KR20160028195A - Method for preparing solid probiotic coating composition with excellent acid-resistance and stability - Google Patents

Abstract

The present invention relates to a process for the preparation of a coating composition comprising: a) a plasticizer coating step of mixing a powder containing a probiotic with a liquid plasticizer, b) mixing the plastic polymer powder to form a seeded matrix, and c) And a curing step of forming a hard coat layer on the surface of the probiotic coating composition.
The method for preparing a probiotic coating solid composition according to the present invention can be manufactured at a temperature of 60 ° C or less without using water or an organic solvent in the process so that a probiotic coated solid composition having improved acid resistance and stability can be produced without killing probiotic bacteria There are advantages. Therefore, the present invention can be effectively used to develop a probiotic product having an excellent rate of in-vitro engraftment in vivo in the field of foods and medicines containing probiotics.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a method for preparing a probiotic coating solid composition having improved acid resistance and stability,

The present invention relates to a method for producing a probiotic coating solid composition having improved acid resistance and stability.

Probiotics is defined as a microbial agent or microbial component that helps balance microbes in the intestine, as opposed to antibiotics, which means antibiotics. Bibiidobacterium genus ) And lactic acid bacteria such as those belonging to the genus Lactobacillus (Lactobacillus genus). Probiotics, represented by lactic acid bacteria, are defined as live microorganisms that provide health benefits to humans and animals when consumed. These lactic acid bacteria occupy most of the probiotics and have been recognized as representative foods, health functional foods, and foods that have a formal effect in Korea [Korean Journal of Food Science and Nutrition, 24 (4): 817-832, 2011]. Numerous studies have shown that intestinal microflora compositions such as hypersensitivity growth syndrome, antibiotic-induced diarrhea, or inflammatory bowel disease are different from those of normal people, and it has been shown that the ingestion of lactic acid bacteria may improve the disease [Gastroenterology, 133: 24-33, 2007]. Clinical studies have also shown improvements in immunological diseases such as atopy and allergies. Thus, probiotics have a wide range of efficacy from traditional well-known diarrhea or constipation like constipation improvement to immune disease [Gastroenterology, 122: 44-54, 2002].

However, despite the usefulness of these probiotics, oral administration to the body has been limited by its use due to degradation by acid and ascorbic acid. Thus, it is only a small amount of about one millionth that lactic acid bacteria survive and reach the intestine. To overcome this problem, the food and pharmaceutical industries have used lactic acid bacteria more than 10 times higher in order to increase the number of lactic acid bacteria in the intestines. However, this is not a fundamental solution, but a very wasteful measure. Thereby increasing the price of the product and increasing the burden on the purchasing consumer.

In addition, moisture and temperature conditions in the air have a great influence on the storage and distribution stability of the lactic acid bacteria product and the processing stability in the manufacturing process of the secondary product, and it is very important to secure the stability accordingly. Therefore, in order to develop a high-quality lactic acid bacteria product, the first step is to enhance the acid resistance upon ingestion, and it is required to secure high processing stability and distribution storage stability. Despite the numerous scientific evidence on the functionality of probiotics, the question of whether the products distributed in the market actually have this functionality has been raised by many scholars, and these questions are not only the composition of the strains contained in the product, Because the survival rate differs from the notation. Therefore, the high acid resistance of the lactic acid bacteria consumed and the maintenance of stability during distribution are very important factors to ensure the functionality and quality of the probiotic product, and this technology can be directly related to the competitiveness of the product [KSBB Journal, 25: 303-310 , 2010].

The manufacturing process of probiotic powder raw materials is largely made by fermentation of lactic acid bacteria, recovery of cells, and drying process. Currently, the coating technologies possessed by probiotic companies are mainly coating techniques in the lactic acid culture fermented concentrate phase. These methods include techniques such as emulsion, co-precipitation, extrusion curing, and adsorption techniques, which are used to prepare a solid powder in a dried state by spray drying or freeze-drying after coating in water [ Journal of Controlled Release, 162: 56-67, 2012]. These coating techniques are used to reduce the cost of additional coating processes after lactic acid bacterial powdering, but they are weak in the hardness of the coating film due to physical adsorption or curing on the surface and weak in acid resistance and bile resistance, And is therefore not sufficient to effectively utilize its efficacy [Journal of Microbiology and Biotechnology, 20 (10): 1367-1377, 2010].

Therefore, in order to increase the practical intestinal transplantation efficiency after administration to an animal, an additional processing step such as a coating which imparts acid resistance to probiotic bacteria is indispensable in the step of preparing into a final dosage form.

The additional coating process of the probiotic formulation includes coating the formulated semi-finished product of the capsule and tablet with conventional conventional processes such as pan coating. However, this method is not applicable when developing a single formulation that simultaneously contains the probiotics to be delivered to the intestine as well as the components acting on the stomach (eg, digestive enzymes, antacids, texturizers, etc.). Accordingly, the development of a technique for efficiently coating probiotic powder has been required, and so on in the art has been continuously studied.

In order to coat the lactic acid bacteria powder, techniques such as spray drying coating, wet and dry granulation, oil / water coating, and compression coating are used. However, since there is a problem of bacterial death due to solvent and heat used in the coating process, There is a limit to the application of [ Encapsulation Technology to Protect Probiotic Bacteria , Probiotics, Prof. Everlon Rigobelo (Ed.), 501-540, 2012]. As can be seen from the report documents and the comparative experiments of the present invention, the existing probiotic powder coating process using solvent and heat is practically inefficient since it causes practically 90% or more bacterial death. In order to solve such a problem, development of a technique of coating at a low temperature without using a solvent has been attempted by a small number of scientists. Such a technique has been proposed as a dry coating (European Journal of Pharmaceutics and Biopharmaceutics, 47 (1): 51-59, 1999). This technique is a technique known in the United States Patent No. 5837291, and the coating technique is a technique of coating an enteric polymer using a plasticizer without using a solvent. However, this technique has a problem in that yield and coating efficiency are very poor when the probiotic powder is applied to a coating, and a raw material made of a seed larger than a certain size is required for coating [Journal of Food Engineering, 71: 223- 230, 2005]. As described in the example of U.S. Patent No. 5837291, the seed preparation for coating is proposed to be produced by the wet granulation method using water, which has the problem that can cause the death of probiotics as mentioned above. In addition, since the injectable solid polymer should be sprayed into the coater simultaneously with the spraying of the plasticizer, the manufacturing process is long and additional equipment investment is required for spraying the solid polymer. Thus, there is still a limit in industrial application for coating probiotics.

International Patent Publication No. WO 2000-074499 discloses a technique of coating with molten high laureate canola oil, which can be carried out at a low temperature and is more effective than a general vegetable oil coating but has a very low hardness of the coating film, It is difficult to help.

International Patent Publication No. WO 2001/068808 discloses a technique of dissolving fat having a melting point of 60 ° C or higher and spray coating it, and this technique requires an additional step of melting fat at 65 to 120 ° C, There is a disadvantage that the melt may be clumped in the nozzle and the flow and spraying speed of the melt is slow, which is not efficient as the process time becomes longer.

International Patent Publication No. WO 2006/122965 discloses a manufacturing method in which a flake is formed through mixing with a medium-long chain fatty acid by a primary coating and extruded to form a seed, followed by further coating with an alginic acid aqueous solution by a secondary coating . However, this patent does not reveal whether the lactic acid bacteria can actually be delivered safely into the intestines since there is no result of acid resistance evaluation showing that it is practically possible to deliver intestines. Also, as can be seen from the examples, when a primary coating is blended, it is impossible to form a complete physical barrier. Therefore, when the secondary coating is sprayed with an aqueous solution, water may penetrate into the interior of the primary coating upon contact with the primary coating. Which may cause the death of probiotics during the process. Further, when the extrusion process is applied to produce the seed after the first coating process, additional extrusion equipment is required and the process can be troublesome.

International Patent Publication No. WO2006 / 077038 discloses a technique similar to that described above, wherein a first coating is carried out at a low temperature using a low-melting fatty acid as a primary coating, and then a second and a multi-coating are applied to a coating solution with a polymer aqueous solution . This patent also has the problem that since the primary coating is not a complete coating, the probability of the solvent coming into contact with the bacteria during the secondary coating process is increased, resulting in the death of the lactic acid bacteria during the coating process

International Patent Publication No. WO 2010/103201 describes a method of granulating by extruding steam at a temperature of 60 to 85 캜. This technology is applicable only to granules having a particle size of 800 to 1200 μm, and has a problem in that it causes the death of bacteria during the process because of using high temperature steam.

Under these circumstances, the inventors of the present invention have recognized the necessity of research on coating technology for improving the acid resistance and stability of probiotics which are worthy of use in economic terms, and as a result of intensive research, they have completed the present invention.

It is an object of the present invention to provide a probiotic coating solid composition capable of efficiently coating a probiotic powder without the use of a solvent at a low temperature without the killing of the probiotics so as to be stable in the body and reach a high survival rate in the course of distribution and storage To provide a method to do so. It is another object of the present invention to provide a method for producing an effective probiotic coating solid composition which can improve the seed production process, which is a problem of existing dry coatings, and can perform seeding and coating processes at the same time.

The present invention has been completed to solve the above-mentioned technical problems, and the present invention will be described below.

According to the present invention,

a) a plasticizer coating step of mixing the probiotic powder with a liquid plasticizer;

b) a seed forming step of mixing the plastic polymer powder; And

c) curing step to form a coating film

Wherein the whole process is carried out at a temperature of 60 DEG C or less without using a solvent in the process. The present invention also provides a probiotic coating solid composition having improved acid resistance and stability during storage, and a process for producing the same.

The present invention enables probiotics coating through a combination of a liquid plasticizer and a plastic polymer without using a solvent and heat in order to minimize the death of the probiotics during the process, The advantage is that efficient process time can be utilized without facility investment. In addition, even if a process of using a solvent in an additional coating process is performed, bacterial death can be prevented during the process, so that it can be said to be a highly valuable technology capable of solving the problems of conventionally known probiotic coating techniques.

Further, when the probiotic coating solid composition is prepared according to the production method of the present invention, it has an advantage of securing an improved storage stability, acid resistance and intestinal reaching ratio.

1 is a schematic view showing a process for preparing a probiotic coating solid composition according to the present invention.
2 is a graph showing the stability results of the solid composition of lactobacillus acidophilus-coated solid composition prepared according to Example 2-5 of the present invention at room temperature at 25 ° C for 1 month
Fig. 3 is a graph showing the results of stability at 40 ° C for one month of the solid composition of lactobacillus acidophilus coatings prepared according to Example 2-5 of the present invention
4 is a graph showing the stability of the solid composition of Bifidus spp. Prepared according to Example 2-5 of the present invention at an acceleration of 40 ° C for one month.

According to the present invention,

a) a plasticizer coating step of mixing the probiotic powder with a liquid plasticizer;

b) a seed forming step of mixing the plastic polymer powder; And

c) curing step of forming a coating film at a temperature of 60 DEG C or lower

To a process for preparing a probiotic coated solid composition having improved acid resistance and stability during storage.

The production method according to the present invention can be illustrated by a schematic diagram of the method for producing a probiotic coating solid composition provided in FIG. 1, but the present invention is not limited thereto.

Specifically, the step a) is a process for primarily coating a liquid plasticizer on the surface of the powder containing probiotics. The acid resistance and stability of the probiotics can be improved only by the step a), and the plasticizer coated as described above functions to plasticize the plastic polymer to be added in the step b).

The probiotics usable in step a) refer to all solid-state microorganisms, and any microorganism may be used, and may also be carried out using a mixture of one or more microorganisms. The microorganism used in the production method of the present invention is preferably a food, a health functional food or a pharmaceutical product which is administered to an animal and can be delivered to the gastrointestinal tract. Examples of the microorganism include Streptococcus, Lactococcus, Enterococcus, Lactobacillus, Lactic acid bacteria such as Pediococcus sp., Luccorno sp., Bissella spp., And Bifidobacterium spp., Or mixtures thereof can be used.

Also, "plasticizer" used in step a) of the present invention generally refers to a compound used to plasticize or soften components of a pharmaceutical composition, such as polymers or binders. Preferably, the plasticizer is a liquid plasticizer which is liquid or viscous at room temperature, but is not limited thereto. Further, preferably, the liquid plasticizer may be added in an amount of 0.01 to 10 parts by weight based on 1 part by weight of the plastic polymer.

Suitable plasticizers generally include low molecular weight polymers, oligomers, copolymers, oils, organic small molecules, low molecular weight polyols having aliphatic hydroxyl groups, ester-type plasticizers, glycol ethers, polypropylene glycols, multiblock polymers, Molecular weight polyethylene glycols, citrate ester-type plasticizers, triacetin, propylene glycol and glycerin. Specific examples thereof include ethylene glycol, 1,2-butylene glycol, 2,3-butylene glycol, styrene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol and other polyethylene-glycol compounds, Propyleneglycol monoethyl ether, propyleneglycol monocaprylate, sorbitol lactate, ethyl lactate, butyl lactate, ethyl glycolate, acetyl triethanol glycol monoethyl ether, ethylene glycol monoethyl ether, Butyl citrate, triethyl citrate, dibutyl sebacate, diethyl phthalate, dibutyl phthalate, acetyl triethyl citrate, tributyl citrate and allyl glycolate. It is also possible to use oils, for example fixed oils such as peanut oil, sesame oil, cottonseed oil, corn oil, olive oil, fatty acids such as oleic acid, stearic acid and isostearic acid, fatty acid esters such as ethyl oleate, isopropyl myristate, And acetylated fatty acid glycerides. Furthermore, mineral oil and petrolatum, or mixtures thereof, may also be used as other additional plasticizers without the addition or addition of pharmaceutically suitable surfactants, suspending agents or emulsifiers, such as polysorbates, sorbitan fatty acids, sucrose esters can do. All such plasticizers are generally commercially available and combinations of one or more of the above-mentioned plasticizers may be used in the present invention, preferably triethyl citrate, triacetin, diethyl phthalate, dibutyl phthalate, polysorbate 80 , Sorbitan monooleate, propylene glycol, propylene glycol monocaprylate, oleic acid, olive oil, carnauba wax, polyethylene glycol 400, glycerin, diacetyl monoglyceride, glyceryl monooleate, mineral oil and dimethylpolysiloxane Lt; / RTI >

In a preferred embodiment, at least one selected from the group consisting of a mucoadhesive polymer, a liposoluble base, and a conductive agent may be added to the mucoadhesive material simultaneously with the probiotic powder in step a).

The term "mucoadhesive substance" means a substance having adherence to the gastrointestinal mucosa.

Among the above mucoadhesive substances, the mucoadhesive polymer has a sufficient viscosity in the presence of water and / or body temperature to be adhered to the intestinal mucosa. The natural or synthetic hydrophilic homopolymer, the modified or unmodified hydrophilic homopolymer, And hydrogels. Examples of such mucoadhesive macromolecules include polycarboxylates, hyaluronan, chitosan, alginates such as sodium alginate, pectin, xanthan gum, poloxamer, cellulose derivatives, polyvinyl acetate and polyvinylpyrrolidone . The modification may include crosslinking, neutralization, hydrolysis and all or partial esterification. In addition, the mucoadhesive polymer may include, but is not limited to, exopolysaccharides.

Mucoadhesive lipids among the above mucoadhesive substances refer to substances capable of improving the rate of adhesion due to hydrogen bonding and / or hydrophobic interaction with mucosal components. Typical examples of mucoadhesive lipids include fatty acids containing from 14 to 22 carbon atoms and salts thereof; Higher alcohols containing from 16 to 22 carbon atoms; Glycerol fatty acid esters such as monoglycerides, diglycerides and triglycerides of the fatty acids; Cottonseed oil, soybean oil, olive oil, animal origin oil And other oils such as hydrogenated oils; Waxes; Hydrocarbons; And phospholipids, but are not limited thereto.

The conductive agent in the mucosal adhesion material refers to a substance that can be changed by changing the charge and charge distribution on the surface of the inner layer in a cross-linking mechanism that proceeds in the intestinal mucosal layer. This can improve the adhesion rate by controlling the interaction with the mucosa. These interactions have a temporary effect on the nature of physical entanglement, especially electrostatic interactions, due to covalent and non-covalent interactions, including hydrogen bonding and hydrophobic interactions with lactic acid bacteria and intestinal membranes.

Such conductive agents are substances which are ionized in aqueous or oily solvents and are, for example, salts, ionic surfactants, charged amino acids, charged proteins or peptides, or charged substances (cationic, anionic, or amphoteric ). Suitable salts include any salt form of sodium, potassium, magnesium, calcium, aluminum, silicon, copper, manganese, zinc, tin or similar elements and preferably includes the salt form of magnesium, calcium and zinc. Suitable ionic surfactants include sodium dodecyl sulfate, magnesium lauryl sulfate and similar surfactants. Suitable charged amino acids include lysine, arginine, histidine, aspartate, glutamate, glycine, cysteine, tyrosine. In addition, derivative materials having an amino acid and lactic acid structure may be included, but are not limited thereto.

Further, in the step a), one or more additives selected from the group consisting of a disintegrant, a pH adjuster, an antioxidant and a stabilizer may be further mixed.

The disintegrating agent may be selected from the group consisting of calcium carboxymethylcellulose, sodium starch glycolate, corn starch, hydroxypropyl starch, partially alpha starch, low-substituted hydroxypropylcellulose, croscarmellose calcium, croscarmellose sodium , And crospovidone may be mentioned.

The pH adjuster refers to a substance that adjusts the internal environment of the coated solid composition to an appropriate pH, and in the present invention, it refers to a basic substance. Suitable pH adjusters include, but are not limited to, inorganic acid neutralization components, basic amino acids, and the like.

Specific examples of inorganic antacid components include magnesium silicate, synthetic aluminum silicate, magnesium aluminosilicate, magnesium metasilicate aluminate, magnesium oxide, magnesium hydroxide, aluminum hydroxide, calcium hydrogenphosphate hydrate, anhydrous calcium hydrogenphosphate, magnesium carbonate, sodium hydrogencarbonate , Calcium carbonate, hydrotalcite, dihydroxyaluminum sodium carbonate, bismuth carbonates, aluminum phosphates, borosilicate, seaweed, and calcined sand. Specific examples of basic amino acids include arginine, lysine, histidine, and other basic amino acids and derivatives thereof, but they are not limited thereto

All of these pH adjusting agents are generally usable in food or pharmaceuticals, and the above-mentioned pH adjusting agents may be used in the present invention as a combination of one or more kinds.

Examples of the antioxidant include, but are not limited to, cysteine, cysteine hydrochloride, tocopherols and their salts and esters thereof, butylhydroxynonone, butylhydroxyanisole, bithianhydroxytoluene, propyl gallate, octyl gallate, , Tertiary butyl hydroxyquinone, fumaric acid, malic acid, ascorbic acid, and salts thereof.

The stabilizer may be selected from the group consisting of dipotassium edetate, dodosidic edethate, edetate calcium disodium, edetic acid, fumaric acid, malic acid, maltol, sodium edetate and trisodium edetate, , And the like.

Further, in step b), the probiotic powder, which is coated with the liquid plasticizer in the step a), is uniformly mixed with the plastic polymer, and then the plastic polymer on the surface of the powder is plasticized while contacting with the liquid plasticizer to be agglomerated, .

The fibrillated polymer of the present invention generally includes, but is not limited to, a polymer having a glass transition temperature of 30 to 100 DEG C and a solid state at room temperature.

Specifically, the temperature at which the plasticizer is plasticized through combination with a liquid plasticizer is 60 DEG C or less. And more specifically exhibits low solubility or erosion in an acidic environment of pH 5 or lower, no solubility or erosion, and shows solubility and / or erosion at pH 5 or higher. Further, preferably, in step b), 0.01 to 10 parts by weight of the plastic polymer may be used per 1 part by weight of the probiotic powder.

Suitable examples of such a plastic polymer include anionic cellulose derivatives, anionic vinyl resins, anionic acrylic resins and combinations thereof. Anionic cellulosic derivatives include cellulose esters, cellulose esters and cellulose ester-ethers, especially alkylcelluloses, hydroxyalkylcelluloses, hydroxyalkylalkylcelluloses and acylcelluloses. Specifically, cellulose acetate phthalate, cellulose acetate trimellitate, hydroxypropylmethylcellulose phthalate and hydroxypropylmethylcellulose acetate succinate are included. Anionic vinyl resins include polyvinyl esters that are esterified with di- or tricarboxylic acids, examples of which include polyvinyl acetate phthalate. Anionic acrylic resins include polymers and copolymers of acrylic acid or alkyl acrylic acids, especially copolymers of acrylic acid or methacrylic acid with alkyl acrylates or alkyl methacrylates. Specifically, poly (methacrylic acid-co-ethylacrylate) and poly (methacrylic acid-co-methyl methacrylate) are included.

Other suitable plastic polymers include ethylcellulose, keratin, phenyl salicylate, acetotannine, shellac, gelatin, crosslinked gelatin, ion exchange resins, glycerin ester fatty acids, poloxamer, fatty acids and waxes, Alone or in combination may be used. Modifications include, but are not limited to, cross-linking, neutralization, hydrolysis and all or partial esterification. Preferably, the plastic polymer is selected from the group consisting of hydroxypropylmethylcellulose acetate succinate, hydroxypropylmethylcellulose phthalate, shellac, methacrylic acid methyl methacrylate copolymer, cellulose acetate phthalate, polyvinylacetate phthalate and carnauba wax One or more selected.

Also, in step c), the seeds prepared in step b) are cured at a temperature of 60 DEG C or less in order to increase the firmness of the coating film and make the particle surface of the coating composition smoother and even. Preferably, the temperature inside the coater can be performed within 35 to 45 占 폚. The curing time may vary depending on the type and mixing ratio of the liquid plasticizer and the plastic polymer, but may be, for example, 30 minutes to 2 hours, but not limited thereto.

In a preferred embodiment, during the curing step of step c), a plasticizer and / or a glidant may be added.

The plasticizer is the same as defined with respect to the plasticizer used in step a).

The lubricant may be selected from the group consisting of talc, magnesium stearate, calcium stearate, colloidal silica, stearic acid, wax, hydrogenated oil, polyethylene glycol, sodium benzoate, sodium stearyl fumarate and metal oxides, ≪ / RTI > The above-mentioned metal oxides include at least one component selected from the group consisting of aluminum oxide, titanium dioxide, magnesium oxide, zinc oxide, zirconium dioxide, and silicon dioxide.

As a further preferred embodiment, the manufacturing method according to the present invention may further comprise the step of forming an additional coating layer after the curing process of step c). Particularly, when the probiotic coating solid composition is prepared by the production method according to the present invention, the prepared probiotics-coated solid composition has a solid shielding film and is resistant to solvents and heat. Therefore, And oil-based coating, it is possible to stably carry out the additional coating without killing the probiotic due to the solvent used in the additional coating process or the drying heat.

Various coating methods for such additional (secondary) coatings include, but are not limited to, water, oil, and dry coating methods commonly used in the art. Secondary coating compositions can be prepared via liposoluble, enteric, waterproofing agents and combinations thereof. Preferably, water, ethanol, acetone, propyl alcohol, and a mixed organic solvent of at least one of the organic solvents may be used as the solvent for the water-based coating or oil-based coating. In a preferred embodiment, the secondary coating layer may be formed by charging 0.01 to 10 parts by weight of the coating material with respect to 1 part by weight of the primary coating prepared in the step c), or the secondary coating layer may be formed, 0.01 to 10 parts by weight relative to 1 part by weight of the primary coating is added and mixed to form a secondary coating layer and then the coating is applied to the secondary coating in an amount of 0.01 to 10 parts by weight per 1 part by weight of the amount of the coating- By weight of a liquid plasticizer.

The probiotic coating solid composition prepared by the technique provided by the present invention not only provides a protective film against oxygen, humidity and heat during the manufacturing process using the probiotic powder, but also can ensure the stability of the lactic acid bacteria during storage and shelf life. In addition, when administered into the body of an animal, it avoids intestinal risk factors such as gastric acid and makes it possible to reach the field with high survival rate.

Furthermore, the coating technique of the present invention can be applied not only to probiotics but also to other active substances whose stability is deteriorated by changes in temperature, solvent and pH.

The present invention will be described in more detail with reference to the following examples. However, these examples are provided only for the understanding of the present invention, and the scope of the present invention is not limited to these examples in any sense.

< Manufacturing example  1 to 33>

In order to judge the applicability of the manufacturing process capable of seed formation and coating of the matrix type without use of the solvent proposed in the present invention, it is determined whether or not the liquid plasticizer and the plastic polymer material form an agglomeration body of a substance to be coated Granulation and coating were judged. Dextrin powder having an average particle size of about 20 um was used as a core material instead of probiotic powder and a liquid plasticizer as shown in Table 1 and a plastic polymer material were combined and applied through the following three steps a) to c) The manufacturability of the composition was evaluated.

a) plasticizer coating step

The applicability of the plasticizer coating step of mixing the a) probiotic powder with the liquid plasticizer of the above-mentioned composition of the present invention was evaluated.

Specifically, 2 kg of dextrin was put into a high-speed mixer (High Speed Mixer, P 1/6 type, Diosna Dierks & Sohne GmbH, Germany), 150 g of a liquid plasticizer as shown in Table 1 was injected and mixed at room temperature for 5 minutes (Agitator Speed: 300 rpm, Chopper Speed: 500 rpm). After the completion of the mixing, physical properties and microscopic observation were performed to determine whether the dextrin powder was wetted, coated, and formed with aggregates. O when the aggregates were formed, and X when no aggregates were formed.

b) Seed formation  step

The applicability of the seed forming step of mixing the b) plastic powder of the above-mentioned composition of the present invention was evaluated.

Specifically, 250 g of various plastic polymer materials as shown in Table 1 were put into a high-speed stirrer and then mixed at room temperature for 3 minutes (Agitator Speed: 300 rpm, Chopper Speed : 500 rpm). After completion of the mixing, adhesion of the plastic polymer to the surface of the aggregate particles and seed formation were observed through a microscope. When the formation was observed, it was indicated as O, and when it was not formed, it was indicated as X.

c) Curing step

C) the applicability of the curing step of forming a coating film at a temperature of 60 DEG C or less was evaluated.

Specifically, a seed matrix composed of dextrin, a liquid plasticizer, and a plastic polymer formed through a seed forming step was cured in an oven at 40 ° C for 30 minutes, and then the surface of the particles was observed with a microscope to determine whether a uniform coating film was formed O when formed, and X when not formed.

Based on these results, it was possible to select a combination capable of forming granules in the combination of the compositions prepared.

Manufacturing example Liquid plasticizer Plastic polymer a) a plasticizer
Whether coating b) Aggregate
Presence or absence of formation c) Curing
Whether One Triethyl citrate Hydroxypropylmethylcellulose acetate succinate O O O 2 Polyethylene glycol 400 O O O 3 Diacetyl monoglyceride (Myvacet) O O O 4 Medium chain triglyceride (Mygliol) O O O 5 Polyoxyl 40 Hydrogenated Castor Oil O O O 6 Macrogol glycerol ricinoleate (Cremophore) O O O 7 Glyceryl monooleate O O O 8 glycerin O O O 9 Mineral oil O O O 10 Dimethylpolysiloxane O O O 11 Propylene glycol O O O 12 Polysorbate 80 (Twin 80) O O O 13 Sorbitan monooleate 80 (span 80) O O O 14 Olive oil O O O 15 Triethyl citrate Stearic acid O O O 16 Stearic alcohol O O O 17 Polyethylene glycol 6000 O O O 18 Ethyl cellulose O O O 19 Polyvinyl acetate O O O 20 Hydroxypropylmethylcellulose phthalate O O O 21 Cellulose acetate phthalate O O O 22 Shellack O O O 23 Methacrylic acid methyl methacrylate copolymer (Eudragit L) O O O 24 Dimethylaminoethyl methacrylate Copolymer of methyl methacrylate (Eudragit E) O O O 25 Ethyl methacrylate Trimethylammonium ethyl methacrylate copolymer (Eudragit RS) O O O 26 Sucrose esters of fatty esters O O O 27 Glyceryl behenate (Compritol 888 ATO) O O O 28 Glyceryl monostearate (Geleol) O O O 29 Stearyl polyoxyl glyceride (Gelucire 50/13) O O O 30 Mono / diglyceride capric acid (Campmul MCM) O O O 31 Polyethylene-polypropylene glycol copolymer (Poloxamer 188) O O O 32 Polyethylene-polypropylene glycol copolymer (Poloxamer 407) O O O 33 Carnauba wax O O O

< Example  1>

Probiotics  Preparation of coating solid compositions: Application of liquid plasticizers and plastic polymers

The liquid plasticizer and the plastic polymer candidate materials, which are considered to be easily applicable to the present invention, were selected through preliminary studies conducted in the above preparation examples and applied to actual probiotic powder to prepare a coating solid composition.

A raw material powder containing Lactobacillus acidophilus strain of Cell Biotech Co., Ltd. and bifidobacterium animilis subsp. Lactis (Probio-Tec BB-12) of Christian Hansen was used in Table 1 1, a part of a liquid plasticizer and a plastic polymer capable of forming a seed without using a solvent was selected as shown in Table 2 below to prepare a probiotic-coated solid composition. 2 kg of probiotic powder (average particle size about 20 μm) was added to a high-speed stirrer, 150 g of a liquid plasticizer was added, and the mixture was mixed at room temperature for 5 minutes. 250 g of plastic polymer was added thereto and then mixed at room temperature for 3 minutes (Agitator Speed: , Chopper Speed: 500 rpm). After seed formation was confirmed by microscopic observation, the seeds were transferred into a fluidized bed coater (Uni-Glatt, Glatt GmbH, Germany) and cured while floating in the process conditions shown in Table 2 below. At this time, the inlet air pressure, the pressure inside the fluidized bed coater, and the filter shaking interval were adjusted according to the situation depending on the suspension of the granules and the curing condition of the coating composition during the process.

Example Liquid plasticizer Plastic polymer Fluidized bed coater process conditions 1-1 Triethyl citrate Eudragit L100 - Inlet air temperature: 50 ℃
- Coater inner temperature: 38 ~ 40 ℃
- Inlet air pressure: 30 to 40 mBar
- Coater inner pressure: 10 ~ 15 mBar
- Filter shaking interval: 5 seconds every minute
- Process time: 30 minutes 1-2 Triethyl citrate Shellack 1-3 Triethyl citrate Hydroxypropylmethylcellulose phthalate 1-4 Triethyl citrate Cellulose acetate phthalate 1-5 Triethyl citrate Carnauba wax 1-6 Triethyl citrate Hydroxypropylmethylcellulose acetate succinate (HPMCAS-LF) 1-7 Glyceryl monooleate (GMO) 1-8 Diacetyl monoglyceride (Myvacet) 1-9 Mineral oil 1-10 Polyethylene glycol 400 1-11 Triethyl citrate: Diacetyl monoglyceride

< Example  2>

Probiotics  Preparation of coating solid compositions: Probiotics  Composition ratio of powder, liquid plasticizer and plastic polymer and optimization of process temperature

As the composition of Example 1-6, triethyl citrate as a liquid plasticizer and hydroxypropylmethyl cellulose acetate succinate (HPMCAS-LF) as a plastic polymer were selected to optimize the combination ratio of the composition, A probiotic coated solid composition was prepared with various composition ratios as well. Except that the ratio between the liquid plasticizer and the plastic polymer as set forth in Table 3 and the temperature inside the fluidized-bed coater during the curing process were changed.

Example HPMCAS weight ratio to probiotics Weight ratio of liquid plasticizer to HPMCAS Fluidized bed coater
Internal air temperature (℃) 2-1 0 0 30 2-2 0 0 40 2-3 0 0 50 2-4 0.62 0.76 40 2-5 0.36 0.63 40 2-6 0.62 0.63 50 2-7 0.1 0.5 40 2-8 0.36 0.76 50 2-9 0.1 0.76 40 2-10 0.36 0.5 30 2-11 0.36 0.76 30 2-12 0.36 0.5 50 2-13 0.1 0.63 30 2-14 0.1 0.63 50 2-15 0.62 0.63 30 2-16 0.62 0.5 40 2-17 0.62 0.63 40

< Comparative Example  1>

Flow Coating

In order to compare the preparation process of a non-solvent-free probiotic coating solid composition according to the present invention with the coating solid composition prepared by a flow coating method using an organic solvent which has been used in the prior art, the following coating process is compared For example. 2 kg of probiotics raw material powder (average particle size of about 20 μm) was sprayed at a rate of 20 ml per minute with 10% hydroxypropylmethylcellulose phthalate ethanol solution while being suspended in a fluidized bed coater and coated with 15% of the mass of probiotic powder. The fluidized bed coating conditions were the same as those in Table 2 of Example 1 above.

< Comparative Example  2>

Water-based coating

The following water-based coating process was performed for the comparison of the coated solid composition prepared by the water-based coating method using water, which has been conventionally used, with the manufacturing process of the non-solvent-free probiotic coating solid composition according to the present invention.

2 kg of the probiotics raw material powder (average particle size of about 20 μm) was coated with 15% of the mass of the probiotic powder while spraying the dispersion of Eudragit L30 D-55 at a rate of 20 ml per minute while floating in a fluidized bed coater. The fluidized bed coating conditions were the same as those in Table 2 of Example 1 except that the internal drying temperature was set at 50 캜.

< Example  3>

Additional Secondary Coating: Dry Coating

10% by weight of hydroxypropylmethyl cellulose acetate succinate (HPMCAS-LF) solid state powder was added to the primary coating solid composition prepared in Example 2-5, followed by mixing in a fluidized bed coater. Thereafter, a mixture of 63% triethyl citrate and diacetyl monoglyceride relative to the weight of the added hydroxypropylmethyl cellulose acetate succinate (HPMCAS-LF) was sprayed with a liquid plasticizer to prepare secondary coating granules. The fluidized bed coating conditions were the same as those in Table 2 of Example 1 above.

< Example  4>

Additional Secondary Coating: Flow Coating

The primary coating solid composition prepared in Example 2-5 was suspended in a fluidized bed coater while spraying a 10% hydroxypropylmethylcellulose phthalate ethanol solution at a rate of 20 ml per minute to obtain 10% of the primary coating solid composition Hydroxypropylmethylcellulose phthalate was coated to prepare a second coating solid composition. The fluidized bed coating conditions were the same as those in Table 2 of Example 1 above.

< Example  5>

Additional Secondary Coating: Waterborne Coating

While the primary coating solid composition prepared in Example 2-5 was suspended in a fluidized bed coater while spraying the dispersion of Eudragit L30 D-55 at a rate of 20 ml per minute, the weight of the primary coating solid composition Secondary coated granules were prepared by coating Eudragit L 100-55. The fluidized bed coating conditions were the same as those of Comparative Example 1.

< Example  6>

Addition of liposoluble base

0.15 part by weight of glycerin monooleate was added as a mucoadhesive liposoluble base material to the probiotic raw material powder in the step a) of Example 2-5 to prepare a coated solid composition.

< Example  7>

Mucosa Attachment  Addition of polymer

In Example 6, 1 part by weight of sodium alginate as a mucoadhesive polymer was added to 1 part by weight of the raw material of probiotics in the process a) to prepare a coating solid composition.

< Example  8>

Conductive agent addition

The coating solid composition was prepared by adding 1 part by weight of calcium carbonate as a mucoadhesive conductive agent in 1 part by weight of the probiotic raw material powder in the process of a) of Example 7 above.

< Example  9>

Tableting

The primary coated solid composition prepared in Example 2-5 was tableted under the prescription and tableting conditions shown in Table 4 below.

division ingredient Input (mg / tablet) chief ingredient The probiotic coated solid composition of Example 2 170 Excipient dextrin 37.5 Excipient Microcrystalline cellulose 37.5 Excipient Calcium silicate 25 Binder Low-substituted hydroxypropylcellulose 22.5 Disintegration Crospovidone 5 Lubricant Magnesium stearate 10 Tablet weight 307.5 ± 10 Hardness 8 ± 2 kgf Disintegration Water, 1 liquid, 2 liquids 10 minutes or less

< Test Example  1>

In process Probiotic bacteria Survival rate  evaluation

In order to observe the killing rate of probiotic bacteria among the processes of Examples 1 to 9 and Comparative Examples 1 and 2 according to the present invention, in vitro experiments were carried out as follows. As a control, the coated granules of the present invention and the flow-through and water-based coating probiotic solid solids prepared in Comparative Examples 1 and 2 were compared.

20 mg of the raw material powder before coating and the same amount of the coating solid composition were homogeneously dispersed in 10 mL of sterilized PBS buffer (pH 6.5), and then stepwise diluted by a 10-fold dilution method. After the final culture, the dilution ratio was adjusted so that the number of colonies in the medium was 30 or more and 300 or less.

Lactobacillus acidophilus was prepared by adding 1.0 mL of the test solution to each of three Petri dishes, adding 20 mL of the MRS transformation medium, mixing well, coagulating at room temperature, applying 10 mL of the MRS modified medium thereto, Lt; / RTI &gt; Lactobacillus bifidus is prepared by adding 1.0 mL of the test solution to each of three Petri dishes, adding 20 mL of BS agar medium, mixing well, coagulating at room temperature, applying 10 mL of BS agar medium and incubating at 37 ° C for 72 hours . The number of colonies was counted by visual observation and the number of colonies of lactobacillus acidophilus and lactobacillus bifidus in 1 g was calculated by multiplying the number of colonies by the dilution factor.

Here, the survival rate of 0% means that the viability was reduced to less than 1/1000 compared to the initiation time when the amount of bacteria was determined by the dilution method. 100% of the survival rate was 100% Respectively.

Survival rate during process (%) Acid resistance (Log ₁₀ CFU) Disintegration sample Lactobacillus acidophilus Bifidus Lactobacillus acidophilus Bifidus pH 1.2 pH 6.8 Commencement of raw material 100 100 9.9 10.9 Example 1-1 100 100 7.2 8.3 X O 1-2 100 100 5.2 6.3 X O 1-3 100 100 7.9 8.8 X O 1-4 100 100 7.8 8.6 X O 1-5 100 100 5.2 6.2 X O 1-6 100 100 8.0 8.9 X O 1-7 100 100 8.0 9.0 X O 1-8 100 100 8.2 9.2 X O 1-9 100 100 7.4 8.2 X O 1-10 100 100 5.9 6.8 X O 1-11 100 100 8.2 9.2 X O 2-1 100 100 4.9 6.0 X O 2-2 37.2 67.9 - - X O 2-3 1.0 29.8 - - X O 2-4 100 100 8.4 9.5 X O 2-5 100 100 8.4 9.5 X O 2-6 85 72 8.7 9.7 X O 2-7 54 75 - - X O 2-8 91 87 8.5 9.5 X O 2-9 100 100 7.6 8.8 X O 2-10 100 100 8.3 9.3 X O 2-11 100 100 8.4 9.5 X O 2-12 91 87 8.4 9.5 X O 2-13 100 100 8.0 8.9 X O 2-14 4.2 30.8 8.5 9.6 X O 2-15 100 100 8.5 9.5 X O 2-16 100 100 8.5 9.5 X O 2-17 100 100 8.4 9.5 X O 3 100 100 8.5 9.6 X O 4 87 92 9.0 10.2 X O 5 100 100 8.7 9.7 X O 6 100 100 8.5 9.5 X O 7 100 100 8.6 9.6 X O 8 100 100 8.6 9.7 X O 9 100 100 8.5 9.5 X O Comparative Example One 14.8 37.8 8.3 9.4 X O 2 36.3 46.2 8.1 9.3 X O Control group One 5.0 - O O 2 - 5.9 O O

As can be seen from the results of Table 5, it was found that the bacterial mortality was greatly influenced by the composition ratio of the probiotic powder, the liquid plasticizer and the plastic polymer, and the temperature during the process, It is possible to inhibit bacterial death due to temperature rise during the coating process. In addition, the probiotic coating method according to the embodiment of the present invention is clearly effective in preventing bacterial death in the process as compared to conventional processes such as the flow-through and water-based coating process of the comparative example.

< Test Example  2>

Acid resistance  evaluation

The killing rate of the probiotics-coated solid composition prepared according to the example of the present invention in acidic aqueous solution of pH 1.2 was observed. The acid resistance of Lactobacillus acidophilus (Control 1) and Bifidobacterium animilis subsp. Lactis (Probio-Tec BB-12, Control 2) of Cell Biotech was evaluated as a control.

The number of bacteria that were exposed to artificial stomach fluid was calculated by the dilution medium method of Test Example 1 above. Then, an in vitro test for evaluating the acid resistance in an acidic aqueous solution having a pH of 1.2 was carried out as follows. 20 mg of the probiotic raw material powder before coating and the same amount of probiotics were shaken in 30 ml of 0.1 N HCl pH 1.2 aqueous solution at 37 ° C for 30 minutes at 200 rpm and then 10 ml of 0.3 M Na ₃ PO ₄ solution was added to adjust pH Was adjusted to 6.8 and shaken at 200 rpm for 10 minutes at 37 ° C to be completely disintegrated. The number of lactobacillus acidophilus and bifidus bacteria survived by the dilution medium of Test Example 1 was calculated.

The acid-fastness values shown in Table 5 were obtained from log values of survival probiotics number (Log ₁₀ CFU).

As can be seen from the results of this acid resistance test, the coating solid composition prepared through the present invention It has excellent acid resistance compared to other company probiotics raw material powders.

Also, when comparing Example 2-5 with Example 9, it can be seen that there is no difference in acid resistance before and after tableting. This result shows that the coating film of the coating solid composition prepared according to the present invention does not break down by the static pressure and maintains the same even in tablets, so that there is no decrease in acid resistance, and thus it can be effectively developed as tablets.

< Test Example  3>

Disintegration  exam

The disintegration test of the provitics-coated solid composition prepared in the examples of the present invention was carried out in the aqueous solutions of pH 1.2 and 6.8 according to the disintegration test method in the Korean Pharmacopoeia General Test Methods. In each condition, when the disintegration was observed within 30 minutes, it was marked as O and when it was not disintegrated, it was marked as X. As can be seen from the results of Table 5, the coating solid composition prepared according to the present invention shows that disintegration is delayed under acidic conditions and rapidly disintegrated within 30 minutes under basic conditions.

< Test Example  4>

Stability evaluation

The tablets containing the probiotics-coated solid composition prepared in Example 9 of the present invention were packaged in HDPE bottles and stored at 25 ° C and 40 ° C for 1 month at room temperature to evaluate the stability of the probiotic bacteria. The measurement of the number of bacteria was carried out in the same manner as in Test Example 1. The raw material powder of probiotics before coating was tableted by the same method as in Example 9 to prepare tablets and used as a control. It can be seen that the stability of the probiotic can be certainly improved by using the technique of the present invention through the stability test results shown in Figs. 2, 3 and 4.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that such detail is solved by the person skilled in the art without departing from the scope of the invention. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (18)

a) a plasticizer coating step of mixing the probiotic powder with a liquid plasticizer;
b) a seed forming step of mixing the plastic polymer powder; And
c) Curing step to form a uniform coating film
Lt; RTI ID = 0.0 &gt; of &lt; / RTI &gt; the solid composition. The method according to claim 1,
Characterized in that the plasticizer is a liquid plasticizer having a liquid or viscous property at room temperature.
A process for the preparation of a probiotic coated solid composition. 3. The method of claim 2,
Wherein the liquid plasticizer is selected from the group consisting of triethyl citrate, triacetin, diethyl phthalate, dibutyl phthalate, polysorbate 80, sorbitan monooleate, propylene glycol, propylene glycol monocaprylate, oleic acid, olive oil, carnauba wax, polyethylene Glycol 400, glycerin, diacetyl monoglyceride, glyceryl monooleate, mineral oil, and dimethyl polysiloxane.
A process for the preparation of a probiotic coated solid composition. The method according to claim 1,
Wherein at least one selected from the group consisting of a mucoadhesive substance, a pH adjuster, an antioxidant and a stabilizer is added in step a)
A process for the preparation of a probiotic coated solid composition. The method according to claim 1,
Wherein the plastic polymer has a glass transition temperature between 30 and 100 DEG C and is in a solid state at room temperature.
A process for the preparation of a probiotic coated solid composition. 6. The method of claim 5,
Characterized in that said plastic polymer is soluble or erodible in an acidic environment of pH 5 or below and exhibits low solubility or erosion and is soluble or erodible at pH 5 or above.
A process for the preparation of a probiotic coated solid composition. 6. The method of claim 5,
Wherein the plastic polymer is selected from the group consisting of ethyl cellulose, hydroxypropyl methyl cellulose acetate succinate, hydroxypropyl methyl cellulose phthalate, cellulose acetate phthalate, polyvinylacetate phthalate, shellac, methacrylic acid methyl methacrylate copolymer, methacrylic acid ethyl acrylate copolymer, (Meth) acrylic acid methyl methacrylate, trimethyl ammonium ethyl methacrylate, methyl (meth) acrylate, methyl (meth) acrylate, ethyl (meth) acrylate,
A process for the preparation of a probiotic coated solid composition. The method according to claim 1,
0.01 to 10 parts by weight of a plastic polymer is added to 1 part by weight of the probiotic powder.
A process for the preparation of a probiotic coated solid composition. 3. The method of claim 2,
And 0.01 to 10 parts by weight of a liquid plasticizer relative to 1 part by weight of the plastic polymer.
A process for the preparation of a probiotic coated solid composition. The method according to claim 1,
Characterized in that steps a) to c) are carried out at a temperature of &lt; RTI ID = 0.0 &gt; 60 C &
A process for the preparation of a probiotic coated solid composition. The method according to claim 1,
Characterized in that in step c) the temperature range within the coater is performed at 35 to &lt; RTI ID = 0.0 &gt; 45 C. &lt;
A process for the preparation of a probiotic coated solid composition. a) a plasticizer coating step of mixing the probiotic powder with a liquid plasticizer;
b) a seed forming step of mixing the plastic polymer powder;
c) a curing step to form a uniform coating film; And
d) forming an additional coating layer
Lt; RTI ID = 0.0 &gt; of &lt; / RTI &gt; the solid composition. 13. The method of claim 12,
Characterized in that the additional coating step of step d) is carried out by one or more methods selected from the group consisting of aqueous,
A process for the preparation of a probiotic coated solid composition. 13. The method of claim 12,
Wherein the additional coating step of step d) comprises charging 0.01 to 10 parts by weight of a plastic polymeric coating material based on 1 part by weight of the primary coating prepared by step c). 13. The method of claim 12,
Characterized in that the further coating step of step d) is carried out by dry coating.
A process for the preparation of a probiotic coated solid composition. 13. The method of claim 12,
The additional coating step of step d) comprises: charging and mixing 0.01 to 10 parts by weight of the plastic polymer coating material per 1 part by weight of the primary coating prepared by step c); And
(D) adding 0.01 to 10 parts by weight of a liquid plasticizer to 1 part by weight of the charged amount of the plastic polymer coating material used in step (d).
A process for the preparation of a probiotic coated solid composition. 16. A probiotic coated solid composition prepared by the method of any one of claims 1-16. 17. A product comprising the probiotic coated solid composition of claim 17.

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