An intestinal-targeted fecal microbiota transplantation capsule and its preparation method

Abstract

The present invention discloses an intestine-targeted fecal microbiota transplantation capsule and a preparation method thereof, which relates to the technical field of microbial medicine; the intestine-targeted fecal microbiota transplantation capsule is composed of fecal bacteria inclusion nanoparticles, a composite gel microcapsule material, a chitosan and calcium chloride mixed solution; the fecal bacteria inclusion nanoparticles are composed of fecal bacteria liquid, hydroxypropyl-β-cyclodextrin, zein, and xanthan gum; the composite gel microcapsule material is composed of sodium alginate, arabinoxylan, and high-methoxyl pectin; by encapsulating the fecal bacteria inclusion nanoparticles in the composite gel microcapsule material, the prepared intestine-targeted fecal microbiota transplantation capsule improves the storage stability of the flora under multiple protections, and has gastrointestinal anti-digestibility, effectively reducing the degradation and inactivation of the flora by gastric acid and digestive juices, and maintaining the relatively complete entry of the flora into the colon target site to play a role, significantly improving the effectiveness of intestinal disease treatment.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial medicine, and specifically refers to an intestinal-targeted fecal microbiota transplantation capsule and a preparation method thereof. Background Art

[0002] Fecal microbiota transplantation is a method of transplanting the microbiota in the feces of healthy people into the intestines of patients, so as to restore the normal structure and function of the intestinal microbiota of the patients and treat related intestinal diseases; fecal microbiota transplantation can effectively restore the normal intestinal microbiota balance of patients, improve the resistance of the immune system, reduce the inflammatory response, and promote the recovery of intestinal function. Fecal microbiota transplantation is highly effective in treating various intestinal diseases, especially for those patients who are ineffective with traditional treatment methods. Moreover, fecal microbiota transplantation can also reduce the recurrence rate for some recurrent intestinal diseases and has a good therapeutic effect; currently, the main routes of fecal microbiota transplantation are through colonoscopy infusion, enema, nasointestinal tube infusion, and oral fecal capsules, etc. Colonoscopy infusion, enema, and nasointestinal tube infusion are not easy to use and difficult to widely promote due to certain invasiveness, while oral capsules are a relatively convenient way of administration.

[0003] Currently, the existing technologies mainly have the following problems:

[0004] When oral capsules are stored, they are easily affected by external environments such as temperature, resulting in a decrease in the activity of the microbiota. And after entering the human body, under the influence of the upper digestive tract such as the stomach and small intestine, the activity and quantity of the microbiota are also reduced, making it difficult to reach the lower digestive tract such as the colon to play a role, which is not conducive to the effectiveness of the treatment effect. Summary of the Invention

[0005] In view of the above situation, in order to overcome the defects of the existing technologies, the present invention provides an intestinal-targeted fecal microbiota transplantation capsule, which comprises the following components in parts by weight: 20-30 parts of fecal bacteria inclusion nanoparticles, 30-50 parts of composite gel microcapsule materials, and 5-10 parts of a mixed solution of chitosan and calcium chloride.

[0006] The fecal bacteria inclusion nanoparticles comprise the following components in parts by weight: 10-20 parts of fecal bacteria solution, 5-10 parts of hydroxypropyl-β-cyclodextrin, 5-10 parts of zein, and 3-5 parts of xanthan gum.

[0007] The composite gel microcapsule materials comprise the following components in parts by weight: 10-12 parts of sodium alginate, 2-5 parts of arabinoxylan, and 4-8 parts of high-methoxyl pectin.

[0008] The preparation method of the fecal bacteria inclusion nanoparticles specifically comprises the following steps:

[0009] (1)Add 40 - 50 g of the donor's fecal sample to 250 mL of normal saline, stir evenly, and filter successively through sieves with pore sizes of 1 - 2 mm and 0.25 - 0.5 mm to remove food residues and particulate matter. Centrifuge the collected liquid at a rotational speed of 2000 - 3000 rpm for 10 - 15 min, remove the supernatant, and add a trehalose solution with a mass fraction of 2 - 5%. Stir evenly. The trehalose solution provides nutrients for the growth of the flora and is conducive to maintaining the vitality of the flora, obtaining a fecal bacteria solution;

[0010] (2)Dissolve hydroxypropyl - β - cyclodextrin and zein in 50 mL of water, and perform ultrasonic treatment. Control the ultrasonic temperature at 4°C, the ultrasonic power at 150 - 350 W, and the ultrasonic time at 5 - 15 min. The complex particles of hydroxypropyl - β - cyclodextrin and zein obtained by ultrasonic treatment are evenly dispersed and have a small particle size. Hydroxypropyl - β - cyclodextrin covers the surface of zein, forming a multi - cavity structure. Then add a xanthan gum solution with a mass fraction of 1 - 2%. The addition of xanthan gum enhances the adhesiveness, which is then conducive to the adhesion to the colon when entering the lower digestive tract. After stirring evenly, add the fecal bacteria solution described in step (1), and continue to stir while adding. The mixture is placed in a shaker for an inclusion reaction. Control the temperature at 30 - 35°C and the rotational speed at 100 - 300 rpm. The reaction solution is freeze - dried at a drying temperature of - 40°C to - 30°C for 5 - 10 h to obtain fecal bacteria inclusion nanoparticles;

[0011] Preferably, in step (1), the selection criteria for the donor are healthy adults aged 18 - 35 years old who are not pregnant, with good eating habits and lifestyles, no history of using drugs that affect the intestinal flora with antibiotics in the past 6 months, no obvious abnormalities in laboratory tests, and no history of contact with epidemic areas in the past 3 months. Strict donor selection criteria are conducive to providing diverse flora;

[0012] Preferably, in step (2), the mass fraction of the hydroxypropyl - β - cyclodextrin solution is 2 - 8%, and the mass fraction of the zein solution is 2 - 8%. Hydroxypropyl - β - cyclodextrin reduces the surface hydrophobicity of zein, which is conducive to dissolution and release in the colon. Zein enhances the shielding effect of hydroxypropyl - β - cyclodextrin and has excellent acid and heat resistance.

[0013] The present invention also provides a preparation method of an intestinal - targeted fecal microbiota transplantation capsule, which specifically includes the following steps:

[0014] S1. Add 10 - 20 g of pectin into 1000 mL of anhydrous methanol solution which contains 0.1 mol / L hydrochloric acid. Stir and disperse it for 10 - 12 h under the water bath condition of 50 - 60 °C. After filtration, wash the pectin with 95% ethanol solution until no chloride ions are detected, then dry it. By methyl esterifying the free carboxyl groups in the pectin molecules, due to the electron-withdrawing effect of adjacent carbonyl groups, the glycosidic bond has strong resistance to acidic environment, and the high methoxyl groups endow the modified pectin with strong interfacial activity, which can form a thicker interface to resist the degradation of bile salts on the flora. Thus, it has digestive stability in the gastrointestinal tract and can be used as an ideal biological carrier for colon-targeted delivery systems, obtaining high-methoxyl pectin;

[0015] S2. Dissolve sodium alginate and the high-methoxyl pectin obtained in step S1 in 100 mL of water. In the solution, the mass fraction of sodium alginate is 10 - 12%, and the mass fraction of high-methoxyl pectin is 4 - 8%. Add 2 - 5 g of arabinoxylan. As a prebiotic, arabinoxylan can promote the proliferation of beneficial intestinal bacteria, improve the quantity and structure of intestinal flora, and is not digested and absorbed by the body's digestive enzyme system. It is only degraded by specific pentosan hydrolase after entering the colon, effectively enhancing the anti-digestibility of the composite gel microcapsule material in the gastrointestinal tract. Stir to obtain the composite gel microcapsule material;

[0016] S3. Add the fecal bacteria inclusion nanoparticles into the composite gel microcapsule material obtained in step S2, stir evenly, then add 10 - 20 mL of the chitosan and calcium chloride mixed solution for crosslinking. Adjust the pH to 5.0 - 5.5, control the temperature at 30 - 35 °C, and the reaction time is 30 - 40 min. The bridging structure of calcium ions and polysaccharide molecules promotes the gelation and encapsulation of polysaccharides, increasing the stability of polysaccharides. Chitosan can form an electrostatic interaction with negatively charged sodium alginate, making the surface binding more compact, further enhancing the mechanical strength and stability, and at the same time preventing the water absorption and swelling of sodium alginate in intestinal juice, having higher gastrointestinal digestive stability. Then pour it into a mold and cool and solidify at a temperature of 10 - 20 °C for 10 - 30 min to obtain the intestinal-targeted fecal microbiota transplantation capsule;

[0017] Preferably, in step S3, in the chitosan and calcium chloride mixed solution, the mass fraction of chitosan is 0.5 - 1%, and the mass fraction of calcium chloride is 0.5 - 1%. Chitosan and calcium chloride act as a composite crosslinking agent.

[0018] The beneficial effects obtained by the present invention are as follows:

[0019] The present invention prepares an intestinal-targeted fecal microbiota transplantation capsule by encapsulating fecal microbiota inclusion nanoparticles in a composite gel microcapsule material, while the fecal microbiota inclusion nanoparticles are embedded in the composite gel microcapsule structure. Under multiple protections, the storage stability of the microbiota is improved, and it has gastrointestinal anti-digestibility, effectively reducing the degradation and inactivation of the microbiota by gastric acid and digestive juices, and maintaining the relatively complete entry of the microbiota into the colon target site to play a role, significantly improving the effectiveness of intestinal disease treatment; in the fecal microbiota inclusion nanoparticles, hydroxypropyl-β-cyclodextrin covers the surface of zein, forming a complex with a multi-cavity structure, increasing the inclusion amount and encapsulation integrity of the fecal microbiota, being beneficial to enhancing the stability and total viability of the bacterial cells. Zein enhances the acid resistance and heat resistance of the complex, reducing both the adverse effects of high temperature on the activity of the bacterial cells and the digestive effect of gastric acid on the bacterial cells. Xanthan gum enhances the adhesion of the complex to the colon. The shielding effect of hydroxypropyl-β-cyclodextrin forms a sustained release for the release of the fecal microbiota, and also enhances the hydrophilicity of the complex, thus being beneficial to dissolving and releasing the microbiota in the colon and improving the treatment effect of intestinal diseases; in the composite gel microcapsule material, sodium alginate, arabinoxylan, and high-methoxyl pectin form a hydrogel three-dimensional network structure, which can block the influence of the external environment on the fecal microbiota inclusion nanoparticles, improving the storage stability of the bacterial cells. High-methoxyl pectin increases the digestive stability of the composite gel microcapsule material in the gastrointestinal tract. Arabinoxylan is degraded by specific pentosan hydrolases only after entering the colon, further enhancing the stability in the gastrointestinal tract and endowing colon-targeted release; the embedding of the fecal microbiota inclusion nanoparticles improves the mechanical properties of the composite gel microcapsule material, while the composite gel structure improves the dispersibility and stability of the fecal microbiota inclusion nanoparticles, protecting zein from being decomposed by digestive enzymes. Therefore, the two cooperate to further enhance the storage stability and gastrointestinal anti-digestibility, and better play the therapeutic role of fecal microbiota transplantation in intestinal diseases; the fecal microbiota inclusion nanoparticles and the composite gel microcapsule material increase the cross-linking density under the action of a composite cross-linking agent of chitosan and calcium chloride, thereby enhancing the stability of the delivery system; the present invention makes an intestinal-targeted fecal microbiota transplantation capsule with fecal microbiota inclusion nanoparticles, a composite gel microcapsule material, chitosan, and calcium chloride, which has excellent storage stability, and after entering the human body, effectively reduces the degradation of the microbiota by gastric acid and digestive juices, enters the colon target site for release to play a role, and improves the effectiveness of intestinal disease treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a scanning electron micrograph of the fecal microbiota inclusion nanoparticles prepared in Example 1 of the present invention;

[0021] Figure 2 It is a scanning electron micrograph of the composite gel microcapsule material prepared in Example 1 of the present invention;

[0022] Figure 3 It is the graph of viable bacteria counts during the storage period for Examples 1-4 and Comparative Examples 1-3 of the present invention;

[0023] Figure 4 It is the graph of viable bacteria counts after simulated gastrointestinal digestion for Examples 1-4 and Comparative Examples 1-3 of the present invention;

[0024] Figure 5 It is the graph of treatment effective rates for Examples 1-4 and Comparative Examples 1-3 of the present invention. Detailed implementation manners

[0025] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments; based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0026] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the described content can be applied to the present invention. The preferred implementation methods and materials described herein are only for illustration purposes and do not limit the content of this application.

[0027] The experimental methods in the following embodiments are all conventional methods unless otherwise specified; the test materials used in the following embodiments are all obtained from commercial channels unless otherwise specified.

[0028] The sources of the reagents used in the examples are as follows:

[0029] Chitosan CAS No: 9012-76-4, brand Innochem, product number A41618;

[0030] Calcium chloride CAS No: 10043-52-4, brand Innochem, product number A01289;

[0031] Hydroxypropyl-β-cyclodextrin CAS No: 128446-35-5, brand Innochem, product number A90470;

[0032] Zein CAS No: 9010-66-6, brand Sigma-aldrich, product number Z3625-500G;

[0033] Xanthan gum CAS No: 11138-66-2, brand Innochem, product number A07292;

[0034] Sodium alginate, CAS No: 9005-38-3, brand Innochem, product number A02570;

[0035] Arabinoxylan, CAS No: 9040-27-1, brand Yuanye, product number S33528-1g;

[0036] Pectin, CAS No: 9000-69-5, brand Tci, product number P0024;

[0037] Trehalose, CAS No: 99-20-7, brand Innochem, product number A52221;

[0038] Anhydrous methanol, CAS No: 67-56-1, brand Innochem, product number A63792;

[0039] Hydrochloric acid, CAS No: 7647-01-0, brand Innochem, product number A04558;

[0040] Anhydrous ethanol, CAS No: 64-17-5, brand Innochem, product number G00004.

[0041] Example 1

[0042] This example presents an intestinal-targeted fecal microbiota transplantation capsule, which includes the following components in parts by weight: 30 parts of fecal bacteria inclusion nanoparticles, 50 parts of composite gel microcapsule materials, and 10 parts of a chitosan and calcium chloride mixed solution.

[0043] Fecal bacteria inclusion nanoparticles include the following components in parts by weight: 20 parts of fecal bacteria solution, 10 parts of hydroxypropyl-β-cyclodextrin, 10 parts of zein, and 5 parts of xanthan gum.

[0044] Composite gel microcapsule materials include the following components in parts by weight: 12 parts of sodium alginate, 5 parts of arabinoxylan, and 8 parts of high-methoxyl pectin.

[0045] The preparation method of fecal bacteria inclusion nanoparticles specifically includes the following steps:

[0046] (1)Add 50 g of the donor's fecal sample to 250 mL of normal saline and stir evenly. The inclusion criteria for the donor are healthy adults aged 18 - 35 years old, with good eating habits and lifestyle, no history of using antibiotics that affect the intestinal flora in the past 6 months, no obvious abnormalities in laboratory tests, and no history of contact with epidemic areas in the past 3 months. Strict donor selection criteria are conducive to providing diverse flora. Filter successively through sieves with pore sizes of 1 mm and 0.25 mm to remove food residues and particulate matter. Centrifuge the collected liquid at a speed of 3000 rpm for 15 min, remove the supernatant, and add a 5% trehalose solution. Stir evenly. The trehalose solution provides nutrients for the growth of the flora and is conducive to maintaining the vitality of the flora, obtaining a fecal bacteria solution;

[0047] (2)Dissolve hydroxypropyl - β - cyclodextrin and zein in 50 mL of water. The mass fraction of the hydroxypropyl - β - cyclodextrin solution is 8%, and the mass fraction of the zein solution is 8%. Hydroxypropyl - β - cyclodextrin reduces the surface hydrophobicity of zein, which is conducive to dissolution and release in the colon. Zein enhances the shielding effect of hydroxypropyl - β - cyclodextrin and has excellent acid and heat resistance. Perform ultrasonic treatment, control the ultrasonic temperature at 4 °C, the ultrasonic power at 350 W, and the ultrasonic time at 15 min. The complex particles of hydroxypropyl - β - cyclodextrin and zein obtained by ultrasonic treatment are evenly dispersed and have a small particle size. Hydroxypropyl - β - cyclodextrin covers the surface of zein, forming a multi - cavity structure. Then add a 2% xanthan gum solution. The addition of xanthan gum enhances the adhesiveness and is thus conducive to the adhesion to the colon when entering the lower digestive tract. After stirring evenly, add the fecal bacteria solution described in step (1), and continue to stir while adding. The mixed solution is placed in a shaker for an inclusion reaction, control the temperature at 35 °C and the rotation speed at 300 rpm. The reaction solution is freeze - dried at a drying temperature of - 40 °C for 10 h to obtain fecal bacteria inclusion nanoparticles.

[0048] This example provides a preparation method of an intestinal - targeted fecal microbiota transplantation capsule, which specifically includes the following steps:

[0049] S1. Add 20 g of pectin to 1000 mL of anhydrous methanol solution, where the anhydrous methanol solution contains 0.1 mol / L of hydrochloric acid. Stir and disperse at 60 °C in a water bath for 12 h. After filtration, wash the pectin with a 95% ethanol solution until no chloride ions are detected, and then dry. By methyl - esterifying the free carboxyl groups in the pectin molecule, due to the electron - withdrawing effect of adjacent carbonyl groups, the glycosidic bond has strong resistance to acidic environments, and the high methyl - ester group endows the modified pectin with strong interfacial activity, which can form a relatively thick interface to resist the degradation of the flora by bile salts. Therefore, it has digestive stability in the gastrointestinal tract and can be used as an ideal biological carrier for a colon - targeted delivery system to obtain high - methoxyl pectin;

[0050] S2, dissolving sodium alginate and the high methoxy pectin described in step S1 in 100 mL of water, wherein the mass fraction of sodium alginate is 12%, the mass fraction of high methoxy pectin is 8%, and adding 5 g of arabinoxylan. As a prebiotic, arabinoxylan can promote the proliferation of beneficial intestinal bacteria, improve the number and structure of intestinal flora, and is not digested and absorbed by the digestive enzyme system in the body. It is degraded by specific pentosan hydrolases after entering the colon, thereby effectively enhancing the digestion resistance of the composite gel microcapsule material in the gastrointestinal tract. Stirring to obtain a composite gel microcapsule material;

[0051] S3. Add the fecal inclusion complex nanoparticles to the composite gel microcapsule material described in step S2, stir evenly, and then add 20 mL of a mixed solution of chitosan and calcium chloride for cross-linking. In the mixed solution of chitosan and calcium chloride, the mass fraction of chitosan is 1%, and the mass fraction of calcium chloride is 1%. Chitosan and calcium chloride act as composite cross-linking agents. The pH is adjusted to 5.5, the temperature is controlled at 35°C, and the reaction time is 40 minutes. The bridging structure of calcium ions and polysaccharide molecules promotes the gelation and encapsulation of polysaccharides and increases the stability of polysaccharides. Chitosan can form an electrostatic effect with negatively charged sodium alginate to make the surface more tightly bound, further enhance the mechanical strength and stability, and at the same time prevent the water absorption and expansion of sodium alginate in the small intestinal fluid, which has higher gastrointestinal digestion stability. Pour it into a mold, cool and solidify at a temperature of 10°C and a reaction time of 30 minutes to obtain an intestinal targeted fecal transplant capsule.

[0052] In this example, the prepared fecal bacteria inclusion compound nanoparticles and composite gel microcapsule materials were subjected to scanning electron microscopy to observe their microscopic morphology. Figure 1 This is a SEM image of the fecal bacteria inclusion complex nanoparticles prepared in Example 1, magnified 1000 times. Figure 2 This is a SEM image of the composite gel microcapsule material prepared in Example 1 magnified 400 times. Figure 1 The fecal bacteria inclusion compound nanoparticles prepared in this embodiment present a block structure, such as Figure 2 The composite gel microcapsule material prepared in this embodiment is a polysaccharide gel carrier.

[0053] Example 2

[0054] This embodiment proposes an intestinal targeted fecal microbiota transplantation capsule, comprising the following components in parts by weight: 20 parts of fecal microbiota inclusion complex nanoparticles, 30 parts of composite gel microcapsule materials, and 5 parts of a mixed solution of chitosan and calcium chloride.

[0055] The fecal bacteria inclusion complex nanoparticles include the following components in parts by weight: 10 parts of fecal bacteria liquid, 5 parts of hydroxypropyl-β-cyclodextrin, 5 parts of zein, and 3 parts of xanthan gum.

[0056] Composite gel microcapsule material, comprising the following components in parts by weight: 10 parts of sodium alginate, 2 parts of arabinoxylan, and 4 parts of high-methoxyl pectin.

[0057] Method for preparing fecal bacteria inclusion nanoparticles, specifically comprising the following steps:

[0058] (1) Add 40 g of fecal samples from the donor to 250 mL of normal saline and stir evenly. The inclusion criteria for the donor are healthy adults aged 18 - 35 years old, non-pregnant, with good eating habits and lifestyle, no history of using drugs that affect the intestinal flora with antibiotics in the past 6 months, no obvious abnormalities in laboratory tests, no history of contact with epidemic areas in the past 3 months. Strict donor selection criteria are beneficial to providing diverse flora. Filter successively with sieves with pore sizes of 2 mm and 0.5 mm to remove food residues and particulate matter. Centrifuge the collected liquid at a centrifugation speed of 2000 rpm for 10 min, remove the supernatant, and add a trehalose solution with a mass fraction of 2%. Stir evenly. The trehalose solution provides nutrients for the growth of the flora and is beneficial to maintaining the vitality of the flora, obtaining fecal bacteria liquid;

[0059] (2) Dissolve hydroxypropyl-β-cyclodextrin and zein in 50 mL of water. The mass fraction of the hydroxypropyl-β-cyclodextrin solution is 2%, and the mass fraction of the zein solution is 2%. Hydroxypropyl-β-cyclodextrin reduces the surface hydrophobicity of zein and is beneficial to the dissolution and release in the colon. Zein enhances the shielding effect of hydroxypropyl-β-cyclodextrin and has excellent acid resistance and heat resistance. Perform ultrasonic treatment, control the ultrasonic temperature at 4 °C, the ultrasonic power at 150 W, and the ultrasonic time at 5 min. The composite particles of hydroxypropyl-β-cyclodextrin and zein obtained by ultrasonic treatment are evenly dispersed and have a small particle size. Hydroxypropyl-β-cyclodextrin covers the surface of zein, forming a multi-chamber structure. Then add a xanthan gum solution with a mass fraction of 1%. The addition of xanthan gum enhances the adhesiveness and is thus beneficial to the adhesion to the colon when entering the lower digestive tract. After stirring evenly, add the fecal bacteria liquid described in step (1), and continue to stir while adding. Place the mixed liquid in a shaker for inclusion reaction, control the temperature at 30 °C, the rotation speed at 100 rpm. Freeze-dry the reaction solution at a drying temperature of -30 °C for 5 h to obtain fecal bacteria inclusion nanoparticles.

[0060] This example provides a method for preparing an intestinal-targeted fecal microbiota transplantation capsule, specifically comprising the following steps:

[0061] S1. Add 10 g of pectin to 1000 mL of anhydrous methanol solution which contains 0.1 mol / L hydrochloric acid. Stir and disperse it for 10 h under the condition of a 50 °C water bath. After filtration, wash the pectin with 95% ethanol solution by mass until no chloride ions are detected. Dry it. Through the methylation modification of the free carboxyl groups in the pectin molecules, due to the electron-withdrawing effect of adjacent carbonyl groups, the glycosidic bond has strong resistance to acidic environment, and the high methoxyl groups endow the modified pectin with strong interfacial activity, which can form a thicker interface to resist the degradation of bile salts on the flora. Therefore, it has digestive stability in the gastrointestinal tract and can be used as an ideal biological carrier for colon-targeted delivery systems, obtaining high-methoxyl pectin;

[0062] S2. Dissolve sodium alginate and the high-methoxyl pectin obtained in step S1 in 100 mL of water. In the solution, the mass fraction of sodium alginate is 10%, and the mass fraction of high-methoxyl pectin is 4%. Add 2 g of arabinoxylan. As a prebiotic, arabinoxylan can promote the proliferation of beneficial intestinal bacteria, improve the quantity and structure of intestinal flora, and is not digested and absorbed by the body's digestive enzyme system. It is only degraded by specific pentosan hydrolases after entering the colon, effectively enhancing the anti-digestibility of the composite gel microcapsule material in the gastrointestinal tract. Stir to obtain the composite gel microcapsule material;

[0063] S3. Add the fecal bacteria inclusion nanoparticles to the composite gel microcapsule material obtained in step S2, stir evenly, and then add 10 mL of a mixed solution of chitosan and calcium chloride for crosslinking. In the mixed solution of chitosan and calcium chloride, the mass fraction of chitosan is 0.5%, and the mass fraction of calcium chloride is 0.5%. Chitosan and calcium chloride act as a composite crosslinking agent. Adjust the pH to 5.0, control the temperature at 30 °C, and the reaction time is 30 min. The bridging structure of calcium ions and polysaccharide molecules promotes the gelation and encapsulation of polysaccharides, increasing the stability of polysaccharides. Chitosan can form an electrostatic interaction with negatively charged sodium alginate, making the surface bind more tightly, further enhancing the mechanical strength and stability, and at the same time preventing the water absorption and swelling of sodium alginate in intestinal juice, having higher gastrointestinal digestive stability. Then pour it into a mold, cool and solidify at 20 °C for 10 min to obtain an intestinal-targeted fecal microbiota transplantation capsule.

[0064] Example 3

[0065] This example proposes an intestinal-targeted fecal microbiota transplantation capsule, which includes the following components in parts by weight: 25 parts of fecal bacteria inclusion nanoparticles, 40 parts of composite gel microcapsule material, and 7.5 parts of a mixed solution of chitosan and calcium chloride.

[0066] The fecal bacteria inclusion nanoparticles include the following components in parts by weight: 15 parts of fecal bacteria solution, 7.5 parts of hydroxypropyl-β-cyclodextrin, 7.5 parts of zein, and 4 parts of xanthan gum.

[0067] The composite gel microcapsule material comprises the following components in parts by weight: 11 parts of sodium alginate, 3.5 parts of arabinoxylan, and 6 parts of high-methoxyl pectin.

[0068] A preparation method of fecal bacteria inclusion nanoparticles specifically comprises the following steps:

[0069] (1) Add 45 g of the fecal sample of the donor to 250 mL of normal saline and stir evenly. The selection criteria for the donor are healthy adults aged 18 - 35 years old, with good eating habits and lifestyles, no history of drug use affecting the intestinal flora with antibiotics in the past 6 months, no obvious abnormalities in laboratory tests, no history of contact with epidemic areas in the past 3 months. Strict donor selection criteria are conducive to providing diverse flora. Filter successively with sieves with pore sizes of 1.5 mm and 0.3 mm to remove food residues and particulate matter. Centrifuge the collected liquid at a centrifugation speed of 2500 rpm for 12.5 min, remove the supernatant, and add a trehalose solution with a mass fraction of 3.5%. Stir evenly. The trehalose solution provides nutrients for the growth of the flora and is conducive to maintaining the vitality of the flora, obtaining fecal bacteria liquid;

[0070] (2) Dissolve hydroxypropyl-β-cyclodextrin and zein in 50 mL of water. The mass fraction of the hydroxypropyl-β-cyclodextrin solution is 5%, and the mass fraction of the zein solution is 5%. Hydroxypropyl-β-cyclodextrin reduces the surface hydrophobicity of zein and is conducive to dissolution and release in the colon. Zein enhances the shielding effect of hydroxypropyl-β-cyclodextrin and has excellent acid resistance and heat resistance. Perform ultrasonic treatment, control the ultrasonic temperature at 4 °C, the ultrasonic power at 250 W, and the ultrasonic time at 10 min. The composite particles of hydroxypropyl-β-cyclodextrin and zein obtained by ultrasonic treatment are evenly dispersed and have a small particle size. Hydroxypropyl-β-cyclodextrin covers the surface of zein, forming a multi-cavity structure. Then add a xanthan gum solution with a mass fraction of 1.5%. The addition of xanthan gum enhances the adhesiveness and is thus conducive to the adhesion to the colon when entering the lower digestive tract. After stirring evenly, add the fecal bacteria liquid described in step (1), and continue to stir while adding. Put the mixed liquid into a shaker for inclusion reaction, control the temperature at 32.5 °C, the rotation speed at 200 rpm. Freeze-dry the reaction solution at a drying temperature of -35 °C for 7.5 h to obtain fecal bacteria inclusion nanoparticles.

[0071] This embodiment provides a preparation method of an intestine-targeted fecal microbiota transplantation capsule, specifically comprising the following steps:

[0072] S1. Add 15 g of pectin to 1000 mL of anhydrous methanol solution, where the anhydrous methanol solution contains 0.1 mol / L of hydrochloric acid. Stir and disperse it for 11 h under the condition of a 55°C water bath. After filtration, wash the pectin with 95% ethanol solution by mass until no chloride ions are detected, and then dry it. Through the methylation modification of the free carboxyl groups in the pectin molecules, due to the electron-withdrawing effect of adjacent carbonyl groups, the glycosidic bond has strong resistance to acidic environments, and the high methoxyl groups endow the modified pectin with strong interfacial activity, which can form a thicker interface to resist the degradation of bile salts on the flora. Therefore, it has digestive stability in the gastrointestinal tract and can be used as an ideal biological carrier for colon-targeted delivery systems, obtaining high-methoxyl pectin;

[0073] S2. Dissolve sodium alginate and the high-methoxyl pectin obtained in step S1 in 100 mL of water. In the solution, the mass fraction of sodium alginate is 11%, and the mass fraction of high-methoxyl pectin is 6%. Add 3.5 g of arabinoxylan. As a prebiotic, arabinoxylan can promote the proliferation of beneficial intestinal bacteria, improve the quantity and structure of the intestinal flora, and is not digested and absorbed by the body's digestive enzyme system. It is only degraded by specific pentosan hydrolases after entering the colon, effectively enhancing the anti-digestibility of the composite gel microcapsule material in the gastrointestinal tract. Stir to obtain the composite gel microcapsule material;

[0074] S3. Add the fecal bacteria inclusion nanoparticles to the composite gel microcapsule material obtained in step S2, stir evenly, and then add 15 mL of a chitosan and calcium chloride mixed solution for crosslinking. In the chitosan and calcium chloride mixed solution, the mass fraction of chitosan is 0.75%, and the mass fraction of calcium chloride is 0.75%. Chitosan and calcium chloride act as a composite crosslinking agent. Adjust the pH to 5.3, control the temperature at 32.5°C, and the reaction time at 35 min. The bridging structure of calcium ions and polysaccharide molecules promotes the gelation and encapsulation of polysaccharides, increasing the stability of polysaccharides. Chitosan can form an electrostatic interaction with negatively charged sodium alginate, making the surface bind more tightly, further enhancing the mechanical strength and stability, and at the same time preventing the water absorption and swelling of sodium alginate in intestinal juice, having higher gastrointestinal digestive stability. Then pour it into a mold and cool and solidify at a temperature of 15°C for 20 min to obtain an intestinal-targeted fecal microbiota transplantation capsule.

[0075] Example 4

[0076] This example presents an intestinal-targeted fecal microbiota transplantation capsule, which includes the following components in parts by weight: 30 parts of fecal bacteria inclusion nanoparticles, 50 parts of composite gel microcapsule material, and 5 parts of chitosan and calcium chloride mixed solution.

[0077] The fecal bacteria inclusion nanoparticles include the following components in parts by weight: 20 parts of fecal bacteria solution, 10 parts of hydroxypropyl-β-cyclodextrin, 10 parts of zein, and 5 parts of xanthan gum.

[0078] Composite gel microcapsule material, comprising the following components in parts by weight: 12 parts of sodium alginate, 5 parts of arabinoxylan, and 8 parts of high-methoxyl pectin.

[0079] Preparation method of fecal bacteria inclusion nanoparticles, specifically comprising the following steps:

[0080] (1) Add 50 g of fecal samples from donors to 250 mL of normal saline and stir evenly. The inclusion criteria for the donors are healthy adults aged 18 - 35 years old, non-pregnant, with good eating habits and lifestyles, no history of using drugs that affect the intestinal flora with antibiotics in the past 6 months, no obvious abnormalities in laboratory tests, no history of contact with epidemic areas in the past 3 months. Strict donor selection criteria are conducive to providing diverse flora. Filter successively with sieves with pore sizes of 1 mm and 0.25 mm to remove food residues and particulate matter. Centrifuge the collected liquid at a centrifugation speed of 3000 rpm for 10 min, remove the supernatant, and add a 5% trehalose solution by mass fraction and stir evenly. The trehalose solution provides nutrients for the growth of the flora and is conducive to maintaining the vitality of the flora, obtaining fecal bacteria liquid;

[0081] (2) Dissolve hydroxypropyl-β-cyclodextrin and zein in 50 mL of water. The mass fraction of the hydroxypropyl-β-cyclodextrin solution is 8%, and the mass fraction of the zein solution is 8%. Hydroxypropyl-β-cyclodextrin reduces the surface hydrophobicity of zein and is conducive to dissolution and release in the colon. Zein enhances the shielding effect of hydroxypropyl-β-cyclodextrin and has excellent acid resistance and heat resistance. Perform ultrasonic treatment, control the ultrasonic temperature at 4 °C, the ultrasonic power at 350 W, and the ultrasonic time at 5 min. The composite particles of hydroxypropyl-β-cyclodextrin and zein obtained by ultrasonic treatment are evenly dispersed and have a small particle size. Hydroxypropyl-β-cyclodextrin covers the surface of zein, forming a multi-cavity structure. Then add a 2% xanthan gum solution by mass fraction. The addition of xanthan gum enhances the adhesiveness and is thus conducive to the adhesion to the colon when entering the lower digestive tract. After stirring evenly, add the fecal bacteria liquid described in step (1), and continue to stir while adding. The mixed solution is placed in a shaker for inclusion reaction, control the temperature at 35 °C and the rotation speed at 300 rpm. The reaction solution is freeze-dried, the drying temperature is -40 °C, and the drying time is 5 h, obtaining fecal bacteria inclusion nanoparticles.

[0082] This embodiment provides a preparation method of an intestinal-targeted fecal microbiota transplantation capsule, specifically comprising the following steps:

[0083] S1. Add 20 g of pectin to 1000 mL of anhydrous methanol solution, where the anhydrous methanol solution contains 0.1 mol / L of hydrochloric acid. Stir and disperse it in a water bath at 60 °C for 10 h. After filtration, wash the pectin with ethanol solution with a mass fraction of 95% until no chloride ions are detected. Dry it. Through the methylation modification of the free carboxyl groups in the pectin molecules, due to the electron-withdrawing effect of adjacent carbonyl groups, the glycosidic bond has strong resistance to acidic environments, and the high methoxyl groups endow the modified pectin with strong interfacial activity, which can form a thicker interface to resist the degradation of bile salts on the flora. Therefore, it has digestive stability in the gastrointestinal tract and can be used as an ideal biological carrier for colon-targeted delivery systems to obtain high-methoxyl pectin;

[0084] S2. Dissolve sodium alginate and the high-methoxyl pectin described in step S1 in 100 mL of water. In the solution, the mass fraction of sodium alginate is 12%, and the mass fraction of high-methoxyl pectin is 8%. Add 5 g of arabinoxylan. As a prebiotic, arabinoxylan can promote the proliferation of beneficial intestinal bacteria, improve the quantity and structure of the intestinal flora, and is not digested and absorbed by the body's digestive enzyme system. It is only degraded by specific pentosan hydrolases after entering the colon, effectively enhancing the anti-digestibility of the composite gel microcapsule material in the gastrointestinal tract. Stir to obtain the composite gel microcapsule material;

[0085] S3. Add the fecal bacteria inclusion nanoparticles to the composite gel microcapsule material described in step S2, stir evenly, and then add 10 mL of a chitosan and calcium chloride mixed solution for crosslinking. In the chitosan and calcium chloride mixed solution, the mass fraction of chitosan is 0.5%, and the mass fraction of calcium chloride is 0.5%. Chitosan and calcium chloride act as a composite crosslinking agent. Adjust the pH to 5.5, control the temperature at 35 °C, and the reaction time is 30 min. The bridging structure of calcium ions and polysaccharide molecules promotes the gelation and encapsulation of polysaccharides, increasing the stability of polysaccharides. Chitosan can form an electrostatic interaction with negatively charged sodium alginate, making the surface bind more tightly, further enhancing the mechanical strength and stability, and at the same time preventing the water absorption and swelling of sodium alginate in intestinal juice, with higher gastrointestinal digestive stability. Then pour it into a mold and cool and solidify at a temperature of 10 °C for 10 min to obtain the intestinal-targeted fecal microbiota transplantation capsule.

[0086] Comparative Example 1

[0087] This comparative example provides an intestinal-targeted fecal microbiota transplantation capsule, which is different from Example 1 in that the fecal bacteria inclusion nanoparticles do not contain hydroxypropyl-β-cyclodextrin, zein, and xanthan gum; the preparation method of the fecal bacteria inclusion nanoparticles does not include step (2); the preparation method of the intestinal-targeted fecal microbiota transplantation capsule is the same as that of Example 1.

[0088] Comparative Example 2

[0089] This comparative example provides an intestine-targeted fecal microbiota transplantation capsule, which is different from that of Example 1 in that the intestine-targeted fecal microbiota transplantation capsule does not contain the composite gel microcapsule material; the preparation method of the fecal microbiota inclusion complex nanoparticles is the same as that of Example 1; the preparation method of the intestine-targeted fecal microbiota transplantation capsule does not include steps S1 and S2.

[0090] Comparative Example 3

[0091] This comparative example provides an intestine-targeted fecal microbiota transplantation capsule, which is different from that of Example 1 in that the intestine-targeted fecal microbiota transplantation capsule does not contain hydroxypropyl-β-cyclodextrin, zein, xanthan gum, and the composite gel microcapsule material; the preparation method of the fecal microbiota inclusion complex nanoparticles does not include step (2); the preparation method of the intestine-targeted fecal microbiota transplantation capsule does not include steps S1 and S2.

[0092] Experimental Example 1

[0093] Storage stability experiment

[0094] Test samples: The intestine-targeted fecal microbiota transplantation capsules prepared in Examples 1-4 and Comparative Examples 1-3.

[0095] Test method: 5 g of the test samples were collected in sterile glass bottles and stored in an environment at 25 °C for 8 w. Samples were taken every 2 w, and the viable cell count (lgCFU / g) in the samples was determined by the plate counting method at the 0th, 2nd, 4th, 6th, and 8th w.

[0096] Figure 3Graph of viable bacteria counts during the storage period for Examples 1-4 and Comparative Examples 1-3; as shown in the figure, the viable bacteria counts of Examples 1-4 at 0, 2, 4, 6, and 8 weeks were 9.5-10 lgCFU / g, 9.2-9.9 lgCFU / g, 8.9-9.8 lgCFU / g, 8.6-9.7 lgCFU / g, and 8.2-9.5 lgCFU / g, respectively, all >8 lgCFU / g, indicating good storage stability; the viable bacteria counts of Comparative Examples 1-3 at 0, 2, 4, 6, and 8 weeks were 9.5-10 lgCFU / g, 8.3-9.4 lgCFU / g, 6.9-8.6 lgCFU / g, 5.4-7.6 lgCFU / g, and 3.8-6.5 lgCFU / g, respectively, and the viable bacteria counts at the 8th week were all ≤6.5 lgCFU / g, indicating poor storage stability; the fecal bacteria inclusion complex nanoparticles of Comparative Example 1 did not contain hydroxypropyl-β-cyclodextrin, zein, and xanthan gum, and could not form a heat-resistant multi-chamber structure complex, nor was it conducive to improving the mechanical properties of the composite gel microcapsule material, weakening the multiple protection effects on fecal bacteria and resulting in poor storage stability; the intestinal-targeted fecal microbiota transplantation capsules of Comparative Example 2 did not contain the composite gel microcapsule material and could not form a hydrogel three-dimensional network structure, and thus could not block the adverse effects of the external environment on the fecal bacteria inclusion complex nanoparticles, resulting in poor storage stability; the intestinal-targeted fecal microbiota transplantation capsules of Comparative Example 3 did not contain hydroxypropyl-β-cyclodextrin, zein, xanthan gum, and the composite gel microcapsule material, neither had an inclusion structure nor a hydrogel structure, and could not form a multiple protection barrier for fecal bacteria, resulting in poor storage stability.

[0097] Experimental Example 2

[0098] Gastrointestinal anti-digestibility experiment

[0099] Test samples: Intestinal-targeted fecal microbiota transplantation capsules prepared in Examples 1-4 and Comparative Examples 1-3.

[0100] Test method: Preparation of simulated gastric digestive fluid: Add pepsin at 3.2 mg / mL to a mixed solution containing 2 mg / mL NaCl and 7 mL / L HCl, and adjust its pH to 2.0 for use;

[0101] Preparation of simulated intestinal digestive fluid: Weigh 36.7 g of calcium chloride and 218.7 g of sodium chloride, dissolve them in distilled water and make up the volume to 1 L for use;

[0102] Add 5 g of the test sample to 9.9 mL of simulated gastric digestive fluid, and place it in a water bath at 37 °C and 100 r / min for shaking for 2 h. After gastric digestion, adjust the pH value of the mixture to 7.0 with 0.25 mol / L NaOH. Add 1.5 mL of simulated intestinal digestive fluid, 180 mg of bile salt (dissolved in 3.5 mL of 5 mmol / L PBS buffer, pH = 7.0), 60 mg of lipase, and 60 mg of trypsin (dissolved in 2.5 mL of 5 mmol / L PBS buffer, pH = 7.0). Continue to place the mixture in a water bath at 37 °C and 100 r / min for shaking for 2 h. Take the digestive fluids after gastric and intestinal digestions respectively, add a 0.5% sodium citrate solution to dissolve the capsules, dilute with normal saline, and coat them on MRS medium. After aerobic culture at 37 °C for 48 h, perform colony counting (lgCFU / g).

[0103] Figure 4 Figure showing the viable cell counts after simulated gastrointestinal digestion for Examples 1-4 and Comparative Examples 1-3; as shown in the figure, the viable cell counts after simulated gastrointestinal digestion for Examples 1-4 are 8.6 - 9.7 lgCFU / g and 8.3 - 9.4 lgCFU / g respectively, indicating strong gastrointestinal anti-digestibility; the viable cell counts after simulated gastrointestinal digestion for Comparative Examples 1-3 are 5.2 - 7.6 lgCFU / g and 5.0 - 7.3 lgCFU / g respectively, indicating weak gastrointestinal anti-digestibility; the fecal bacteria inclusion complex nanoparticles in Comparative Example 1 do not contain hydroxypropyl-β-cyclodextrin, zein, and xanthan gum, cannot form inclusion of fecal bacteria, do not have sustained release property, and cannot exert the acid resistance of zein, thus increasing the release in the gastrointestinal tract and resulting in weak gastrointestinal anti-digestibility; the intestinal-targeted fecal microbiota transplantation capsules in Comparative Example 2 do not contain composite gel microcapsule materials, lack the resistance of the gel structure composed of polysaccharide molecules to gastrointestinal digestion, do not have colon-targeted release property, and at the same time, cannot protect zein, increasing the decomposition effect of digestive enzymes on it, resulting in weak gastrointestinal anti-digestibility; the intestinal-targeted fecal microbiota transplantation capsules in Comparative Example 3 do not contain hydroxypropyl-β-cyclodextrin, zein, xanthan gum, and composite gel microcapsule materials, cannot embed the fecal bacteria inclusion complex nanoparticles into the composite gel microcapsule materials, and thus cannot synergistically exert anti-digestibility against the gastrointestinal tract, resulting in weak gastrointestinal anti-digestibility.

[0104] Experimental Example 3

[0105] Therapeutic efficacy experiment

[0106] Test samples: The intestinal-targeted fecal microbiota transplantation capsules prepared in Examples 1-4 and Comparative Examples 1-3.

[0107] Test method: 140 patients with intestinal diseases suffering from slow transit constipation of different degrees, aged 30 - 60 years old, were selected. According to the order of the patients' admission time for treatment, the random number table method was used to determine the patients' enrollment. They were divided into 7 groups, with 20 patients in each group. Each group took the corresponding test sample. The administration method was once in the morning and once in the evening, one pill each time. After taking it for 90 days, the effects were observed. At the same time, the treatment effect was judged according to the following criteria, and the treatment effective rate (%) was calculated. Specifically:

[0108] Effective: The constipation symptoms completely disappeared. The constipation symptoms changed from severe to moderate, from moderate to mild, and from mild to normal. Specifically, the frequency of spontaneous defecation increased, ≥3 times per week, and abdominal distension and abdominal pain were relieved to varying degrees;

[0109] Ineffective: The constipation symptoms did not improve significantly. Specifically, the frequency of spontaneous defecation did not increase, <3 times per week, and abdominal distension and abdominal pain were not relieved.

[0110] The formula for calculating the treatment effective rate is as follows:

[0111] Treatment effective rate (%) = Number of effective patients / 20 × 100%

[0112] Figure 5 It is the graph of the treatment effective rate results of Examples 1 - 4 and Comparative Examples 1 - 3; as shown in the figure, the treatment effective rate of Examples 1 - 4 is 80 - 95%, indicating good treatment effectiveness; the treatment effective rate of Comparative Examples 1 - 3 is 45 - 70%, indicating poor treatment effectiveness; the fecal bacteria inclusion complex nanoparticles in Comparative Example 1 do not contain hydroxypropyl-β-cyclodextrin, zein, and xanthan gum, and cannot form a complex by encapsulating fecal bacteria, weakening the dual protection effect with the composite gel microcapsule material, which is not conducive to maintaining the activity of bacteria in the gastrointestinal tract, reducing the quantity and vitality of the effective flora entering the colon, resulting in poor treatment effectiveness; the intestinal-targeted fecal microbiota transplantation capsules in Comparative Example 2 do not contain the composite gel microcapsule material and do not have colon targeting, which is not conducive to the release of fecal bacteria inclusion complex nanoparticles in the colon, resulting in poor treatment effectiveness; the intestinal-targeted fecal microbiota transplantation capsules in Comparative Example 3 do not contain hydroxypropyl-β-cyclodextrin, zein, xanthan gum, and the composite gel microcapsule material, and cannot form an inclusion complex and a protective barrier of the gel structure for fecal bacteria, and cannot maintain the stability of bacteria in the gastrointestinal tract, and cannot be targeted and released at the colon site, resulting in a significant reduction in the quantity and activity of the flora entering the colon, resulting in poor treatment effectiveness.

[0113] The above experimental results show that the storage stability, gastrointestinal anti-digestibility and therapeutic efficacy of Examples 1-4 of the present invention are significantly better than those of the samples of Comparative Examples 1-3. Among them, Example 1 using the fecal bacteria inclusion complex nanoparticles and the composite gel microcapsule material has better storage stability, gastrointestinal anti-digestibility and therapeutic efficacy. The fecal bacteria inclusion complex nanoparticles have a multi-cavity structure complex, which increases the inclusion amount and encapsulation integrity of fecal bacteria, is beneficial to enhancing the stability and total viability of the bacteria. Zein enhances the acid resistance and heat resistance of the complex, reduces the adverse effects of gastric acid on the bacteria, xanthan gum enhances the adhesion of the complex to the colon, and the shielding effect of hydroxypropyl-β-cyclodextrin forms a sustained release for the release of fecal bacteria, which is beneficial to dissolve and release the flora in the colon, and improves the therapeutic effect of intestinal diseases; the composite gel microcapsule material forms a hydrogel three-dimensional network structure, which can block the influence of the external environment on the fecal bacteria inclusion complex nanoparticles, improves the storage stability of the bacteria, high-methoxyl pectin increases the digestive stability of the composite gel microcapsule material in the gastrointestinal tract, arabinoxylan is degraded by specific pentosan hydrolases only after entering the colon, further enhancing the stability in the gastrointestinal tract and endowing colon-targeted release properties.

[0114] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made in these embodiments without departing from the principles and spirit of the present invention.

[0115] The present invention and its embodiments have been described above. Such description is not restrictive. What is shown in the drawings is only one of the embodiments of the present invention, and the actual application is not limited thereto. Generally speaking, if those of ordinary skill in the art are inspired by it and design similar ways and embodiments to this technical solution without creative efforts without departing from the purpose of the present invention, they should all fall within the protection scope of the present invention.

Claims

1. An intestinal targeted fecal microbiota transplantation capsule, characterized in that: The intestinal targeted fecal microbiota transplantation capsule comprises the following components in parts by weight: 20-30 parts of fecal microbiota inclusion nanoparticles, 30-50 parts of composite gel microcapsule materials, and 5-10 parts of chitosan and calcium chloride mixed solution; the fecal microbiota inclusion nanoparticles comprise the following components in parts by weight: 10-20 parts of fecal microbiota liquid, 5-10 parts of hydroxypropyl-β-cyclodextrin, 5-10 parts of zein, and 3-5 parts of xanthan gum; the composite gel microcapsule material comprises the following components in parts by weight: 10-12 parts of sodium alginate, 2-5 parts of arabinoxylan, and 4-8 parts of high methoxyl pectin; The method for preparing the fecal bacteria inclusion compound nanoparticles specifically comprises the following steps: (1) Add 40-50 g of the donor's fecal sample to 250 mL of normal saline, stir evenly, and filter using sieves with pore sizes of 1-2 mm and 0.25-0.5 mm in sequence to remove food residues and particulate matter. Centrifuge the collected liquid at a speed of 2000-3000 rpm for 10-15 min, remove the supernatant, add 2-5% mass fraction of trehalose solution, and stir evenly to obtain fecal bacteria liquid; (2) Dissolve hydroxypropyl-β-cyclodextrin and zein in 50 mL of water, perform ultrasonic treatment, control the ultrasonic temperature at 4°C, the ultrasonic power at 150-350 W, and the ultrasonic time at 5-15 min, then add a xanthan gum solution with a mass fraction of 1-2%, stir evenly, and then add the fecal bacteria solution described in step (1), continue stirring while adding, and place the mixed solution in a shaker for inclusion reaction, control the temperature at 30-35°C, the rotation speed at 100-300 rpm, and freeze-dry the reaction solution at a drying temperature of -40°C to -30°C for 5-10 h to obtain fecal bacteria inclusion complex nanoparticles; The preparation method of the intestinal targeted fecal microbiota transplantation capsule specifically comprises the following steps: S1. Add 10-20 g of pectin to 1000 mL of anhydrous methanol solution containing 0.1 mol / L of hydrochloric acid, stir and disperse in a water bath at 50-60° C. for 10-12 h, filter, wash the pectin with a 95% ethanol solution until no chloride ions are detected, and dry to obtain high methoxy pectin. S2, dissolving sodium alginate and the high methoxy pectin described in step S1 in 100 mL of water, wherein the mass fraction of sodium alginate is 10-12%, the mass fraction of high methoxy pectin is 4-8%, adding 2-5 g of arabinoxylan, stirring, and obtaining a composite gel microcapsule material; S3. Add the fecal bacteria inclusion complex nanoparticles to the composite gel microcapsule material described in step S2, stir evenly, then add 10-20 mL of a mixed solution of chitosan and calcium chloride for cross-linking, adjust the pH to 5.0-5.5, control the temperature to 30-35°C, and react for 30-40 min. Pour the mixture into a mold, cool and solidify at a temperature of 10-20°C for 10-30 min to obtain an intestinal targeted fecal bacteria transplant capsule.

2. A method for preparing the intestinal targeted fecal microbiota transplantation capsule according to claim 1, characterized in that: The specific steps include: S1. Add 10-20 g of pectin to 1000 mL of anhydrous methanol solution containing 0.1 mol / L of hydrochloric acid, stir and disperse in a water bath at 50-60° C. for 10-12 h, filter, wash the pectin with a 95% ethanol solution until no chloride ions are detected, and dry to obtain high methoxy pectin. S2, dissolving sodium alginate and the high methoxy pectin described in step S1 in 100 mL of water, wherein the mass fraction of sodium alginate is 10-12%, the mass fraction of high methoxy pectin is 4-8%, adding 2-5 g of arabinoxylan, stirring, and obtaining a composite gel microcapsule material; S3, adding the fecal bacteria inclusion complex nanoparticles to the composite gel microcapsule material described in step S2, stirring evenly, then adding 10-20 mL of a mixed solution of chitosan and calcium chloride for cross-linking, adjusting the pH to 5.0-5.5, controlling the temperature to 30-35°C, the reaction time to 30-40 min, then pouring into a mold, cooling and curing at a temperature of 10-20°C for 10-30 min, and obtaining an intestinal targeted fecal bacteria transplantation capsule; The method for preparing the fecal bacteria inclusion compound nanoparticles specifically comprises the following steps: (1) Add 40-50 g of the donor's fecal sample to 250 mL of normal saline, stir evenly, and filter using sieves with pore sizes of 1-2 mm and 0.25-0.5 mm in sequence to remove food residues and particulate matter. Centrifuge the collected liquid at a speed of 2000-3000 rpm for 10-15 min, remove the supernatant, add 2-5% mass fraction of trehalose solution, and stir evenly to obtain fecal bacteria liquid; (2) Dissolve hydroxypropyl-β-cyclodextrin and zein in 50 mL of water, perform ultrasonic treatment, control the ultrasonic temperature at 4°C, the ultrasonic power at 150-350 W, and the ultrasonic time at 5-15 min, then add a xanthan gum solution with a mass fraction of 1-2%, stir evenly, and then add the fecal bacteria solution described in step (1), continue stirring while adding, and place the mixed solution in a shaker for inclusion reaction, control the temperature at 30-35°C, the rotation speed at 100-300 rpm, and freeze-dry the reaction solution at a drying temperature of -40°C to -30°C for 5-10 h to obtain fecal bacteria inclusion complex nanoparticles.

3. The method for preparing the intestinal targeted fecal microbiota transplantation capsule according to claim 2, characterized in that: In step S3, in the mixed solution of chitosan and calcium chloride, the mass fraction of chitosan is 0.5-1%, and the mass fraction of calcium chloride is 0.5-1%.

4. The method for preparing the intestinal targeted fecal microbiota transplantation capsule according to claim 3, characterized in that: In step (1), the inclusion criteria for the donors are: healthy non-pregnant adults aged 18-35 years, with good eating habits and lifestyle, no history of antibiotic use that affects intestinal flora in the past 6 months, no obvious abnormalities in laboratory tests, and no history of contact with epidemic areas in the past 3 months.

5. The method for preparing the intestinal targeted fecal microbiota transplantation capsule according to claim 4, characterized in that: In step (2), the mass fraction of the hydroxypropyl-β-cyclodextrin solution is 2-8%, and the mass fraction of the zein solution is 2-8%.

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