Compositions for treating ulcerative colitis and methods of making the same

Abstract

The application discloses a composition for treating ulcerative colitis and a preparation method thereof, and belongs to the field of biological medicines, and the composition comprises 60-80 parts of a temperature-sensitive gel, 1-10 parts of a usnea extract, 10-15 parts of a plant fermentation product, 1-5 parts of butyrate and derivatives thereof, 1-10 parts of microencapsulated clostridium butyricum and 1-10 parts of a plant extract. The temperature-sensitive gel is a chitosan / sodium beta-glycerophosphate temperature-sensitive gel, the plant fermentation product is a clostridium butyricum fermentation product of zizania aquatica, nymphaea alba and cabomba caroliniana, the microencapsulated clostridium butyricum is clostridium butyricum subjected to microencapsulation and encapsulation treatment, the plant extract is an extract of coptis chinensis, radix ginseng, panax notoginseng and dracaena cantoniensis, and the butyrate and derivatives thereof are sodium butyrate. The application solves the problems of low response rate and large side effects in the prior art due to the lack of physical repair barriers, single target and the dependence on hormone and biological agent drugs. The application has the potential to prepare a medicine for treating ulcerative colitis.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to compositions for treating ulcerative colitis and methods for preparing the same. Background Technology

[0002] Ulcerative colitis (UC) is an organic disease characterized by chronic diarrhea and rectal bleeding. Approximately 30% of patients also have non-intestinal conditions such as primary sclerosing cholangitis. Conventional treatment involves oral mesalazine or mesalazine suppositories. Moderate to severe cases require oral corticosteroids or vedelizumab to relieve symptoms. However, long-term use of corticosteroids can lead to complications such as osteoporosis and diabetes, while biologics like vedelizumab increase the risk of tumors and thrombosis. These side effects severely limit the long-term use of these medications.

[0003] Furthermore, despite continuous advancements in the treatment of ulcerative colitis, the highest drug response rate is only 30-60%. Five years after diagnosis, approximately 20% of UC patients are hospitalized, and about 7% undergo colectomy. Twenty years after diagnosis, patients have a 4.5% risk of developing colon cancer, which is about 1.7 times higher than the general population, and their average lifespan is about 5 years shorter than that of people without the disease.

[0004] Therefore, there is an urgent need for a drug to treat ulcerative colitis in order to solve the above problems. Summary of the Invention

[0005] Therefore, the present invention provides a composition for treating ulcerative colitis and a method for preparing the same, in order to solve the problems of low response rate and large side effects caused by the lack of physical repair barrier, single target and dependence on hormones and biological agents in the prior art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: According to a first aspect of the invention, a composition for treating ulcerative colitis is provided, the composition comprising the following components in parts by weight: 60-80 parts of thermosensitive gel, 1-10 parts of *Opuntia ficus-indica* extract, 10-15 parts of plant ferment, 1-5 parts of butyrate and its derivatives, 1-10 parts of microencapsulated *Clostridium butyricum*, and 1-10 parts of plant extract; all components except the thermosensitive gel are dispersed in a phosphate buffer solution after preparation, and the mass components are based on the dry weight of the solute; the mass-volume ratio of the sum of the masses of all components except the thermosensitive gel to the phosphate buffer solution is (0.5-1) g: 1 mL.

[0007] Furthermore, the thermosensitive gel is a chitosan / β-glycerophosphate sodium thermosensitive gel, the plant fermentation product is the butyric acid Clostridium fermentation product of water chestnut, water caltrop and water shield, the microencapsulated butyric acid Clostridium is butyric acid Clostridium that has undergone microencapsulation treatment, the plant extract is the extract of Coptis chinensis, Codonopsis pilosula, Panax notoginseng and Bletilla striata, and the butyrate and its derivatives are sodium butyrate.

[0008] Furthermore, the mass ratio of the butyric acid clostridium fermentation products of water chestnut, water caltrop and water shield in the plant fermentation product is 2:1:1; the mass ratio of the extracts of Coptis chinensis, Codonopsis pilosula, Panax notoginseng and Bletilla striata in the plant extract is 3:3:1:3.

[0009] According to a second aspect of the present invention, a method for preparing a composition for treating ulcerative colitis is provided, the method comprising the following steps: S1. Preparation of plant fermentation products: After washing, drying and crushing the water chestnuts, water lilies and water shield into powder of 80-100 mesh, they were mixed with Clostridium butyricum suspension at a mass-to-volume ratio of 1 g: 5 mL, anaerobic incubation at 37℃ for 24-48 h, filtered and autoclaved at 121℃ for 10 min, and freeze-dried to obtain the fermented products of water chestnuts, water lilies and water shield respectively. S2. Preparation of temperature-sensitive gel solution: Under ice bath conditions, sodium β-glycerophosphate solution was slowly added dropwise to chitosan solution, pH was adjusted to 7.0-7.2, and stirred for 30-60 min to obtain thermosensitive gel solution; S3. Preparation of microencapsulated Clostridium butyricum: β-cyclodextrin was dissolved in phosphate buffer and stirred at 100℃ and 200-400rpm for 10-30min. Then, gellan gum and sodium alginate were added and stirred for another 10min to obtain microcapsule wall material solution. After autoclaving at 121℃ for 10min and cooling, it was mixed with Clostridium butyricum suspension and stirred at 50-100rpm for 30min to mix evenly. The mixture was then freeze-dried to obtain microencapsulated Clostridium butyricum. S4. Preparation of the composition: Sodium butyrate, rock tripe extract, plant extract, plant ferment, and microencapsulated Clostridium butyricum were uniformly mixed and then dispersed in phosphate buffer. The mass-to-volume ratio of each component mixture to phosphate buffer was (0.5-1) g: 1 mL, resulting in an active substance suspension. Mix the temperature-sensitive gel solution and the active substance suspension, and stir at 30-50 rpm for 10 min at room temperature to obtain the composition.

[0010] Furthermore, the preparation of the Clostridium butyricum suspension in step S1 includes the following steps: Clostridium butyricum was inoculated into RCM medium at a volume fraction of 5% and incubated anaerobically at 200 rpm and 37°C for 24-36 h. After incubation, the culture was centrifuged at 4°C, and the precipitate was washed with physiological saline and then dispersed with phosphate buffer to a viable count of 1 × 10⁻⁶. 7 -1×10 8 CFU / mL, to obtain Clostridium butyricum suspension.

[0011] Further, in step S2, the concentration of the sodium β-glycerophosphate solution is 0.56 g / mL, the chitosan solution is a chitosan-dilute hydrochloric acid solution with a concentration of 0.03 g / mL, the volume ratio of the sodium β-glycerophosphate solution to the chitosan solution is 1:3, and the degree of deacetylation of chitosan in the chitosan solution is ≥80%.

[0012] Furthermore, the viable count of the Clostridium butyricum suspension in step S3 is 1 × 10⁻⁶. 7 -1×10 8 The concentrations of β-cyclodextrin, gellan gum, and sodium alginate in the microcapsule wall material solution in phosphate buffer were 50 mg / mL, 30 mg / mL, and 30 mg / mL, respectively, and the volume ratio of the microcapsule wall material solution to the Clostridium butyricum suspension was 3:1.

[0013] According to a third aspect of the invention, the use of the composition for treating ulcerative colitis in the preparation of a medicament for treating ulcerative colitis is provided.

[0014] Furthermore, the drug is in the form of a gel, which is liquid at room temperature and gels at body temperature; the administration method is rectal administration, by pushing the gel into the sigmoid colon, where a temperature-sensitive gel is formed in situ.

[0015] The present invention has the following advantages: This invention utilizes a chitosan / β-glycerophosphate sodium thermosensitive gel to form a gel in situ within the colon. This liquid enema delivery enhances drug coverage while simultaneously creating a physical barrier to cover the ulcer surface, isolating intestinal irritants and providing a scaffold for colonic repair. Extracts from Coptis chinensis, Codonopsis pilosula, Panax notoginseng, and Bletilla striata synergistically exert anti-inflammatory, hemostatic, and healing-promoting functions, and sodium butyrate is added to supplement the nutritional source for colonic epithelial cells. Furthermore, Clostridium butyricum ferments plants high in polysaccharides and dietary fiber, such as water chestnut, water shield, and water lily, to supplement butyrate while hydrolyzing the anti-inflammatory substances into more easily absorbed and utilized small molecule compounds. In addition, this invention incorporates microencapsulated Clostridium butyricum to ensure effective delivery of live bacteria to the colon, restoring the local colonic flora and promoting the repair of colonic epithelial cells. Through the multi-target synergistic effects of physical barrier, anti-inflammatory repair, and flora regulation, the components of this invention effectively improve ulcerative colitis. Detailed Implementation

[0016] The following specific embodiments illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0017] According to a first aspect of the invention, a composition for treating ulcerative colitis is provided, the composition comprising the following components in parts by weight: 60-80 parts of thermosensitive gel, 1-10 parts of *Opuntia ficus-indica* extract, 10-15 parts of plant ferment, 1-5 parts of butyrate and its derivatives, 1-10 parts of microencapsulated *Clostridium butyricum*, and 1-10 parts of plant extract; all components except the thermosensitive gel are dispersed in a phosphate buffer solution after preparation, and the mass components are based on the dry weight of the solute; the mass-volume ratio of the sum of the masses of all components except the thermosensitive gel to the phosphate buffer solution is (0.5-1) g: 1 mL.

[0018] Among them, the thermosensitive gel is a chitosan / β-glycerophosphate sodium thermosensitive gel, the plant fermentation product is the butyric acid Clostridium fermentation product of water chestnut, water caltrop and water shield, the microencapsulated butyric acid Clostridium is butyric acid Clostridium that has been microencapsulated, the plant extract is the extract of Coptis chinensis, Codonopsis pilosula, Panax notoginseng and Bletilla striata, and the butyrate and its derivatives are sodium butyrate.

[0019] Among them, the mass ratio of butyric acid Clostridium fermentation products of water chestnut, water caltrop and water shield in the plant fermentation products was 2:1:1; the mass ratio of extracts of Coptis chinensis, Codonopsis pilosula, Panax notoginseng and Bletilla striata in the plant extracts was 3:3:1:3.

[0020] According to a second aspect of the present invention, a method for preparing a composition for treating ulcerative colitis is provided, the method comprising the following steps: S1. Preparation of plant fermentation products: After washing, drying and crushing the water chestnuts, water lilies and water shield into powder of 80-100 mesh, they were mixed with Clostridium butyricum suspension at a mass-to-volume ratio of 1 g: 5 mL, anaerobic incubation at 37℃ for 24-48 h, filtered and autoclaved at 121℃ for 10 min, and freeze-dried to obtain the fermented products of water chestnuts, water lilies and water shield respectively. S2. Preparation of temperature-sensitive gel solution: Under ice bath conditions, sodium β-glycerophosphate solution was slowly added dropwise to chitosan solution, pH was adjusted to 7.0-7.2, and stirred for 30-60 min to obtain thermosensitive gel solution; S3. Preparation of microencapsulated Clostridium butyricum: β-cyclodextrin was dissolved in phosphate buffer and stirred at 100℃ and 200-400rpm for 10-30min. Then, gellan gum and sodium alginate were added and stirred for another 10min to obtain microcapsule wall material solution. After autoclaving at 121℃ for 10min and cooling, it was mixed with Clostridium butyricum suspension and stirred at 50-100rpm for 30min to mix evenly. The mixture was then freeze-dried to obtain microencapsulated Clostridium butyricum. S4. Preparation of the composition: Sodium butyrate, rock tripe extract, plant extract, plant ferment, and microencapsulated Clostridium butyricum were uniformly mixed and then dispersed in phosphate buffer. The mass-to-volume ratio of each component mixture to phosphate buffer was (0.5-1) g: 1 mL, resulting in an active substance suspension. Mix the temperature-sensitive gel solution and the active substance suspension, and stir at 30-50 rpm for 10 min at room temperature to obtain the composition.

[0021] The preparation of the Clostridium butyricum suspension in step S1 includes the following steps: Clostridium butyricum was inoculated into RCM medium at a volume fraction of 5% and incubated anaerobically at 200 rpm and 37°C for 24-36 h. After incubation, the culture was centrifuged at 4°C, and the precipitate was washed with physiological saline and then dispersed with phosphate buffer to a viable count of 1 × 10⁻⁶. 7 -1×10 8 CFU / mL, to obtain Clostridium butyricum suspension.

[0022] In step S2, the concentration of the sodium β-glycerophosphate solution is 0.56 g / mL, the chitosan solution is a chitosan-dilute hydrochloric acid solution with a concentration of 0.03 g / mL, the volume ratio of the sodium β-glycerophosphate solution to the chitosan solution is 1:3, and the degree of deacetylation of chitosan in the chitosan solution is ≥80%.

[0023] In step S3, the viable count of the Clostridium butyricum suspension is 1×10⁻⁶. 7 -1×10 8 The concentrations of β-cyclodextrin, gellan gum, and sodium alginate in the microcapsule wall material solution in phosphate buffer were 50 mg / mL, 30 mg / mL, and 30 mg / mL, respectively, and the volume ratio of the microcapsule wall material solution to the Clostridium butyricum suspension was 3:1.

[0024] According to a third aspect of the invention, the use of the composition for treating ulcerative colitis in the preparation of a medicament for treating ulcerative colitis is provided.

[0025] The drug is in the form of a gel, which is liquid at room temperature and gels at body temperature. It is administered rectally by pushing the gel into the sigmoid colon, where it forms a temperature-sensitive gel in situ.

[0026] To better illustrate the technical path of the present invention, the following embodiments and comparative examples are provided.

[0027] Example 1 S1. Preparation of plant fermentation products: 10g of water bamboo, 10g of water snowflake, and 10g of water shield were washed, dried, and crushed into 80-mesh powder. Each powder was then mixed with 50mL of Clostridium butyricum suspension (purchased from Shanghai Fuxiang Biotechnology Co., Ltd., ATCC 19398). The mixtures were anaerobic incubated at 37℃ for 36h, filtered, and then autoclaved at 121℃ for 10min. After freeze-drying, the fermented products of water bamboo, water snowflake, and water shield were obtained, respectively. The preparation of Clostridium butyricum suspension includes the following steps: Clostridium butyricum was inoculated into RCM medium at a volume fraction of 5% and incubated anaerobically at 200 rpm and 37°C for 30 h. After incubation, the culture was centrifuged at 4°C and 5000 rpm for 10 min. The precipitate was washed with physiological saline and then dispersed with phosphate buffer to a viable count of 1 × 10⁻⁶. 8 CFU / mL, to obtain Clostridium butyricum suspension; S2. Preparation of temperature-sensitive gel solution: Dissolve 56g of sodium β-glycerophosphate in 100mL of deionized water to obtain a sodium β-glycerophosphate solution. 3g of chitosan (purchased from Wuhan Kemike Biomedical Technology Co., Ltd., degree of deacetylation 85%, CAS: 9012-76-4) was dissolved in 100mL of dilute hydrochloric acid to obtain a chitosan solution. Under ice bath conditions, 33.3 mL of sodium β-glycerophosphate solution was slowly added dropwise to 100 mL of chitosan solution, the pH was adjusted to 7.0-7.2, and the mixture was stirred for 60 min to obtain a thermosensitive gel solution. S3. Preparation of microencapsulated Clostridium butyricum: Dissolve 5g of β-cyclodextrin in 100mL of phosphate buffer, stir at 100℃ and 300rpm for 20min, add 3g of gellan gum and 3g of sodium alginate and stir for another 10min to obtain microcapsule wall material solution; autoclave at 121℃ for 10min and cool, then mix with 33.3mL of Clostridium butyricum suspension, stir at 50rpm for 30min to mix evenly, and freeze dry to obtain microencapsulated Clostridium butyricum; S4. Preparation of the composition: 3g sodium butyrate, 5g rock tripe extract (purchased from Baoji Kosmai Plant Polysaccharide Development Co., Ltd., polysaccharide content 95%, 80 mesh brown powder, hot water extraction), 5g plant extract, 12g plant fermentation product, and 5g microencapsulated Clostridium butyricum were mixed evenly and dispersed in phosphate buffer. The mixture of each component was then mixed with 30mL of phosphate buffer to obtain an active substance suspension. The plant fermentation products include 6g of *Clostridium butyricum* fermentation product from *Zizania latifolia*, 3g of *Nelumbo nucifera* fermentation product from *Nelumbo nucifera*, and 3g of *Brasenia schreberi* fermentation product from *Clostridium butyricum*; the plant extracts include 1.5g of *Coptis chinensis* extract (purchased from Shanyang Lianfeng Biotechnology Co., Ltd., concentration ratio 10:1, brownish-yellow water-soluble concentrated powder, water extraction method), 1.5g of *Codonopsis pilosula* extract (purchased from Shanyang Lianfeng Biotechnology Co., Ltd., concentration ratio 10:1, water extraction method, brownish-yellow powder), 0.5g of *Panax notoginseng* extract (purchased from Shanyang Lianfeng Biotechnology Co., Ltd., concentration ratio 10:1, brownish-yellow powder, water extraction method), and 1.5g of *Bletilla striata* extract (purchased from Shanyang Lianfeng Biotechnology Co., Ltd., concentration ratio 10:1, brownish-yellow powder, water extraction and alcohol precipitation method). Mix 70g of thermosensitive gel solution and active substance suspension in a certain proportion, and stir at 50rpm for 10min at room temperature to obtain the composition.

[0028] Example 2 This embodiment is based on Example 1, except that it includes the following components by weight: 68g thermosensitive gel, 10g rock tripe extract, 5g *Clostridium butyricum* fermentation product of water chestnut, 2.5g *Clostridium butyricum* fermentation product of water caltrop and 2.5g *Clostridium butyricum* fermentation product of water shield, 1g sodium butyrate, 1g microencapsulated *Clostridium butyricum*, 3g *Coptis chinensis* extract, 3g *Codonopsis pilosula* extract, 1g *Panax notoginseng* extract and 3g *Bletilla striata* extract; except for the thermosensitive gel, all other components are based on solute dry weight, and the other specific parameters are the same as in Example 1.

[0029] Example 3 This embodiment is based on Example 1, except that it includes the following components by weight: 68g thermosensitive gel, 1g rock tripe extract, 7.5g *Clostridium butyricum* fermentation product of water chestnut, 3.75g *Clostridium butyricum* fermentation product of water caltrop and 3.75g *Clostridium butyricum* fermentation product of water shield, 5g sodium butyrate, 10g microencapsulated *Clostridium butyricum*, 0.3g *Coptis chinensis* extract, 0.3g *Codonopsis pilosula* extract, 0.1g *Panax notoginseng* extract and 0.3g *Bletilla striata* extract; except for the thermosensitive gel, all other components are based on solute dry weight, and the other specific parameters are the same as in Example 1.

[0030] Example 4 This embodiment is based on Example 1, except that it includes the following components by weight: 69g thermosensitive gel, 5g *Opuntia ficus-indica* extract, 5g *Clostridium butyricum* fermentation product of *Zizania latifolia*, 2.5g *Clostridium butyricum* fermentation product of *Nymphoides peltata* and 2.5g *Clostridium butyricum* fermentation product of *Brasenia schreberi*, 1g sodium butyrate, 10g microencapsulated *Clostridium butyricum*, 1.5g *Coptis chinensis* extract, 1.5g *Codonopsis pilosula* extract, 0.5g *Panax notoginseng* extract and 1.5g *Bletilla striata* extract; except for the thermosensitive gel, all other components are based on solute dry weight, and the other specific parameters are the same as in Example 1.

[0031] Example 5 This embodiment is based on Example 1, except that it includes the following components by weight: 69g thermosensitive gel, 10g rock tripe extract, 6g *Clostridium butyricum* fermentation product of water chestnut, 3g *Clostridium butyricum* fermentation product of water caltrop and 3g *Clostridium butyricum* fermentation product of water shield, 3g sodium butyrate, 5g microencapsulated *Clostridium butyricum*, 0.3g *Coptis chinensis* extract, 0.3g *Codonopsis pilosula* extract, 0.1g *Panax notoginseng* extract and 0.3g *Bletilla striata* extract; except for the thermosensitive gel, all other components are based on solute dry weight, and the other specific parameters are the same as in Example 1.

[0032] Comparative Example 1 This comparative example is based on Example 1, except that the same mass of thermosensitive gel was used instead of the rock tripe extract, while the other specific parameters are the same as in Example 1.

[0033] Comparative Example 2 This comparative example is based on Example 1, except that equal mass of thermosensitive gel is used to replace Coptis chinensis extract, Codonopsis pilosula extract, Panax notoginseng extract and Bletilla striata extract, while the other specific parameters are the same as in Example 1.

[0034] Comparative Example 3 This comparative example is based on Example 1, except that equal mass of thermosensitive gel is used instead of Clostridium butyricum fermentation products of water chestnut, water caltrop, and water shield, and sodium butyrate. All other specific parameters are the same as in Example 1.

[0035] Comparative Example 4 This comparative example is based on Example 1, except that an equal mass of thermosensitive gel is used instead of microencapsulated Clostridium butyricum, while the other specific parameters are the same as in Example 1.

[0036] Test case This invention uses rats as an animal model to verify the therapeutic effect of the composition on ulcerative colitis, specifically including the following steps: Sixty 7-8 week old male SD rats were randomly divided into 12 groups of 5 rats each. One group was randomly selected as the control group, which was fed normal food and water. The other groups were acclimatized for one week and then fed with a 4.5 wt% sodium dextran sulfate solution (purchased from Shanghai Maclean Biochemical Technology Co., Ltd., CAS: 9011-18-1, molecular weight 40000 Da, catalog number: D808272) for 7 days instead of drinking water. They were fed normal food. All groups were fed a 12h / 12h alternating day and night diet, at a constant temperature of 25℃ and humidity of 60%. The rats were weighed daily and fecal occult blood was tested. A positive fecal occult blood test was considered a successful establishment of the model. Administration method: After weighing, rats were anesthetized by intraperitoneal injection of 10wt% chloral hydrate at a dose of 0.3mL / 100g body weight. After anesthesia, a 2mm diameter rubber tube was lubricated with paraffin oil and slowly inserted into the rat rectum for about 8cm. The compositions prepared in each example and comparative example were administered at a dose of 1.5mg / g body weight. After administration, the rats were kept supine with their buttocks facing upward for 20min. The administration was carried out twice a day for 4 consecutive weeks. Rats that were administered 0.4mg / g (based on dry weight of mesalazine) mesalazine solution were used as the positive control group, and the group that was administered an equal volume of phosphate buffer was used as the model group. The blank group was not administered.

[0037] From the date of model establishment, the rats were weighed daily, and the characteristics of their stool and the fecal occult blood / gross blood stool score were observed. The disease activity index (DAI) values ​​of each group of rats were measured. The specific scoring criteria are shown in Table 1, and the results are recorded in Table 2. The total DAI score = weight loss score + stool characteristics score + fecal occult blood / gross blood stool score. After the drug administration was completed, the rats were fasted for 12 hours, sacrificed, and their colons were harvested. The levels of IL-6 and TNF-α in the colon tissue were measured by ELISA, and the results are recorded in Table 3.

[0038] Table 1. DAI Scoring Reference Table

[0039] Table 2. DAI scores of rats in each group

[0040] Table 3. IL-6 and TNF-α levels in rats of each group

[0041] As shown in Table 2, the model group exhibited the highest DAI value, 35 times that of the blank group, indicating successful modeling of ulcerative colitis in rats. The example groups showed good remission of ulcerative colitis, with DAI values ​​of 1.89-2.20, significantly lower than Comparative Examples 1-4, and close to the 1.54 of the positive control group treated with mesalazine. Among them, Example 1, with its balanced component ratios, showed the lowest DAI value among the examples and comparative examples. Compared to Example 1, the DAI values ​​of Comparative Examples 1-4 increased by 41.27%, 45.50%, 48.19%, and 30.16%, respectively, indicating that each component in this invention independently contributes to the treatment of ulcerative colitis, and the absence of any component leads to a significant decrease in efficacy. Comparative Example 4 showed the smallest increase, indicating that the role of microencapsulated Clostridium butyricum in this composition is mainly synergistic assistance and colonization to improve the colonic microbiota environment in the long term, rather than short-term treatment. Comparative Example 3 showed the largest increase, indicating that sodium butyrate and butyrate produced from fermented water chestnut, water shield, and wild rice can effectively provide energy to colonic epithelial cells and promote ulcer repair. Furthermore, fermented water chestnut, water shield, and wild rice also contain a large amount of anti-inflammatory substances, which can help alleviate lesion inflammation. Comparative Example 1, lacking extracts of *Auricularia auricula-judae*, and Comparative Example 2, lacking extracts of *Coptis chinensis*, *Codonopsis pilosula*, *Panax notoginseng*, and *Bletilla striata*, showed a DAI increase of approximately 40% compared to Example 1, indicating that the anti-inflammatory and mucosal repair effects of *Auricularia auricula-judae* significantly contribute to the overall efficacy of the composition. *Coptis chinensis*, *Codonopsis pilosula*, *Panax notoginseng*, and *Bletilla striata* synergistically produced effects of promoting blood circulation, stopping bleeding, promoting tissue regeneration, reducing inflammation, and relieving pain. These results indicate that the components in the composition prepared in this invention exhibit a multi-target synergistic effect of physical barrier, anti-inflammatory repair, and microbial regulation, systematically improving the pathological state of ulcerative colitis. The combined efficacy is superior to any single-target or component-deficient regimen, and is comparable to the therapeutic effect of the commonly used clinical drug mesalazine, but avoids its side effects.

[0042] IL-6 and TNF-α are both typical inflammatory factors in ulcerative colitis. Table 3 shows that the model group exhibited the highest levels of IL-6 and TNF-α, increasing by 97.14% and 175.68% respectively compared to the control group, indicating successful modeling in rats. The positive control group using mesalazine showed a 35.68% and 42.96% decrease in IL-6 and TNF-α levels compared to the model group, indicating that mesalazine can effectively alleviate inflammation in rats. The IL-6 levels in Examples 1-5 were 90.76 ng / L, 92.17 ng / L, 95.33 ng / L, 97.62 ng / L, and 95.99 ng / L, respectively, and the TNF-α levels were 17.94 ng / L, 18.31 ng / L, 19.01 ng / L, 19.33 ng / L, and 19.15 ng / L, respectively, significantly lower than the model group and close to the positive control group, indicating that the composition prepared in these examples can significantly improve ulcerative colitis. Example 1 showed the lowest IL-6 and TNF-α levels among the example groups. Compared with Example 1, the IL-6 levels of Comparative Examples 1-4 increased by 31.82%, 27.17%, 30.84%, and 16.66%, respectively, and the TNF-α levels increased by 25.20%, 29.04%, 37.18%, and 22.35%, respectively. This indicates that the absence of any functional component in this invention leads to a significant rebound in inflammatory factor levels, confirming the independent contribution of each component to alleviating ulcerative colitis. Furthermore, the rebound in inflammatory factors after the absence of each component in the composition was higher than its mass percentage, indicating that the contributions of each component are not simply additive, but rather synergistic through pathway intersection.

[0043] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A composition for treating ulcerative colitis, characterized in that, The composition comprises the following components by weight: 60-80 parts of thermosensitive gel, 1-10 parts of rock tripe extract, 10-15 parts of plant ferment, 1-5 parts of butyrate and its derivatives, 1-10 parts of microencapsulated Clostridium butyricum, and 1-10 parts of plant extract; all components except the thermosensitive gel are dispersed in phosphate buffer after preparation, and the weight of each component is based on the dry weight of the solute; the mass-volume ratio of the sum of the masses of all components except the thermosensitive gel to the phosphate buffer is (0.5-1) g: 1 mL.

2. The composition according to claim 1, characterized in that, The thermosensitive gel is a chitosan / β-glycerophosphate sodium thermosensitive gel; the plant fermentation product is the butyric acid Clostridium fermentation product of water chestnut, water caltrop and water shield; the microencapsulated butyric acid Clostridium is butyric acid Clostridium that has undergone microencapsulation treatment; the plant extract is the extract of Coptis chinensis, Codonopsis pilosula, Panax notoginseng and Bletilla striata; and the butyrate and its derivatives are sodium butyrate.

3. The composition according to claim 2, characterized in that, The mass ratio of the butyric acid Clostridium fermentation products of water chestnut, water caltrop and water shield in the plant fermentation products is 2:1:1; the mass ratio of the extracts of Coptis chinensis, Codonopsis pilosula, Panax notoginseng and Bletilla striata in the plant extracts is 3:3:1:

3.

4. A method for preparing a composition for treating ulcerative colitis, for preparing the composition according to any one of claims 1-3, characterized in that, Specifically, the following steps are included: S1. Preparation of plant fermentation products: After washing, drying and crushing the water chestnuts, water lilies and water shield into powder of 80-100 mesh, they were mixed with Clostridium butyricum suspension at a mass-to-volume ratio of 1 g: 5 mL, anaerobic incubation at 37℃ for 24-48 h, filtered and autoclaved at 121℃ for 10 min, and freeze-dried to obtain the fermented products of water chestnuts, water lilies and water shield respectively. S2. Preparation of temperature-sensitive gel solution: Under ice bath conditions, sodium β-glycerophosphate solution was slowly added dropwise to chitosan solution, pH was adjusted to 7.0-7.2, and stirred for 30-60 min to obtain thermosensitive gel solution; S3. Preparation of microencapsulated Clostridium butyricum: β-cyclodextrin was dissolved in phosphate buffer and stirred at 100℃ and 200-400rpm for 10-30min. Then, gellan gum and sodium alginate were added and stirred for another 10min to obtain microcapsule wall material solution. After autoclaving at 121℃ for 10min and cooling, it was mixed with Clostridium butyricum suspension and stirred at 50-100rpm for 30min to mix evenly. The mixture was then freeze-dried to obtain microencapsulated Clostridium butyricum. S4. Preparation of the composition: Sodium butyrate, rock tripe extract, plant extract, plant ferment, and microencapsulated Clostridium butyricum were uniformly mixed and then dispersed in phosphate buffer. The mass-to-volume ratio of each component mixture to phosphate buffer was (0.5-1) g: 1 mL, resulting in an active substance suspension. Mix the temperature-sensitive gel solution and the active substance suspension, and stir at 30-50 rpm for 10 min at room temperature to obtain the composition.

5. The preparation method according to claim 4, characterized in that, The preparation of the Clostridium butyricum suspension in step S1 includes the following steps: Clostridium butyricum was inoculated into RCM medium at a volume fraction of 5% and incubated anaerobically at 200 rpm and 37°C for 24-36 h. After incubation, the culture was centrifuged at 4°C, and the precipitate was washed with physiological saline and then dispersed with phosphate buffer to a viable count of 1 × 10⁻⁶. 7 -1×10 8 CFU / mL, to obtain Clostridium butyricum suspension.

6. The preparation method according to claim 4, characterized in that, The concentration of the sodium β-glycerophosphate solution in step S2 is 0.56 g / mL, the chitosan solution is a chitosan-dilute hydrochloric acid solution with a concentration of 0.03 g / mL, the volume ratio of the sodium β-glycerophosphate solution to the chitosan solution is 1:3, and the degree of deacetylation of chitosan in the chitosan solution is ≥80%.

7. The preparation method according to claim 4, characterized in that, The viable count of the Clostridium butyricum suspension mentioned in step S3 is 1 × 10⁻⁶. 7 -1×10 8 The concentrations of β-cyclodextrin, gellan gum, and sodium alginate in the microcapsule wall material solution in phosphate buffer were 50 mg / mL, 30 mg / mL, and 30 mg / mL, respectively, and the volume ratio of the microcapsule wall material solution to the Clostridium butyricum suspension was 3:

1.

8. The use of the composition for treating ulcerative colitis in the preparation of a medicament for treating ulcerative colitis, characterized in that, Prepared using the composition according to any one of claims 1-3.

9. The application as described in claim 8, characterized in that, The drug is in the form of a gel, which is liquid at room temperature and gels at body temperature; the administration method is rectal administration, by pushing the gel into the sigmoid colon, where a temperature-sensitive gel is formed in situ.

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