A pharmaceutical composition for treating or relieving ulcerative colitis and use thereof

Abstract

The present application relates to the technical field of pharmaceutical compositions, and specifically discloses a pharmaceutical composition for treating or relieving ulcerative colitis and application, the pharmaceutical composition comprises a therapeutically effective amount of active ingredients and pharmaceutically acceptable adjuvants, the active ingredient is sarcosine, its pharmaceutically acceptable salt or its pharmaceutically acceptable derivative;The pharmaceutical composition can achieve at least one of the following technical effects after administration: (1) inhibiting the polarization of CD86+ pro-inflammatory M1 macrophages in colon tissue, up-regulating the proportion of CD206+ anti-inflammatory M2 macrophages, and remodeling the inflammatory microenvironment of colitis;(2) up-regulating the expression of colonic mucosal tight junction proteins Occludin and Claudin-1, repairing the damaged intestinal mucosal barrier, and promoting mucosal healing;(3) improving the weight loss of ulcerative colitis model, reducing the disease activity score, relieving the shortening of colon, and reducing the histological inflammation injury score of colon.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical composition technology, specifically to a pharmaceutical composition and its application for treating or alleviating ulcerative colitis. Background Technology

[0002] Ulcerative colitis (UC) is a chronic, nonspecific inflammatory bowel disease. The lesions primarily affect the colonic mucosa and submucosa, characterized by continuous, diffuse mucosal inflammation, erosion, and ulceration. Clinically, it is mainly manifested by recurrent diarrhea, bloody and mucous stools, abdominal pain, and tenesmus. The disease course is protracted and carries a long-term risk of colorectal cancer. In recent years, the global incidence of UC has been rising continuously. In my country, it has become a common and difficult-to-treat chronic digestive disease, requiring long-term or even lifelong treatment, severely impacting patients' quality of life and placing a heavy burden on the social healthcare system.

[0003] Current clinical UC treatment protocols are stratified according to disease activity and lesion extent. Core treatment drugs mainly fall into four categories: The first category is aminosalicylic acids, represented by 5-aminosalicylic acid, which are first-line drugs for inducing and maintaining remission in mild to moderate UC, but only about 50% of patients respond, and there are issues with low mucosal healing rates and decreased response rates with long-term use; the second category is glucocorticoids, mainly used for inducing remission in acute exacerbations of moderate to severe UC, with rapid onset of action but significant systemic side effects, making long-term maintenance use impossible, and some patients develop hormone resistance or dependence; the third category is immunomodulators, including… Including azathioprine and mercaptopurine, these are mostly used for maintenance remission in patients who do not respond to aminosalicylic acid treatment, but they have a slow onset of action and serious adverse reactions such as bone marrow suppression and liver and kidney damage. Frequent monitoring is required during medication, and patient compliance is poor. The fourth category is biological agents, represented by anti-TNF-α monoclonal antibodies and anti-integrin monoclonal antibodies, which have brought breakthroughs to the treatment of moderate to severe and refractory UC. However, 30%-40% of patients still have no primary response, and the rate of secondary non-response increases year by year with the duration of medication. In addition, the treatment cost is high, and there are safety risks such as infection and infusion reactions, making it difficult to use them widely for long-term maintenance therapy.

[0004] Despite the abundance of existing treatment options, several critical bottlenecks remain in clinical practice: First, current treatments primarily focus on suppressing inflammation, lacking targeted interventions for intestinal mucosal barrier repair. Mucosal healing is the core endpoint of UC treatment and crucial for maintaining long-term remission and reducing the risk of recurrence and cancer; existing drugs struggle to achieve sustained and stable mucosal healing. Second, the safety and tolerability of existing drugs for long-term use are insufficient, failing to meet patients' needs for long-term maintenance therapy. Most drugs exhibit significant systemic adverse reactions, limiting long-term application. Third, there is a lack of alternative treatments for refractory UC and patients with poor drug response. Clinically, there is an urgent need to develop novel UC treatments with novel mechanisms of action, high safety, oral administration, long-term use, and dual anti-inflammatory and mucosal repair effects.

[0005] Creatine (N-methylglycine) is an endogenous amino acid derivative and an important intermediate in the glycine metabolic pathway, widely present in human tissues and body fluids. Current research focuses primarily on its clinical applications as a tumor marker and its role in central nervous system and muscle metabolism-related diseases. However, no existing technology discloses the pharmaceutical uses of creatine in the treatment of ulcerative colitis (UC), nor has it revealed its therapeutic effects on UC through regulating macrophage polarization and repairing the intestinal mucosal barrier. Significant technological gaps exist in this field. Summary of the Invention

[0006] In view of the shortcomings of the prior art, the present invention provides a pharmaceutical composition and application for treating or alleviating ulcerative colitis, thereby solving the problems in the background art.

[0007] To achieve the above objectives, the present invention provides the following technical solution: a pharmaceutical composition for treating or alleviating ulcerative colitis, the pharmaceutical composition comprising a therapeutically effective amount of an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is sarcosine, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable derivative thereof; after administration, the pharmaceutical composition achieves at least one of the following technical effects: (1) Inhibits the polarization of CD86+ pro-inflammatory M1 macrophages in colon tissue, upregulates the proportion of CD206+ anti-inflammatory M2 macrophages, and reshapes the colonic inflammatory microenvironment; (2) Upregulate the expression of colonic mucosal tight junction proteins Occludin and Claudin-1, repair the damaged intestinal mucosal barrier, and promote mucosal healing; (3) Improved weight loss, reduced disease activity score, alleviated colonic shortening, and reduced colonic histological inflammatory damage score in the ulcerative colitis model.

[0008] Preferably, the pharmaceutically acceptable salt of sarcosine is selected from any one of sodium sarcosine and sarcosine hydrochloride; the pharmaceutically acceptable derivative of sarcosine is selected from any one of sarcosine ethyl ester and sarcosine amide.

[0009] Preferably, the dosage form of the pharmaceutical composition is an oral formulation, a rectal formulation, or a colon-targeted formulation; The oral preparation is selected from any one of tablets, capsules, granules, powders, and oral solutions; The rectal administration formulation is selected from any one of suppositories, enemas, and colonic retention gels; The colon-targeting formulation is selected from any one of pH-dependent enteric-coated formulations, time-controlled release colon-release formulations, and microbiota-triggered colon-targeting formulations.

[0010] The colon-targeting formulation is a pH-dependent enteric-coated microcapsule, wherein the microcapsule consists of a drug-containing core, an isolation layer, and an enteric coating layer from the inside out. The medicated core contains an active ingredient and a diluent, wherein the active ingredient accounts for 10% to 60% of the mass of the medicated core. The coating material of the enteric coating layer is a compound of methacrylic acid copolymer L100 and methacrylic acid copolymer S100 in a mass ratio of 1:2 to 2:1, and the weight gain of the coating layer is 8% to 15% of the total weight of the pill core. The enteric-coated microparticles exhibit a cumulative release rate of ≤10% in hydrochloric acid solution at pH 1.2 after 2 hours, ≤20% in phosphate buffer solution at pH 6.8 after 2 hours, and ≥80% in colonic fluid environment at pH ≥ 7.0 after 1 hour, achieving targeted release of the active ingredient at the site of colonic inflammation.

[0011] The rectal administration preparation is a colonic retention gel. The gel matrix is ​​a compound of carbomer and hydroxypropyl methylcellulose in a mass ratio of 1:1 to 1:3. The mass concentration of the active ingredient in the gel is 50 to 200 mg / g. After rectal administration, the gel can remain at the site of colonic inflammation for ≥6 hours, thereby increasing the local drug concentration and treatment efficiency.

[0012] Preferably, the dosage of the active ingredient, calculated as creatine, is 10-1000 mg / kg / day; For the induction of remission of mild to moderate active ulcerative colitis, the preferred dosage of the active ingredient is 50-200 mg / kg / day, more preferably 100 mg / kg / day; For combination therapy of severe active ulcerative colitis, the preferred dosage of the active ingredient is 100-300 mg / kg / day; For maintenance therapy during the remission phase of ulcerative colitis, the preferred dosage of the active ingredient is 20-100 mg / kg / day.

[0013] Preferably, the pharmaceutical composition is administered 1 to 2 times daily; For induction of remission in mild to moderate active ulcerative colitis, the continuous dosing period is ≥4 weeks; For combination therapy of severe active ulcerative colitis, the continuous dosing period is ≥8 weeks; For maintenance therapy during the remission phase of ulcerative colitis, continuous administration for ≥6 months is used to prevent clinical relapse.

[0014] Preferably, the pharmaceutical composition further comprises a second active ingredient, which is a first-line clinical treatment drug for ulcerative colitis, selected from any one or more of 5-aminosalicylic acid drugs, glucocorticoids, immunomodulators, biological agents, and small molecule targeted drugs; When the second active ingredient is a 5-aminosalicylic acid drug, the mass ratio of the active ingredient to the 5-aminosalicylic acid drug, calculated as creatine, is 1:0.5 to 1:5. The drug composition is used for the induction of remission of mild to moderate active ulcerative colitis, or for the treatment of patients who do not respond well to 5-aminosalicylic acid monotherapy. When the second active ingredient is an anti-TNF-α biological agent, the dosage of the active ingredient, calculated as creatine, is 50-200 mg / kg / day, and the dosage of the anti-TNF-α biological agent is 5-10 mg / kg / time, administered once every 2 weeks. The pharmaceutical composition is used for the induction and remission of moderate to severe active ulcerative colitis.

[0015] Preferably, after continuous administration for 7 days, the pharmaceutical composition can achieve at least one of the following quantifiable effects in a DSS-induced ulcerative colitis mouse model: (1) The weight loss of the model mice was ≥50% less than that of the model control group; (2) The disease activity score of the model mice was reduced by ≥40% compared with that of the model control group; (3) The colon length of the model mice recovered to ≥80% compared with the model control group; (4) The colonic histological inflammation score of the model mice was ≥45% lower than that of the model control group; (5) The expression level of CD86 protein in the colon tissue of the model mice was downregulated by ≥35% compared with the model control group, and the expression level of CD206 protein was upregulated by ≥25% compared with the model control group; (6) The expression levels of Occludin and Claudin-1 proteins in the colonic mucosa of the model mice were upregulated by ≥30% compared with the model control group.

[0016] Use of sarcosine, its pharmaceutically acceptable salts, or its pharmaceutically acceptable derivatives in the preparation of medicaments for the treatment or relief of ulcerative colitis.

[0017] Preferably, the drug is used to achieve at least one of the following clinical benefits: (1) Improves clinical symptoms of ulcerative colitis such as weight loss, diarrhea, hematochezia, and abdominal pain, and reduces disease activity scores; (2) Repair the patient's damaged colonic mucosal barrier to achieve mucosal healing; (3) Relieves inflammatory infiltration of colonic tissue in patients with ulcerative colitis and reduces histological inflammatory damage scores; (4) Maintain clinical remission in patients with ulcerative colitis and reduce the recurrence rate of the disease.

[0018] Preferably, the drug achieves treatment or relief of ulcerative colitis through the following synergistic mechanism: (1) It inhibits the polarization of CD86+ pro-inflammatory M1 macrophages in colon tissue, downregulates the expression of pro-inflammatory factors TNF-α, IL-6 and IL-1β, upregulates the proportion of CD206+ anti-inflammatory M2 macrophages, increases the expression of anti-inflammatory factor IL-10, and reshapes the colonic inflammatory microenvironment; (2) Upregulate the expression of colonic mucosal tight junction proteins Occludin and Claudin-1, reduce intestinal mucosal permeability, repair damaged intestinal mucosal barrier, and promote complete healing of colonic mucosa; (3) Improved weight loss, reduced disease activity score, alleviated colonic shortening, and reduced colonic histological inflammatory damage score in the ulcerative colitis model.

[0019] Preferably, the drug is also used to prepare a drug for treating intestinal inflammatory diseases associated with ulcerative colitis, including ulcerative proctitis, left colitis, pancolitis, and for intestinal mucosal repair and inflammation control after ulcerative colitis surgery.

[0020] This invention provides a pharmaceutical composition and its application for treating or alleviating ulcerative colitis, which has the following beneficial effects: Compared with the prior art, the sarcosine pharmaceutical composition provided by the present invention and its application in the treatment of ulcerative colitis have at least the following significant beneficial effects: First, this invention systematically reveals the therapeutic and alleviating effect of sarcosine on ulcerative colitis, clarifies its dual protective role in inhibiting intestinal inflammation and repairing mucosal damage, provides a novel active ingredient and intervention target for UC treatment, expands the clinical application scenarios of sarcosine, and solves the technical problems of existing UC treatment drugs having single targets and insufficient primary / secondary response rates.

[0021] Second, in vivo experiments of this invention have demonstrated that creatine can exert its therapeutic effect on UC through a synergistic mechanism: on the one hand, it significantly inhibits the polarization of CD86+ pro-inflammatory M1 macrophages in colonic tissue, upregulates the proportion of CD206+ anti-inflammatory M2 macrophages, and then downregulates pro-inflammatory factors such as TNF-α, IL-6, and IL-1β, while increasing the level of the anti-inflammatory factor IL-10, thus precisely reshaping the colonic inflammatory microenvironment and effectively alleviating intestinal inflammation; on the other hand, it significantly upregulates the expression of colonic mucosal tight junction proteins Occludin and Claudin-1, reduces intestinal mucosal permeability, repairs damaged mucosal barriers, and promotes complete healing of the colonic mucosa, breaking through the technical bottleneck that most existing UC treatment drugs can only reduce inflammation and cannot effectively achieve mucosal healing.

[0022] Third, the present invention was validated by a DSS-induced experimental colitis model. Creatine administration can significantly improve the weight loss of model animals, reduce disease activity scores, alleviate colonic shortening, and reduce colonic histological inflammation damage scores. At the same time, it significantly downregulates the expression of the inflammatory marker CD86, comprehensively covering the core efficacy endpoints of UC treatment, providing reliable experimental evidence for clinical translation.

[0023] Fourth, creatine is a human endogenous small molecule metabolite with good biocompatibility and high in vivo safety. It does not have the serious systemic adverse reactions of existing immunosuppressants, glucocorticoids, and biologics. It can be prepared into a variety of commonly used clinical dosage forms, such as oral, rectal, and colon-targeted formulations, which are convenient to administer and have high patient compliance. It can be used as a monotherapy for induction of remission and long-term maintenance therapy of UC, or it can be used in combination with existing first-line drugs. It has a wide range of applications and solves the problems of poor long-term drug safety and low patient compliance of existing UC treatment drugs.

[0024] Fifth, the pharmaceutical composition of the present invention can be used in combination with existing first-line drugs for UC such as sarcosine, 5-aminosalicylic acid, and anti-TNF-α biological agents, which can significantly improve the treatment effect of patients with poor monotherapy response, increase the mucosal healing rate, reduce the dosage and adverse reaction risk of existing drugs, reduce the clinical recurrence of UC, and provide a new combination treatment option for patients with refractory and moderate to severe UC, which has important clinical application value.

[0025] Sixth, this invention addresses the pathological characteristics of UC lesions located in the colonic mucosa by optimizing the design of exclusive formulations such as pH-dependent enteric-coated microcapsules and colonic retention gels. These formulations enable the targeted release of active ingredients at the site of colonic inflammation, significantly increasing the local drug concentration in the lesion, reducing premature release and absorption of drugs in the stomach and small intestine, lowering the risk of systemic exposure, and improving treatment efficiency. This solves the problems of poor colonic targeting of conventional oral formulations and short local retention time of ordinary rectal administration formulations. Attached Figure Description

[0026] Figure 1 A schematic diagram illustrating the role of sarcosine in the treatment of UC. Figure 2 This is a schematic diagram showing the experimental results of sarcosine improving body weight, disease activity, colon length, and histological damage in a DSS-induced colitis model. Figure 3 This diagram illustrates the IHC detection and quantitative analysis of CD86 and CD206 in a DSS-induced colitis model using sarcosine. Detailed Implementation

[0027] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. 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.

[0028] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. The following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Any improvements, substitutions, or combinations made by those skilled in the art under the guidance of the core technical concept of the present invention, without the need for creative effort, are all within the scope of protection claimed by the present invention.

[0029] The sarcosine, sodium sarcosine, sarcosine hydrochloride, sarcosine ethyl ester, and sarcosine amide used in the following embodiments of the present invention were all purchased from regular manufacturers that meet the production standards for pharmaceutical excipients, with a purity ≥99.5%; the methacrylic acid copolymer L100, methacrylic acid copolymer S100, carbomer, hydroxypropyl methylcellulose, 5-aminosalicylic acid, diluents, adhesives, and other excipients used all meet the pharmaceutical grade standards of the Chinese Pharmacopoeia; the experimental animals used were SPF-grade C57BL / 6 male mice, 6-8 weeks old, weighing 18-22g, purchased from a regular experimental animal center; DSS (sodium dextran sulfate) with a molecular weight of 36,000-50,000 was purchased from Sigma-Aldrich; and all other experimental reagents were of analytical grade.

[0030] Example 1: Oral creatine tablets: This example provides an oral creatine tablet for the treatment of ulcerative colitis, with the following formulation: 100g sarcosine, 80g microcrystalline cellulose, 40g lactose, 8g crospovidone, 2g magnesium stearate, and an appropriate amount of 10% povidone K30 ethanol solution were used to make 1000 tablets.

[0031] Preparation method: sarcosine, microcrystalline cellulose, lactose, and crospovidone are passed through an 80-mesh sieve, mixed according to the prescription amount, and 10% povidone K30 ethanol solution is added to make a soft mass. The mass is then granulated through a 20-mesh sieve, dried in a forced-air dryer at 60℃, granulated through an 18-mesh sieve, and magnesium stearate is added and mixed before being compressed into tablets. Each tablet contains 100mg of sarcosine.

[0032] In this embodiment, sarcosine can be replaced by sodium sarcosine, sarcosine hydrochloride, sarcosine ethyl ester, sarcosine amide, etc., to prepare corresponding oral tablets, all of which fall within the protection scope of this invention.

[0033] Example 2: Sarcosine pH-dependent enteric-coated microcapsules: This embodiment provides a colon-targeted microcapsule for treating ulcerative colitis. The microcapsule consists of a drug-containing core, an isolation layer, and an enteric coating layer from the inside out. The specific formulation and preparation method are as follows: Formula for medicated pellet core: 250g sarcosine, 200g microcrystalline cellulose, 50g lactose, and an appropriate amount of 10% povidone K30 aqueous solution, to prepare 1000g blank pellet core, wherein the mass percentage of sarcosine in the medicated pellet core is 25%.

[0034] Preparation method: The drug-containing pellet core is prepared by extrusion-spheronization method. The prescribed amount of raw and auxiliary materials are mixed through an 80-mesh sieve, and a binder is added to make a soft material. The extruder extrudes the material through a sieve with a diameter of 0.8 mm, and the pellets are spheronized by a spheronization machine. The pellets are dried at 40℃ for 24 hours and then sieved to 20~30 mesh for later use.

[0035] Isolation layer coating: Take 1000g of the above-mentioned drug-containing pellet core and coat it using a bottom spray fluidized bed coating method. The isolation layer coating solution is a 2% aqueous solution of hydroxypropyl methylcellulose. The coating weight gain is 3% of the total mass of the drug-containing pellet core. After coating, dry at 40℃ for 2 hours for later use.

[0036] Enteric coating: The coating material is a mixture of methacrylic acid copolymer L100 and methacrylic acid copolymer S100 in a mass ratio of 1:1. The liquid-solid content of the coating is 10%, the solvent is 95% ethanol, and the weight gain of the coating is 10% of the total mass of the pill core. After coating, the mixture is cured at 40°C for 12 hours, sieved into 20-30 mesh enteric coated microspheres, and filled into gelatin hard capsules to obtain the final product. Each capsule contains 100 mg of sarcosine.

[0037] The enteric-coated microcapsules prepared in this embodiment showed the following release rates after testing: a cumulative release rate of 4.2% in hydrochloric acid solution at pH 1.2 after 2 hours, a cumulative release rate of 12.6% in phosphate buffer solution at pH 6.8 after 2 hours, and a cumulative release rate of 89.7% in colonic fluid environment at pH 7.2 after 1 hour, demonstrating that the active ingredients can be released at specific sites of colonic inflammation.

[0038] In this embodiment, the mass ratio of methacrylic acid copolymer L100 to S100 can be adjusted within the range of 1:2 to 2:1, the weight gain of the coating layer can be adjusted within the range of 8% to 15%, and the mass percentage of sarcosine in the drug-containing pellet core can be adjusted within the range of 10% to 60%, all of which can achieve the desired colon-targeted release effect.

[0039] Example 3: Creatine Colonic Retention Gel This embodiment provides a rectal administration gel for treating ulcerative colitis, with the following formulation: 100g sarcosine, 10g carbomer 940, 20g hydroxypropyl methylcellulose K4M, 50g glycerol, appropriate amount of triethanolamine, and purified water added to 1000g. The mass ratio of carbomer to hydroxypropyl methylcellulose in the gelling agent matrix is ​​1:2, and the mass concentration of sarcosine is 100mg / g.

[0040] Preparation method: Carbomer and hydroxypropyl methylcellulose were separately dissolved and dispersed in purified water, then combined and glycerol and the prescribed amount of sarcosine were added. The mixture was stirred evenly, and triethanolamine was added to adjust the pH to 6.0-6.5. Purified water was added to the total volume, and the mixture was defoamed to obtain the final product.

[0041] The colonic retention gel prepared in this embodiment was verified by in vitro rat colonic retention experiments. Its retention time at the site of colonic inflammation can reach 8 hours, significantly improving local drug concentration and therapeutic efficiency. In this embodiment, the mass ratio of carbomer to hydroxypropyl methylcellulose can be adjusted within the range of 1:1 to 1:3, and the mass concentration of sarcosine can be adjusted within the range of 50 to 200 mg / g, all of which fall within the scope of protection of this invention.

[0042] Example 4: Compound tablets of sarcosine and 5-aminosalicylic acid: This embodiment provides a compound oral tablet for treating ulcerative colitis, with the following prescription: 100g of sarcosine, 200g of 5-aminosalicylic acid, 120g of microcrystalline cellulose, 60g of lactose, 12g of croscarmellose sodium, 3g of magnesium stearate, and an appropriate amount of 10% povidone K30 ethanol solution were used to prepare 1000 tablets, in which the mass ratio of sarcosine to 5-aminosalicylic acid was 1:2.

[0043] The preparation method is the same as in Example 1. Each tablet contains 100 mg of sarcosine and 200 mg of 5-aminosalicylic acid. It can be used to induce remission of mild to moderate active ulcerative colitis and to treat patients who do not respond well to 5-aminosalicylic acid monotherapy.

[0044] In this embodiment, the mass ratio of sarcosine to 5-aminosalicylic acid can be adjusted within the range of 1:0.5 to 1:5, and all of these ratios can achieve synergistic therapeutic effects.

[0045] Example 5: Pharmacodynamic experiment of sarcosine in the treatment of ulcerative colitis: Experimental model construction and grouping: SPF-grade C57BL / 6 mice were randomly divided into 4 groups, with 10 mice in each group, as follows: Blank control group (PBS group): Normal drinking water, and daily gavage with an equal volume of PBS buffer; Sarcosine control group: Normal drinking water, daily gavage administration of 100 mg / kg sarcosine; Model control group (PBS+DSS group): The ulcerative colitis model was established by drinking sterile water containing 3% DSS for 7 days. The model was given an equal volume of PBS buffer by gavage daily. The treatment group (Sarcosine+DSS group) was established by drinking sterile water containing 3% DSS for 7 days. At the same time, 100 mg / kg of sarcosine was administered by gavage daily for 7 consecutive days.

[0046] Detection indicators: During the experiment, the body weight, fecal characteristics, and fecal blood loss of mice were recorded daily, and the disease activity index (DAI) score was calculated. On the 7th day of the experiment, mice were sacrificed, the colon was dissected and its length was measured, and tissue from the same part of the distal colon was taken for HE staining and histological inflammation damage scoring. Immunohistochemistry (IHC) was used to detect the protein expression levels of CD86 and CD206 in colon tissue, and Western blotting was used to detect the expression levels of colonic mucosal tight junction proteins Occludin and Claudin-1.

[0047] Experimental Results and Explanations: like Figure 1 The diagram illustrates the concept of sarcosine in the treatment of ulcerative colitis (UC) according to the present invention, clarifying the core intervention logic of the invention: sarcosine is the core active ingredient, administered via oral gavage, rectal administration, or colon-targeted formulation. In this example, a 3% DSS-induced colitis model was intervened with a dose of 100 mg / kg·d for 7 days, ultimately achieving the core efficacy endpoints of reduced weight loss, decreased disease activity, improved colon length, reduced histological damage, and reduced inflammation-related markers (CD86 signal). This fully reveals the core technical effect of the present invention: "sarcosine intervention → improvement of colitis phenotype".

[0048] like Figure 2 The diagram shows the experimental results of creatine improving body weight, disease activity, colon length, and histological damage in a DSS-induced colitis model. Figure 2A shows the weight change trend of mice in each group over 7 days. It can be seen that the weight of mice in the model control group decreased significantly with the extension of the modeling time, while the weight loss of mice in the creatine administration group was 62.3% less than that of the model control group, which can significantly improve the weight loss caused by colitis. Figure 2 B represents the disease activity (DAI) score of each group of mice. The DAI score of the creatine-treated group was 58.7% lower than that of the model control group, which can significantly alleviate colitis-related symptoms such as diarrhea and bloody stools. Figure 2 D represents the statistical results of colon length in each group of mice. The colon of mice in the model control group was significantly shortened, while the colon length of mice in the creatine administration group recovered to 87.2% of that in the model control group, which can significantly alleviate the pathological shortening of the colon caused by colitis. Figure 2 E represents the colonic histological inflammation score of mice in each group. The histological score of mice in the creatine administration group was reduced by 51.4% compared with the model control group, which can significantly reduce colonic mucosal erosion, ulceration and inflammatory cell infiltration, and improve histological damage.

[0049] like Figure 3 The image shows a schematic diagram illustrating the staining and quantitative analysis results of CD86 and CD206 by IHC in a DSS-induced colitis model using sarcosine. Figure 3 A shows the immunohistochemical staining results of CD86 and CD206 in the colon tissue of mice in each group. It can be seen that the number of CD86 positive cells in the colon tissue of the model control group is significantly increased and the number of CD206 positive cells is significantly decreased, while creatine administration can significantly reverse this trend. Figure 3 B represents the quantitative analysis results of CD86 protein expression. The expression level of CD86 protein in the colon tissue of mice in the sarcosine-treated group was downregulated by 42.6% compared with the model control group. Figure 3 C represents the quantitative analysis results of CD206 protein expression. The expression level of CD206 protein in the colon tissue of mice in the sarcosine-treated group was upregulated by 38.9% compared with the model control group.

[0050] The above results confirm that creatine can significantly inhibit the polarization of CD86+ pro-inflammatory M1 macrophages in colonic tissue, upregulate the proportion of CD206+ anti-inflammatory M2 macrophages, and reshape the colonic inflammatory microenvironment.

[0051] Meanwhile, Western blot analysis in this embodiment confirmed that the expression levels of Occludin and Claudin-1 proteins in the colonic mucosa of mice in the sarcosine-treated group were upregulated by 45.3% and 41.8% respectively compared with the model control group, demonstrating that sarcosine can significantly upregulate the expression of tight junction proteins in the colonic mucosa, repair the damaged intestinal mucosal barrier, and promote mucosal healing.

[0052] This embodiment also set up creatine dosage groups of 10 mg / kg, 50 mg / kg, 200 mg / kg, 300 mg / kg and 1000 mg / kg. The results showed that creatine had a significant therapeutic effect on colitis in the dosage range of 10~1000 mg / kg / day, with the 50~200 mg / kg / day dosage range showing the best effect and 100 mg / kg / day being the optimal dosage.

[0053] Example 6: Dosing regimens for different clinical scenarios: Based on the clinical application scenarios of the pharmaceutical composition of the present invention, this embodiment provides the following differentiated dosing regimens: Induction of remission in mild to moderate active ulcerative colitis: The sarcosine oral tablets of Example 1 of this invention were used at a dose of 100 mg / kg / day, divided into two oral doses per day, for a continuous administration period of ≥4 weeks until clinical remission and mucosal healing were achieved. Combination therapy for severe active ulcerative colitis: The compound tablets of Example 4 of this invention are used, with creatine dosage of 200 mg / kg / day, combined with anti-TNF-α biological agent 5 mg / kg / time, intravenously infused once every 2 weeks, for a continuous administration period of ≥8 weeks, to induce clinical remission; Maintenance therapy during remission of ulcerative colitis: Colon-targeted microcapsules of Example 2 of this invention are used at a dosage of 50 mg / kg / day, orally once daily for a continuous period of ≥6 months to maintain clinical remission and reduce the recurrence rate of the disease.

[0054] Example 7: Application of creatine in UC-related intestinal inflammatory disease: This embodiment verifies the therapeutic effect of creatine in ulcerative proctitis and left-sided colitis. Using the colonic retention gel of Example 3, rectal administration at a dose of 100 mg / kg / day, once daily via enema, for 4 consecutive weeks, significantly improved mucosal inflammation and erosion in the rectum and left colon, reducing symptoms such as rectal bleeding and tenesmus. Simultaneously, the pharmaceutical composition of this invention can be used for intestinal mucosal repair and inflammation control after ulcerative colitis surgery. Oral administration at a dose of 50 mg / kg / day can promote postoperative intestinal mucosal healing and reduce the risk of postoperative inflammation recurrence.

[0055] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for treating or alleviating ulcerative colitis, said pharmaceutical composition comprising a therapeutically effective amount of an active ingredient and pharmaceutically acceptable excipients, characterized in that, The active ingredient is sarcosine, its pharmaceutically acceptable salt, or a pharmaceutically acceptable derivative thereof; after administration, the pharmaceutical composition can achieve at least one of the following technical effects: (1) Inhibits the polarization of CD86+ pro-inflammatory M1 macrophages in colon tissue, upregulates the proportion of CD206+ anti-inflammatory M2 macrophages, and reshapes the colonic inflammatory microenvironment; (2) Upregulate the expression of colonic mucosal tight junction proteins Occludin and Claudin-1, repair the damaged intestinal mucosal barrier, and promote mucosal healing; (3) Improved weight loss, reduced disease activity score, alleviated colonic shortening, and reduced colonic histological inflammatory damage score in the ulcerative colitis model.

2. The pharmaceutical composition for treating or alleviating ulcerative colitis according to claim 1, characterized in that, The pharmaceutically acceptable salt of sarcosine is selected from any one of sodium sarcosine and sarcosine hydrochloride; the pharmaceutically acceptable derivative of sarcosine is selected from any one of sarcosine ethyl ester and sarcosine amide.

3. The pharmaceutical composition for treating or alleviating ulcerative colitis according to claim 1, characterized in that, The dosage form of the pharmaceutical composition is an oral formulation, a rectal formulation, or a colon-targeted formulation; The oral preparation is selected from any one of tablets, capsules, granules, powders, and oral solutions; The rectal administration formulation is selected from any one of suppositories, enemas, and colonic retention gels; The colon-targeting formulation is selected from any one of pH-dependent enteric-coated formulations, time-controlled-release colon-release formulations, and microbiota-triggered colon-targeting formulations. The colon-targeting formulation is a pH-dependent enteric-coated microcapsule, wherein the microcapsule consists of a drug-containing core, an isolation layer, and an enteric coating layer from the inside out. The medicated core contains an active ingredient and a diluent, wherein the active ingredient accounts for 10% to 60% of the mass of the medicated core. The coating material of the enteric coating layer is a compound of methacrylic acid copolymer L100 and methacrylic acid copolymer S100 in a mass ratio of 1:2 to 2:1, and the weight gain of the coating layer is 8% to 15% of the total weight of the pill core. The enteric-coated microparticles exhibit a cumulative release rate of ≤10% in hydrochloric acid solution at pH 1.2 after 2 hours, a cumulative release rate of ≤20% in phosphate buffer solution at pH 6.8 after 2 hours, and a cumulative release rate of ≥80% in colonic fluid environment at pH ≥7.0 after 1 hour, achieving targeted release of active ingredients at sites of colonic inflammation. The rectal administration preparation is a colonic retention gel. The gel matrix is ​​a compound of carbomer and hydroxypropyl methylcellulose in a mass ratio of 1:1 to 1:

3. The mass concentration of the active ingredient in the gel is 50 to 200 mg / g. After rectal administration, the gel can remain at the site of colonic inflammation for ≥6 hours, thereby increasing the local drug concentration and treatment efficiency.

4. A pharmaceutical composition for treating or alleviating ulcerative colitis according to claim 3, characterized in that, The dosage of the active ingredient, calculated as creatine, is 10-1000 mg / kg / day. For the induction of remission of mild to moderate active ulcerative colitis, the preferred dosage of the active ingredient is 50-200 mg / kg / day, more preferably 100 mg / kg / day; For combination therapy of severe active ulcerative colitis, the preferred dosage of the active ingredient is 100-300 mg / kg / day; For maintenance therapy during the remission phase of ulcerative colitis, the preferred dosage of the active ingredient is 20-100 mg / kg / day.

5. A pharmaceutical composition for treating or alleviating ulcerative colitis according to claim 4, characterized in that, The drug composition is administered 1 to 2 times daily; For induction of remission in mild to moderate active ulcerative colitis, the continuous dosing period is ≥4 weeks; For combination therapy of severe active ulcerative colitis, the continuous dosing period is ≥8 weeks; For maintenance therapy during the remission phase of ulcerative colitis, continuous administration for ≥6 months is used to prevent clinical relapse.

6. A pharmaceutical composition for treating or alleviating ulcerative colitis according to claim 1, characterized in that, The pharmaceutical composition further comprises a second active ingredient, which is a first-line clinical treatment for ulcerative colitis and is selected from any one or more of 5-aminosalicylic acid drugs, glucocorticoids, immunomodulators, biological agents, and small molecule targeted drugs. When the second active ingredient is a 5-aminosalicylic acid drug, the mass ratio of the active ingredient to the 5-aminosalicylic acid drug, calculated as creatine, is 1:0.5 to 1:

5. The drug composition is used for the induction of remission of mild to moderate active ulcerative colitis, or for the treatment of patients who do not respond well to 5-aminosalicylic acid monotherapy. When the second active ingredient is an anti-TNF-α biological agent, the dosage of the active ingredient, calculated as creatine, is 50-200 mg / kg / day, and the dosage of the anti-TNF-α biological agent is 5-10 mg / kg / time, administered once every 2 weeks. The pharmaceutical composition is used for the induction and remission of moderate to severe active ulcerative colitis.

7. Use of sarcosine, its pharmaceutically acceptable salts or pharmaceutically acceptable derivatives thereof in the preparation of medicaments for the treatment or relief of ulcerative colitis.

8. The application according to claim 7, characterized in that, The drug is intended to achieve at least one of the following clinical benefits: (1) Improves clinical symptoms of ulcerative colitis such as weight loss, diarrhea, hematochezia, and abdominal pain, and reduces disease activity scores; (2) Repair the patient's damaged colonic mucosal barrier to achieve mucosal healing; (3) Relieves inflammatory infiltration of colonic tissue in patients with ulcerative colitis and reduces histological inflammatory damage scores; (4) Maintain clinical remission in patients with ulcerative colitis and reduce the recurrence rate of the disease.

9. The application according to claim 8, characterized in that, The drug treats or alleviates ulcerative colitis through the following synergistic mechanism: (1) It inhibits the polarization of CD86+ pro-inflammatory M1 macrophages in colon tissue, downregulates the expression of pro-inflammatory factors TNF-α, IL-6 and IL-1β, upregulates the proportion of CD206+ anti-inflammatory M2 macrophages, increases the expression of anti-inflammatory factor IL-10, and reshapes the colonic inflammatory microenvironment; (2) Upregulate the expression of colonic mucosal tight junction proteins Occludin and Claudin-1, reduce intestinal mucosal permeability, repair damaged intestinal mucosal barrier, and promote complete healing of colonic mucosa; (3) Improved weight loss, reduced disease activity score, alleviated colonic shortening, and reduced colonic histological inflammatory damage score in the ulcerative colitis model.

10. The application according to claim 9, characterized in that, The drug is also used to prepare a drug for treating intestinal inflammatory diseases associated with ulcerative colitis, including ulcerative proctitis, left colitis, pancolitis, and for intestinal mucosal repair and inflammation control after ulcerative colitis surgery.

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