Use of il13 in combination with ccl22 in the preparation of a medicament for treating acute pancreatitis

Abstract

The application relates to application of IL13 and CCL22 in preparation of a drug for treating acute pancreatitis, and belongs to the technical field of biological medicines. It is found that, in an animal model of acute pancreatitis, joint administration of IL13 and CCL22 can significantly reduce pathological damage of pancreatic tissue compared with single use of IL13 or CCL22. Animal experiment results show that, after intraperitoneal injection of IL13 and CCL22, the activities of serum amylase and lipase and the levels of inflammatory factors IL-6, TNF-alpha and IL-1beta can be effectively reduced, and pancreas edema, necrosis and inflammatory cell infiltration can be relieved. The application provides a new combined drug strategy for treating acute pancreatitis.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to the application of IL13 combined with CCL22 in the preparation of drugs for treating acute pancreatitis. Background Technology

[0002] Acute pancreatitis is a common acute abdominal condition characterized by rapid onset and progression. In severe cases, it can develop into severe acute pancreatitis, accompanied by systemic inflammatory response syndrome and multiple organ failure, resulting in a high mortality rate. Its pathogenesis is a complex pathophysiological process, the core of which involves the abnormal activation of pancreatic enzymes within the pancreas, leading to pancreatic autodigestion and subsequently causing tissue damage, edema, hemorrhage, and even necrosis. Current clinical treatment primarily focuses on symptomatic and supportive care, lacking highly specific and effective drugs. Therefore, finding combination therapy strategies that can effectively inhibit local pancreatic inflammation and reduce tissue damage is of significant clinical importance.

[0003] IL-13 (interleukin-13) and CCL22 (chemokine ligand 22) are both important cytokines involved in immune regulation. IL-13 plays a crucial role in anti-inflammatory responses, inhibiting the production of pro-inflammatory factors and reducing tissue inflammatory damage, while CCL22 mainly participates in the chemotaxis of regulatory T cells, maintaining immune tolerance and suppressing excessive inflammatory responses. Studies have shown that IL-13 participates in regulating epithelial cell repair and mucus secretion, playing an important role in allergic inflammation, while CCL22 has important regulatory functions in autoimmune diseases and chronic inflammation. However, the application of IL-13 combined with CCL22 in the treatment of acute pancreatitis has not yet been reported. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides the application of IL13 combined with CCL22 in the preparation of drugs for treating acute pancreatitis. Compared to using IL13 or CCL22 alone, the combination of IL13 and CCL22 can effectively reduce serum amylase and lipase activity; significantly reduce the levels of inflammatory factors IL-6, TNF-α, and IL-1β; alleviate pancreatic edema, necrosis, and inflammatory cell infiltration; and reduce pathological scores.

[0005] This invention is achieved through the following technical solution:

[0006] The purpose of this invention is to provide the application of IL13 combined with CCL22 in the preparation of drugs for treating acute pancreatitis.

[0007] In one embodiment of the present invention, the dosage of IL13 in the drug is 10 μg / kg to 300 μg / kg, preferably 25 μg / kg to 100 μg / kg;

[0008] And / or, the amount of CCL22 used is 5 μg / kg to 150 μg / kg, preferably 20 μg / kg to 80 μg / kg.

[0009] In one embodiment of the present invention, the drug is administered once daily for 1 to 5 consecutive days.

[0010] In one embodiment of the present invention, the drug dosage form is selected from tablets, capsules, granules, oral liquids, emulsions, dry suspensions, dry extracts, or injections.

[0011] In one embodiment of the invention, the drug further includes a pharmaceutically or pharmacologically acceptable carrier and / or salt.

[0012] In one embodiment of the present invention, the carrier is selected from one or more of the following: disintegrant, diluent, lubricant, adhesive, humectant, flavoring agent, filler, suspending agent, surfactant, and preservative.

[0013] In one embodiment of the invention, the filler is selected from one or more of starch, sucrose, and lactose; the wetting agent includes glycerin; and the surfactant includes hexadecyl alcohol.

[0014] In one embodiment of the present invention, the adhesive is selected from one or more of cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone.

[0015] In one embodiment of the present invention, the disintegrant is selected from one or more of agar, calcium carbonate, and sodium bicarbonate.

[0016] In one embodiment of the invention, the pharmaceutically or pharmacologically acceptable salt is selected from inorganic acid salts and / or organic acid salts; the organic acid salt is selected from alkyl sulfonates and / or aryl sulfonates.

[0017] In one embodiment of the present invention, the administration of IL13 and CCL22 includes intraperitoneal injection.

[0018] This invention treats acute pancreatitis through the synergistic effect of IL-13 and CCL22. The mechanism is as follows: IL-13 exerts its anti-inflammatory effect by activating the STAT6 signaling pathway, promoting macrophage polarization towards the M2 type and facilitating the repair of damaged tissue. CCL22, through its specific binding to its receptor CCR4, recruits regulatory T cells (Tregs) and Th2 cells, enriching them at the site of pancreatic inflammation, reshaping the local inflammatory microenvironment, and simultaneously increasing the number of target cells for IL-13. More importantly, the two form a complete bidirectional positive feedback regulatory loop in the pancreatic injury area: IL-13 stimulates local pancreatic macrophages and dendritic cells to upregulate CCL22 expression; simultaneously, Treg and Th2 cells recruited to the inflammatory site by CCL22 can also secrete IL-13. This creates a stepwise amplification mechanism of IL-13, CCL22, Treg / Th2, and IL-13, continuously inhibiting the inflammatory response, optimizing the inflammatory microenvironment, and promoting pancreatic tissue repair.

[0019] This specific positive feedback synergistic regulatory loop can only be achieved through the combined administration of IL-13 and CCL22. The above-mentioned benign regulatory loop cannot be constructed by administering IL-13 or CCL22 alone, which makes this invention different from conventional single anti-inflammatory cytokine treatment regimens.

[0020] Compared with the prior art, the above-described technical solution of the present invention has the following advantages:

[0021] This invention is the first to propose the use of IL13 combined with CCL22 for the treatment of acute pancreatitis, which has a synergistic effect. The combined drug of this invention can significantly reduce serum amylase and lipase levels, indicating a significant reduction in pancreatic damage. The combined drug of this invention can effectively inhibit the level of inflammatory factors and alleviate pancreatic edema, necrosis and inflammatory infiltration. In HE-stained pancreatic sections of mice with acute pancreatitis after the combined drug of this invention, edema was reduced, inflammatory cell infiltration and necrosis were reduced, indicating that the inflammation was relieved.

[0022] This invention is based on a specific positive feedback coupling mechanism between IL-13 and CCL22. IL-13 can induce target cells to upregulate CCL22 expression, and CCL22, in turn, recruits immune cells expressing CCR4 to the pancreas. These recruited cells are precisely the targets of IL-13, thus forming a positive feedback loop where IL-13 promotes CCL22 production, and CCL22 recruits more target cells, further amplifying the effect of IL-13. This mechanism is significantly different from ordinary combinations of anti-inflammatory factors. Experiments show that the effects of IL-13 or CCL22 alone are limited, while the reduction in inflammatory factor levels after combined administration is significantly better than that after single administration, demonstrating a synergistic effect. These results indicate that the combined use of IL-13 and CCL22 achieves synergistic enhancement through a positive feedback loop, achieving therapeutic effects that cannot be achieved by a single agent. Attached Figure Description

[0023] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0024] Figure 1 This is a statistical chart showing the serum amylase and lipase activities of wild-type mice, acute pancreatitis model mice, and mice in the single-drug and combined-drug groups in this invention.

[0025] Figure 2 This is a statistical chart showing the levels of IL-6, TNF-α, and IL-1β in the serum of mice in each treatment group in this invention.

[0026] Figure 3 This is a statistical chart of HE staining pathological scores of pancreatic tissue from mice in each treatment group in this invention;

[0027] Figure 4 The images show the pathological scores of pancreatic edema, inflammatory cell infiltration, and necrosis in mice of each treatment group in this invention. Detailed Implementation

[0028] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0029] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the materials and reagents used are commercially available.

[0030] Recombinant mouse interleukin-13 (IL13): purchased from TargetMol Chemicals Inc., product code: [Cat#TMPJ-00889], purity ≥98%, lyophilized powder, reconstituted with sterile PBS before use.

[0031] Recombinant mouse chemokine ligand 22 (CCL22, also known as macrophage-derived chemokine MDC): purchased from TargetMol Chemicals Inc., product code: [Cat# TMPY-00305], purity ≥98%, lyophilized powder form, reconstituted with sterile PBS before use.

[0032] C57BL / 6J mice were purchased from Vital Rivers.

[0033] Example 1: Establishment of an acute pancreatitis mouse model and drug administration regimen

[0034] Six- to eight-week-old male C57BL / 6J mice were randomly divided into the following groups (n=6 per group): normal control group (WT), model group (AP), IL13 monotherapy group, CCL22 monotherapy group, and IL13+CCL22 combination group. An acute pancreatitis model was established by intraperitoneal injection of linalool (50 μg / kg, once per hour for a total of 7 times). The normal control group received an equal volume of phosphate-buffered saline (PBS).

[0035] After modeling, each treatment group received an intraperitoneal injection of the corresponding drug: the IL13 monotherapy group received IL13 40 μg / kg, the CCL22 monotherapy group received CCL22 50 μg / kg, the combination group received both IL13 (40 μg / kg) and CCL22 (50 μg / kg), and the normal control group and model group received an equal volume of physiological saline. Administered once daily for two consecutive days.

[0036] Test Example 1: Detection of Serum Amylase and Lipase Activities

[0037] Twelve hours after the last administration, venous blood was collected from mice in each group, serum was separated, and serum amylase and lipase activities were detected using a biochemical analyzer. Results are as follows: Figure 1 As shown, the serum amylase and lipase activities of mice in the model group were significantly higher than those in the normal control group, while the IL13+CCL22 combination group could significantly reduce the activities of these two enzymes, and the effect was better than either single drug group.

[0038] Test Example 2: Detection of Inflammatory Factor Expression in Serum

[0039] Twelve hours after the last administration, venous blood was collected from mice in each group, serum was separated, and the serum levels of IL-6, TNF-α, and IL-1β in each group were measured using an ELISA kit (purchased from Jiangsu Jingmei Biotechnology Co., Ltd.). Results are as follows: Figure 2 As shown, the expression of the above-mentioned inflammatory factors in the combined drug group was significantly lower than that in the model group and the single drug group, indicating that IL13 combined with CCL22 can effectively inhibit the local inflammatory response of the pancreas.

[0040] Test Example 3: Histopathological Evaluation of Pancreatic Tissue

[0041] Pancreatic tissues from mice in each group were collected, fixed in 4% paraformaldehyde, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and the pathological changes of the pancreatic tissues were observed under a light microscope. Pathological scoring was performed according to the degree of edema, necrosis, and inflammatory cell infiltration. Results are as follows: Figure 3 and Figure 4 As shown, the combined drug group showed significantly reduced pancreatic tissue damage, and the pathological score was significantly lower than that of the model group and the single drug group.

[0042] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. Application of IL13 combined with CCL22 in the preparation of drugs for the treatment of acute pancreatitis.

2. The application according to claim 1, characterized in that, The dosage of IL13 in the drug is 10 μg / kg to 300 μg / kg; And / or, the dosage of CCL22 is 5 μg / kg to 150 μg / kg.

3. The application according to claim 1, characterized in that, The drug is administered once daily for 1 to 5 consecutive days.

4. The application according to claim 1, characterized in that, The drug dosage form is selected from tablets, capsules, granules, oral liquids, emulsions, dry suspensions, dry extracts, or injections.

5. The application according to claim 1, characterized in that, The drug also includes pharmaceutically or pharmacologically acceptable carriers and / or salts.

6. The application according to claim 5, characterized in that, The carrier is selected from one or more of the following: disintegrant, diluent, lubricant, adhesive, humectant, flavoring agent, filler, suspending agent, surfactant, and preservative.

7. The application according to claim 6, characterized in that, The filler is selected from one or more of starch, sucrose, and lactose; the wetting agent includes glycerin; and the surfactant includes hexadecyl alcohol.

8. The application according to claim 6, characterized in that, The adhesive is selected from one or more of cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone.

9. The application according to claim 6, characterized in that, The disintegrant is selected from one or more of agar, calcium carbonate, and sodium bicarbonate.

10. The application according to claim 5, characterized in that, The pharmaceutically or pharmacologically acceptable salt is selected from inorganic acid salts and / or organic acid salts; the organic acid salt is selected from alkyl sulfonates and / or aryl sulfonates.

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