Application of wogonin derivative V8 in preparation of medicine for treating pancreatic fibrosis
By using hambasalin derivative V8 to inhibit pancreatic stellate cell activation and inflammatory factors, the problem that cannot prevent pancreatic fibrosis progression in the prior art is solved, and an effective pancreatic fibrosis treatment plan is provided.
Patent Information
- Application Number
- CN202510519203.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-24
- Publication Date
- 2025-07-04
AI Technical Summary
There is currently a lack of effective drugs that can inhibit the activation of pancreatic stellate cells, thereby preventing the progression of pancreatic fibrosis. The existing treatment methods can only alleviate the symptoms and cannot terminate the fibrosis process.
The hambasalin derivative V8 was used to reduce collagen secretion and significantly reduce the level of pancreatic fibrosis by inhibiting the release of inflammatory factors and inhibiting the activation of pancreatic stellate cells.
The hambasalin derivative V8 can significantly inhibit the activation of pancreatic stellate cells, reduce collagen secretion, and reduce the degree of pancreatic fibrosis, providing a new drug choice for the treatment of pancreatic fibrosis.
Smart Images
Figure CN120241704A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biomedicine and relates to the application of wogonin derivative V8 in the preparation of a medicament for treating pancreatic fibrosis. Background Art
[0002] Pancreatic fibrosis (PF) is an important pathological feature of chronic pancreatitis. A large amount of extracellular matrix deposits in pancreatic tissue, ultimately leading to pancreatic exocrine and endocrine insufficiency. So far, the main clinical treatments for pancreatic fibrosis include symptomatic treatment and surgery. Although these treatment measures can relieve the symptoms, they cannot terminate the fibrosis process, and there is no drug specifically for pancreatic fibrosis. Therefore, exploring drugs for treating pancreatic fibrosis has important clinical value and practical significance. Pancreatic stellate cells (PSCs) are key effector cells in the initial stage of pancreatic tissue fibrosis. After acute pancreatic injury and inflammation occur, quiescent PSCs can be activated by various damage-related stimulatory factors and pro-inflammatory factors (such as TGFβ1, PDGF, TNFα, IL 1β, IL 6, etc.) in the tissue microenvironment, transform into myofibroblast-like cells, and secrete extracellular matrix components such as fibronectin (FN) and type I, II, and IV collagens to participate in tissue repair; if PSCs are over-activated, it can cause a large amount of extracellular matrix deposition, ultimately leading to pancreatic tissue fibrosis. Therefore, inhibiting the activation of pancreatic stellate cells can effectively inhibit the progression of pancreatic fibrosis and become an important target for treating pancreatic fibrosis. Research reports show that traditional Chinese medicine compounds, components or monomers, such as Chaihu Guizhi Ganjiang Decoction, oxymatrine, etc., have the effect of inhibiting the activation, proliferation and collagen secretion of pancreatic stellate cells and play an anti-pancreatic fibrosis role. Therefore, based on the advantages of traditional Chinese medicine in anti-organ fibrosis, it is of extremely important scientific significance to deeply explore traditional Chinese medicine for anti-pancreatic fibrosis and its active ingredients.
[0003] Previous studies have shown that compound V8 has good prospects in the treatment of liver fibrosis. The core pathological process of chronic pancreatitis is continuous inflammatory damage to the pancreatic parenchyma, ultimately leading to pancreatic fibrosis. Alcohol, biliary tract diseases, and genetic factors (such as SPINK1 and CFTR gene mutations) are the main inducing factors. Among them, abnormal activation of the NF-κB and Hedgehog pathways can promote inflammation and fibrosis; autophagy defects can exacerbate pancreatic fibrosis damage through oxidative stress. Liver fibrosis and liver inflammation are mainly caused by chronic liver injury (such as viral hepatitis and alcoholic liver disease), which trigger an inflammatory response, activate hepatic stellate cells (HSCs), and lead to excessive deposition of ECM. The involved mechanisms include macrophages promoting the activation of HSCs through cytokines such as IL-4 / IL-13 and the complement system (such as C5a) participating in the progression of liver fibrosis. There are obvious differences in the pathogenic mechanisms between chronic pancreatitis, pancreatic fibrosis, liver fibrosis, and liver inflammation. Currently, the treatment of pancreatic fibrosis focuses on relieving symptoms (pain and exocrine insufficiency), and new therapies focus on probiotics and targeted pathway inhibitors. While the treatment of the liver emphasizes the removal of the cause (such as antiviral treatment) and multi-target intervention with traditional Chinese medicine, combined with antioxidant and immunomodulatory therapies.
[0004] Compound V8 is a derivative of wogonin, an effective active ingredient of the traditional Chinese medicine Scutellaria baicalensis, which has been newly synthesized. Its structural formula is shown below. In terms of structure, V8 introduces a tertiary amine side chain with a hydroxyl group into the flavone mother ring of the original wogonin, making it have better water solubility. Only a small number of previous studies have found that it may have certain anti-tumor potential, but there are no reports on its research in other diseases. Therefore, the role of V8 in the preparation of drugs for preventing and / or treating pancreatic fibrosis has not been reported currently.
[0005] Summary of the Invention
[0006] The purpose of the present invention is to provide the application of wogonin derivative V8 in the preparation of drugs for treating pancreatic fibrosis in view of the above deficiencies of the prior art.
[0007] Another purpose of the present invention is to provide the application of wogonin derivative V8 in the preparation of drugs for treating chronic pancreatitis.
[0008] The purpose of the present invention is achieved through the following technical solutions:
[0009] The application of wogonin derivative V8 in the preparation of drugs for treating chronic pancreatitis.
[0010] As a preference of the present invention, wogonin derivative V8 can inhibit the release of inflammatory factors.
[0011] The application of wogonin derivative V8 in the preparation of drugs for treating pancreatic fibrosis.
[0012] As a preference of the present invention, the pancreatic fibrosis is pancreatic fibrosis caused by choledocholithiasis, metabolic disorder, nutritional disorder, heredity, endocrine abnormality and / or other diseases.
[0013] As a preference of the present invention, wogonin derivative V8 can inhibit the activation of pancreatic stellate cells, weaken the degree of pancreatic cell injury and fibrosis in mice, inhibit the release of inflammatory factors, and significantly reduce the level of pancreatic fibrosis.
[0014] A pharmaceutical composition for treating chronic pancreatitis or pancreatic fibrosis, which is composed of an active ingredient wogonin derivative V8 and a pharmaceutically acceptable excipient.
[0015] As a preference of the present invention, the dosage form of the pharmaceutical composition is tablet, granule, pill, capsule or injection.
[0016] Beneficial effects:
[0017] In the present invention, by constructing a mouse model of inflammation-induced pancreatic fibrosis, it is proved that the treatment with wogonin derivative V8 can significantly inhibit the activation of pancreatic stellate cells, weaken the degree of pancreatic cell injury and fibrosis in mice, inhibit the release of inflammatory factors, and the Western blot experiment finds that the protein level of inflammatory factors in pancreatic stellate cells shows a downward trend after administration. The level of pancreatic fibrosis is significantly reduced. Therefore, it is proposed that wogonin derivative V8 can be used to prepare drugs for improving pancreatic fibrosis-related diseases. Further, through the results of oil red staining and immunofluorescence experiments, it can be found that the role of the wogonin derivative V8 in preventing and / or treating pancreatic fibrosis is through one or both of the following ways: inhibiting the activation of pancreatic stellate cells and reducing collagen secretion. Description of the drawings
[0018] Figure 1 、Effect of DBTC and V8 on the morphology of mouse pancreas
[0019] Figure 2 、Content of α-amylase in mice on the 2nd day (A) and the 28th day (B)
[0020] Figure 3 、Results of H&E staining of mouse pancreatic sections
[0021] Figure 4 、Results of Masson staining of mouse pancreatic sections
[0022] Figure 5 、Results of immunofluorescence experiment
[0023] Figure 6 、Results of oil red staining experiment
[0024] Figure 7 、Results of Western blot experiment of inflammatory factors Detailed implementation mode
[0025] Effect of Example 1 on DPTC-induced primary pancreatitis model in mice
[0026] 1. Experimental materials
[0027] 1.1 Test drugs
[0028] Name: V8, Content: >98%, Batch number: 24102512
[0029] Preparation method: For cell experiments, the drug powder was prepared into a stock solution with a concentration of 30 mM using dimethyl sulfoxide (DMSO) and stored at 80 °C for later use; for animal experiments, the drug was formulated into a suspension with a concentration of 40 mg / mL using a 0.5% sodium carboxymethylcellulose (CMC Na) solution and stored at 4 °C for later use.
[0030] 1.2 Experimental animals
[0031] Source, germline, strain: ICR mice, purchased from Jiangsu Qinglongshan Biotechnology Co., Ltd.
[0032] Week age: 6 - 8 weeks old
[0033] Gender: Male
[0034] 2. Animal experiments
[0035] 2.1 Dibutyltin dichloride (DPTC)-induced pancreatic fibrosis model in mice
[0036] Specific pathogen-free ICR mice (6 - 8 weeks old, body weight: 18 - 20 g) were injected with an 8 mg / mL DPTC solution (dissolved in absolute ethanol: glycerol = 3:2) (8 mL / kg) via the tail vein once a week for 4 weeks. Drug treatment started on the fourth day after the first injection of DPTC and continued for 4 weeks until the end of the experiment.
[0037] The grouping and dose settings of the experimental animals are as follows:
[0038]
[0039] 2.2 DPTC-induced primary pancreatitis model in mice
[0040] 6-week-old ICR mice were injected with DPTC (8 mg / kg) via the tail once a week for 4 weeks. Drug treatment started 4 days after the first injection and continued for 4 weeks until the end of the experiment.
[0041] 2.3 Determination of serum α-amylase content
[0042] Blood was collected from the orbital sinus of mice, about 500 - 1000 μL per mouse. The blood was allowed to stand at room temperature for 30 min, centrifuged at 4000 rpm for 10 min, and the supernatant was collected. The supernatant could be stored at -20 °C. The serum α-amylase content was measured by Wuhan Sevier Biotechnology.
[0043] 2.4 H&E staining
[0044] The mice were sacrificed, and the pancreas was removed surgically and fixed in 4% paraformaldehyde solution for 24 h, then sent to the Pathology and PDX Pharmacodynamic Evaluation Platform of China Pharmaceutical University for paraffin embedding and sectioning. H&E staining and slide reading were completed by the pharmacodynamic platform. The sections were observed and photographed under a microscope. The cell nuclei were blue, and the cytoplasm was pink or red.
[0045] 2.5 Masson staining
[0046] The mice were sacrificed, and the pancreas was removed surgically and fixed in 4% paraformaldehyde solution for 24 h, then sent to the Pathology and PDX Pharmacodynamic Evaluation Platform of China Pharmaceutical University for paraffin embedding and sectioning. Masson staining and slide reading were completed by the pharmacodynamic platform. The sections were observed and photographed under a microscope. Collagen fibers, mucus, and cartilage were blue, muscle fibers, cellulose, and red blood cells were stained red, and cell nuclei were stained blue-black.
[0047] 3. Cell experiments
[0048] 3.1 Immunofluorescence staining
[0049] Cells were fixed with 4% paraformaldehyde solution for 10 minutes, then washed twice with PBS, incubated with blocking and permeabilization solution for 45 minutes, incubated with fluorescent primary antibody overnight, washed three times with PBS, then added fluorescent secondary antibody, and finally blocked with DAPI and photographed under a confocal microscope.
[0050] 3.2 Oil Red O staining of cells
[0051] Cells were fixed with 4% paraformaldehyde solution for 10 minutes, then washed twice with PBS, stained with an appropriate amount of Oil Red O staining solution for 30 minutes, the staining solution was discarded and washed with washing solution, then washed with PBS, and finally an appropriate amount of PBS was added to observe the size of lipid droplets under a microscope.
[0052] 3.3 Western blot
[0053] Cells were collected, lysed with RAPI lysis buffer for 30 minutes, centrifuged at high speed to collect the supernatant, the concentration was detected by BCA method, and then electrophoresis, membrane transfer, antibody incubation, and development were performed.
[0054] 4. Experimental results
[0055] 4.1 V8 inhibits the occurrence and development of pancreatic fibrosis in mice in vivo.
[0056] During the model establishment period, the mental state of the mice in the DPTC-induced model group gradually became listless, their activities decreased, and their ears, limbs, and tails turned yellow. During dissection, it was found that the peritoneal organs of the DPTC-induced model group showed yellowing and there was a large amount of abdominal effusion. When comparing the pancreatic tissues of the mice in each group, it was found that the pancreas of the mice in the DPTC-induced model group was significantly atrophied compared with the blank group, with a yellowish color, and there was no significant difference between the V8 treatment group and the blank group.
[0057] 3.2 DBTC successfully induced pancreatic inflammatory response in mice
[0058] Serum was extracted and the content of α-amylase was measured. The experiment found that on the 2nd day, there were significant differences in the α-amylase measured in the DPTC-induced model group and the V8 treatment group compared with the blank group, and there was no significant difference among the three groups on the 28th day.
[0059] DBTC successfully induced the occurrence and development of chronic pancreatitis in mice, while V8 significantly inhibited the occurrence and development of chronic pancreatitis in mice induced by DBTC.
[0060] H&E staining and Masson staining were performed on pancreatic sections and the morphological analysis of the sections was described. In the blank group, the acinar and islet structures were normal, no obvious atrophy or necrosis was seen, and no obvious edema or bleeding was seen in the interstitium; there were more chronic inflammatory cell infiltrations and a small amount of fibrous tissue hyperplasia in the focal area under the capsule at the edge of the pancreatic tissue. In the DPTC-induced model group, the acini were atrophied, the number of islets decreased, there were a large number of inflammatory cell infiltrations and fibrous tissue hyperplasia in the interstitium, a large number of macrophages were seen in some areas, and the duct epithelial cells were degenerated. In the V8 treatment group, the acinar and islet structures were normal, no obvious atrophy or necrosis was seen, no obvious edema or bleeding was seen in the interstitium, and no inflammatory cells or fibrous tissue were seen; there was a small focal tissue around the pancreatic tissue that had no obvious connection with the pancreas, suspected to be atrophied pancreatic tissue, and inflammatory cell infiltrations and a small amount of fibrous tissue hyperplasia were seen in the interstitium. The immunofluorescence experiment was used to detect the expression level of α-SMA protein and it was found that after administration, the expression level of α-SMA protein decreased. The oil red staining experiment found that the v8 administration group increased the lipid droplet expression of pancreatic stellate cells and inhibited the activation of pancreatic stellate cells. The immunoblotting experiment found that the protein level of inflammatory factors in pancreatic stellate cells showed a downward trend after administration.
Claims
1. Use of wogonin derivative V8 in the preparation of a medicament for treating chronic pancreatitis.
2. The application according to claim 2, characterized in that Wogonin derivative V8 can inhibit the release of inflammatory factors.
3. Use of wogonin derivative V8 in the preparation of a medicament for treating pancreatic fibrosis.
4. The application according to claim 3, characterized in that The pancreatic fibrosis is caused by choledocholithiasis, metabolic disorder, nutritional disorder, heredity, endocrine abnormality and / or other diseases.
5. The application according to claim 3, characterized in that, Wogonin derivative V8 can inhibit the activation of pancreatic stellate cells.
6. The application according to claim 3, characterized in that, Wogonin derivative V8 can reduce the degree of pancreatic cell injury and fibrosis in mice.
7. The application according to claim 3, wherein Wogonin derivative V8 can inhibit the release of inflammatory factors.
8. The application according to claim 3, characterized in that, Wogonin derivative V8 can significantly reduce the level of pancreatic fibrosis.
9. A pharmaceutical composition for treating chronic pancreatitis or pancreatic fibrosis, characterized in that, The pharmaceutical composition is composed of the active ingredient wogonin derivative V8 and pharmaceutically acceptable excipients.
10. The pharmaceutical composition according to claim 9, characterized in that, The dosage form of the pharmaceutical composition is tablets, granules, pills, capsules or injections.