KR102171287B1 - Composition for delivery of probiotics in intestine and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a composition for delivery of intestinal microorganisms and a method for producing the same, according to the present invention, a pH-sensitive inulin polymer into which a carboxyl group is introduced supports probiotics to reduce pH due to gastric acid, and to prepare probiotics from various nutrient-degrading enzymes in the digestive intestine. The composition of the present invention comprising a pH-sensitive inulin polymer and probiotics can be used for oral administration of humans and livestock, and can improve the delivery rate and storage properties in the intestine, and can be safely delivered to the intestine. It can contribute to the stabilization of the flora and enhancing the intestinal immunity. In addition, the composition has an advantage of providing convenience and economics to not only producers but also livestock farmers to be fed by adding to feed, since the composition can increase shelf life by improving shelf life in addition to the protective effect of probiotics.

Description

Composition for delivery of probiotics in intestine and manufacturing method thereof

The present invention relates to a composition for delivery of intestinal probiotics and a method for preparing the same.

Due to the ban on the use of antibiotics, there is a need to develop an effective antibiotic substitute, and interest in the use of probiotics is increasing recently. This is because probiotics are generally considered safe and present health benefits to the host. Probiotics are used as a substitute for antibiotics and as an anti-inflammatory agent. Overdose of antibiotics and synthetic antibiotics can cause side effects such as the production of antibiotic-resistant bacteria and antibiotic-related diarrhea. Probiotics produce antibacterial molecules (e.g., lactic acid and bacteriocin) and enzymes, and as a result, antimicrobial effect against pathogens, suppression of pathogen colonization, immune system regulation, and nutrient absorption enhancement, are in the spotlight as an effective antibiotic alternative. Receiving. Among probiotics, Lactobacillus is known to exhibit excellent antibacterial activity against Salmonella spp. and Escherichia coli., which are major pathogens of livestock. Lactobacillus inhibits weight loss, improves feed intake, and improves the ability of animals to grow. Studies have been reported that Lactobacillus reuteri isolated from pig feces showed a high antibacterial effect against K88-positive E. coli and Salmonella enterica subsp.

Probiotics that provide beneficial effects to the host as an alternative to therapeutic agents and/or antibiotics have targeted the intestine as a major space for delivery. Among the many candidates, antibiotics are replaced by feeding probiotics to maintain a healthy gut microbiota based on research findings that gut microbiota is deeply associated with various diseases ranging from inflammation, obesity, diabetes, atopy, and cancer as well as its antibacterial ability. And trying to overcome antibiotic-resistant bacteria. Because the intestine has a neutral pH, the delivery time is long, and the activity of the host enzyme is low, the intestinal-specific drug delivery system can increase the bioavailability of probiotics. However, oral delivery of probiotics is very difficult because probiotics are destroyed in severe acidic conditions of the stomach or cause cell death. Therefore, the safe delivery of probiotics through the stomach and into the intestines can be an important aspect of exhibiting therapeutic effects to the host.

Recently, polymeric delivery systems have gained popularity in the delivery of biological substances, proteins, genes and chemotherapy due to their ability to deliver drugs to target sites. Among the many strategies for oral delivery of probiotics, hydroxypropyl methylcellulose phthalate (HPMCP), hydroxypropyl methyl cellulose acetated succinate, cellulose acetate phthalate (CAP). A pH-sensitive polymer such as) can support probiotics and is not released under acidic pH conditions due to deprotonated carboxylic acid, thus protecting probiotics under severe gastric conditions.

On the other hand, inulin is found in many natural resources (eg, chicory root, Jerusalem artichoke, leek, onion) and is therefore used as an industrial prebiotic source. Inulin is composed of a fructose polymer linked by a β (2 → 1) bond containing a glucosyl residue at the end of the chain. Due to the β (2 → 1) binding of inulin, the intestinal microbiota ferment inulin to produce short chain fatty acids (SCFAs), which can induce the growth of beneficial microorganisms, and the intestinal microbiota ( microbiome) and can affect the host immune system. In addition, there is an increasing interest in the use of inulin as an adjuvant or drug delivery system. Interestingly, delta inulin in particulate form showed the ability to enhance immune activity as a vaccine against influenza and hepatitis B.

The present invention protects probiotics at low pH by developing new pH-sensitive formulations such as phthalyl inulin (PI) to protect probiotics from severe gastric conditions, and allows them to be well released at neutral pH of the small intestine and at the same time storage properties. It is intended to provide a technology that improves and maintains the number of probiotics for a long period of time.

Korean Patent Registration No. 10-1370143 (registered on Feb. 26, 2014)

It is an object of the present invention to provide a composition for delivery of probiotics in the intestine and a method for preparing the same.

Another object of the present invention is to provide a formulation for oral administration containing the composition as an active ingredient.

In order to achieve the above object, the present invention is a pH-sensitive polymer formed by introducing a carboxyl group into the hydroxyl group of a polysaccharide; And probiotics, wherein the probiotics are supported on a pH-sensitive polymer.

In addition, the present invention provides a formulation for oral administration containing the composition as an active ingredient.

In addition, the present invention is a first step of dissolving inulin in an organic solvent and adding an organic acid and a catalyst to prepare a pH-sensitive inulin polymer into which a carboxyl group is introduced; A second step of pulverizing the freeze-dried probiotics into fine powder; It provides a method for preparing a composition for delivery of intestinal probiotics comprising; a third step of mixing the pH-sensitive inulin polymer into which the carboxyl group of the first step is introduced and the powdered probiotics of the second step.

According to the present invention, the pH-sensitive inulin polymer into which a carboxyl group has been introduced protects probiotics from low pH caused by gastric acid and various nutrient-degrading enzymes in the digestive intestine by supporting probiotics and can be safely delivered to the intestine. The composition of the present invention comprising polymers and probiotics can be used for oral administration of humans and livestock, can improve intestinal delivery and storage properties, and can contribute to stabilization of intestinal microflora and enhancing intestinal immunity. In addition, the composition has an advantage of providing convenience and economics to not only producers but also livestock farmers to be fed by adding to feed, since the composition can increase shelf life by improving shelf life in addition to the protective effect of probiotics.

Figure 1 relates to the structure and properties of phthalyl inulin (PI), (A) shows the chemical reaction of PI and (B) NMR spectrum results of PI.
2 is a result of confirming the viability of Lactobacillus luteri (LR) according to pressure conditions after purification under different pressure conditions (5, 10 and 15 KP).
3 is a result of confirming the swelling ratio after incubating PI tablets carrying Lactobacillus luteri prepared under different pressure conditions (5, 10 and 15 KP) in simulated gastric juice (SGF) for 2 hours.
Figure 4 is a Lactobacillus luteri-supported PI purification in (A) simulated gastric juice to which pepsin is added (pH 2.0) and (B) simulated gastric juice to which pepsin is not added (pH 2.0) at 37°C for 2 hours. This is the result of confirming the viability of Lactobacillus luteri after incubation.
5 is a result of confirming the decomposition time according to the pressure conditions after culturing PI tablets carrying Lactobacillus luteri prepared under different pressure conditions in PBS (pH 6.8).
Figure 6 is a Lactobacillus luteri (A) release cells number and ( B) This is the result of checking the number of viable cells.
7 is a result of confirming the storage stability of Lactobacillus luteri after culturing PI tablets carrying Lactobacillus luteri for 6 months under a temperature condition of 4°C.

Probiotics are very difficult to have activity because they are easily killed by low pH due to gastric acid and various nutrient-degrading enzymes in the digestive intestine and are delivered alive to the intestine.The inventors of the present invention believe that when probiotics are administered orally, the delivery rate and storage properties of the digestive tract In order to increase the inulin, a pH-sensitive polymer was prepared by introducing a hydrophobic moiety having a carboxyl group (-COOH) in inulin, and a preparation capable of safely delivery to the intestine was prepared by carrying probiotics on the polymer, thereby completing the present invention.

Inulin has been widely used as a prebiotic for decades, and inulin itself is difficult to be used in oral colon drug delivery systems, but has a water-soluble property, so it is a drug delivery system for nasal, parenteral, intravenous and subcutaneous administration. Is used in Therefore, strategies for reducing the solubility of inulin in water by mixing a hydrophobic coating material such as Eudragit or by binding a hydrophobic moiety have been attempted. In the present invention, it was confirmed that the water solubility of inulin was reduced by binding a phthalyl group to inulin, and probiotics were protected under low pH conditions after purification.

Accordingly, the present invention includes a pH-sensitive polymer and probiotics formed by introducing a carboxyl group into a hydroxyl group of a polysaccharide, and the probiotics provide a composition for delivery of intestinal probiotics, characterized in that the probiotics are supported on the pH-sensitive polymer.

The polysaccharide may be selected from the group consisting of inulin, fluran, dextran, and starch, but it is specified that the polysaccharide is not limited thereto.

The carboxyl group may be introduced from an organic acid selected from the group consisting of phthalic acid, glutamic acid, succinic acid, and mercaptosuccinic acid, but is not limited thereto.

The probiotics may be selected from the group consisting of Lactobacillus luteri and Lactobacillus salivarius, but are not limited thereto.

The composition protects probiotics from low pH of gastric acid and nutrient-degrading enzymes in the digestive intestine, increases the delivery rate and storage of probiotics in the intestine, and can stabilize the intestinal microbial flora and improve intestinal immunity.

In addition, the present invention provides a formulation for oral administration containing the composition as an active ingredient.

The composition can be formulated and used in the form of oral dosage forms such as tablets, powders, granules, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injectable solutions.

In addition, the present invention is a first step of dissolving inulin in an organic solvent and adding an organic acid and a catalyst to prepare a pH-sensitive inulin polymer into which a carboxyl group is introduced; A second step of pulverizing the freeze-dried probiotics into fine powder; It provides a method for preparing a composition for delivery of intestinal probiotics comprising; a third step of mixing the pH-sensitive inulin polymer into which the carboxyl group of the first step is introduced and the powdered probiotics of the second step.

An organic solvent of the first step is N, N - may be selected from the group consisting of dimethylformamide (N, N -dimethylformamide) and dimethyl sulfoxide (dimethyl sulfoxide), and specifies a not limited thereto.

The organic acid of the first step may be selected from the group consisting of phthalic acid, glutamic acid, succinic acid, and mercaptosuccinic acid, but it is specified that the organic acid is not limited thereto.

The catalyst of the first step may be selected from the group consisting of sodium acetate and dimethylaminopyridine, but is not limited thereto.

It should be noted that the first step may be performed under nitrogen gas at 35 to 45° C. for 20 to 30 hours, but is not limited thereto.

The pH-sensitive inulin polymer into which the carboxyl group of the third step is introduced and the powdered probiotic may be mixed in a weight ratio of 1:1 to 10:1, but it is specified that the present invention is not limited thereto.

It should be noted that the composition may be administered orally, but is not limited thereto.

When the composition of the present invention is a pharmaceutical composition, for administration, it may include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above-described active ingredients. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oils.

The pharmaceutical compositions of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injectable solutions according to a conventional method. . Specifically, when formulated, it may be prepared using diluents or excipients such as fillers, weight agents, binders, wetting agents, disintegrants, and surfactants that are commonly used. Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc. in addition to the active ingredient. Further, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. It can be prepared by adding various excipients, such as wetting agents, sweetening agents, fragrances, preservatives, and the like, in addition to oral liquids and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and tasks. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like may be used. As a base for suppositories, witepsol, macrosol, Tween 61, cacao butter, laurin, glycerol gelatin, and the like can be used.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the severity of the disease, the form of the drug, and the time, but may be appropriately selected by a person skilled in the art, and the daily dosage of the composition is preferably It is 0.001 mg/kg to 50 mg/kg, and it can be administered once to several times a day as needed.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example 1: Materials

Lactobacillus reuteri LRT18 (LR, KCTC3594) was isolated by referring to a previous paper (Microb Ecol. 2017 Oct;74(3):709-721). All materials and chemicals used in the experiments were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise specified. MRS (developer: De Man, Rogosa and Sharpe) medium and MRS agar for bacterial culture were purchased from BD Difco (Sparks, MD, USA).

Example 2: Synthesis of phthalyl inulin (PI)

Phthalyl inulin was synthesized by referring to a previous paper (Sci Rep. 2018 Apr 12;8(1):5878). Briefly, 1 g of inulin (MW: 5000 g/mol) was dissolved in 5 mL of N,N -dimethylformamide , and 2.0 g of phthalic anhydride was added to the solution, 5% (w/v) sodium acetate was used as a catalyst. The reaction was carried out at 40° C. for 24 hours under nitrogen gas. Thereafter, PI was dialyzed in cold water for 24 hours, freeze-dried, and stored at -20°C until use in the experiment. The binding of the phthalyl group to the PI was confirmed by 600 MHz ¹ H-NMR spectroscopy (AVANCE600, Bruker, Germany).

Example 3: Preparation of tablets

The LR culture was collected by incubating for 24 hours in MRS medium at 37°C and centrifuging. The obtained cells were washed 3 times with a phosphate buffer solution, and suspended in 10% skim milk. Thereafter, the cells were frozen at -20°C for 12 hours and freeze-dried under reduced pressure. Freeze-dried probiotics were pulverized into fine powder and stored at 4°C until use in experiments. Tablets were prepared at room temperature by direct compression using a single press. For purification, a mixture of LR and PI (weight ratio of LR to PI = 1:1) is filled into a 4 mm diameter die and purified under different pressures of 3 to 10 kilopascals (KP) on a flat surface. Was prepared.

Example 4: Analysis of probiotic (LR) viability and digestion time of tablets

The viability of LR in tablets was analyzed in colony forming units (CFU). Briefly, the tablets were broken and dispersed in 1 mL of a phosphate buffer solution (PBS, pH 7.2), and then the serially diluted suspension was dropped onto an MRS agar plate and incubated at 37°C to count LR colonies. In addition, the tablet was transferred to 5 mL PBS (pH 6.8), and the complete disintegration time was measured.

Example 5: Analysis of the expansion ratio of tablets

After transferring the tablet to 5 mL of simulated gastric fluid (SGF), the swelling ratio was calculated using the following equation.

[Calculation 1]

Q = (Ms-Md) / Md

The expansion ratio is Q, Md is the mass of the tablet in the dried state, and Ms is the mass of the tablet in the expanded state. At the beginning of the experiment, excess water outside the tablet was removed.

Example 6: Stability analysis of tablets with or without pepsin in simulated gastric juice (SGF)

The stability analysis of the tablet was performed by referring to the previous paper (J Microbiol Biotechnol. 2017 Apr 28;27(4):739-746). Simulated gastric juice with or without pepsin (1000 units/mL) was prepared with PBS adjusted to pH 2.0 with 1 M HCl. Purified and LR powder was transferred to 5 mL of simulated gastric juice (with or without pepsin), and the viability of LR was incubated at 37°C at 100 rpm and colonies were counted at the end of the incubation time (0, 30, 60, 90, 120 minutes). And analyzed.

Example 7: Viability analysis of tablets by sequential exposure in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) medium

The cell viability of LR in the tablets sequentially exposed to simulated gastric juice and simulated intestinal fluid was analyzed by the following method. After incubating the tablets in 5 mL of simulated gastric fluid at 37°C and 100 rpm for 2 hours, the tablets were quickly transferred to 5 mL of simulated intestinal fluid (SIF) (PBS, pH 6.8), and then at 37°C and 100 rpm. Incubated for 4 hours. Viability was analyzed by counting viable cells in the supernatant as well as non-degraded tablets at each culture time.

Example 8: Tablet stability analysis

The tablets were stored at 4° C. for up to 6 months for stability analysis. Cell viability was counted monthly by the method described above.

Example 9: Statistical Analysis

Data are presented as the mean±SEM of three independent experiments. Statistical significance between PI and other groups was analyzed using one-way ANOVA and Tukey's test (* p <0.05, ** p <0.01, *** p <0.001).

Experimental Example 1: Synthesis and Characterization of PI

In order to prepare a pH-sensitive polymer, a phthalyl group was introduced into the inulin as an ester bond between the hydroxyl of inulin and the carboxylic acid of phthalic acid. The reaction scheme of PI synthesis is shown in Figure 1A. After synthesizing PI, the degree of phthalyl group in PI was measured through ¹ H-NMR analysis.

As shown in Figure 1B, it was confirmed that the fifth proton of inulin appears at 3.8 ppm, and the proton of the phthalyl group in PI appears at 7.4 to 7.7 ppm. Based on the integration of protons in both inulin and phthalyl groups, it was confirmed that the degree of phthalyl group in PI was 36.4 mol%.

Experimental Example 2: Analysis of the effect of pressure on LR viability and purification properties

In order to analyze the viability of LR according to pressure, the viability of LR was measured after purification. As shown in Figure 2, the pressure used did not affect the viability of LR in the tablet.

In order to analyze the protective effect of LR in acidic gastric conditions, the swelling ratio of the tablet and the viability of LR were measured in simulated gastric juice. As shown in Figure 3, it was confirmed that the expansion ratio of the tablets prepared according to different pressures was very low in the simulated gastric juice. In addition, the tablet did not completely disintegrate within 2 hours under the above conditions. In particular, it showed the smallest expansion ratio in the tablet group manufactured with the highest pressure (15 KP) among the groups.

Next, the viability of LR was measured in simulated gastric juice (with or without pepsin). Tablets and free probiotics (powder) prepared at 5, 10 and 15 KP pressure were immersed in simulated gastric juice for 2 hours. As a result, as shown in FIG. 4, it was confirmed that the viability of free probiotics rapidly decreased under simulated gastric juice conditions and particularly in the presence of pepsin. However, the PI purification with LR protected the death of probiotics from simulated gastric juice with or without pepsin. The viability of LR between purified and free LR showed a significant difference in simulated gastric juice after 2 hours in the presence of pepsin (FIG. 4B ), suggesting that PI purification could protect LR under severe gastric conditions. In addition, high pressure increased the viability of LR under gastric conditions. The LR viability of the tablets prepared at 15 KP pressure was significantly higher than the free LR in the simulated gastric juice to which pepsin was not added at 30 minutes (Fig. 4A). In simulated gastric juice to which pepsin was added, the difference in viability of LR between free LR and supported LR in tablets prepared at 15 KP pressure was significantly higher at 60 minutes (Fig. 4B). Moreover, the viability of LR supported on tablets prepared at 15 KP pressure was significantly higher than LR supported on tablets prepared at 5 KP or 10 KP pressure, especially after 2 hours in the presence of pepsin, which means that the high pressure is higher than that of probiotics in the stomach. To better protect

Experimental Example 3: LR release and cell viability analysis from PI tablets carrying LR in simulated gastric juice and simulated intestinal fluid

The release of LR and cell viability from PI tablets carrying LR in simulated gastric juice and simulated intestinal fluid were analyzed by sequentially immersing the tablets in simulated gastric juice and simulated intestinal fluid. As a result of calculating the LR released from the tablet in the simulated gastric juice and the simulated intestinal fluid, as shown in Fig.6A, no viable cells were observed in the simulated gastric juice, and the tablet prepared at 5 KP and 10 KP pressure in the simulated intestinal fluid was 15 KP. It was confirmed that viable cells were released faster than tablets prepared with pressure. Tablets with higher pressure delayed the release of LR in the tablets compared to the other two tablets. However, after 5 hours, almost all of the probiotics were released in all groups of tablets.

The viability of LR was analyzed by continuously exposing the tablets to simulated gastric juice and simulated intestinal fluid. As shown in FIG. 6B, it was confirmed that the viability of LR decreased with time when the tablet was exposed to simulated gastric juice. The viability of LR was slightly changed in the simulated intestinal fluid after 5 hours, but more viable cells were maintained in tablets prepared at higher pressure than lower pressure. The viability of LR in tablets prepared at 15 KP pressure after 7 hours in simulated gastric juice and simulated intestinal fluid was significantly higher than that of the other two tablets.

In conclusion, due to the pH sensitivity of PI, it was confirmed that PI tablets protect LR from severe gastric conditions and release LR in intestinal conditions.

Experimental Example 4: LR viability analysis of PI tablets carrying LR for long-term storage

The stable viability of probiotics is a major indicator for industrialization of probiotics. Therefore, after culturing the PI tablets carrying LR for 6 months under a temperature condition of 4°C, the storage stability of LR was confirmed. This temperature was chosen because most probiotic products recommend storage in the refrigerator. As shown in Figure 7, the stability of LR for 6 months was calculated as the survival CFU per tablet. After 3 months, the free probiotics in powder form rapidly decreased, but it was confirmed that the LR viability in tablets was remarkably stable after 6 months. In particular, tablets prepared at higher pressure (10 and 15 KP) showed significantly higher viable cells at the end of 6 months than the 5 KP tablets. Overall, PI-based purification can stabilize the viability of LR, and high pressure during purification can increase the stability of cells.

As the specific parts of the present invention have been described in detail above, it is obvious that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereto for those of ordinary skill in the art. Therefore, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

The scope of the present invention is indicated by the claims to be described later, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims (14)

A pH-sensitive polymer formed by introducing a carboxyl group into the hydroxyl group of a polysaccharide; And probiotics; wherein the polysaccharide is inulin, the carboxyl group is introduced from an organic acid selected from the group consisting of phthalic acid, glutamic acid, and succinic acid, and the probiotics are supported on a pH-sensitive polymer for delivery of intestinal probiotics Tablet composition. delete delete The tablet composition for delivery of intestinal probiotics according to claim 1, wherein the probiotics are selected from the group consisting of Lactobacillus luteri and Lactobacillus salivarius. The tablet composition of claim 1, wherein the composition protects probiotics from low pH of gastric acid and nutrient decomposition enzymes in the digestive intestine. The tablet composition for delivery of intestinal probiotics according to claim 1, wherein the composition increases intestinal delivery and storage, stabilizes intestinal microflora and improves intestinal immunity. A formulation for oral administration containing the tablet composition according to any one of claims 1, 4, 5 and 6 as an active ingredient. A first step of dissolving inulin in an organic solvent and adding an organic acid and a catalyst to prepare a pH-sensitive inulin polymer into which a carboxyl group is introduced;
A second step of grinding the freeze-dried probiotics into fine powder;
A third step of mixing the pH-sensitive inulin polymer into which the carboxyl group of the first step is introduced and the powdered probiotics of the second step; and
The organic acid of the first step is a method of producing a tablet composition for delivery of intestinal probiotics, characterized in that it is selected from the group consisting of phthalic acid, glutamic acid, and succinic acid. 10. The method of claim 8, wherein the organic solvent of the first step is N, N - intestinal probiotics for transmission, characterized in that selected from the group consisting of dimethylformamide (N, N -dimethylformamide) and dimethyl sulfoxide (dimethyl sulfoxide) Method for producing a tablet composition. delete The method of claim 8, wherein the catalyst in the first step is selected from the group consisting of sodium acetate and dimethylaminopyridine. The method of claim 8, wherein the first step is performed under nitrogen gas at 35 to 45°C for 20 to 30 hours. The method for preparing a tablet composition for delivery of intestinal probiotics according to claim 8, wherein the pH-sensitive inulin polymer and powdered probiotics into which the carboxyl group of the third step is introduced are mixed in a weight ratio of 1:1 to 10:1 . The method of claim 8, wherein the composition is administered by oral administration.

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