Pharmaceutical composition for preventing and treating small intestinal mucosal barrier damage and its application

Abstract

The present application provides a pharmaceutical composition for preventing and treating small intestinal mucosal barrier damage and its application. The pharmaceutical composition includes sanguinarine and curcumin; the mass ratio of the sanguinarine to the curcumin is (0.33~3.3): (20~80). The present application has found through a large number of studies that, compared with the use of sanguinarine or curcumin alone, the use of the two in a specific ratio can reduce the dosage of sanguinarine, thereby reducing the toxicity of sanguinarine and saving raw material costs; at the same time, the pharmaceutical composition of the present application has a good therapeutic effect on small intestinal mucosal barrier damage. The pharmaceutical composition provided in the present application provides a new solution for the prevention and treatment of small intestinal mucosal barrier damage in clinical practice.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and in particular to a pharmaceutical composition for the prevention and treatment of small intestinal mucosal barrier damage and its application. Background Technology

[0002] Nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, diclofenac, indomethacin, and celecoxib are widely used to treat pain and inflammation in a variety of chronic diseases, such as rheumatoid arthritis and osteoarthritis. Adverse reactions to NSAIDs include serious gastrointestinal complications such as bleeding, ulceration, and perforation. Studies have shown that NSAIDs can also cause small bowel lesions, including various types of mucosal damage such as petechiae, erythema, erosion, and ulceration; therefore, the use of NSAIDs is highly limited.

[0003] Inflammatory bowel disease (IBD) includes ulcerative colitis (UC) and Crohn's disease (CD). The former primarily affects the colon, while the latter affects both the large and small intestines. The small intestine, as a key target for the prevention and treatment of human intestinal diseases and for animal growth, has become a topic of increasing interest.

[0004] The small intestinal mucosal barrier comprises physical, chemical, immune, and biological barriers. The physical barrier, primarily composed of a monolayer of small intestinal epithelial cells (IECs) and tight junctions (TJs), plays a crucial role in NSAID-induced intestinal injury. Small intestinal mucosal barrier (SIMB) damage is considered a common complication of stroke. Goldstein et al. reported that 55% of healthy volunteers developed small intestinal mucosal rupture two weeks after taking naproxen; Matsumoto et al.'s research indicated that approximately 51% of patients developed intestinal ulcers after taking NSAIDs. Similarly, 68%–75% of healthy volunteers who ingested sustained-release diclofenac for two consecutive weeks developed intestinal ulcers.

[0005] Studies have shown that damage to the small intestinal mucosal barrier, especially endothelial cells, is the main pathological change induced by nonsteroidal anti-inflammatory drugs (NSAIDs), with inflammatory stress playing a crucial role. Therefore, preventing and treating small intestinal mucosal inflammation is a primary strategy for reducing intestinal damage in NSAID users. Reports have indicated that misoprostol, rebamipide, rifaximin, colchicine, omeprazole, and probiotics can all alleviate NSAID-induced intestinal damage. However, the therapeutic effects of these drugs on NSAID-induced small intestinal lesions are limited.

[0006] Therefore, traditional technologies still need improvement. Summary of the Invention

[0007] Based on this, one or more embodiments of this application provide a pharmaceutical composition for preventing and treating small intestinal mucosal barrier damage and its application. The technical solution includes:

[0008] According to a first aspect of this application, a pharmaceutical composition for preventing and treating small intestinal mucosal barrier damage is provided, comprising sanguinarine and curcumin; wherein the mass ratio of sanguinarine to curcumin is (0.33~3.3):(20~80).

[0009] In one embodiment, the mass ratio of sanguinarine to curcumin is (1~3.3):(20~80).

[0010] According to a second aspect of this application, the use of the above-described pharmaceutical composition in the preparation of a medicament for the prevention and treatment of small intestinal mucosal barrier damage is provided.

[0011] In one embodiment, the small intestinal mucosal barrier damage includes small intestinal mucosal barrier damage caused by nonsteroidal anti-inflammatory drugs.

[0012] In one embodiment, the nonsteroidal anti-inflammatory drug includes at least one of aspirin, diclofenac, indomethacin, and celecoxib.

[0013] In one embodiment, the drug comprises the pharmaceutical composition and pharmaceutically acceptable excipients.

[0014] In one embodiment, the dosage form of the drug is a tablet, capsule, granule, pill, injection, or sustained-release formulation.

[0015] In one embodiment, the pharmaceutically acceptable excipient includes at least one of a diluent, binder, disintegrant, lubricant, and humectant.

[0016] In one embodiment, the drug satisfies at least one of the following (1) to (5):

[0017] (1) The diluent is selected from at least one of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate and calcium carbonate;

[0018] (2) The adhesive is selected from at least one of starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone and polyethylene glycol;

[0019] (3) The disintegrant is selected from at least one of starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium carboxymethyl cellulose, sodium carboxymethyl starch, polyoxyethylene, sorbitol, fatty acid ester and sodium dodecyl sulfonate;

[0020] (4) The lubricant is selected from at least one of talc, silica, stearate, tartaric acid, liquid paraffin, and polyethylene glycol; and

[0021] (5) The wetting agent is selected from at least one of water, ethanol and isopropanol.

[0022] According to a third aspect of this application, a medicament for preventing and treating damage to the small intestinal mucosal barrier is provided, comprising the above-described pharmaceutical composition and pharmaceutically acceptable excipients.

[0023] Compared with traditional technologies, this application has the following advantages:

[0024] This application, through extensive research, has found that, compared to using sanguinarine or curcumin alone, combining them in a specific ratio can reduce the dosage of sanguinarine, thereby reducing its toxicity to the user and saving on raw material costs. Simultaneously, the pharmaceutical composition of this application exhibits good therapeutic effects on small intestinal mucosal barrier damage. The pharmaceutical composition provided by this application offers a new approach for the prevention and treatment of small intestinal mucosal barrier damage in clinical practice. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the specific embodiments of this application or the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0026] Figure 1 This is a line graph showing the rate of change in body weight of rats from day 1 to day 8 after drug treatment.

[0027] Figure 2 This is a general anatomical diagram of the small intestine of rats after drug treatment;

[0028] Figure 3 This is a partial anatomical diagram of the small intestine of a rat after drug treatment;

[0029] Figure 4 A statistical chart showing the CMDI scores of rats after drug treatment;

[0030] Figure 5 This is an H&E staining image of rat jejunal tissue after drug treatment;

[0031] Figure 6 A statistical chart showing the TDI scores of rats after drug treatment;

[0032] Figure 7The image shows the results of Western blot analysis of AKR1B10, Nrf-2, p-p65, and ZO-1 in rat jejunal tissue.

[0033] Figure 8 A statistical graph showing the relative protein expression levels of AKR1B10 in rats after treatment with different drugs;

[0034] Figure 9 A statistical graph showing the relative protein expression levels of Nrf-2 in rats after treatment with different drugs;

[0035] Figure 10 A statistical graph showing the relative expression levels of p-p65 protein in rats after treatment with different drugs;

[0036] Figure 11 A statistical graph showing the relative protein expression levels of ZO-1 in rats after treatment with different drugs;

[0037] Figure 12 A statistical graph showing the LDH content in the jejunal tissue of rats after treatment with different drugs;

[0038] Figure 13 A statistical graph showing the serum LDH content in rats after treatment with different drugs;

[0039] Figure 14 A statistical graph showing the IL-1β content in the jejunal tissue of rats after treatment with different drugs;

[0040] Figure 15 A statistical graph showing the serum IL-1β levels in rats after treatment with different drugs;

[0041] Figure 16 A statistical graph showing the IL-6 content in the jejunal tissue of rats after treatment with different drugs;

[0042] Figure 17 A statistical graph showing the serum IL-6 levels in rats after treatment with different drugs;

[0043] Figure 18 A statistical graph showing the TNF-α content in the jejunal tissue of rats after treatment with different drugs;

[0044] Figure 19 This is a statistical graph showing the serum TNF-α content in rats after treatment with different drugs. Detailed Implementation

[0045] To make the above-mentioned objectives, features, and advantages of this application more apparent and understandable, a detailed description of specific embodiments of this application is provided. Many specific details are set forth in the following description to provide a thorough understanding of this application. However, this application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of this application. Therefore, this application is not limited to the specific embodiments disclosed below.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. Unless otherwise specifically stated, all raw materials, reagents, instruments, and equipment used in this application are commercially available or can be prepared by existing methods.

[0047] In a first aspect, this application provides a pharmaceutical composition for preventing and treating damage to the small intestinal mucosal barrier, comprising sanguinarine and curcumin; the mass ratio of sanguinarine to curcumin is (0.33~3.3):(20~80).

[0048] Sanguisorbine has certain anti-inflammatory and antioxidant effects and is effective in treating intestinal inflammation, but it is highly toxic and has a narrow safe dosage range. Curcumin has weaker anti-inflammatory, antioxidant, and intestinal protective effects than sanguisorbine, but it has advantages such as a wider safe dosage range and lower price.

[0049] Combining sanguinarine and curcumin in a specific ratio reduces the dosage of sanguinarine, keeping the amount within a safe range and saving on medication costs, while also providing good anti-inflammatory and antioxidant effects. The applicant's research found that combining 1.0 mg / kg sanguinarine with 40 mg / kg curcumin achieves superior efficacy compared to using 3.3 mg / kg sanguinarine alone. Therefore, the pharmaceutical composition of this application can achieve higher therapeutic effects within the safe dosage range of sanguinarine.

[0050] Understandably, the mass ratio of sanguinarine to curcumin in this application is “(0.33~3.3):(20~80)”, which can take the minimum and maximum values ​​of the range (0.33~3.3):(20~80), as well as every value between such minimum and maximum values. Specifically, this includes, but is not limited to, the point values ​​in the embodiments and the following point values: 0.33:20, 0.33:30, 0.33:40, 0.33:50, 0.33:60, 0.33:70, 0.33:80, 0.5:20, 0.5:30, 0.5:40, 0.5:50, 0.5:60, 0.5:70, 0.5:80, 0.8:20, 0.8:30, 0.8:40, 0.8:50, 0.8:60, 0.8:70, 0.8:80, 1:20, 1:3 0, 1:40, 1:50, 1:60, 1:70, 1:80, 2:20, 2:30, 2:40, 2:50, 2:60, 2:70, 2:80, 3:20, 3:30, 3:40, 3:50, 3:60, 3:70, 3:80, 3.3:20, 3.3:30, 3.3:40, 3.3:50, 3.3:60, 3.3:70 or 3.3:80; or a range consisting of any two of these values, such as (0.5~2.5):(30~70).

[0051] In some alternative embodiments, the mass ratio of sanguinarine to curcumin is (1~3.3):(20~80).

[0052] Further optionally, the mass ratio of sanguinarine to curcumin is (1~3):(20~80).

[0053] Further optionally, the mass ratio of sanguinarine to curcumin is 1:(20~80).

[0054] Alternatively, the mass ratio of sanguinarine to curcumin is 1:40.

[0055] Understandably, by controlling the mass ratio of sanguinarine and curcumin within the specific range described above in this application, the amount of sanguinarine used can be reduced, thereby increasing the safe dosage range of the pharmaceutical composition and saving the preparation cost of the pharmaceutical composition, while also exhibiting excellent anti-inflammatory and antioxidant effects and excellent therapeutic effects on small intestinal mucosal barrier damage.

[0056] A second aspect of this application provides the use of the above-described pharmaceutical composition in the preparation of a medicament for the prevention and treatment of small intestinal mucosal barrier damage.

[0057] In some embodiments, small intestinal mucosal barrier damage includes small intestinal mucosal barrier damage caused by nonsteroidal anti-inflammatory drugs.

[0058] In some specific embodiments, the nonsteroidal anti-inflammatory drug includes at least one of aspirin, diclofenac, indomethacin, and celecoxib.

[0059] In some of these embodiments, the drug comprises the pharmaceutical composition of any of the above embodiments and pharmaceutically acceptable excipients.

[0060] Understandably, in this application, "medicine" includes any agent, compound, composition, or mixture that provides physiological and / or pharmacological effects in vivo or in vitro, and often provides beneficial effects. The scope of the physiological and / or pharmacological effects produced by a "medicine" in vivo is not particularly limited; it may have systemic effects or only local effects. The activity of the "medicine" is not particularly limited; it may be an active substance that can interact with other substances or an inert substance that does not interact with other substances.

[0061] In some embodiments, the drug may be a liquid formulation or a solid formulation. Liquid formulations refer to formulations containing a liquid phase, and non-limiting examples include solutions, injections, suspensions, emulsions, etc. Non-limiting examples of solid formulations include tablets, capsules, granules, pills, etc.

[0062] In some of these embodiments, depending on the method of administration, the preparation can be an oral medication, injection, drops, patch, tube feeding formulation, etc.

[0063] In some embodiments, the excipients selected may vary depending on the dosage form.

[0064] In this application, "excipients" include, but are not limited to, mannitol, sorbitol, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, cysteine ​​hydrochloride, thioglycolic acid, methionine, vitamin C, disodium EDTA, sodium calcium EDTA, carbonates, acetates, phosphates or aqueous solutions of monovalent alkali metals, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acids, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and its derivatives, alginate, gelatin, polyvinylpyrrolidone, glycerol, Tween 80, agar, calcium carbonate, calcium bicarbonate, surfactants, polyethylene glycol, cyclodextrin, phospholipids, kaolin, talc, calcium stearate, and magnesium stearate.

[0065] In this application, "pharmaceuticalally acceptable excipient" means a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. As used herein, the term "pharmaceuticalally acceptable carrier" includes buffers compatible with drug administration, sterile water for injection, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption delay agents, and the like. Each substance must be "pharmaceuticalally acceptable" in the sense of compatibility with other components in the formulation and harmlessness to the patient. Suitable examples include, but are not limited to: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch, potato starch and substituted or unsubstituted β-cyclodextrins; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn... Rice oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0066] In some embodiments, "acceptable excipients" are selected from at least one of diluents, adhesives, wetting agents, disintegrants, and lubricants.

[0067] Specifically, the diluent is selected from at least one of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, and calcium carbonate; the binder is selected from at least one of starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, and polyethylene glycol; the disintegrant is selected from at least one of starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium, sodium carboxymethyl starch, polyoxyethylene, sorbitol, fatty acid ester, and sodium dodecyl sulfonate; the lubricant is selected from at least one of talc, silica, stearate, tartaric acid, liquid paraffin, and polyethylene glycol; and the wetting agent is selected from at least one of water, ethanol, and isopropanol.

[0068] In some specific embodiments, the above-described drug is a tablet. To formulate the above-described drug into tablets, various excipients known in the art can be used as pharmaceutically acceptable excipients. Specifically, since the above-described drug is a tablet, pharmaceutically acceptable excipients include one or more of diluents, binders, wetting agents, disintegrants, and lubricants.

[0069] In some specific embodiments, the aforementioned drug is a capsule or soft capsule, directional capsule, or fast-dissolving capsule, or other special capsule formulation. To formulate the aforementioned drug into capsules or soft capsules, directional capsules, or fast-dissolving capsules, the aforementioned stem cell exosomes can be mixed with diluents, flow aids, etc., and then the resulting mixture can be directly placed into hard capsules, soft capsules, or special capsules. Alternatively, in other embodiments, the aforementioned stem cell exosomes can be first mixed with one or more of a diluent, binder, and disintegrant to form particles, microspheres, microspheres, liposomes, etc., before being placed into hard capsules, soft capsules, or special capsules.

[0070] In some specific embodiments, the above-described drug is an injectable preparation. To prepare the above-described drug into an injectable preparation, water, ethanol, isopropanol, propylene glycol, polyethylene glycol, or mixtures thereof can be used as solvents, and an appropriate amount of pharmaceutically acceptable excipients commonly used in the art can be added before use.

[0071] A third aspect of this application provides a medicament for treating small intestinal mucosal barrier damage, comprising the pharmaceutical composition of the first aspect of this application and pharmaceutically acceptable excipients.

[0072] Understandably, pharmaceutically acceptable excipients include those defined in the second aspect of this application.

[0073] A fourth aspect of this application provides a method for treating small intestinal mucosal barrier injury, the method employing a pharmaceutical composition of the first aspect of this application to treat small intestinal mucosal barrier injury.

[0074] In some specific embodiments, the small intestinal mucosal barrier damage is caused by nonsteroidal anti-inflammatory drugs.

[0075] In some examples, nonsteroidal anti-inflammatory drugs include at least one of aspirin, diclofenac, indomethacin, and celecoxib.

[0076] The present application will be further described below with reference to specific embodiments and comparative examples, but should not be construed as limiting the scope of protection of the present application.

[0077] Example 1:

[0078] I. Provide animal models

[0079] The animal model used in this embodiment is SPF-grade SD male rats, purchased from Hunan Slack Jingda Experimental Animal Co., Ltd., with the license number GYS-201905230001. Approved by the Ethics Committee of Hunan University of Chinese Medicine, the experimental animals are housed in the Experimental Animal Center of Hunan University of Chinese Medicine, and the license number for the experimental unit is SYXK (Xiang) 2019-0009.

[0080] The feeding conditions are as follows: temperature 22°C - 24°C, humidity 60% - 65%, day-night alternation, free diet and water intake.

[0081] II. Reagent preparation

[0082] a) Prepare 7.5 mg / kg indomethacin (Indo) solution: Based on the total number N and total body weight X of the rats on the day, weigh (X÷N÷1000×7.5 mg / kg)×(N + 5) indomethacin powder; dissolve the above indomethacin powder in a mixed solution (by volume percentage, containing 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline) to obtain the indomethacin stock solution.

[0083] Dilute the indomethacin stock solution to (N + 5) mL with normal saline, and calculate the volume of the solution for subcutaneous injection according to the body weight of each rat. Assume the body weight of each rat is X0 and the average body weight of the rats is X mean , then the injection volume for each rat = 1 mL×(X0÷X mean ), so that the final concentration of indomethacin injected into the rats is 7.5 mg / kg.

[0084] b) Prepare 1.0 mg / kg sanguinarine (SA) solution: Weigh (X÷N÷1000×1.0 mg / kg)×(N + 5) sanguinarine powder; dissolve the sanguinarine powder in DMSO solution to obtain the sanguinarine stock solution. Dilute the stock solution with 0.5 w / v% CMC-Na solution and make up to (N + 5) mL, and calculate the volume of the solution for gavage according to the body weight of each rat. Assume the body weight of each rat is X0 and the average body weight of the rats is X mean , then the gavage volume for each rat = 1 mL×(X0÷X mean ), so that the final concentration of sanguinarine gavaged into the rats is 1.0 mg / kg.

[0085] Prepare 0.33 mg / kg and 3.3 mg / kg sanguinarine solutions using the same principle.

[0086] c) Preparation of a 20 mg / kg curcumin (Cur) solution: Weigh (X÷N÷1000×20 mg / kg)×(N+5) mg of curcumin powder; dissolve the curcumin powder in DMSO solution to obtain a curcumin stock solution. Dilute the stock solution with 0.5 w / v% CMC-Na solution and bring the volume to (N+5) mL. Calculate the volume of solution to be administered by gavage based on the weight of each rat. Assume the weight of each rat is X0, and the average weight of the rats is X. mean Therefore, the gavage volume for each rat = 1 mL × (X0 ÷ X mean This was done so that the final concentration of curcumin administered to the rats via gavage was 20 mg / kg.

[0087] Sanguisorbine solutions of 40 mg / kg and 80 mg / kg were prepared using the same principle.

[0088] d) Preparation of a 1.0 mg / kg sanguisorbin + 20 mg / kg curcumin mixed solution: Weigh (X÷N÷1000×1.0 mg / kg)×(N+5) mg of sanguisorbin powder and (X÷N÷1000×20 mg / kg)×(N+5) mg of curcumin powder, dissolve them in DMSO solution to obtain a stock solution. Dilute the stock solution with 0.5 w / v% CMC-Na solution and bring the volume to (N+5) mL. Calculate the volume of solution to be administered by gavage based on the weight of each rat. Assume the weight of each rat is X0, and the average weight of the rats is X. mean Therefore, the gavage volume for each rat = 1 mL × (X0 ÷ X mean ).

[0089] III. Mouse Treatment Methods

[0090] Seventy-two 6-week-old SD rats were selected and, after acclimatization (body weight controlled at approximately 240 g), were randomly divided into 12 groups of 6 rats each. The rats were treated according to the methods shown in Table 1.

[0091] The treatment method for the drug administration group is as follows: SA solution, Cur solution, SA+Cur mixed solution or berberine (Ber) were administered for prophylaxis for 3 days, followed by two subcutaneous injections of indomethacin at a dose of 7.5 mg / kg, with an interval of 24 hours between each injection, to establish the model; at the same time, SA, Cur, SA+Cur mixed solution or Ber were administered, and the drug was continuously administered by gavage until day 8, once a day by gavage.

[0092] Berberine can inhibit the activation of signaling pathway proteins such as TLR4, MyD88, and NF-κB, protect the tight junction protein structure between intestinal epithelial cells from damage, and increase the expression level of TJ protein, thus repairing intestinal mucosal mechanical barrier dysfunction to a certain extent. In this example, rats were administered 60 mg / kg berberine by gavage as a positive control group to verify the therapeutic effect of the sanguinarine + curcumin mixed administration group.

[0093] Table 1

[0094]

[0095] IV. Experimental Results

[0096] (1) Rate of change in body weight

[0097] The volume change rate of rats in each treatment group was continuously recorded during the gavage administration period. The volume change rate was calculated as follows: (body weight on day N after gavage administration - body weight on day 0 after gavage administration) / body weight on day 0 after gavage administration × 100%. The volume change rate results for each treatment group on days 1-8 after gavage administration are shown in Table 2 and [Table data missing]. Figure 1 As shown.

[0098] Table 2

[0099]

[0100] As shown in the table above, the rats experienced a sharp decline in body weight after indomethacin-induced weight loss, and each treatment group was able to slow down this weight loss trend to varying degrees. Among them, the combination of SA (1.0 mg / kg) + Cur (20 mg / kg) significantly accelerated the weight gain of rats on the 7th day after administration.

[0101] (2) Macro observation

[0102] Twenty-four hours after the last administration, rats were anesthetized with chloral hydrate, and the abdominal cavity was opened. Macroscopic examination was used to assess and record intestinal fluid accumulation, redness, adhesions, and necrosis. Results were as follows: Figure 2 and Figure 3 As shown.

[0103] As shown in the figure, indomethacin treatment induced small intestinal edema, severe adhesions, and difficulty in dissection in rats, with some cases also accompanied by necrotic lesions. Both the treatment groups and the control group showed varying degrees of improvement; specifically, the improvement effects of the SA (1.0 mg / kg) + Cur (20 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg), SA (1.0 mg / kg) + Cur (80 mg / kg), SA (3.3 mg / kg) + Cur (20 mg / kg), and SA (3.3 mg / kg) + Cur (40 mg / kg) treatment groups were significantly better than those of SA alone and Cur alone, similar to the improvement results of the berberine treatment group; indicating that the pharmaceutical composition of this application can effectively improve the symptoms of small intestinal mucosal barrier damage.

[0104] (3) CMDI score

[0105] Subsequently, jejunal tissue was harvested, rinsed with ice-cold phosphate-buffered saline (PBS), and longitudinally incised along the mesentery. The degree of jejunal mucosal ulceration, erosion, hyperplasia, and adhesions was observed, and the colonic mucosal damage index (CMDI) for each treatment group was recorded. The evaluation criteria for CMDI are shown in Table 3, and the evaluation results are shown in Tables 4 and 5. Figure 4 As shown ( Figure 4 In the graph, “*” indicates a significant difference (P < 0.05); “**” indicates an extremely significant difference (P < 0.001).

[0106] Table 3

[0107]

[0108] Table 4

[0109]

[0110] As shown in the table above, all treatment groups significantly improved the small intestinal mucosal barrier damage induced by indomethacin. Among them, the mixed treatment groups of SA (0.33 mg / kg) + Cur (80 mg / kg), SA (1.0 mg / kg) + Cur (20 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg) and SA (1.0 mg / kg) + Cur (80 mg / kg) showed better improvement than the single treatment groups.

[0111] (4) H&E staining and TDI score

[0112] Twenty-four hours after the last administration, histological analysis and TDI scoring were performed on the jejunal tissue of rats in each treatment group. The tissue was routinely fixed and paraffin-embedded, then sectioned and stained with Hematoxylin and eosin (H&E). Histological evaluation and photographic recording were performed under an optical microscope. The H&E staining results are as follows: Figure 5 As shown.

[0113] The TDI score is determined based on indicators such as epithelial damage, ulcer depth, lymphocyte infiltration, edema, and inflammatory cell (such as neutrophils and eosinophils) infiltration. The TDI scoring criteria mainly include inflammatory cell infiltration, infiltration depth, and intestinal ulcer depth.

[0114] No inflammatory cell infiltration scores 0; mild inflammatory cell infiltration scores 1; severe inflammatory cell infiltration scores 2. Infiltration depth scores are as follows: infiltration into the mucosal layer scores 1; infiltration into the mucosa and submucosa scores 2; infiltration into the entire colonic layer scores 3. Intestinal ulcer depth scores are as follows: no ulcer scores 0; epithelial ulcer scores 1; lamina propria mucosal ulcer scores 2; mucomuscularis mucosae ulcer scores 3.

[0115] The TDI score results are shown in Table 5 and Figure 6 As shown ( Figure 6 In the graph, “*” indicates a significant difference (P < 0.05); “**” indicates an extremely significant difference (P < 0.001).

[0116] Table 5

[0117]

[0118] Depend on Figure 5 and Figure 6 It was found that rats treated with indomethacin exhibited significant small intestinal mucosal damage, including edema, inflammatory cell infiltration, microvilli shortening, shedding, and ulceration; histopathological scores also showed a significant increase in the Tissue Injury Index (TDI). All treatment groups significantly reduced mucosal damage and improved TDI; among them, SA (0.33 mg / kg) + Cur (80 mg / kg), SA (1.0 mg / kg) + Cur (20 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg), and SA (1.0 mg / kg) + Cur (80 mg / kg) showed superior improvement compared to the single-drug groups.

[0119] (5) Protein blot analysis

[0120] Twenty-four hours after the last administration, 100 mg of tissue was extracted from the rat jejunum, washed with PBS at 4°C, and then homogenized in RIPA lysis buffer with a protease inhibitor (100:1) as described above. The mixture was centrifuged at 16000 g for 15 minutes, and the supernatant was collected. Protein quantification was performed using a BCA kit (manufacturer: Elabscience; product model: E-BC-K318-M). Bromophenol blue loading buffer was added, and the mixture was denatured at 100°C for 10 minutes. Western blot was performed by SDS-PAGE gel electrophoresis. The target protein was separated by 6%–10% SDS-PAGE electrophoresis, transferred to a PVDF membrane, and blocked with 5% milk or 5% BSA (prepared in TBST) for 1 hour. Primary antibodies corresponding to AKR1B10, Nrf-2, p-p65, and ZO-1 were added and incubated overnight at 4°C. The next day, secondary antibodies corresponding to the primary antibody host were incubated at 37°C for 1 hour, followed by the addition of developing solution for development. The results are as follows: Figure 7 As shown in the figure. The grayscale values ​​of the target protein bands were calculated using ImageJ software. The statistical results of the relative expression levels of AKR1B10, Nrf-2, p-p65, and ZO-1 are shown in the figure. Figure 8 , Figure 9 , Figure 10 and Figure 11 As shown ( Figures 8-11 In the graph, “*” indicates a significant difference (P < 0.05); “**” indicates an extremely significant difference (P < 0.001).

[0121] Western blot analysis showed that the expression of AKR1B10, ZO-1, and Nrf2 proteins in the small intestine tissue of rats induced by indomethacin was significantly decreased, while the expression of AKR1B10, ZO-1, and Nrf2 proteins increased to varying degrees in the treatment groups. Specifically, the expression of AKR1B10 protein was significantly enhanced in the SA (1.0 mg / kg) + Cur (20 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg), SA (1.0 mg / kg) + Cur (80 mg / kg), and SA (3.3 mg / kg) + Cur (20 mg / kg) treatment groups; the enhancement effect on ZO-1 was similar across all treatment groups. Nrf2 showed significant improvement in the SA (1.0 mg / kg) + Cur (40 mg / kg) and SA (1.0 mg / kg) + Cur (80 mg / kg) administration groups. Considering the relative expression level of Nrf2 protein and cost, the SA (1.0 mg / kg) + Cur (40 mg / kg) combination was preferred. pp65 levels increased sharply in the model group, and all mixed administration groups showed more significant improvement compared to single administration. Among them, the SA (1.0 mg / kg) + Cur (40 mg / kg) administration group showed the best improvement.

[0122] (6) Determination of lactate dehydrogenase (LDH)

[0123] 24 hours after the last administration, 100 mg of jejunal tissue was collected, washed with ice-cold PBS, ground at low temperature, centrifuged, and the supernatant was balanced. The LDH level in the colonic tissue was calculated, and the results are as follows. Figure 12 As shown ( Figure 12 In the graph, “*” indicates a significant difference (P < 0.05); “**” indicates an extremely significant difference (P < 0.001).

[0124] Blood was collected from the abdominal aorta, serum was separated, and the solution was incubated at 4°C for 30 minutes. The solution was then centrifuged at 1000 g, and the supernatant was collected. The serum LDH level was calculated, and the results are as follows: Figure 13 As shown ( Figure 13 The asterisk (*) indicates a significant difference (P < 0.05).

[0125] LDH = (Measured OD value - Control OD value) / (Standard OD value - Blank OD value) × Standard concentration of test sample / Protein concentration.

[0126] Depend on Figure 12 and Figure 13It was found that indomethacin-induced LDH levels in the small intestine and serum of rats increased significantly, with varying degrees of improvement observed in all treatment groups. Compared with single administration, the SA (0.33 mg / kg) + Cur (40 mg / kg), SA (1.0 mg / kg) + Cur (20 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg), SA (3.3 mg / kg) + Cur (20 mg / kg), and SA (3.3 mg / kg) + Cur (40 mg / kg) treatment groups showed more significant reductions in serum LDH levels; the SA (1.0 mg / kg) + Cur (40 mg / kg) and SA (3.3 mg / kg) + Cur (20 mg / kg) treatment groups showed more significant reductions in serum LDH levels.

[0127] (7) Determination of the levels of IL-1β, IL-6 and TNF-α

[0128] Twenty-four hours after the last administration, 100 mg of rat jejunal tissue was accurately weighed and added to 1000 μL of pre-cooled RIPA lysis buffer. Steel balls were added, and the mixture was homogenized three times at 4°C using a tissue homogenizer, each time for 45 seconds, with a 20-second interval. After centrifugation at 7000 g for 10 min, the supernatant was collected to obtain the small intestinal tissue sample.

[0129] Take a whole blood sample and place it at room temperature for 1 hour (or overnight at 2℃~8℃), then centrifuge it at 1000×g for 20 minutes at 2℃~8℃, collect the supernatant, and obtain the plasma sample.

[0130] The levels of TNF-α, IL-6, and IL-1β in small intestinal tissue and plasma were detected using ELISA kits (manufactured by Elabscience; product models E-EL-R2856, E-EL-R0015, and E-EL-R0012, respectively).

[0131] The results of the detection of TNF-α levels in small intestinal tissue and plasma are as follows: Figure 14 and Figure 15 As shown ( Figures 14-15* indicates a significant difference (P < 0.05); ** indicates an extremely significant difference (P < 0.001). Indomethacin-induced TNFα levels in the small intestine and serum of rats significantly increased, with varying degrees of improvement in all treatment groups. Compared with SA alone and Cur alone, the following treatment groups showed superior reduction in tissue TNFα: SA (0.33 mg / kg) + Cur (40 mg / kg), SA (0.33 mg / kg) + Cur (80 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg), SA (1.0 mg / kg) + Cur (80 mg / kg), SA (3.3 mg / kg) + Cur (20 mg / kg), and SA (3.3 mg / kg) + Cur (40 mg / kg). Compared with SA alone and Cur alone, the SA (0.33 mg / kg) + Cur (80 mg / kg), SA (1.0 mg / kg) + Cur (20 mg / kg), SA (3.3 mg / kg) + Cur (20 mg / kg) and SA (3.3 mg / kg) + Cur (40 mg / kg) groups showed a superior effect in reducing serum TNFα.

[0132] The results of the detection of IL-6 levels in small intestinal tissue and plasma are as follows: Figure 16 and Figure 17 As shown ( Figures 16-17 * indicates a significant difference (P < 0.05); ** indicates an extremely significant difference (P < 0.001). Indomethacin-induced IL-6 levels in the small intestine and serum of rats significantly increased, with varying degrees of improvement in all treatment groups. Compared with SA and Cur alone, the SA (0.33 mg / kg) + Cur (40 mg / kg), SA (0.33 mg / kg) + Cur (80 mg / kg), SA (1.0 mg / kg) + Cur (20 mg / kg), SA (1.0 mg / kg) + Cur (40 mg / kg), SA (1.0 mg / kg) + Cur (80 mg / kg), and SA (3.3 mg / kg) + Cur (80 mg / kg) treatment groups showed a more significant reduction in tissue IL-1β levels. Regarding the reduction effect of serum IL-6, the reduction effect of each mixed administration group was better than that of the single administration group. Among them, the SA (1.0 mg / kg) + Cur (80 mg / kg) administration group showed the most significant reduction effect on serum IL-6.

[0133] The results of the detection of IL-1β levels in small intestinal tissue and plasma are as follows: Figure 18 and Figure 19 As shown ( Figures 18-19* indicates a significant difference (P < 0.05); ** indicates an extremely significant difference (P < 0.001). Indomethacin-induced IL-1β levels in the small intestine and serum of rats significantly increased, with varying degrees of improvement in all treatment groups. Compared to SA and Curl alone, the SA (1.0 mg / kg) + Cur (80 mg / kg) and SA (0.33 mg / kg) + Cur (80 mg / kg) treatment groups showed a more significant reduction in IL-1β levels in the small intestine. Regarding the reduction in serum IL-1β, all combined treatment groups showed better reductions than the single-treatment groups, with the SA (0.33 mg / kg) + Cur (80 mg / kg), SA (1.0 mg / kg) + Cur (80 mg / kg), and SA (3.3 mg / kg) + Cur (20 mg / kg) treatment groups showing the most significant reductions in serum IL-1β.

[0134] In summary, this application combines sanguinarine with curcumin to achieve superior anti-inflammatory and antioxidant effects while reducing the dosage of sanguinarine, thereby preventing or treating damage to the small intestinal mucosal barrier.

[0135] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0136] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this application should be determined by the appended claims.

Claims

1. A pharmaceutical composition for preventing and treating small intestinal mucosal barrier damage, characterized in that: The invention comprises sanguinarine and curcumin; the mass ratio of the sanguinarine to the curcumin is 1:(20-80), and the dosage of the sanguinarine is 1 mg / kg.

2. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for preventing and treating small intestinal mucosal barrier damage.

3. The use according to claim 2, characterized in that: The small intestinal mucosal barrier damage includes small intestinal mucosal barrier damage caused by non-steroidal anti-inflammatory drugs.

4. The use according to claim 3, characterized in that: The non-steroidal anti-inflammatory drug includes at least one of aspirin, diclofenac, indomethacin and celecoxib.

5. The use according to any one of claims 2 to 4, characterized in that: The medicine comprises the pharmaceutical composition and pharmaceutically acceptable excipients.

6. The use according to any one of claims 2 to 4, characterized in that: The dosage form of the drug is tablet, capsule, granule, pill, injection or sustained-release preparation.

7. The use according to claim 5, characterized in that: The pharmaceutically acceptable excipients include at least one of a diluent, a binder, a disintegrant, a lubricant and a wetting agent.

8. The use according to claim 7, characterized in that: The drug satisfies at least one of the following (1) to (5): (1) The diluent is at least one selected from the group consisting of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate and calcium carbonate; (2) The adhesive is selected from at least one of starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone and polyethylene glycol; (3) The disintegrant is at least one selected from the group consisting of starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, polyoxyethylene, sorbitol, fatty acid esters and sodium dodecyl sulfate; (4) the lubricant is at least one selected from talc, silicon dioxide, stearate, tartaric acid, liquid paraffin and polyethylene glycol; and (5) The wetting agent is selected from at least one of water, ethanol and isopropanol.

9. A drug for preventing and treating small intestinal mucosal barrier damage, characterized in that: The invention comprises the pharmaceutical composition according to claim 1 and pharmaceutically acceptable excipients.

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