KR100740989B1 - Spherical anti-inflammatory and anti-cancer enzyme granules and method for preparing the same - Google Patents

Abstract

The present invention relates to spherical anti-inflammatory and anti-cancer enzyme granules containing anti-inflammatory and anti-cancer enzymes and a method for producing the same.

The present invention provides a spherical anti-inflammatory salt having excellent enzyme stability and appearance by preparing granules by preparing an aqueous dispersion of enzyme / organic solvent stabilized against the change of enzymatic activity over time and spraying it on a spherical core to grow particles. An anticancer enzyme granule and a method for producing the same are provided.

The present invention has the effect of providing a spherical anti-inflammatory and anti-cancer enzyme granules having excellent appearance such as ease of control of granule size and high sphericity, while increasing stability against aging of enzyme activity over time.

Spherical, anti-inflammatory and anti-cancer enzymes, granules, stability

Description

Spherical anti-inflammatory and anti-cancer enzyme granules and method for preparing the same}

1 is a cross-sectional view of a granule (A) coated with an anti-inflammatory and anti-cancer enzyme layer on a spherical core and a granule (B) formed by further adding a coating layer to the anti-inflammatory and anti-cancer enzyme layer according to a preferred embodiment of the present invention.

Figure 2 is a graph showing the stability of the enzyme by the pH at 4 ℃ and 40 ℃ to determine the optimum pH for enzyme stability according to Example 1 of the present invention,

3 is a photograph showing a comparison of the granules (A) of the anti-inflammatory and anti-cancer enzymes prepared according to Example 1 of the present invention and the granules (B) prepared according to Comparative Example 1 according to the related art.

Figure 4 is a graph showing the ratio of the weight of each particle size to the total weight of the anti-inflammatory, anti-cancer enzyme granules prepared according to Example 1 of the present invention,

Figure 5 is a graph showing a comparison of the activity of the remaining enzymes with time-dependent changes of the ceratiopeptidase granules prepared according to a preferred embodiment of the present invention and the ceratiopeptidase granules prepared according to the conventional method.

The present invention relates to a spherical anti-inflammatory and anti-cancer enzyme granules and a method of manufacturing the same, and more particularly, by changing the pH-controlled anti-inflammatory and anti-cancer enzyme aqueous dispersions to form a anti-inflammatory and anti-cancer enzyme layers by spraying and adsorbing an aqueous dispersion of pH-controlled The present invention relates to a spherical anti-inflammatory and anti-cancer enzyme granule having improved enzyme stability, easy to control the size of granules in the manufacturing process, and improved high sphericity, and a method of manufacturing the same.

In general, enzyme is a biocatalyst whose protein is the main skeleton, and has a relatively good reaction specificity, and the reaction is gentle and continuous so that the reaction is relatively easy to control, and it has many effects such as non-toxicity, low toxicity, and environmental friendliness. It has advantages. Enzymes have been widely used in the food industry such as brewing, fermentation, bakery, confectionery, and throughout the industry from leather, textile processing, and detergents.In recent years, enzymes are used for bio-conversion, biochip, and various disease diagnosis. The application is gradually expanding, and in particular, enzymes are used in digestive agents, anti-inflammatory agents, and the like, and especially in the West, they are being widely used in the anti-inflammatory and anti-cancer agents.

However, in spite of the numerous advantages and wide application fields that come from the use of enzymes, the use of the enzymes is limited due to instability due to external conditions. In particular, unlike in the industrial field using enzymes for chemical reactions and processing, in the pharmaceutical industry, the enzyme activity prescribed at the time of manufacture should be kept constant until the patient's dose. More precise control is required.

For the above reasons, oral enzymes are limited to tablets as formulations that are more favorable for stability than other formulations, and also for cosmetic reasons, even when trying to fill the capsule with stability problems. It is limited to opaque capsules.

Many reports and efforts have been made on improving the stability of enzymes, but most of them are simple addition of stabilizers, for example, polymers such as gelatin, polyalkylene glycols (Korean Patent Application No. 10-1986-0000415), albumin ( Korean Patent Application No. 10-1986-0002879), 4-substituted-phenyl-boronic acid (Korean Patent Application No. 10-1997-7009376), polysaccharide (Korean Patent Application No. 10-2001-0084980) and many others, or encapsulation ( Korean Patent Application Nos. 10-1993-0008473; 10-2002-0073433) and immobilization (Korean Patent Application No. 10-1992-7000925) and the like are known, but the altitude and purpose of the invention to be achieved in the present invention are known. There can be different.

In addition, although the prior art related to the production of enzyme granules (Korean Patent Application No. 10-1994-0701087) is known, it is also an enzyme for the detergent industry, with the aim of low dust formation ability and stability and sustainability in application as a detergent. The high level of the invention to be achieved in the present application and the purpose as an oral anti-inflammatory and anticancer enzyme can be said to be different.

As a spherical enzyme granulation method, a method of firstly mixing and associating each enzyme raw material and then extruding it through a mesh of a specific size and manufacturing it using a spheronizer is generally well known. It has a disadvantage of time and labor consumption, such as having to go through a phased work process.

Recently, by using a fluidized bed process (fluidized bed process) while flowing the raw material in a certain (chamber) by spraying the binder liquid to agglomerate to a certain size and at the same time spherical through the rotation of the device, Although the method of producing granules by simplifying the process into one process is widely used for convenience of work, there is a limitation in obtaining high uniformity and sphericity of granularity in the manufacturing process.

Thus, in the present invention, in order to achieve a high particle size uniformity and sphericity on the granulation, a spherical core of a certain size is flowed and sprayed with an organic solvent aqueous dispersion in which enzymes are dissolved together with a binder and a stabilizer, and particles are sprayed. By growing, a method for producing anti-inflammatory and anticancer granules having high particle size uniformity and sphericity was devised.

In conclusion, as an oral anti-inflammatory and anti-cancer enzyme granule to be achieved in the present invention, there is no technology that simultaneously improves the stability, size control, and spherical degree of enzymatic activity over time.

Accordingly, an object of the present invention is to develop oral anti-inflammatory and anti-cancer enzyme granules having a high sphericity and excellent enzymatic stability and formulation advantages, and a method for preparing the same. It is to provide an anti-inflammatory, anti-cancer enzyme granules prepared by coating.

However, dissolution of anti-inflammatory and anti-cancer enzymes in water and organic solvents for high roundness, and the use of an aqueous organic solvent dispersion of enzymes, affects the structural denaturation of enzymes in aqueous solution. Occurs. This means that the enzyme has a stable pH range that shows optimum activity depending on the type, and is associated with ionizing groups in the molecule where the hydrogen ion concentration (pH) in the aqueous solution affects the three-dimensional structural characteristics of the enzyme. This is because it greatly affects the enzymatic activity and stability of [Effects of pH on Enzymes, Methods in Enzymology, 183-235].

In addition, the stability of the enzyme activity can be changed by the interaction of different enzymes. As a specific example, in the case of protease, it is possible to directly hydrolyze itself or another enzyme, thereby seriously lowering the stability of the enzyme to be degraded.

In addition, the properties and stability of the hydrolyzate in aqueous solution according to pH are both individual and complex, so that even if the information on the individual properties of the enzyme is difficult to predict in terms of enzyme activity and stability. In addition, anti-inflammatory and anti-cancer enzymes that contain heterogeneous enzymes, most of which are not homogeneous, are more difficult to secure enzyme activity and stability. Moreover, hydrogen ion concentration is closely related to formulation stability by moisture absorption even after preparation of granules.

In addition, the activity and stability of the enzyme can be influenced by factors of temperature affecting the structural denaturation and pH change of the pore enzyme.

Accordingly, the present inventors use one or at least two selected anti-inflammatory and anti-cancer enzymes in preparing an organic solvent aqueous dispersion of anti-inflammatory and anti-cancer enzymes, and the single pH range in which the anti-inflammatory and anti-cancer enzymes exhibit optimal activity ( When an aqueous solution is prepared by mixing an organic solvent in an anti-inflammatory and anti-cancer enzyme solution dissolved in a controlled buffer solution within one type of enzyme) or in a commonly included pH range (two or more types of enzymes), The present invention has been completed by demonstrating the minimization of the loss of anti-inflammatory and anti-cancer enzyme activity caused by the change.

In order to achieve the above object, the present invention is a spherical core for molding; And it provides an oral spherical anti-inflammatory, anti-cancer enzyme granules comprising an anti-inflammatory, anti-cancer enzyme layer coated on the core.

In addition, in order to achieve another object, the present invention (1) trypsin (trypsin) and its precursor (trypsinogen), chymotrypsin and its precursor (chymotrypsinogen), elastase, serathiopeptidase, streptoto Kinase (streptokinase), streptodornase (streptodornase), bromelain, papain, deoxyribonuclease, ribonuclease, lysozyme, pancreatin ), At least one anti-inflammatory and anticancer enzyme selected from the group consisting of pancreatic protease, semi-alkaline protease, and the like,

Solubilizing the anti-inflammatory and anti-cancer enzymes by dissolving them together with a controlled buffer solution within a pH range in which the selected anti-inflammatory and anti-cancer enzymes maintain optimal activity and a pharmaceutically acceptable carrier;

(2) preparing an aqueous anti-inflammatory, anti-cancer enzyme organic solvent by mixing the pharmaceutically acceptable organic solvent with the received anti-inflammatory and anti-cancer enzymes; And

(3) a method for preparing oral spherical anti-inflammatory and anti-cancer enzyme granules comprising spraying and adsorbing the prepared aqueous anti-inflammatory and anti-cancer enzyme organic solvents to a molding core having a predetermined size to grow particles. to provide.

Furthermore, the present invention provides an anti-inflammatory and anti-cancer enzyme capsule filled with the oral spherical anti-inflammatory and anti-cancer enzyme granules described above as an active ingredient.

Hereinafter, the present invention will be described in detail.

At this time, if there is no other definition in the technical terms and scientific terms used, it has a meaning commonly understood by those of ordinary skill in the art.

In addition, repeated description of the same technical configuration and operation as in the prior art will be omitted.

The present invention relates to a single pH range in which one or at least two or more anti-inflammatory and anti-cancer enzymes exhibit optimal activity at -10 to 60 ° C, more preferably at 0 to 50 ° C. Species) or commonly contained within a pH range (in the case of two or more enzymes) in a controlled buffer solution, mixed with an organic solvent to prepare an aqueous organic solvent dispersion of anti-inflammatory and anticancer enzymes, and the aqueous dispersion By growing particles by spraying and adsorbing onto spherical cores, we have developed granules of oral spherical anti-inflammatory and anti-cancer enzymes that are uniform and easy to adjust in size and have excellent sphericity. . In this case, the lower the temperature, the more favorable the activity and stability of the enzyme in the preparation of the organic solvent aqueous dispersion of the enzyme whose pH is controlled, but the production cost such as the equipment cost due to the maintenance of low temperature increases and the organic solvent in the fluidized bed granulation and coating process The problem of prolonging drying time due to the lowering of the volatilization rate is caused, and the temperature rises excessively, the particle growth and drying effect of the anti-inflammatory and anti-cancer enzyme granules are excellent, but the activity and stability of the enzyme are deteriorated due to the degeneration of the enzyme. In this invention, it is preferable to manufacture in the range which is not lower than -10 degreeC and does not exceed 60 degreeC. More preferably, the preparation of the organic solvent aqueous dispersion of the enzyme is prepared in the range of 0 ° C. to 10 ° C. where no additional costly additional cooling device is required, and the fluidized bed granulation and coating process is made in the temperature range of 30 to 50 ° C.

However, in the production method according to the present invention, the temperature range may not coincide with the temperature range according to the optimum temperature of the desired anti-inflammatory and anticancer enzymes, and thus may be out of the temperature range in some cases.

The anti-inflammatory and anti-cancer granules according to the present invention are prepared by the following preferred process. However, in order to achieve the objective of this invention, if the method which can form the anti-inflammatory and anti-cancer enzyme layer disperse | distributed with the organic solvent to the spherical core for shaping | molding may be manufactured by replacing all or part of the following detailed processes.

First step: preparing an aqueous solution of anti-inflammatory and anti-cancer enzymes

As shown in Fig. 1, at least one anti-inflammatory and anti-cancer enzyme selected according to the use is selected from the pH range of the optimal activity under process temperature conditions for the selected anti-inflammatory and anti-cancer enzymes (if the enzyme is two or more). A common pH range of each optimum pH range) is mixed and solubilized in a controlled buffer solution.

That is, as a specific method of determining an optimal pH range for stability in the granulation process and over time, it is possible to maintain (control) by adding a salt capable of buffering at the pH range. Determination of the pH range of can be determined simply by measuring the activity at the optimum pH of each enzyme after a certain period of pre-incubation of any anti-inflammatory and anticancer enzyme (s) at pH conditions at each process temperature. . More specifically, as an example of determining an optimal pH range, as shown in FIG. 2, in the case of ceratiopeptidase, an interval of 1.0 at a predetermined pH range and preferably in a range of pH3.0 to 10.0 ) Pre-incubation at a predetermined temperature (within the process temperature range), preferably 1 to 48 hours, more preferably 4 to 10 hours for a certain time in the range of -10 ℃ to 60 ℃, the optimum active pH The activity is measured under the condition 9.0 to determine the pH range that maintains the maximum activity in common. And the control of their pH value uses a group consisting of a weak acid and its base or a weak base and the base acid, and select a salt whose pKa value is located in the center of the pH range determined above to determine the appropriate amount, preferably the concentration of the salt. It can be achieved by adding between 0.001 ~ 0.5 M more preferably between 0.01 ~ 0.2 M on the basis. This is because when the salt concentration is too low, the pH buffering action is not sufficiently performed. If the salt concentration is too high, it may affect the solubility and stability of the enzyme due to salting-out phenomenon. As described above, when the anti-inflammatory and anti-cancer enzyme aqueous solution is controlled to the optimum pH for stability, not only can the enzyme stability in the manufacturing process be optimally obtained, but also moisture is absorbed over time even after being formulated into granular form. It provides the effect of bringing the pH of anti-inflammatory and anti-cancer enzyme absorbents into optimal stabilization conditions.

In addition to the selected anti-inflammatory and anti-cancer enzymes, the aqueous solution of the anti-inflammatory and anti-cancer enzymes may be selected and used in a known range that can be commonly used for oral use in the pharmaceutical related process and food additives. . Specific examples include abnormal protein series such as gelatin, albumin, gluten, polyglutamine, polylysine, cellulose, nitrocellulose, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, cellulose such as methyl cellulose and ethyl cellulose, gum arabic. At least one carrier selected from the group consisting of vegetable gums such as agar, sodium alginate, konjac mannan, polyvinyl alcohol, starch, dextran, and Eudragit E-100 (trade name) can be used. However, the binder of the present invention is not limited to a constant binder. In addition, conventionally known for use in pharmaceuticals, sustained-release polymers, fillers, stabilizers, anti-coagulants, lubricants, lubricants, disintegrants, wetting agents, fragrances, emulsifiers, preservatives, etc. can be used to further include various additives for the purpose of additional functionality Specifically, in the case of sustained-release polymers, enteric polymers including hydroxypropyl methylcellulose phthalate, high viscosity hydroxypropyl methylcellulose, and the like are used. To improve the stability of anticancer enzymes, suitable anti-inflammatory and anticancer enzyme substrates and their substances may be used.

Second step: anti-inflammatory and anticancer enzymes Organic solvent Water dispersion  Produce

In the step (1), the organic solvent is mixed with an aqueous solution of anti-inflammatory and anti-cancer enzymes of which pH is controlled to prepare an aqueous dispersion of the anti-inflammatory and anti-cancer enzyme organic solvents.

At this time, the organic solvent used in the present invention is preferably a boiling point (boiling point) lower than the water, it is possible to use a mixture of two or more solvents. More preferably, the organic solvent standards of the International Conference on Harmonization (ICH) are selected as those having low human toxicity, but one or two or more solvents are selected as organic solvents having a boiling point of 85 ° C. or lower. Use it. In addition, the mixing ratio of the solvent on the basis of the organic solvent / water is preferably added between 0.2 ~ 0.8, more preferably between 0.3 ~ 0.6. This is because when the addition ratio of the organic solvent is lower than a certain level or higher than a certain level, the effect of lowering the boiling point is small compared to the volume increase of the increased enzyme dispersion due to the addition of the organic solvent. This is because it may undermine the stability of the enzyme and the uniformity of the product. More specifically, Class 3 (class 3; ethanol, acetic acid, acetone, 1-propanol, 2-propanol, etc.) and class 2 (class 2; acetonitrile, chloroform, methanol) as defined by the International Pharmaceutical Standards Council (ICH). , Dichloromethane, etc.) can be selected from among the organic solvents.

Third step: granulation of anti-inflammatory and anti-cancer enzymes

Oral spherical anti-inflammatory and anti-cancer enzyme granules are prepared by spraying and adsorbing the anti-inflammatory and anti-cancer enzyme aqueous dispersion prepared in the above step (2) to a spherical core of a specific or predetermined size to grow the particles.

At this time, the anti-inflammatory and anti-cancer enzyme aqueous dispersion is preferably sprayed and adsorbed onto the core while flowing the spherical core using a fluidized bed process, and then dried to grow the particles into monodispersion. This is because monodisperse growth of the particles can improve the sphericity while making the granules fine.

In addition, the spherical core used in the present invention can be prepared by mixing, coalescing, extruding and spheronizing together with a pharmaceutically acceptable polymer such as sugars and poly vinyl alcohol. It can be purchased and used commercially. In this case, the size of the core to be used in the pharmaceutical form is preferably between 200 ~ 1,400㎛ diameter, more preferably 350 ~ 1,000㎛ diameter core size is preferred. When the size of the core is smaller than a certain range, the cores are more likely to combine to form dimers, trimers, and polymers. This is because the fluidized bed process itself, which sprays and adsorbs the anti-inflammatory and anti-cancer enzyme aqueous dispersions, is not made smoothly or prevents uniform particle growth.

Meanwhile, in the present invention, the anti-inflammatory and anti-cancer enzyme granules prepared through the above process may be prepared as a coated granule having a function on the anti-inflammatory and anti-cancer enzyme granules prepared through the above process through an additional coating process in addition to the granule manufacturing process. Can be.

4th process: coating process

By forming a coating layer made of a separate polymer in the anti-inflammatory and anti-cancer enzyme layers of the anti-inflammatory and anti-cancer enzyme granules prepared above, the functionality and stability of the anti-inflammatory and anti-cancer enzymes are further improved.

1 is a granulation agent (A) coated with an anti-inflammatory and anti-cancer enzyme layer on a spherical core and a coating layer (B) respectively formed on the anti-inflammatory and anti-cancer enzyme layer in accordance with a preferred embodiment of the present invention to facilitate the present invention. It is sectional drawing which shows the granule briefly.

(2) coating

As shown in Figure 1 (B), in the present invention, the coating layer is applied by spraying the aqueous dispersion of the polymer on the anti-inflammatory and anti-cancer enzyme layer (2) from the granules of the anti-inflammatory and anti-cancer enzyme prepared above and dried to form a coating layer Say (3).

In this case, it is preferable to use an enteric polymer composed of pharmaceutically acceptable natural or synthetic polymers that are not solubilized in an environment of pH 5.0 or less, and specifically, hydroxypropylmethyl cellulose phthalic acid, cellulose acetate phthalic acid, methacrylic acid. At least one polymer selected from the group consisting of a methyl methacrylate copolymer and a methacrylic acid-ethyl methacrylate copolymer is used. This is because anti-inflammatory and anti-cancer enzymes act in the intestine and show drug efficacy, so that enteric polymers that are decomposed in the intestine are required. However, in addition to the polymer described above, any polymer that can achieve the object of the present invention can be used.

On the other hand, the spherical anti-inflammatory and anti-cancer enzyme granules according to the present invention is a conventionally known method of capsules prepared by filling the granules in capsules or tablets prepared by tableting or the like for the purpose of anti-inflammatory, anti-cancer, thrombolysis, etc. It can be applied widely as a dosage agent for use.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the present invention is not limited to the following examples, and it will be apparent to those skilled in the art that various changes or modifications can be made within the spirit and scope of the present invention.

Example 1

Cerathiopeptidase Enteric Granules

1) pH control

Serratia marcescens ) 1g each of the concentrate of the culture enzyme was dissolved in 100 mL of buffer solutions of pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0, respectively to prepare an enzyme solution. Each of these was allowed to stand for 8 hours in a constant temperature water bath at 4 ± 0.5 ° C. and 40 ± 0.5 ° C., and then their activities were measured according to the Cerapeptase standard and the quantification method of the test method. In this way the optimum pH pH 6.0 was determined (see FIG. 2).

Serratia marcescens ) 10 g of the enzyme concentrate, 3 g of hydroxy propyl methyl cellulose, 3 g of talc, and 1 g of polyethylene glycol # 6000 were stirred and dissolved in 300 g of 20 mM maleic acid buffer (pH 6.0).

(2) Preparation of anti-inflammatory enzyme organic solvent dispersion

150 g of ethanol and 150 g of acetone were added to the anti-inflammatory enzyme solution prepared by the above step (1), respectively, and stirred at 4 ° C. to prepare an aqueous anti-inflammatory and anti-enzyme organic solvent dispersion.

(3) particle growth process

Sucrose core with 95% or more in diameter (Φ) 500 ~ 600㎛, 350g inlet temperature 40 ℃, spray rate 10 using a fluidized bed granulator (Glatt GmbH, Germany) Particles were grown by spraying an aqueous anti-inflammatory and anti-cancer enzyme organic solvent prepared by the above procedure (2) under the condition of mL / min. This finally gave 363 g (99% yield) of the ceratopeptidase spherical granules.

(4) Enteric coating process

While 363 g of the ceratipeptidase spherical granules obtained above were flowed in a fluidized bed coater, 100 g of Kollicoat aqueous dispersion (Kollicoat MAE 30DP, BASF), 6.0 g of talc, and 5.0 g of polyethylene glycol # 6000 were dissolved in 1,000 g of purified water and sprayed with a coating solution. Coated. This process yielded about 471 g (99% yield) of ceratopeptidase enteric granules.

(5) surface modification process

For the ceratopeptidase enteric granules described above, an additional coating process was performed to prevent formulation static electricity and to improve lubricity and to improve aesthetics.

16 g of stearic acid, colorant (red food coloring # 1), 1.5 g, titanium oxide, 1.5 g were dissolved in 400 g of ethanol / acetone (1/1) while spraying the enteric granules, 471 g, in a fluidized bed coater and coated with a coating solution. It was. Finally, about 486 g (99% yield) of ceratopeptidase enteric granules were obtained.

[Comparative Example 1]

Serratia marcescens ) 360g enzyme mixture was prepared by mixing 10 g of the culture enzyme concentrate and 350 g of sucrose, 3.0 g of hydroxy propyl methyl cellulose, 3.0 g of talc, and 1.0 g of polyethylene glycol # 6000 in 600 g of purified water. A spray solution was prepared by mixing with an: ethanol: acetone (w: w: w, 2: 1: 1) solution. 360 g of the enzyme mixture was sprayed while flowing in a fluidized bed granulator to obtain a ceratiopeptidase granule, and then the ceratopeptidase enteric granules obtained by performing steps (4) and (5) in Example 1 were compared. Example 1 was set.

Example 2

Trypsin, chymotrypsinogen, ribonucleases enteric granules

(1) pH control

Try 1g each of trypsin (Seravac, South Africa), chymotrypsinogen (Seravac, South Africa), and ribonuclease (Seravac, South Africa) at pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0. The enzyme solution was prepared by dissolving in 100 mL of buffer solution. Each of these was measured for trypsin, chymotrypsinogen, and ribonuclease activity at pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0 conditions to determine the pH conditions that showed the greatest activity as follows. Decided. (Trypsin pH 7.0, chymotrypsinogen pH 8.0, and ribonuclease pH 7.0) Again, 1 g of trypsin, chymotrypsinogen, and ribonuclease were dissolved together in buffers prepared at each pH, and the temperature was 4 After standing for 8 hours in a constant temperature bath of ± 0.5 ℃ and 40 ± 0.5 ℃, the activity was measured at pH conditions showing the activity of trypsin, chymotrypsinogen, and ribonuclease maximum determined in the above experiment. In this way, an optimum pH 4.0 which is commonly satisfied was determined (see FIG. 3).

Trypsin 10.0g, chymotrypsinogen 23.0g, ribonuclease 4.7g, 2.9g hydroxy propyl methyl cellulose, 2.8g talc, and 1.9g polyethylene glycol # 6000 500g 20mM citrate buffer It was dissolved in (pH 4.0) by stirring.

(2) Preparation of anti-inflammatory and anti-cancer enzyme organic solvent dispersion

250 g of ethanol and 250 g of acetone were added to the anti-inflammatory and anti-cancer enzyme solution prepared by the above step (1), respectively, and stirred at 4 ° C. to prepare an aqueous anti-inflammatory and anti-cancer enzyme organic solvent.

(3) particle growth process

Sucrose core with 95% or more in diameter (Φ) 500 ~ 600㎛, 326.3g inlet temperature 40 ℃, spray rate using a fluidized bed granulator (Glatt GmbH, Germany) Particles were grown by spraying an aqueous anti-inflammatory and anticancer organic solvent dispersion prepared by the above process (2) under the condition of 10 mL / min. Thus, about 368 g (99% yield) of spherical anti-inflammatory and anticancer granules containing trypsin, chymotrypsinogen, and ribonucleases were finally obtained.

(4) Enteric coating process

Trypsin, chymotrypsinogen, ribonuclease-containing spherical anti-inflammatory and anticancer granules, 100 g of Kollicoat MAE 30DP (BASF), talc 6.0 g, and polyethylene glycol # while flowing in a fluidized bed coater 5.0 g of 6000 was dissolved in 1,000 g of purified water and sprayed with a coating solution prepared for coating. This process yielded about 475 g (99% yield) of trypsin, chymotrypsinogen, and ribonuclease enteric granules.

(5) surface modification process

For the trypsin, chymotrypsinogen, and ribonuclease enteric granules, an additional coating process was performed to prevent the static electricity and to improve lubricity and to improve aesthetics.

The above enteric granules, 475g of the stearic acid 18g, colorant (yellow food coloring # 4, 1.5g and red food coloring # 1, 0.1g), 1.6g dissolved in 400g of ethanol / acetone (1/1) while flowing in a fluidized bed coater The coating was sprayed with the prepared coating solution. Finally, about 490 g (99% yield) of trypsin, chymotrypsinogen, and ribonuclease enteric granules were obtained.

Example 3

Capsule Manufacturing

According to the preparation method of the tablets according to the Korean Pharmacopoeia General Regulations, 500g each of the granules of Example 1 and Example 2 was filled into gelatin transparent capsules of No. 1 size to prepare capsules. Each capsule was filled with about 500 mg per capsule to obtain about 1,000 capsules each.

Experimental Example

(1) sphere

In this example, the granules prepared according to Example 1 and Comparative Example 1 were observed under a microscope, and the uniformity and sphericity of the granules were visually checked and numerically determined.

In addition, the sphericality was taken, then measured the perimeter (Pimeter) and the cross-sectional area (A) of the shape of the granules taken to quantify the sphericality (S) by the following formula (Fig. 4) Reference).

[Equation 1]

Sphericity (S) = (4 × π × A) / Pm ²

Here, A is a cross section of the cut surface of the granules, and Pm represents a perimeter of the cut surface of the granules, which means that the closer to 1, the closer the spherical shape is.

As a result, it was found that the sphericity 0.972 ± 0.021 (mean ± standard deviation) of Example 1 prepared according to the present invention was very excellent compared to the sphericity 0.868 ± 0.082 of Comparative Example 1 prepared by the conventional method. .

(2) Granule size distribution

The anti-inflammatory and anti-cancer enzyme granules prepared according to Example 1 were filtered through standard meshes of different sizes, and the respective weights of the filtered particles were measured to evaluate the distribution ratio and uniformity of each granule.

As a result, as shown in Figure 4, the total particle size of the particles produced in the range of 30 sieves (600 ㎛) ~ 20 sieves (850 ㎛) of the total occupied more than 98%, as defined by the General Regulations of the Korean Pharmacopoeia The granular size of the granules passed through the '12 sieve (1,680㎛) and the remaining 14 sieve (1,410㎛) is less than 5% of the total amount, and passing through the 45 sieve (350㎛) When it is 15% or less, it is suitable. '

(3) stability

In order to determine the enzyme stability of the enteric granules of ceratipeptidase prepared by Example 1 and Comparative Example 1, respectively, with the change over time, it was left at a temperature of 40 ± 0.5 ° C and a relative humidity of 75%. Serathiopeptidase activity was measured to confirm the degree of stability.

As a result, as shown in Figure 5, it can be seen that the stability of the ceratiopeptidase granules according to the present invention is superior to the conventional granules.

As described above, the anti-inflammatory and anti-cancer enzyme granules of the present invention select a pH range that is optimal for the stability of the anti-inflammatory and anti-cancer enzyme components to prepare an aqueous dispersion and spray-dry it to a predetermined core to grow the particles. By providing high particle size uniformity and sphericity, it was possible to have excellent enzyme stability and formulation advantages.

Claims (12)

(1) Trypsin and its precursors (trypsinogen), chymotrypsin and its precursors (chymotrypsinogen), elastase, serratiopeptidase, streptokinase, streptokinase (streptodornase), bromelain, papain, deoxyribonuclease, ribonuclease, lysozyme, pancreatin, pancreatic protease, At least one anti-inflammatory and anti-cancer enzyme selected from the group consisting of semi-alkaline proteases, Solubilizing the anti-inflammatory and anti-cancer enzymes by dissolving them together with a controlled buffer solution within a pH range in which the selected anti-inflammatory and anti-cancer enzymes maintain optimal activity and a pharmaceutically acceptable carrier; (2) preparing an aqueous anti-inflammatory, anti-cancer enzyme organic solvent by mixing the pharmaceutically acceptable organic solvent with the received anti-inflammatory and anti-cancer enzymes; And (3) oral spherical anti-inflammatory salt comprising the step of spraying and adsorbing the prepared anti-inflammatory, anti-cancer enzyme organic solvent aqueous solution to a predetermined size (core) to grow the particles coated with the anti-inflammatory, anti-cancer enzyme layer Preparation of anticancer enzyme granules. The method of claim 1, The molding spherical core is a method for producing oral spherical anti-inflammatory anticancer enzyme granules, characterized in that consisting of a pharmaceutically acceptable sugar and a polymer. The method of claim 1, The molding spherical core has an average diameter of 200 ~ 1,400㎛ method for producing oral spherical anti-inflammatory anticancer enzyme granules. The method of claim 1, Oral spherical anti-inflammatory anti-cancer enzyme granules, characterized in that it further comprises the step of forming a coating layer by spraying a coating liquid made of an enteric polymer to the formed particles of step (3). delete delete The method of claim 4, wherein The enteric polymer is a hydroxypropyl methyl cellulose phthalic acid, cellulose acetate phthalic acid, methacrylic acid-methyl methacrylate copolymer or methacrylic acid-ethyl methacrylate copolymer for the preparation of oral spherical anti-inflammatory and anticancer enzyme granules Way. delete delete The method of claim 1, When the selected anti-inflammatory and anti-cancer enzymes are at least heterogeneous, the anti-inflammatory and anti-cancer enzymes are solubilized by using a buffer solution controlled to a common pH range among the respective pH ranges in which the anti-inflammatory and anti-cancer enzymes exhibit optimal activity. Method for preparing spherical anti-inflammatory and anticancer granules for oral use. The method of claim 1, A method for producing oral spherical anti-inflammatory and anticancer enzyme granules, which is prepared at a temperature of -10 to 40 ° C. Oral spherical anti-inflammatory and anticancer granules prepared by the method of any one of claims 1 to 4, 7, 10 and 11.

Images