medium may be a mammalian cell, insect cell, plant cell or bacterial culture medium.

In the method, the granulating step,

i') quantitatively feeding from the hopper of the roller compactor to the screw feeder;

ii') rolling the powder particles by a screw feeder to form a compact;

iii') forming a flake by pressing the compacted material on the upper and lower rolls; and

It may include; iv') the flakes are crushed by a stainless steel strip to form granules of a certain size.

In the above method, in the roller compactor, when the powder raw material is put into the raw material hopper, the raw material is quantitatively supplied by the screw feeder, the supplied raw material is constantly supplied by the screw feeder in the hopper, and the flow of the screw is quantitatively A uniform condensed product is created, the roll is operated up and down, and a flake (cake) is created by pressing the powder supplied uniformly. will do

In the method, the additive is preferably any one or two or more selected from the group consisting of saponin, hydroxypropyl cellulose (HPC) and polyvinyl pyrrolidone, and various additive materials in addition to the additive can be applied to granulation.

In the above method, the content of the additive is preferably 0.005 to 0.01 weight ratio relative to the powder medium content, and more preferably 0.0075 to 0.01 weight ratio.

In the above method, it is preferable that the screw speed of the roller compactor is 20 to 70 rpm, the roller speed is 2 to 5 rpm, or the number of screen meshes is 5 to 30 mesh, and the screw speed is 30 to 60 rpm , the roller speed is 3 ~ 5 rpm, or the number of screen mesh is more preferably 10 ~ 30 mesh (mesh).

In addition, the present invention provides a granulated medium prepared by the granulation methods according to the present invention.

Specifically,

The present invention relates to a granulated medium having an average particle size of 200 to 350 μm and a solubility of 95 to 100%, prepared according to the fluidized bed granulation method according to the present invention; or

It provides a granulated medium, which is prepared according to the dry granulation method according to the present invention, has an average particle size of 500 to 900 μm and a solubility of 98 to 100%.

Hereinafter, the present invention will be described in more detail with reference to the following examples, experimental examples or drawings.

However, the description of the following examples, experimental examples or drawings is only intended to specifically describe specific embodiments of the present invention, and is not intended to limit or interpret the scope of the present invention to the contents described therein.

< Example 1> Fluid bed granulation

A fluid bed granulator (Midi-Glatt (Germany)) was prepared and granulation was performed under the following conditions.

① Preparation of solution

- RPMI 1640 medium powder: dissolved in 10g/100ml or 5g/100ml.

② Fluid bed equipment operation

- Medium Powder (RPMI 1640) : 150g

- Temperature: 50, 60, 70℃

- Solution feeding rate: 2.5 ~ 10 ml/min

- Feeding time: 40 minutes

- Air pressure (fluidization condition): 0.4 ~ 1.0 bar

- Spray pressure (solution injection): 1.5 bar or 0.5 bar

- 30 minutes to dry

③ Experimental conditions for each sample

Sample solution concentration
(g/ml) Additive type additive concentration
(g/ml) final fluid pressure
(bar) One 0.05 - - 0.34 2 0.05 saponins 0.01 0.45 3 0.05 Polyethylene glycol 0.01 0.62 4 0.05 Polyvinyl pyrrolidone 0.01 0.58

< Example 2> dry granulation

A roller compactor (model: RC-60 (Korea, Seoul Hi-Tech)) was prepared and granulation was performed under the following conditions.

① Powder input

② Roller compactor equipment operation

- Medium Powder (RPMI 1640) : 200g

- Compression pressure: 7 MPa (fixed, maximum laboratory equipment)

- Screw speed: 20 ~ 50 rpm

- Roller speed: 3 ~ 5 rpm

- Screen : 16, 20 mesh

- In general, the higher the screw speed and roller speed, the higher the compression density.

③ Separate granulated and non-granulated ones using a 30 mesh (600 um) sieve

Sample Additive type Additive content
(g) screw speed
(rpm) roller speed
(rpm) screen watch One - - - - 16 mesh 2 Hydroxypropyl cellulose (HPC) 2 - - 16 mesh 3 - - 27.4 3.2 16 mesh 4 - - 20 5 16 mesh 5 - - 50 3.7 16 mesh 6 chaponin 1.5 50 3.7 16 mesh 7 Hydroxypropyl cellulose (HPC) 1.5 50 3.7 16 mesh 8 Polyvinyl pyrrolidone 1.5 50 3.7 16 mesh 9 - - 50 3.7 20 mesh

< comparative example 1> dry granulation

By preparing a spray dryer, RPMI 1640 medium powder was granulated by a conventionally known method.

No. Granulation type Additive type screw speed roller speed screen watch One fluidized bed - 2 saponins 3 polyethylene glycol 4 Polyvinyl pyrrolidone 5 roller compactor - 27.4 3.2 16 mesh 6 - 20 5 16 mesh 7 - 50 3.7 16 mesh 8 chaponin 50 3.7 16 mesh 9 Hydroxypropyl cellulose (HPC) 50 3.7 16 mesh 10 Polyvinyl pyrrolidone
(K30) 50 3.7 16 mesh 11 - 50 3.7 20 mesh 12 spray drying SP No. 1 13 SP No. 2

< Experimental example 1> Particle size analysis

The particle size of each of the granulated media prepared in Examples and Comparative Examples was measured using a particle size analyzer.

Looking at the distribution of the volume particle size according to the granulation method, the average particle size is 200 ~ 350 μm when using a fluidized bed granulator, and 500 ~ 900 μm in the case of using a roller compactor. When used, it was in the form of a lump, but it was easily broken and showed a remarkably low particle size value (FIG. 1).

Looking at the distribution of the volume particle size according to the process conditions of the granulation method, it was confirmed that there is a particle size deviation depending on the type of additive, and the particle size varies depending on the extrusion density and mesh size of the screw/roller of the roller compactor (Fig. 2).

< Experimental example 2> Solubility analysis

The solubility of each of the granulated media prepared in Examples and Comparative Examples was measured.

Specifically, 0.5 g of granular powder medium was dissolved in 100 ml distilled water for 0, 1, and 5 minutes, filtered through a 1 μm filter paper filter, all undissolved powder was removed, and completely dried at 60°C. Solubility was evaluated by comparing the weight of the powder obtained after dissolving for 0, 1, and 5 minutes at the weight of the existing filter paper filter and drying.

S: solubility

Ma : Amount of medium powder obtained after filtering and drying the dissolved medium for n minutes (g)

Mi : Initial weight of powder medium (g)

As a result of analyzing the solubility according to the granulation method, when using a fluid bed granulator and using a roller contactor, overall solubility was higher than 95%, whereas in the case of using a spray dryer, it was dissolved in lumps for about 1 minute. The solubility was remarkably low to such an extent that it did not dissolve (FIG. 3).

As a result of analyzing the solubility of each additive type when using a fluidized bed granulator, when polyvinyl pyrrolidone (K30) was added, the solubility was 98.5%, indicating the highest solubility (FIG. 4).

As a result of analyzing the solubility for each process condition in the case of using a roller compactor, when the mesh size was 20, the particle size size was small and showed higher solubility, and when chaponin was added, it showed the highest solubility at 98.8% (FIG. 5).

< Experimental example 3> Check the level of dust generation

The degree of dust generation was checked before and after granulation of the granulated medium prepared in the above Example.

As a result, the powder form before granulation had very severe powder flying, which resulted in poor workability and loss, whereas there was almost no dust generation after granulation (FIG. 6).

The present invention can be usefully used for cell culture preparation, cell culture development related to future biopharmaceutical production, automated medium production, medium safety, and the like.

Claims (10)

a) preparing a powder medium aqueous solution by dissolving the powder medium in a solvent; and
b) supplying the powder medium aqueous solution and additives to a fluidized bed granulator for granulation;
The additive is any one or two or more selected from the group consisting of saponin, polyethylene glycol and polyvinyl pyrrolidone,
Method of granulation of powder medium.
According to claim 1,
The granulating step is,
i) fluidizing the powder particles by high-pressure hot air rising from the bottom of the vessel of the fluidized bed granulator;
ii) the granulation fluid is sprayed onto the powder particles by a bottom spray to shape the granules by a surface tension effect; and
iii) drying the granules; characterized in that it comprises,
Method of granulation of powder medium.
According to claim 1,
The concentration of the additive is a granulation method of a powder medium, characterized in that 1/6 to 1/4 of the concentration of the aqueous solution of the powder medium.
According to claim 1,
The final fluid pressure of the fluid bed granulator is a granulation method of a powder medium, characterized in that 0.45 ~ 0.65 bar.
a') mixing the powder medium and additives; and
b') supplying a mixture of the powder medium and additives to a roller compactor to granulate;
The additive is
Any one or two or more selected from the group consisting of saponins, hydroxypropyl cellulose (HPC) and polyvinyl pyrrolidone (Polyvinyl pyrrolidone),
Method of granulation of powder medium.
6. The method of claim 5,
The granulating step is,
i') quantitatively feeding from the hopper of the roller compactor to the screw feeder;
ii') rolling the powder particles by a screw feeder to form a compact;
iii') forming a flake by pressing the compacted material on the upper and lower rolls; and
iv') flakes are crushed by a stainless steel strip to form granules of a certain size; characterized in that it comprises,
Method of granulation of powder medium.
6. The method of claim 5,
The content of the additive is a granulation method of a powder medium, characterized in that 0.005 ~ 0.01 weight ratio compared to the content of the powder medium.
6. The method of claim 5,
The screw speed of the roller compactor is 20 to 70 rpm, the roller speed is 2 to 5 rpm, and the number of screen meshes is 5 to 30 mesh.
Prepared according to the method of any one of claims 1 to 4,
Volume of average particle size The particle size is 200 ~ 350 μm,
solubility of 95 to 100%,
Granulated medium.
Prepared according to the method of any one of claims 5 to 8,
Volume of average particle size The particle size is 500 ~ 900 μm,
solubility of 98 to 100%,
Granulated medium.

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